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ABSTRACT

Shallow waters are globally important, for their uses (e.g. fisheries, water purification,
water for human consumption), amenity values and ecological importance, despite their
generally small size and volume. Shallow lakes apparently exist in either of two
alternative states, one dominated by aquatic plants and characterized by clear water, the
other turbid and often considered undesirable. Either state can be found across a wide
range of nutrient concentrations. It is postulated that each state is stabilized by a suite of
mechanisms. One of the mechanisms stabilizing the plant-dominated state may be the
grazing of cladocerans living in tight or loose association to plants.

Little Mere (Cheshire) is a small (2.3 ha) and very shallow lake (average depth 0.7 m) in
North-West England, which received a poor standard effluent prior to 1991. In that
year, sewage was diverted for treatment elsewhere. The lake seems to have stabilized,
about ten years after diversion in June 1991, with aquatic plants, both nymphaeids and
submerged, covering almost its entirety. Lily beds now cover circa 40 % of the total
surface area. In Little Mere, top-down processes (i.e. cladoceran herbivory and fish
predation on zooplankton) are thought to be more important than bottom-up forces (i.e.
nutrient limitation) in the regulation of the maximum size of algal crops. However, little
is known on the specific mechanisms stabilizing the plant-dominated state. Indeed,
occasionally high levels of water clarity are maintained in this lake even in the absence
of significant populations of the common open-water cladoceran Daphnia.

The main hypothesis examined in this study is that plant-associated cladocerans are
important grazers helping to maintain clear water in Little Mere during the growing
season (April to November). The lake was intensively sampled for Cladocera and other
variables related to chemistry and phytoplankton during the period April 1998-April
2000. Samples were collected at five sites across the lake, three of them located within
lily beds, the other two over submerged plant beds of mixed composition. Specific
sampling techniques were developed for floating lily leaves, petioles, submerged plants
and water.

Significant horizontal differences were identified for most cladoceran species, both
open-water and plant-associated, chydorid periphyton scrapers and filter-feeders.
Daphnia and Ceriodaphnia were significantly more abundant in lily beds than in more
open water in both growing seasons, suggesting these beds are an effective refuge against
fish predation. Size-structure and egg-ratio data support this contention. Specific
differences with respect to micro-habitat association in lily beds (i.e. floating lily beds vs
petioles) were also identified. Spatial distributions of most cladocerans were very
heterogeneous, particularly in the case of chydorids. Patchiness was driven by resource
availability and intraspecific competition when populations were dense, but fish
predation was apparently the main regulator when numbers were scarce (i.e. in growing
season 1999). Egg-ratio models were examined for Daphnia and Simocephalus vetulus
(O.F.Muller), a plant-associated cladoceran. These models were difficult to apply to the
latter species, probably as a result of large spatial heterogeneity.

In situ feeding experiments using a dual radio-isotope technique, labelling planktonic


and periphytic algae with H-3 and C-14, respectively, were conducted during one
growing season in Little Mere, to gauge measures of feeding ´intensity´ for the main
cladoceran species in the lake. Sida crystallina (O.F.Muller), a plant-associated
cladoceran, was found to be an efficient filterer (though not as efficient as Daphnia
hyalina L.). Periphyton inhibited its filtering, but less so in young than adults.
Ceriodaphnia and, especially, Simocephalus vetulus, were relatively inefficient filter-
feeders in Little Mere.

Measures of areal plant biomass and filtering rates (ml d-1 ind.-1), in conjunction with
density estimates for the main cladoceran species (both open water and plant-associated)
were used to calculate measures of grazing rate (% lake volume filtered per day), for
filter-feeders in Little Mere. Consideration of the multiple sources of error in these
estimates showed that they are a valid method for comparison of different large
components of the grazer ´community´ of a lake such as Little Mere. The comparison
between Daphnia and the main plant-associated filterers (Simocephalus vetulus and Sida
crystallina) showed that the former species is the main grazer during most of the
growing season with grazing rates often above 100 %d-1. However, Sida crystallina was a
significant filter-feeder both at the beginning and end of growing seasons, and
sometimes the main grazer in the lake, with grazing rates occasionally exceeding 500
%d-1. These impacts were short-lived, although the duration of peaks was much larger
in 1999 than in 1998, and very localized in space. Simocephalus vetulus had a maximum
grazing rate of 15-30 %d-1 in both years, but generally its impact is considered small,
albeit not negligible when considered together with the other grazers.

Mesocosm experiments were conducted in each of the growing seasons. Treatments were
nutrient loadings (nitrogen and phosphorus) and applied fish densities (three-spined
sticklebacks, Gasterosteus aculeatus L.). Fish predation was found to be the main
regulator of the grazing impact of Daphnia. Herbivory by Daphnia could be sufficient to
control the size of algal populations in Little Mere, too. A threshold of less, but close to,
4 gm-2 (or circa 3 fry m-2) was the maximum fish density that allowed Daphnia
populations to persist in enclosures. In the lake this threshold may be larger than 4gm-2,
given lily beds act as refuge for the zooplankton against fish predation.

Plant-associated filter-feeders are a by no means negligible grazer ‘community’ in plant-


dominated lakes such as Little Mere, with a large plant cover. Generally,
biomanipulation success can be aided by their buffering of negative effects on plant-bed
persistence. Indirect effects on nutrient cycling in plant beds by chydorid periphyton
scrappers may be another important link, although scarce data on periphyton ingestion
rates and natural periphyton concentrations hamper the testing of this idea. In
conclusion, plant-associated microcrustacea are important components of plant beds
and merit further study. These studies need intensive sampling, to capture effects that
are often short-lived (i.e. less than two weeks), and efforts should be distributed
spatially, as conclusions may vary substantially if spatial heterogeneity is not taken into
account.
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302
CHAPTER 1

General introduction
CHAPTER 1

1.1.- Introduction.

Human life is directly related to freshwater availability. The distribution of human


populations is closely related to the distribution of a freshwater supply, in the form of lakes,
rivers or underground sources of water. Water is very scarce. Only about 0.014 % of the blue
planet’s water is fresh and easily available (i.e. excluding ice and ground waters),
furthermore, only a small part of this amount is potable. More than half of this water is in
lakes, mostly in large deep lakes. However, in many areas of the world, both in the temperate
zone and in tropical areas (Howard-Williams & Lenton 1975), shallow waters are an
important source of water for human consumption too. Interest in shallow lakes also rests on
their economic importance as fisheries or for their amenity values. However, their rich
diversity and role as reservoirs for flora and fauna in the face of human development are
perhaps the main reasons why we should endeavour to better understand and protect these
ecosystems.

The proportionally large area of sediment-water interface and, often, the growth of aquatic
plants, make shallow lakes a particularly complex kind of lake (Moss et al. 1997; see Table
1.1). Surfaces of sediments or plants, with their great opportunities for interactions, are also
where life can display its maximal diversity (Fryer 1968). Equally, aquatic plants can be
thought to be the backbone of a suite of inter-linked mechanisms stabilising the state of
shallow lakes (see Figure 1.1).

It seems a good start to define the object of study; what is meant by ‘shallow lake’?. Indeed it
is conventional wisdom we cannot talk about anything fruitfully if we first do not agree on the
meaning of our words (Plato-Socrates, IV century B.C.). Shallow lakes can be characterised
by three main features.

First, they are not permanently thermally stratified during summer. Heat from the sun is
mostly absorbed by the top one or two metre layers of water in temperate lowland lakes (this
depth will vary with latitude and incident radiation). Cold water is denser and will, therefore,
tend to sink in relation to warmer water. In shallow lakes the entire water column is frequently
mixed, even during the summer. Because of the complete mixing, internal cycling at the
sediment-water interface is more efficient than in deep lakes, at least during the growing
season. Both these factors, mixing and efficient cycling, determine that littoral areas of deep

2
GENERAL INTRODUCTION

lakes and shallow lakes as a whole (effectively a whole-lake ‘littoral area’) are ‘virtual
hothouses’ (Wesenberg-Lund 1912, in Lindeman 1942), absorbing more radiant energy per
unit volume than more pelagial environments. Last, but not least, the bottom of shallow
waters has a considerable coverage of aquatic plants (see Moss 1990, 1995). The extent of this
cover will vary a lot with trophy, depth profile (e.g. Duarte, Kalff & Peters 1986; Gasith &
Hoyer 1997), sediment type, average water temperature and length of the growing season (for
the classic review of these factors see Spence 1982). Then, by ‘shallow’ we mean the group of
temperate lakes with a mean depth of around 3 metres or less.

Figure 1.1. Plants are the centre of a suite of stabilizing mechanisms of the state of shallow temperate lake
ecosystems. Plus and minus symbols on the connecting arrows represent the impact of an increase in the
source factor (i.e. start of the arrow) on the variable in question (end of arrow). Carp and bream may
cause inorganic turbidity through sediment resuspension due to their bottom-feeding habit (e.g. Meijer et
al. 1990). From Stephen (1997).

In general, aquatic plants will have an important structuring role (Jeppesen, Sondergaard &
Christofferson 1997). In shallow lakes there are many processes simultaneously at work,
mediated or directly driven by plants (see Figure 1.1). Many of these appear to stabilise the
plant-dominated state, maintaining clear water and an adequate light climate for the plants. A
good classification of these multiple mechanisms (Table 1.1) could be as follows (modified
from Gasith & Hoyer 1997):

3
CHAPTER 1

1) Limnological effects: changes in physico-chemistry of soil and water. In Table 1.1:


mechanisms 1, 2, 5, 6, 7, 9 and 10.
2) Production and processing of organic matter and nutrient cycling. Table 1.1: 6 and 8.
3) Effects on biotic interactions and community structure related to plants providing a
structured habitat for links in the food web. Table 1.1: 3 and 4.

Table 1.1.- List of possible stabilizing-buffering mechanisms of plant-dominated lakes.

FEEDBACK STABILIZING MECHANISMS IN


‘PLANT-DOMINATED’ SHALLOW LAKES
1) Nutrient uptake from the plant parts in the water. A percentage volume infested (P.V.I.) of 40 % has
been put forward as a minimum for macrophytes being able to clarify the whole lake hypothetically
isolated from any other contributory factor (Howard-Williams 1981; Canfield et al. 1984; Schriver et
al. 1995).
2) Higher denitrification rates of microbial biota in plant beds (see, for example, Karjalainen et al.
1998).
3) Lower fish zooplanktivory in the ‘refugium’ afforded against visual predators (e.g. Timms & Moss
1984). Zooplankton migrate in and out of lily beds in response to fish predation (e.g. Lauridsen &
Buenk 1996). Similar patterns are observed among planktivorous fish in response to piscivory (Persson
& Eklov 1995). It is not known how vertical migration can influence predation in relation to the refuge
effect afforded by different plant densities.
4) Plant-associated microcrustacean communities can have an impact on both phytoplankton (filter-
feeders such as Sida crystallina and Simocephalus, but also facultative microfiltrators such as
Eurycercus; Figure 1.4), and impacted phytoplankton or periphyton (the ‘scraper community’). They
are apparently ‘immune’ to fish predation (e.g. Irvine, Balls & Moss 1990; Beklioglu & Moss 1995;
1996).
5) Faster sinking rates of phytoplankton in the quieter waters of plant beds (non-motile algae species).
6) Periphyton growth on plant surfaces can sequester considerable amounts of nutrients otherwise
available to phytoplankton (Wetzel 1964; Hansson 1992). However, abundant periphyton growth has
been related to macrophyte decline and switches to an algae-dominated state (Phillips et al. 1978;
Howard-Williams 1981, Bronmark & Weisner 1992, but see Balls et al. 1989).
7) Less resuspension of sediment, particularly if flocculent and in large lakes. Plants stabilize it (see, for
example, Barko & James 1991).
8) Nutrient cycling: upon senescence, plants break up into phosphorus-rich organic particles which, via
bacterial decomposition and chemical leaching, release phosphorus (Landers 1982). This phosphorus
may be taken up in part by detritivores (e.g. Eurycercus) but mostly goes to sediment.
9) ‘Allelopathy’: plant species possibly secrete compounds that inhibit the growth of phytoplankton
(Wium-Andersen et al. 1982; Forsberg et al. 1990; Gross 1999)
10) Shading: plant beds create an unfavourable light climate to phytoplankton growth (Frodge et al.
1990).

Under certain circumstances, overcoming enough of these mechanisms will lead to a lake
becoming dominated by planktonic algal growth. Interest in how phytoplankton crops are
maintained in lakes is a very applied issue with strong economic and even political
implications (see Lehman 1986; Peters 1986; Shapiro 1990). For example, water treatment for
human consumption is more expensive in a eutrophicated lake because of the need to remove
large algal crops. Blue-green algae, which can sometimes develop as nutrient concentrations
increase, are sometimes toxic for cattle. Amenity values may be lost as water transparency

4
GENERAL INTRODUCTION

decreases and aquatic plants die back or do not grow in the unfavourable light climate of
algae-dominated lakes.

Initially, plant loss during eutrophication was thought to be a direct consequence of nutrient
enrichment and shading by large algal biomass (ref.). Evidence is growing that rejects the idea
that eutrophication in shallow lakes is a linear process solely controlled by nutrients. In
shallow lake ecosystems at least, there seem to be two different possible states (Holling 1973;
May 1977) (see Figure 1.2), particularly with respect to aquatic plant coverage and water
transparency (Balls et al. 1989; Scheffer 1989; Moss 1990).

In shallow lakes one of these alternative states is the ‘plant-dominated’ state, stabilized by
multiple mechanisms, both biological and physico-chemical (see Table 1.1). The alternative,
and often undesired state (see Moss 1986) is dominated by algal growth. Both states can be
found over a wide range of phosphorus concentrations (Figure 1.2).

Total phosphorus concentration (micrograms per litre)

25 50 100 1000

Alternative states of plant or plankton dominance

Clear water Clear water, dominance by taller plants, stabilized by buffers Clear water with
dominance sparser plants
by plants
PLANT DOMINANCE

REVERSE SWITCHES
FORWARD SWITCHES
(BIOMANIPULATION)

Turbid water, dominance by phytoplankton algae stabilized by buffers Possibly unique


phytoplankton
dominance
PHYTOPLANKTON DOMINANCE (highest conc.)

Increasing stability of phytoplankton dominance

Increasing stability of plant dominance

Figure 1.2. Diagram representing the two stable states in shallow lakes, which are possible across a very
wide range of total phosphorus concentrations. A ‘switch’ towards either state may occur if the
mechanisms apparently stabilizing each state are overcome. See text for details. From Moss et al. 1996.

When the stabilizing mechanisms are finally overcome (often due to human influences), the
system ‘snaps’ and the algal community takes over, leading to a ‘phytoplankton-dominated’

5
CHAPTER 1

state, characterized by high algal biomass and low transparency. This can happen through
excessive plant management, damage to the plants by boat propellers or pesticides leaching
into the water. Often the exact mechanism/s causing the switch are unknown. However, they
all have in common the removal or death of large amounts of aquatic plant growth. Rarely,
‘catastrophic’ climatic events may be determinant, such as a freak hot summer reducing
greatly water levels, or an extremely cold winter freezing the lake down to the sediment (see,
for example, Blindow, Hargeby & Andersson 1997). In both cases the resting parts of plants
cannot sprout back next season. Light is often the limiting factor to aquatic plant growth,
hence the correlations between plant coverage and lake depth (Chambers & Kalff 1985;
Canfield et al. 1985). In very turbid water, vegetation cannot develop at the start of the
growing season (i.e. April to October). Conversely, once plant growth is established, high
water transparency is maintained and buffered from change.

Perhaps the best evidence for two possible states comes from the observation of the
coexistence of the two states in the same body of water. In the English Norfolk Broads,
Hoveton Great Broad and Hudsons Bay (Timms & Moss 1984) are two connected lakes with
areas of similar depth and that receive comparable nutrient loads. Fish can freely move
between these two basins, but while in Hudsons Bay there was lily growth and extremely
clear water one summer (less than 5 µg l-1 chlorophyll-a), Hoveton Great Broad was very
turbid. In this case it was the grazing of zooplankton, Daphnia and perhaps other, plant-
associated, grazers, that kept phytoplankton densities low in Hudsons Bay. It was postulated
this occurred because they were predated less in the lily beds, which somehow interfered with
fish predation.

When a lake switches to an algae-dominated system, restoration may be possible by re-


establishing the buffers of the plant-dominated state (see Figure 1.2). In lakes with sufficient
plant coverage and density, for instance, the plant beds are thought to be used as refugia for
herbivorous zooplankton against fish predation (Timms & Moss 1984). It has been suggested
that these refugia could be provided artificially (Shapiro 1990), though tests with netting, rope
and twigs have had mixed success (Irvine, Moss & Stansfield 1990). On the other hand,
restoration is facilitated if those mechanisms perpetuating the undesired state are removed
first (Shapiro 1990). In addition, the desired switch may be easier to effect when nutrient

6
GENERAL INTRODUCTION

loads are reduced. Though this has not been experimentally tested yet, there is circumstantial
evidence for this being true (e.g. Benndorf 1990).

Biomanipulation: lake restoration and management using the structure of biological


communities.

The switch may be provoked artificially by manipulating the biological community of the lake
(i.e. ‘biomanipulation’). When predation pressure on the herbivores is released by eliminating
their predators, the increased grazing pressure on phytoplankton results in a marked increase
in water transparency (Shapiro & Wright 1984; Shapiro 1990). The same effect can be
accomplished in many ways (see Benndorf 1988), though it is usually by the mass removal of
zooplanktivorous fish (Briand & McCauley 1978; Shapiro & Wright 1984; Carpenter et al.
1987; Post & McQueen 1987; Threlkeld 1988; Lampert 1988; McQueen et al. 1990;
Lammens et al. 1990; Jeppesen et al. 1990; Kasprzak 1995). The ratio of piscivorous to
planktivorous fish can be increased, as a way to maintain naturally low planktivore densities
(e.g. Shapiro & Wright 1984). On the other hand, Edmondson and Abella (1988) suggested
enhancing the reproductive capacity of herbivorous populations by providing their
requirements in addition to removing their main mortality causes.

SMALL-SHALLO W LAKES

LARGE-DEEP LAKES

Figure 1.3.- Representation of the differential response of different types of lake to biomanipulation. In
the extreme, biomanipulated large deep lakes are said to be more stable, as are their biological
communities, than shallower, and smaller lakes. The broken line expresses the manipulated state (from
Gophen 1995).

Nutrient control and biomanipulation as restoration measures developed in parallel. Predation


is an important force in the shaping of ecosystem structure (e.g. Hrbacek et al. 1961, Brooks
& Dodson 1965, Carpenter et al. 1987). Biomanipulation, understood as the deliberate change
in the lake fish communities, is a relatively old technique (e.g. Thereinen and Helm 1954).
However, it has had a broad use only from the 1980s. For some workers, though, grazing was

7
CHAPTER 1

an additional factor explaining algal biomass in different lakes (e.g. Schindler 1978; Canfield
et al. 1984). For other workers, by contrast, transparency was mainly controlled by the food
web and the emphasis was on providing the best conditions for the success of the
manipulation (e.g. Benndorf 1988, 1990; Edmondson & Abella 1988; McQueen 1990).

With internal loading (e.g. Jeppesen et al. 1991; Phillips et al. 1994; Stephen et al. 1997) or
when nutrient control plans are in place, recovery may be speeded up by manipulation of the
food web. However, a combination of these strategies is probably the most cost-effective
approach (Benndorf 1988). Moreover, nutrient-load threshold limits, which are often specific
to each lake, should not be exceeded if biomanipulation is to be successful (e.g. Benndorf
1990; DeBernardi & Giussani 1995).

Knowledge of both the actors and plays enacted in, especially, shallow lakes is still very
fragmentary. Trophic links and indirect effects are not uncommonly discovered during and
after biomanipulation, as opposed to during the planning step (Meijer et al. 1994; Lammens et
al. 1990 and references therein). There are gaps in our knowledge of many of the mechanisms
buffering the plant-dominated state. Much is known about nitrogen transformations (e.g.
Howard-Williams 1981), though little about the relative contribution of plant uptake when in
competition with algae. Some plant species are thought to produce compounds that may
inhibit algal growth, or allelopathic compounds (Gross 1999) but the effectiveness of
allelopathy in situ (e.g. Forsberg et al. 1990) is not understood

On the other hand, the relative importance of nutrients and fish predation (i.e. ‘bottom-up’ and
‘top-down’ processes, respectively) explaining water transparency remains a fundamental
question in Aquatic Ecology.

8
GENERAL INTRODUCTION

Figure 1.4.- From left to right, top to bottom: Sida crystallina, an ovigerous Simocephalus vetulus, an
ephippial Simocephalus vetulus, Eurycercus lamellatus (ovigerous), an adult Daphnia hyalina and an
ephippial Daphnia hyalina. The first three species are the larger-bodied plant-associated Cladocera.

Little attention has been paid to the contributory role that microcrustacean grazers other than
Daphnia species, particularly plant-associated genera such as Sida and Simocephalus (see
Figure 1.4), may have in biomanipulation success in eutrophic shallow lakes. There have been
many studies investigating particular aspects of their biology. For example, spatial
distribution (e.g. Whiteside 1974; Whiteside, Williams & White 1978; Campbell, Clark &
Kosinski 1982; DiFonzo & Campbell 1988), feeding and other behaviour (e.g. Cannon 1921;
Smirnov 1962, 1964; Shan 1968; Szlauer 1973; Fairchild 1981), habitat preference and
general ecology (e.g. Fryer 1968; Quade 1969; Goulden 1971; Whiteside 1974; Paterson
1993), population dynamics of species (e.g. Sharma & Pant 1982; Fairchild 1983; Perrin
1989), effects of fish predation on their abundance (Fairchild 1982), or specific feeding rates
(Downing & Peters 1980; Downing 1981; Pant & Sharma 1982), have been studied.
Strikingly, and as far as I am aware, the issue of their trophic role has not been addressed. The
scarcity in studies has been highlighted repeatedly (e.g. Irvine, Balls & Moss 1990; Moss
1990; Beklioglu & Moss 1995, 1996; Jeppesen et al. 1997; Stephen 1997; Stephen, Moss &
Phillips 1998). Two factors explain the relative lack of quantitative work on this community.
Methodologies have never been standardized in the same way as they have in pelagic research
(Edmondson & Winberg 1971; Bottrell et al. 1976) and there are considerable difficulties of

9
CHAPTER 1

interpretation because of the patchy distributions of these species in plant beds (Irvine, Balls
& Moss 1990). Results are consequently hardly comparable across studies, let alone lakes.
Nevertheless, some very ingenious sampling methods have been devised, often for particular
cases and purposes (e.g. Whiteside 1975, Campbell, Clark & Kosinski 1982). Refer to section
3.1 for a brief review of sampling methods.

This study aims to contribute to filling this gap by quantifying the grazing impact of the plant-
associated microcrustacean community (Cladocera) of a plant-dominated shallow lake. A
subsidiary aim is to look at the extent to which conventional models, i.e. nutrient loading and
open water herbivory, explain transparency.

1.2.- Outline of research.

In particular years and within periods of the growing season (i.e. April to October), low
chlorophyll-a concentration and high water transparency cannot be accounted for by Daphnia
grazing in the experimental lake, Little Mere, in Cheshire (UK) (see Chapter 2 for details).
There are other processes in operation related to plant cover, either removing phytoplankton
faster than it is produced (‘top-down’ alternative control, but not by Daphnia), or mechanisms
depleting nutrient resources otherwise available to algal growth (‘bottom-up’ control
mediated by plants, directly or indirectly).

The null working hypothesis is that ‘the plant-associated microcrustacea community


(Cladocera) has NO significant effect on water clarity in the experimental lake’. To test this
the grazing rate of the plant-associated Cladocera community has been estimated in order to
compare it with that of more ‘open-water’ cladoceran species, particularly Daphnia spp. This
rate is expressed in percentage of volume of lake water (litres) filtered per day by the
populations of each species in given areas centred around the sites sampled (i.e. % day-1; see
Map in section 4.2).

In order to estimate grazing rates (% day-1), the following variables were estimated (see
section 4.2.3 for details):

a) Densities of species, both in number of individuals per gram of plant (plant-associated)


and per litre of lake water (‘open-water’).

10
GENERAL INTRODUCTION

b) Plant biomass across areas (grams plant m-2 lake bottom)


c) Filtering rates (filter-feeders) or ingestion rates (periphyton scrapers) for each species (per
individual: µl water day-1 or µg periphyton day-1).
d) Plant cover and depth profile of the lake.

The relative role of nutrients and grazing in the control of algal biomass (particularly in
summer months) remains an open question. A controlled mesocosm experiment running
separately during two growing seasons was conducted in Little Mere (Cheshire), testing both
nutrients levels and zooplanktivorous fish (hence grazer densities) and their effects on algal
biomass and water transparency.

1.3.- Thesis layout.

Chapter 2 to introduces the experimental lake, Little Mere. Chapter 3 describes the densities
of the cladoceran species in Little Mere. It is divided into four: distribution of vegetation,
zooplankton (i.e. ‘open-water’ species), periphyton scrapers in lily beds, and lily-associated
filter-feeders. Patterns of abundance of the main zooplankton species are examined through
seasonal size-structure of the populations of the main filter-feeders, Daphnia spp. and
Simocephalus vetulus. Two simple egg-ratio models have been calculated for these two
species.

In Chapter 4, field abundances of the main grazers (Chapter 3) are combined with clearance
rate estimates, to estimate grazing rates. Clearance rates are from short-term feeding
experiments made in '99 using radioactively-labelled foods.

Chapter 5 investigates the issue of whether nutrients of grazing controls the summer
phytoplankton in Little Mere, using the results of two separate enclosure (mesocosm)
experiments..

Chapter 6 brings together the information presented in the previous chapters to defend the
thesis that plant-associated Cladocera are by no means negligible grazers in Little Mere,
particularly during April to October.

11
CHAPTER 2

The experimental lake: Little Mere. A summary of its


recent history.
CHAPTER 2

The experimental lake: Little Mere

The experimental lake, Little Mere (53°20’N, 2°24’W; 50 metres above sea level; NGR
SJ733823) is a small, shallow fertile lake situated in Cheshire, North West England (details in
Table 2.1). It belongs to a group of over 100 lakes (area <1 ha to about 50 ha), the West
Midland Meres (Figure 2.4). Most of them have originated from the deposition of glacial
moraines in a low-lying area at the end of the last glaciation (circa 13,000 B.P.) (Sinker
1962). Little Mere, however, was formed by damming in the 18th century the outflow stream
of another (natural) lake located just upstream (Figure 2.3; Stephen, Moss & Phillips 1998).

Table 2.1. Main physical features of Little Mere.

Surface area1 0.028 km2 (2.8 ha)


Catchment area1 3.2 km2
Depth (max./mean)2 2.8 m / 0.7 m
Retention time/ 11-146 days
Flushing rate3 9-0.5 % day-1
Approximate volume4 21 x 103 m3
1
Carvalho (1994); 2Carvalho (1994) and own measurements
3
Stephen (1997); 4Stephen (1997) and own measurements

Little Mere is immediately surrounded by a narrow fringe of mixed woodland and partly by
private housing. The relatively large catchment area is occupied by a golf-course and
agricultural land, as can be seen from an aerial view taken in 1992 (Figure 2.1).

Figure 2.1. Aerial photograph of Little Mere taken in 1992. Mere Mere can be seen to the right. A golf-
course is located on its west bank (further away in the photograph). A road separates the lake from
pasture fields, on the other bank. Large nymphaeid beds can be seen in the lake, after a period when
nutrient loads from a sewage treatment works located closer to the golf-course discharged large amounts
of organic sewage directly into it (see also Figure 2.5).

14
RECENT HISTORY OF LITTLE MERE

The North West Midland Meres have been described as Britain’s naturally eutrophic lakes
(Reynolds & Sinker 1976). Moreover, there is paleolimnological evidence for blue-green
algal blooms having occurred thousands of years ago (McGowan et al. 1999). Deposits of
apatite, a phosphorus-rich mineral, are present in the glacial drift (Reynolds 1979; but see
Moss et al. 1997). Nitrogen-to-phosphorus ratios are low in many cases during most of the
year (frequently less than 5, suggesting nitrogen rather than phosphorus-limitation; see Figure
2.2). Moss et al. (1994) found that in the West Midland Meres the relative importance of
grazing versus nutrients in the maintenance of algal crops was a function of depth. In deeper
meres (i.e. deeper than 3 metres) it was nitrogen which limited phytoplankton growth, while
in the shallower meres inverse correlations between peak chlorophyll-a concentrations and
cladoceran density (ind l-1), particularly of Daphnia species, were taken as indicative of top-
down regulation of algae.

Little Mere. TDIN:TP during the period 1990-1999 Rostherne Mere. TDIN:TP during the period 1990-1999
100 25
160
90

80 20

70

60 15
TDIN:TP

TDIN:TP

50

40 10
30

20 5
10

0 0
ene-90 ene-91 ene-92 ene-93 ene-94 ene-95 ene-96 ene-97 ene-98 ene-99 ene-00
ene-90 ene-91 ene-92 ene-93 ene-94 ene-95 ene-96 ene-97 ene-98 ene-99 ene-00

Figure 2.2. TDIN:TP ratios in Little Mere (left) and Rostherne Mere (right) in the period 1990 to 1999.
Large seasonal oscillations are evident in Little Mere. In Rostherne Mere ratios are maintained below 5
during most of the year.

15
CHAPTER 2

Figure 2.3. Map of the group of linked lakes Little Mere, Mere Mere and Rostherne Mere. The three lakes
have been monitored for the past ten years (i.e. since 1990), for basic limnological information (water
chemistry, temperature, phytoplankton and zooplankton) every three weeks or less. The location of the
sampling station for this monitoring in each lake is shown in the map. Two sewage treatment works were
located on the west bank of Little Mere and on Rostherne brook, both operating until June 1991 (see text
for details).

16
RECENT HISTORY OF LITTLE MERE

Figure 2.4- Little Mere, Mere Mere and Rostherne Mere are located in the northern part of a plain which
includes well over 100 lakes of varying depth and size in the West Midlands (England). The estuary of the
river Mersey and Liverpool city lie to the north of this map. Mean annual conductivities (µS cm-1), mean
winter dissolved inorganic nitrogen concentrations (nitrate-N plus ammonium-N) (mg l-1) and mean
annual total phosphorus concentrations (µg l-1) are shown in that order, below the names of a selection of
meres. Asterisks indicate lakes known to be polluted by stock or sewage effluent. Shaded areas show relief
above sea level. The map is orientated in a north to south direction. Scaled to 1:400,000. From Moss et al.
(1997).

17
CHAPTER 2

Pollution history of Little Mere.

Until June 1991 this lake was being used effectively as an oxidation pond for the sewage
effluent of a treatment plant located on one of its banks (Carvalho 1994). There was an ample
supply of phosphorus, particularly during the growing season (April to October). This plant
had been operating since ca 1932, overloaded at three times its design capacity. Indeed,
considerable ammonium and phosphate concentrations were measured at the outflow of the
lake before diversion (Carvalho 1994). Sewage was finally diverted from this plant for
treatment elsewhere in June 1991.

There is no information on the biology of Little Mere prior to 1990. The limnology of the
group of three lakes has been followed since that year, providing an excellent opportunity to
follow the differential response of a shallow lake (Little Mere), and a deep one (Rostherne
Mere, Zmax= 28 m) to nutrient load reduction. In addition, Mere Mere, upstream, has never
received effluent nutrient loads and diffuse inputs are probably unchanged from pre-diversion
levels. Mere Mere, thus, can act for comparison with Little Mere (see Carvalho et al. 1995).

After diversion of the sewage effluent, Rostherne Mere, which received inputs from the Little
Mere works through the Rostherne Brook inflow as well as from a smaller plant discharging
directly into the brook (see Figure 2.3), has responded only weakly to the reduction in loads.
Load reduction on the lake is manifest from measurements made at the inflow itself (Carvalho
et al. 1995). The lack of response in the lake is surprising considering this input accounted for
71 % of the nutrient budget (Moss et al. 1997). Two possibilities are that either the nutrient
budget is flawed, missing out important inputs or grossly underestimating those identified, or
that there are mechanisms of internal release of the phosphorus accumulated in the sediment
during the high load periods (Moss et al. 1997).

In contrast, Little Mere responded almost immediately (see Figure 2.5), with a drastic
reduction in measured total phosphorus and spring chlorophyll-a concentrations in the water.
Reducing conditions in the pre-diversion period were reflected in extremely high
concentrations of ammonia (i.e. 5-10 mg l-1). Since diversion, both ammonia and nitrate levels
have been virtually undetectable, particularly during growing seasons (April to October). It is
possible nitrogen becomes limiting to phytoplankton growth during summer months. What is
not known is if limitation comes about through algal growth or if other processes such as, for
example, plant uptake from the water, are important.

18
RECENT HISTORY OF LITTLE MERE

Conductivity and pH, on the other hand, have not experienced significative variations in
relation to sewage diversion or in the nine years after diversion (see Figure 2.6). Water levels
have been relatively stable, with minima generally in June-July-August, months when both
inflow and outflow run dry. Despite the large potential for algal growth, phytoplankton
density has remained low and measured chlorophyll-a concentrations have seldom exceeded
about 30-50 µg l-1 (see Figure 2.5).

Mean Total Phosphorus and chlorophyll-a


4000 (growing seasons 1990-1999) 100
3500 90
Total Phosphorus

micrograms chl-a/litre
80
micrograms TP / litre

3000 Chlorophyll-a
70
2500 60
2000 50
1500 40
30
1000
20
500 10
0 0
1990

1991

1992

1993

1994

1995

1996

1997

1998

1999

Figure 2.5. Diversion of sewage input from June 1991 had a clear effect on mean total phosphorus
concentrations. Chlorophyll-a values are very variable among years and do not seem to follow changes in
TP concentration. Plotted are growing season (i.e. April to October) means with one standard error at
either side of the mean.

Conductivity and pH in Little Mere during the period 1990-1999


10.5 pH 700
conductivity
10
600
Conductivity (microS cm-1)

9.5
500
9

8.5 400
pH

8 300
7.5
200
7
100
6.5

6 0
ene-90
abr-90
jul-90
nov-90
mar-91
jun-91
sep-91
ene-92
abr-92
jul-92
nov-92
feb-93
may-93
ago-93
nov-93
mar-94
jul-94
ene-95
jun-95
nov-95
abr-96
jul-96
nov-96
may-97
nov-97
abr-98
sep-98
feb-99
jul-99
dic-99

Figure 2.6. Conductivity (µS cm-1) and pH in Little Mere over the period 1990-1999. The pH has remained
quite stable, in a range of values between 7 to 8 in most dates, with occasional peaks relating to a larger
photosynthetic activity during periods of high algal biomass. Conductivity was generally low, with values
generally around 400 µS cm-1.

19
CHAPTER 2

Changes in the biota over the past ten years in Little Mere.

Prior to diversion, Little Mere preserved a surprisingly large cover of lily growth, even with
total phosphorus (TP) concentrations in the order of mg l-1 (see Figure 2.5). Plant coverage,
particularly nymphaeids, expanded considerably, judging from an aerial photograph taken in
1992 (see Figure 2.1), soon after diversion, to the circa 40 % of total surface found in 1996
(Beklioglu & Moss 1996) and at present (Table 2.2 & section 4.2.2). Submerged vegetation
covered around 30 to 70 % of the lake bottom in the first period after diversion (1991-1993),
mainly the fine-leaved Potamogeton berchtoldii Fieber, with some scattered patches of
Elodea canadensis Michaux (Beklioglu & Moss 1996). Plant dominance seems to have
switched annually between the former species and Callitriche hermaphroditica L. (Stephen,
Moss & Phillips 1998). Submerged plant coverage has increased to near 100 % in the past
three years. Lately (i.e. 1998 and 1999), Ceratophyllum demersum L. has been clearly more
abundant than in previous years, in combination with dense and heterogeneously distributed
patches of C. hermaphroditica (see Table 2.2 and Appendix-D).

Table 2.2. Submerged aquatic flora in Little Mere in the period 1998-99. Percentage cover and species
composition of the submerged aquatic flora are shown for both years, separately for the periods between
the beginning and middle, and middle and end of the growing season (April to October). See text for
details.

Maximum macrophyte cover c98 %


Maximum lily cover c40 %
Aquatic flora 1998 Beginning-Middle of growing season C; P; E; Cal; Lt
composition* Middle-End of growing season C+; Cal+; Lt; E; P
(submerged only) 1999 Beginning-Middle of growing season C+; Cal+; E; Lt; P
Middle-End of growing season C+; Cal; Lt; E; P
+
*Key for abbreviations: C = sparse Ceratophyllum demersum; C abundant; P = sparse Potamogeton
berchtoldii; Cal = sparse Callitriche hermaphroditica; Lt = sparse Lemna trisulca; E = sparse Elodea
canadensis. Sparse/abundant refer to coverage, not percentage-volume-infested (PVI) or biomass.

The zooplankton community has changed considerably from year to year. Before diversion
(i.e. 1990), the oxygen-depleted conditions produced by decomposition of the poorly treated
sewage precluded many fish from living in the lake (Carvalho 1994). The virtual absence of
predation pressure allowed large populations of the large-bodied, and otherwise vulnerable,
Daphnia magna Strauss to develop. This species can live under very low O2 concentrations
thanks to the production of haemoglobin, a very efficient carrier of oxygen (Carvalho 1984).
Extremely low chlorophyll-a concentrations were measured during summer 1990 (i.e. less
than 5 µg l-1 chlorophyll-a; see Figure 2.7) despite the huge potential for algal growth
reflected in nutrient concentrations, suggesting intense herbivory by this large Daphnia

20
RECENT HISTORY OF LITTLE MERE

species (Carvalho 1994). In the two summers immediately after diversion (i.e. 1991 and
1992), the peak size of populations of Daphnia magna became smaller as the lake
reoxygenated and fish reinvaded from Mere Mere upstream, intensifying predation on their
populations (Carvalho 1994). Average summer chlorophyll-a concentrations increased
slightly as the Daphnia magna populations were replaced by the smaller and less efficient
Daphnia species (mainly D. hyalina L.). Water clarity was generally maintained.

Despite the re-entrance of fish, predominantly of perch (Perca fluviatilis L.), following
diversion (Beklioglu & Moss 1996), significant, though decreased, populations of D.magna
were maintained for some time, perhaps because of the presence of sufficient plant bed cover.
These plant beds, it is suggested, acted as ‘refuge’ for the zooplankton against fish predation.
Indeed, as Daphnia magna populations were replaced by the smaller Daphnia species,
significantly higher numbers were found in lily beds than in more open waters. In contrast,
submerged plants, especially the fine-leaved P.berchtoldii, did not seem to constitute an
effective refuge (Beklioglu & Moss 1996; Moss, Kornijow & Measey 1998). The role of
aquatic plants as stabilizers of the clear-water state in Little Mere has been repeatedly
emphasized (e.g. Carvalho 1994; Moss et al. 1994; Beklioglu & Moss 1996; Stephen et al.
1998; Moss et al. 1998). The exact mechanisms mediated by plants (or caused by the plants
themselves) that are relevant in Little Mere, though, are not known.

Summer peak Daphnia density against chlorophyll-a


500 30
450
total Daphnia/litre 25
micrograms chl-a/litre

400
chlorophyll-a
Total Daphnia/litre

350
20
300
250 15
200
10
150
100
5
50
0 0
1990 1991 1992 1993 1994 1995 1996 1997 1998 1999

Figure 2.7. Spring-early summer maxima (columns) of Daphnia spp (ind l -1) plotted against the co-
occurring value of chlorophyll-a (µg l-1; line). Top-down effects are generally evident prior to 1996. See
text for details.

21
CHAPTER 2

In general, top-down control by Daphnia seems to have been the most powerful influence on
phytoplankton (Stephen, Moss & Phillips 1998). When few Daphnia were present,
chlorophyll-a values were considerably higher, for example during 1995 (see Figure 2.7). In
1996, on the other hand, low and unexplained values of chlorophyll-a cannot be accounted for
by Daphnia grazing.

Although, the ecosystem seems to have stabilized in a clear water plant-dominated state, there
is considerable variation in chlorophyll-a among years (see Figure 2.5). Likewise, there is
large variability in the seasonal behaviour of Daphnia spp populations from year to year. For
example, in 1996 there was no Daphnia summer peak at all, and in early summer 1995 and,
especially, during the summer of 1999, the open-water grazer community was dominated by
small cladocerans such as Ceriodaphnia spp and Bosmina longirostris. The size of peaks, and
perhaps the extent of top-down control by Daphnia, varies considerably among years too.

The remarkable resilience of the lake even in the absence of grazers such as Daphnia spp.
suggests other buffering mechanisms of the plant-dominated system are in operation. The
expansion of rooted macrophyte stands has been related to this resilience (Beklioglu & Moss
1996). What is not known is what are the relative roles of direct mechanisms mediated by
plants, for example the nutrient uptake by plants in competition with planktonic algae, and
indirect ones, for instance the grazing of phytoplankton by the plant-associated cladoceran
genera (Sida and Simocephalus; see Figure 1.4).

Plant-associated genera (Eurycercus lamellatus O.F.Muller, Sida crystallina O.F.Muller,


Simocephalus spp O.F.Muller/Koch) were first detected in 1993, two years after diversion
(Carvalho 1994), though sampling had been strictly planktonic (i.e. only in ‘open waters’;
Figure 2.3). In 1993 these species constituted 21 % of total zooplankton numbers, while in
1994 none were recorded and in 1995 only 2 % were of plant-associated species (Stephen
1997). Only open waters were routinely sampled, and it is possible low densities of these
genera were simply due to the lack of suitable habitat around the sampling station (see, for
example, Table 2.3). It seems, however, expansion particularly of lily bed coverage, but also
submerged vegetation (reported in Beklioglu & Moss 1995, 1996; see Table 2.2) has been
accompanied by a growing role for these plant-associated grazers.

22
RECENT HISTORY OF LITTLE MERE

Table 2.3. Diversity of cladoceran species found in Little Mere in the period April 1998-April 2000.
Frequency (number of sampling occasions when detected of a total of 36) and relative abundance of each
species during the sampling period are also indicated.

Species/genera Frequency Relative abundance in Little Mere


(number of sampling (1998-2000)
occasions when detected;
total: 30)
Acroperus harpae Very scarce to scarce (16) 0-209 ind g-1 DW lily
Alona spp Scarce (29) 0-450 ind g-1 DW lily
Alonella spp Very scarce (29) Occasional
Bosmina longirostris Very abundant (22) 0-1264 ind l-1
Ceriodaphnia spp Very abundant (25) 0-1358 ind l-1
Chydorus spp Very abundant (29) 0-418 ind g-1 DW lily
Daphnia cucullata Very scarce Occasional
Daphnia hyalina-longispina Very abundant (28) 0-406 ind l-1
Daphnia pulex Very scarce Occasional
Daphnia magna Very scarce (occasional) Occasional
Diaphanosoma brachyurum Very scarce (occasional) Occasional
Eurycercus lamellatus Scarce to medium (15) 0-76 ind g-1 DW submerged plant
Graptoleberis testudinaria Scarce to medium (28) 0-551 ind g-1 DW lily
(occasionally abundant)
Peracantha truncata Abundant (27) 0-380 ind g-1 DW lily
Pleuroxus denticulatus Very scarce (occasional) Occasional
Polyphemus pediculus Abundant (25) 0-2700 ind g-1 DW lily
Scapholeberis mucronata Abundant (27) 0-270 ind g-1 DW lily
Sida crystallina Abundant (20) 0-1731 ind g-1 DW lily
Simocephalus vetulus Abundant (27) 0-386 ind g-1 DW lily
Simocephalus spp Very scarce (occasional) Occasional

Thus, in particular years and within periods of the growing season (i.e. April to October), low
chlorophyll-a concentration and high water transparency cannot be accounted for by Daphnia
grazing in Little Mere (see Figure 2.7). There are other processes in operation related to plant
cover, either removing phytoplankton faster than it is produced (‘top-down’ control, but not
by Daphnia), or mechanisms depleting nutrient resources otherwise available to algal growth
(‘bottom-up’ control mediated by plants, directly or indirectly).

23
CHAPTER 3

Population and community dynamics of the plant-


associated Cladocera in Little Mere over two years.
CHAPTER 3

3.1. Introduction.

Intensive and detailed studies of the plant-associated cladoceran community are few.
Sampling techniques introduce a large amount of error when sampling from plants. The same
sampling techniques have not been widely used very often, making comparability a problem.
Thus, few patterns have emerged from sampling studies on this community (Scheffer 1998).

High levels of spatial heterogeneity, particularly for periphyton scrapers, both in numbers and
species composition, are expected across short distances because the animals very often live
in close association with the plant surfaces (Fryer 1968; Quade 1969; Whiteside 1970;
Fairchild 1981). Because of large variability, a higher sampling effort is needed to detect
effects over ‘noise’ (Downing 1986). One solution may be to use surrogates (for example,
macrophyte species composition and/or biomass; Downing 1986) that are ‘well correlated’
with population density (i.e. r>0.7; Cassie, in Edmondson & Winberg (eds.) 1971). A
mathematical relationship was found by regression analysis, allowing subsequent use of the
‘easier-to-sample variable’ as a predictor of animal density, instead of directly sampling for
the animals (Downing & Cyr 1985; Downing 1986).

±10 cm.

±100 cm.

±1 cm.
± 15 cm.

Figure 3.1. ‘Pattern’ sampler design. A number of inverted funnels facing down towards the lake bottom
are held in a frame, as shown. Each funnel leads to a small bottle containing all animals caught while
migrating upwards. This sampling method is only useful for relatively homogeneous substrates such as
Chara plant beds and mud habitats. In the photo, this sampler is used to assess the small-scale distribution
of chydorid populations in the mud habitat of Lake Naardermeer (The Netherlands).

In the search for better sampling methods two basic approaches have been followed. An
example of the first, a non-interference method, is given by Whiteside (1974). He used square
funnels fitted head downwards in a frame that can be positioned over the plant bed. As the
animals migrate upwards at night, they are collected in the funnel which leads to a plastic
collecting bottle (see Fig. 3.1). In this way, detailed information can be obtained on both

26
PLANT-ASSOCIATED CLADOCERA DYNAMICS

density and close spatial distribution (“pattern”, in Whiteside’s terms; see Whiteside &
Williams 1975; Whiteside, Williams & White 1978). However, Paterson (1993) showed in his
study of the littoral microcrustacea of a small oligotrophic lake, that estimates of cladoceran
and cyclopoid density obtained using funnel traps are only about a tenth of estimates from
traditional cores. Moreover, he could not find evidence of vertical migration, although he
suggested this may be a lake-specific effect of low edible-phytoplankton concentrations and
relatively high night-oxygen concentrations in the plant beds. This method has not been
widely used and is inferior to the second approach.

The second approach to sampling these populations involves taking a representative “section”
of the plant-animal system, with a “jaw” or box sampler, generally around the stem. Many
types of these samplers have been used (see review by Downing & Cyr 1985) varying in the
way the plant is collected (e.g. mesh bags, Vuille 1991).

These variations in method across researchers add to those difficulties due to the nature of the
samples themselves. Samples are of the biota associated with a living plant surface of diverse
characteristics (e.g. age, amount of periphyton, quality of this periphytic community and plant
species). The complexity of this system means that the size of the sample is more of an issue
than in samples taken in more “open” waters (i.e. zooplankton samples). In addition, the
scales of change in population density may vary with the animal species in question,
depending on their habitat preference and, linked with this, their feeding mode (Fryer 1968).
For example, Sida crystallina, a filter-feeder, will probably have a larger spatial scale of
change (a ‘coarser grain’; see, for example, Fairchild 1981) than Graptoleberis testudinaria
(e.g. Quade 1969), a periphyton scraper, which lives in a finer-grain habitat. The latter species
may be more dependent on leaf quality (i.e. species composition of the periphytic community,
proximity to substrate, light climate, presence and abundance of macroinvertebrates, etc.),
which will most likely change in the space of centimetres, rather than metres.

Spatio-temporal scales of change have often been totally ignored in modelling. For example,
Edmondson’s (1960) original egg-ratio model assumes populations are distributed uniformly
across space. However, Elster & Schwoerbel (1970) have found differences of more than a
factor of 100 in population numbers when sampling in different stations in the same lake.
Furthermore, animals may migrate horizontally in a diel cycle in response to predators or
“shore-avoidance”, Gliwicz & Rykowska 1992; Davies 1985; Lauridsen & Buenk 1996).

27
CHAPTER 3

Fairchild (1981) found 30 metres to be the limit of representativeness of point samples of


population density of Sida crystallina. He also found juveniles and adult individuals to exhibit
different swimming behaviours.

Vertical migration may also be important. Densities obtained using common destructive
methods tend to refer to either only one part of the plant, generally close to the water surface,
or to the whole plant block, ignoring potential vertical differences in density (e.g. Downing &
Cyr 1985; Downing 1986). Densities may even change with different plant parts (DiFonzo
and Campbell 1988).

With respect to sampling frequency and intensity, a compromise must be reached between
what is practical within the time and resources available (Downing & Cyr 1985; Peters &
Downing 1984), and what level of accuracy is satisfactory given the purpose of the
investigation. A large literature was generated during the 1970s and early 1980s
communicating advances and criticisms on zooplankton population modelling, based on the
egg-ratio method (Edmondson 1960, 1968, 1974). The repeated census of a population yields
counts that can then be related to population growth using this method. The egg-ratio is the
number of eggs born on average by females in the population. Because cladoceran species are
usually parthenogenetic, no correction is needed to account for unequal sex ratios in the
population; it is effectively composed of females, at least during episodes of population
growth.

Some of the egg-ratio equations have helped to direct sampling programmes, so important
properties of populations were taken into account depending on the aim of each study. Seitz
(1979) assessed the influence of the variances of birth and death estimates on the accuracy of
different models by comparing the correlation coefficients between “true” values, and the
calculated values given by them. He found Paloheimo’s model to be valid if size-selective
predation was not to be expected (Paloheimo 1974). Gabriel et al. (1987) showed the
sampling interval can strongly influence reliability of estimates, by comparing different
formulae in computer simulations of growth. Indeed, faster growing populations reach stable
age distribution faster and, therefore, a more frequent sampling will be required. Keen &
Nasaar (1981) suggest frequency of sampling should be chosen such that 95 % confidence
intervals, calculated for birth rate, overlap for successive collections. The general assumption
of all egg-ratio equations is that rates are constant (i.e. the population is growing

28
PLANT-ASSOCIATED CLADOCERA DYNAMICS

exponentially). If births and deaths were “time-dependent” and could not be well
approximated by an intensive sampling effort (Lynch 1982), or when not all members of a
population are expected to experience similar temperatures and, thus, have different
developmental periods (Hall 1964; Prepas & Rigler 1978), then estimates of growth rate, birth
and death, cannot be reliably provided by egg-ratio methods. “Substrate-associated”
cladocerans such as chydorids, and some other species of the order such as Simocephalus spp
or Sida crystallina, may have growth rates dependent on very variable habitat conditions, such
as periphyton abundance and composition, or availability of plant substrate. In such cases
doubt remains whether egg-ratio methods are applicable.

In this chapter, the distribution and seasonality of cladoceran species over a two-year period
in Little Mere are closely examined. Size-structure changes and a population dynamics model
using the egg-ratio method are then examined for the main filter-feeders in the lake.
Distribution patterns of species, emerging from the analysis of data on densities across the
lake, are complemented with an analysis of the small-scale variability of these densities.
Results are discussed in relation to both potential regulatory forces of distribution patterns
(e.g. fish predation, food availability), and ‘habitat preference’ of different cladoceran species.

The main aims for this chapter are

1) To describe sampling techniques used, particularly for plant-associated Cladocera.

2) To present population sizes of main grazers in Little Mere so that, in conjunction with
measures of ‘feeding intensity’ (see Chapter 4), grazing rates can be estimated for both the
‘open water Cladocera community’ and the plant-associated. The ultimate aim is to compare
the trophic role of Daphnia and other, plant-associated species, in the maintenance of clear
water.

3) To show spatio-temporal differences in population densities and dynamics of cladoceran


species in Little Mere and discuss reasons for these.

29
CHAPTER 3

3.2. Materials and Methods.

3.2.1. Sampling programme description.

Lily beds and more open waters (i.e. with only submerged vegetation growth) were sampled
with different techniques (section 3.2.1). Samples were taken of floating lily leaves and of
petioles (5 replicates) at three sites in the lake (for Map see Fig. 3.2). No samples were taken
of the submerged leaves of yellow lilies (Nuphar lutea). This was forced by time constraints.
Uncertainty remains whether this habitat holds important reservoirs of grazers. Nevertheless,
samples were taken of petioles reaching close to the bottom (see section 3.2.4 for details).
Both leaves and petioles and the water around them were collected using purpose-designed
samplers (section 3.2.3).

4
3 2
1
5
Figure 3.2. Map of Little Mere with the outline of lily bed growth, generally growing close to the shoreline
and sampling sites location. Sites 1, 2 and 5 are located within lily beds while sites 3 and 4 are over
submerged plant beds (relatively ‘open waters’).

At the other two sites, with only submerged vegetation (generally beds of Potamogeton
berchtoldii, Ceratophyllum demersum mixed with Elodea canadensis, or rooted and half-
floating Callitriche hermaphroditica), samples were taken at surface level and at arm’s length
depth (i.e. about 30 cm deep).

Generally two 10-l depth-integrated samples of the water in the plant beds were also collected
at the five sampling sites. For open water grazers (i.e. mainly Daphnia spp, Ceriodaphnia spp
and Bosmina longirostris) units were calculated by extrapolating the number of litres sampled
to the volume of water in a hypothetical column of base 1 m2 and height the depth around the
sampling site (see depth map in Figure 4.14).

30
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Densities are first expressed per unit of plant biomass sampled (i.e. number gram-1 of leaf,
petiole or submerged plant part). Extrapolation for plant-associated grazers required
independent estimates of areal plant biomass across sites. Details of plant surveys conducted
to estimate these are given in Chapter 4.

Little Mere was intensively sampled from April 1998 to April 2000. However, there is
information on mainly open-water cladocerans sampled at a central location in the lake for a
longer period (from 1990 onwards; see Figure 2.3) as part of an ongoing monitoring
programme of the system of three lakes (see Chapter 2). During the intensive programme,
samples were taken generally every week during the growing season (April to October) in
both years, and generally every two to three weeks the rest of each year.

In addition to animal samples, a 1-litre sample of unfiltered water and another of water
filtered through a 25 µm nylon mesh were taken at each site, in order to estimate chlorophyll-
a concentrations and suspended solid parameters, all related to the food concentrations in the
water experienced by the biota. Cladocerans can probably only ingest particles smaller than
100 µm, depending on shape, and more likely smaller than 25 µm (Burns 1968b). The
fraction sampled represents most of the “more edible” algae available to these grazers.
Generally temperature was also measured at each site at the surface and half a metre down.
Only one reading for the whole lake was taken outside the growing season (i.e. November to
March). Developmental times vary with temperature, influencing birth rates as inputs of egg-
ratio models (Bottrell 1974, Paloheimo 1974). Temperature is also thought to influence
filtering rates (e.g. Mourelatos & Lacroix 1990).

Sampling was stratified (sensu Cassie, in Edmondson 1971, e.g. Lampert & Taylor 1984).
From the outset a decision was taken to sample both lily beds and submerged plant beds.
Further stratification of sampling involved selecting the lily beds to sample on the basis of
‘size’ and location in the lake. One lily bed sampled grows in very shallow water (average
depth 0.5 m) dominated by yellow lily (Nuphar lutea). A second, deeper, bed sampled
(average depth approximately 0.7 m) is a monospecific white-lily stand (Nymphaea alba), and
the third bed selected contains a mixture of both lily species. See Table 3.1 for details.

31
CHAPTER 3

Table 3.1. Size (m2) and relative proportions (%) of different lily-bed types in Little Mere, Cheshire
(survey July ’98). Each type of bed is represented by one sampling site, as shown.

Lily bed type


Yellow water lily White water lily Mixed yellow & TOTAL
(Nuphar lutea) (Nymphaea alba) white water lilies
Surface area (m2) 4,600 1,100 5,200 10,900
% of total lily bed area 42 10 48 100
% of Little Mere 16.8 4 19.2 40
Sampling site code number 2 1 5

The number of sampling sites was limited to five in order to intensify replication per site. It
was felt a larger number of sites would not add significantly to the coverage, while it was
paramount to monitor variation due to sampling methodology and the biology of species as
much as possible within time constraints. The number of replicates taken was set by time
constraints. Indeed, sampling more replicates would perhaps include differences related to
different timing of sampling within each sampling occasion. Sampling generally began at
around midday and was completed in about 3-4 hours during the growing seasons.

3.2.2. Sample processing and information recorded.

Samples from plants (i.e. leaves, petioles and parts of submerged vegetation of various
species) were washed of any attached animals into a bucket by means of a strong jet of water
from a bottle, using a hand to help detach tightly associated animals. Animals retained by
meshes were carefully washed into the bucket, too. The plant part was then bagged for further
processing in the laboratory. The water and animals in the bucket were strained through a fine
mesh (Nytex 50 µm) to remove most of the water, and the animals were washed into a sample
bottle. These samples were briefly narcotized with chloroform (2 minutes) and preserved with
absolute ethanol (in at least 70 ethanol : 30 sample proportions). Each sample bottle was kept
in the bag with its corresponding plant part.

Once in the laboratory, plant parts were placed on labelled pieces of aluminium foil and left to
dry in a dessicator oven (T=60°C, 24 hours). A series of variables was recorded for each plant
part, as described in Table 3.2.

32
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.2. Variables recorded for the plant parts of samples taken during a sampling programme for
microcrustacea in Little Mere, Cheshire, during growing seasons ’98 and ‘99.

PLANT PART Variable name Codes adopted for Meaning of codes


each variable
LEAF Species y/w Yellow/white lily

Approximate age 1/2/3 Young/Mature/Senescent

Degree of attack by 0/1/2/3/4 Not eaten at all/Very slightly eaten/Quite


macroinvertebrates eaten/Very much eaten/Heavily eaten
PETIOLE Length 8-79 cm -
‘Redness’ 0/1/2/3 Light green/Mature green/Reddish/Very red
SUBMERGED PLANT PART Approximate age 1/2/3 Young/Mature/Senescent

Species C/E/P/Cal. Ceratophyllum demersum


Elodea canadensis
Potamogeton berchtoldii
Callitriche hermaphroditica

All animals were counted under a dissecting microscope at 30X magnification. To concentrate
the sample, most of the liquid was decanted, the residue left to settle and further liquid
removed using a syringe. Five millilitres at a time of this concentrate were placed in a groove
cut in a plastic disc. This circle could be rotated on a platform fixed to the base of the
microscope. On the few occasions when the sample was extremely dense or a lot of sediment
had been inadvertently collected, the sample was thoroughly mixed and a subsample taken.
Either 100+ individuals of the commonest species were counted, when the sample was not too
dense, or at least three subsamples counted and values averaged, when the sample contained a
lot of sediment.

Carapace lengths of at least 30 individuals of each of the largest species (i.e. Daphnia spp–
generally Daphnia hyalina-longispina–, Simocephalus vetulus, Sida crystallina and
Eurycercus lamellatus) were measured, using an ocular micrometer (max. precision 10 µm).

3.2.3. Construction of samplers.

Floating lily leaves were sampled using a small rectangular box (capacity ~3 litres) from
which the plastic bottom had been removed and replaced by a 50 µm Nylon mesh (see Fig.
3.3). A commercially available water-resistant cement glue was used to attach the mesh to the
edges. To enhance adhesion these were made rough by filing.

Sampling the petioles of the lilies posed many problems. It was necessary to take the stem
itself along with the water around it by cutting it as close to the bottom as possible without

33
CHAPTER 3

disturbing the very flocculent sediment of the lake. This had to be done quickly and with
minimal disturbance, both to avoid losing any animals swimming away from around the
petiole, and to minimize unwanted effects on subsequent sampling of other nearby petioles.
After some trials it was decided to attach a pair of shears to a 10 cm-diameter plastic tube
enclosing the stem so the blades could be remotely closed from the boat (see Fig. 3.4 and
section 3.2.4).

To sample the microfauna associated to the submerged vegetation, particularly outside the lily
beds, a decision had to be taken to restrict this part of sampling to the period when maximal
or near-maximal growth was attained. The sampler consisted of a short cylinder (30 cm-long)
from which large windows had been removed and replaced by fine mesh (50 µm). The piece
of tube was cut in half lengthwise and hinged. Two windows were cut out on either side and
covered with the fine mesh (see Fig. 3.5). Both ends on either half were closed with Perspex
semi-circles so closure was as water-tight as possible. To aid this, very thin strips of closed-
pore neoprene were sealed on closing edges (original design by Kórnijow, U. of Lublin,
Poland).

The zooplankton among lily beds cannot be easily sampled with traditional methods such as
nets or depth bottles. This is because of the shallow depth (often less than one metre) and the
physical obstacle of the plant growth itself. Swarming behaviour of many zooplankton
species, perhaps more frequent in more heterogeneous environments (Folt et al. 1993),
reduces the representativeness of single samples using traditional methods, too (Wiebe &
Holland 1968). Thus, a 1-m tube was used to collect a depth-integrated sample by covering
the surface end with a bung as it was lifted and retrieving the contents into a bucket (see Fig.
3.6). Generally around 4 or 5 tubefuls were sufficient to collect a 10-l sample (see section
3.2.4, also).

Methods for areal plant biomass samples and estimates are explained in Chapter 4.

3.2.4. Trades-off of sampling techniques used.

Lily floating leaf sampler.

Floating leaves were sampled using a small box (see Fig. 3.3), collecting the leaf from
beneath it as it was cut off the petiole with a pair of scissors.

34
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Extreme spatial heterogeneity of plant-associated microcrustacea distributions (e.g. Whiteside


1975) is a major sampling problem (Frost et al. 1988). Multiple small samples allow a
measure of very small-scale variability that is missed by larger samples but cover only a small
part of the population. In practice, leaves were taken as the unit ‘patch’ and the size and shape
of the sampler was guided by the range of sizes of leaves.

Figure 3.3. Sampler for floating lily leaves (capacity ~3 litres). The photograph on the right also shows the
filter (50 µm) and bottle used to concentrate and wash the collected animals into the sample bottle. The
ruler in the picture is 50 cm long.

Some disturbance during sampling is inevitable. A record was kept of the order in which
samples were collected, and leaf samples, involving relatively low disturbance, were collected
before petiole samples. However, it is difficult to know whether the repeated disturbance (i.e.
between sampling dates) in a small area around the sampling buoy used to fix the location,
had a significant effect on actual densities (see Fig. 4.23).

The sampler allowed water but not animals to flow through the mesh. The flow of water into
the box facilitated leaf collection, but at the price of letting in a variable amount of water
containing pelagic animals. While this is not thought to be important, particularly compared
with other sources of variability (e.g. spatial heterogeneity of species’ distributions), densities

35
CHAPTER 3

of pelagic animals (Daphnia, Ceriodaphnia and Bosmina) were gauged from tubes rather than
from leaf samples.

Petiole sampler (Figure 3.4).

Each sample was taken by enclosing the petiole with the tube, cutting the petiole close to the
bottom. The top end of the tube when full of water was then covered with a bung and the
contents of the tube (i.e. petiole and surrounding water) emptied into a bucket for processing
(see section 3.2.2).

Figure 3.4. Lily petiole sampler. The sampler was made of a 1-m long PVC tube attached by means of a
conveniently shaped piece of wood to a pair of shears. These were placed so the cutting parts closed
around the bottom end of the tube to cut the lily petiole, but without interfering with the retrieving of the
water contents. A stopper is used to cover the surface end. The ruler in the picture is 50 cm long. See text
for details.

36
PLANT-ASSOCIATED CLADOCERA DYNAMICS

‘Kórnijow’ jaw sampler (see Figure 3.5).

Samples were taken by submerging the sampler and carefully enclosing the plant, then cutting
it. The sample was put in a bucket for further processing (see section 3.2.2). The main
limitation was access to sample plants growing beyond arm’s length. Depth differences in
animal densities were examined by comparing samples at surface level and at about 30-60 cm
depth (i.e. arm’s length). In addition, samples were taken continuously throughout the
sampling period using the tube sampler (see Fig. 3.6), partially offsetting the drawback of not
sampling submerged plants during dates with little growth.

Figure 3.5. Sampler for submerged vegetation. The ruler in the picture is 50 cm long. Also shown in the
picture are the mesh and bottle used to collect the animals in the sample. The concentrated sample was
then washed into a 300-ml bottle for preservation (see text for details). Based on an original design by
Ryszard Kornijow (U. of Lublin, Poland).

Tube sampler (Figure 3.6).

A depth-integrated sample was taken by repeatedly submerging the tube, covering one end of
the tube with a bung, and retrieving the contents into a bucket. Taking 4 or 5 tubefuls made
the sample less sensitive to bias due to aggregations of animals. Two such samples were taken
per site.

37
CHAPTER 3

Figure 3.6. Tube sampler. A depth-integrated zooplankton sample was collected by submerging a tube
and covering the surface end with a rubber bung. A 10-litre sample was made up of 4-5 tubefuls. See text
for details.

3.2.5. Statistical analyses.

1. Spatio-temporal patterns in animal densities:

A) Open-water species.

Data from each of the two years’ growing seasons (April to end of October) were separately
analyzed. The significance of site differences in density (individuals/litre) of open water
species (i.e. Daphnia spp, Bosmina longirostris, Ceriodaphnia spp, Cyclops spp,
Eudiaptomus gracilis and total copepod nauplii) was analyzed by a univariate ANOVA. The
F-statistic was used to assess the magnitude and significance of date and site effects and their
first-order interactions in relation to density of each species. To identify the main sources of
differences among the levels of a variable (i.e. among sites or between dates) when these were
statistically significant, Tukey honestly significant difference (HSD) tests were
simultaneously run on the data.

Analysis of variance (ANOVA) is robust to departures from normality, although the data must
be at least symmetric (Kinnear & Gray 1997). Density data were plotted on an imaginary axis
to make sure the overall distribution fitted this assumption. On the other hand, ANOVA
assumes variances are equal across means (i.e. variance ‘homogeneity’). To investigate this
assumption, plots of observed by standardized residual plots were generated for the dependent
variable (i.e. density of the given species). Density data were logarithm-transformed when any

38
PLANT-ASSOCIATED CLADOCERA DYNAMICS

of these two conditions were not satisfactorily fulfilled (i.e. when Levene’s test hypothesis for
equality of variances was rejected). Only densities larger than 1 individual/litre were included
in analyses. This is because large numbers of low values may lead to spuriously narrow errors
for predicted means, therefore overestimating the significance of differences between these
(Bailey 1992).

In summary tables, F-tests evaluating the significance of main effects and first-order
interactions are shown.

B) Plant-associated species:

To investigate temporal patterns in animal densities (individuals gram-1 of lily leaf or petiole)
within each year, across sites with lilies, and between lily leaves and petioles, I used a three-
factor ‘mixed’ ANOVA with repeated measures. The dependent variable (density) is
considered in this ‘mixed’ approach to be the response to the levels of the ‘within-subjects’
factors (i.e. ‘site’ and ‘leaf-petiole’; Norusis 1994). ‘Date’ is the grouping variable (i.e. the
‘between-subjects’ factor). Replicates can be considered ‘repeated measures’ because the
levels of factors (e.g. for ‘site’, 1, 2 and 5, the three lily-bed sites, analyzed in this section) do
not change among replicates. Thus, the power of the test can be increased considerably as
degrees of freedom increase (five-fold as there are five replicates). In addition, sampling error
is explicitly included in tests. In this type of analysis, ‘date’ is the grouping variable, and
differences due to location (‘site’) or type of sample (leaf or petiole) are analyzed within each
date.

Analogously to the analysis of open-water species densities, data were plotted to make sure
the overall distribution was at least approximately symmetric. For each species’ density data,
plots of observed values against residuals of their fit to the ANOVA model were examined to
check the homogeneity of variance assumption of ANOVA. Density data were logarithm-
transformed when any of these two conditions were not satisfactorily fulfilled (i.e. when
Levene’s test hypotheses for equality of variances was rejected).

Comparisons were made only on non-zero data, by strictly excluding all zero-values found on
leaves in the three lily-bed sites (Table 3.3). This is because large numbers of low values may
lead to spuriously narrow errors for predicted means, therefore overestimating the

39
CHAPTER 3

significance of differences between these (Bailey 1992). Exceptionally, data for Eurycercus
lamellatus, Chydorus spp and Graptoleberis testudinaria in growing season 1999 and
Acroperus harpae, Eurycercus lamellatus and Sida crystallina in growing season 1998, were
so scarce as to ‘soften’ the criterion of exclusion in order to perform a valid analysis. Thus,
data included allowed the comparison of zero values with non-zero values, but never the
comparison of zero-values with zero-values. The biological interpretation of this procedure is
that comparisons are made only during periods of species presence. Absence of a species
throughout the lake cannot inform about differences in density between sites or between
leaves and petioles, thus warranting the exclusion of these data.

Table 3.3. Number of samples and proportion of total number (%) collected during the sampling
programme (April-October of 1998 and 1999) that were included in statistical analyses (see text for details
on exclusion criteria).

Number of samples & 1998 1999


% of total number N % N %
Acroperus harpae 198 55.0 0 0
Alona sp 150 41.7 120 22.2
Chydorus spp 168 46.7 282 52.2
Eurycercus lamellatus 222 61.7 96 17.6
Graptoleberis testudinaria 150 41.7 252 46.7
Peracantha truncata 150 41.7 102 18.9
Polyphemus pediculus 90 25.0 138 25.6
Scapholeberis mucronata 90 25.0 150 27.8
Sida crystallina 126 35.0 108 20.0
Simocephalus vetulus 114 31.2 216 40.0

In addition, for the full interpretation of repeated measures ANOVA the validity of the F-
statistic used must be assured (Norusis 1994). A conservative adjustment of F-ratio degrees of
freedom was sometimes necessary (Greenhouse-Geisser’s correction).

ANOVA tables are given for main factor effects (date, site and leaf/petiole) and all their
possible interactions. Changes across dates and interactions can also be visually examined in
plots. Within each main factor (except date), levels were compared using the deviation
method. This method compares the mean of each level to the mean of all of the levels (i.e. the
grand mean). The Bonferroni correction, based on Student’s t-statistic, was applied to adjust
the observed significance level for the fact that multiple comparisons are made. Multiple
comparisons between dates used Tukey’s Honestly Significant Difference (HSD) test.

40
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Differences arising between each year’s patterns are compared by testing with a t-test
(α=0.05) the significance of differences between grand means, site means and means for leaf
and petiole densities in both years.

Data from submerged plants merit a separate section (see section 3.3.3). Comparisons
between data for lilies and submerged plants were made by separately examining plots of
density in both types of vegetation. Densities expressed on a number per gram basis are very
much confounded by other factors unrelated to plant biomass (for example, plant growth
structure; see Discussion). Indeed, this problem arises also when comparing densities (in
ind.gram-1 of lily part) on lily leaves and petioles. The key variable in this case may be
available attachment surface rather than biomass (see Discussion for details). Attachment area
is related to plant weight in a different manner in leaves and petioles (i.e. there is much less
surface per unit weight in petioles than in leaves). On the other hand, calculation of densities
per m2 of lake bottom introduces probably much larger errors (see Chapter 4), reducing the
sensitivity of tests to the questions posed. Errors in calculation are also time-dependent. Thus,
data analyzed are in numbers per gram units. Apparent differences are discussed in relation to
these methodological difficulties.

2. Submerged vegetation and plant-associated Cladocera.

Data were densities (ind g-1 of submerged plant) of the main plant-associated Cladocera
species found in samples, i.e. Acroperus harpae, Alona sp, Chydorus spp, Eurycercus
lamellatus, Graptoleberis testudinaria, Peracantha truncata, Polyphemus pediculus,
Scapholeberis mucronata, Sida crystallina and Simocephalus vetulus.

Differences between years in mean densities of top and bottom samples separately were
compared statistically using t-tests (0.05 significance level). Analogously, t-tests were also
used to assess the significance of differences between means of top and bottom samples
within each year.

3. Size-structure of populations of Daphnia spp and Simocephalus vetulus.

Size-structure changes were examined for populations of the two most common species in
Little Mere, Daphnia spp (mainly Daphnia hyalina-longispina; see Table 2.3 for details) and
Simocephalus vetulus.

41
CHAPTER 3

To examine differences between years, plots of percentages of the total population in each
size-class were plotted for each date. In the case of Daphnia the comparison between sites
was done by grouping lily-bed sites in one plot-series, and open-water sites in a separate
series. For Simocephalus vetulus, only size data from lily leaf samples could be used.
Densities on petioles were too low to allow any size-structure analysis. A detailed time-series
for site 2 is shown for both species. Site 2 was the lily-bed site where Simocephalus vetulus
was more abundant in both years (see section 3.3.2 for details). Size-structure changes for
Daphnia at site 2 are shown in order to compare these with changes observed in
Simocephalus populations at the same site.

4. Egg-ratio models for Daphnia spp and Simocephalus vetulus.

Finite and instantaneous birth rates were calculated for Simocephalus vetulus and Daphnia
spp (mainly Daphnia hyalina-longispina; see Table 2.3 for details) in the two growing
seasons sampled. Data on number of eggs per individual and animal size in replicate samples
were pooled and the following parameters calculated, for each date:

NT Total number of individuals in the sample.

Novigerous Number of ovigerous individuals in the sample.

% ovigerous Percentage of ovigerous individuals in the sample.

Eggs (total) Total number of eggs in ovigerous individuals in each sample.

Brood size Mean number of eggs in ovigerous individuals in each sample.

Egg-ratio Calculated as Eggs(total in sample) / NT.

‘b’ Instantaneous birth rate, i.e. b = D-1 * Ln (Egg-ratio + 1)


(Edmondson 1968; Paloheimo 1974).

‘D’ is the egg developmental period, estimated from regression equations


relating it to water temperature (see Table 3.4).

‘B’ Finite birth rate (i.e. number of newborn per individual per day),
i.e. B = (1/r) * b * (er – 1), where ‘r’ is:

‘r’ the average growth rate in the sampling period t Æ t+1, i.e.
r = (1/∆t) * Ln (Nt+1 / Nt)

‘d’ Instantaneous death rate, i.e. d = b – r.

42
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.4. Egg developmental period (D, in days) at different temperatures for Simocephalus vetulus
(Bottrell 1975) and Daphnia spp (Bottrell et al. 1976).

Temperature (°C) Simocephalus vetulus Daphnia spp


(LnD=3.68-0.16T+0.0019T2) (LnD=3.4+0.22LnT-0.34LnT2)
13 7.2 days 5.5 days
17 4.9 4.0
19 4.1 2.9
21 3.5 2.3

Temperature at surface at five sites (1998) Temperatures at surface at five sites (1999)
25 25

20 20
degrees centigrade

degrees centigrade
15 15

10 10

5 5

0 0

5-May

13-May

19-May

26-May

15-Jul

22-Jul

5-Aug

15-Aug

27-Sep

8-Oct
18-Jun

25-Jun

2-Jul

10-Jul

29-Jul

17-Aug

2-Sep

17-Sep

7-Oct

Figure 3.7. Instantaneous temperatures recorded during the relevant dates in growing seasons ’98 and ’99
across sampling sites in Little Mere (Cheshire). Day-night differences were never of more than about 3º.
For a Map with site locations, see Fig. 3.2

Measured animals were those first encountered when counting a well-mixed sample (see
section 3.2.2 for details). For Daphnia spp at least 30 individuals were measured per replicate
sample (1 or 2 replicates, depending on dates, see section 3.2.2) where possible. For
Simocephalus vetulus at least 50 individuals were measured per replicate (N=5 replicates)
where possible. Only samples from lily leaves were analyzed, as they generally contained
larger densities of this species (see section 3.3.2). Information is presented only for dates with
at least 20 measured individuals, for both species.

5. Small-scale distribution of plant-associated species.

Spatial distribution is sometimes measured by an index relating mean abundance to the


variance of the mean estimate (‘mean crowding’ (MC), Lloyd 1967; see Equation 1).

MC = Mean density + [( s2 – 1 ) / mean density] Equation 1.

where ‘s’ is the standard error of mean density.

Unfortunately, there is a general trend to increased variance as means increase (Samuels


1991). Thus, when ‘mean crowding’ is standardized by dividing it by the mean, a measure of

43
CHAPTER 3

‘patchiness’ is generated that is quite independent of sampling variability (Lloyd 1967; see
Equation 2).

‘Patchiness’ = MC / Mean density Equation 2.

Lily leaf density data (ind g-1DW leaf) was used to calculate these two indices whenever mean
density was more than 1 ind g-1. Density values lower than 1 ind g-1 would lead to
inordinately large values of mean crowding and patchiness, impossible to interpret
biologically.

3.3. Results.

3.3.1. Zooplankton.

In general, much higher densities of Daphnia were found in the first year, while the second
year it was the smaller species, such as Bosmina longirostris and Ceriodaphnia sp that were
most abundant (Table 3.6). Copepods were also more abundant in the second year,
particularly cyclopoids. Cyclops spp was about twice as abundant in the second year. Total
copepod nauplii (i.e. both cyclopoid and calanoid) presented similar densities in both years.

There was considerable heterogeneity in Daphnia spp densities across the lake in both years.
(Fig. 3.8). Higher densities were generally found in the lily beds than in open waters in both
years (1998: F=5.4, p<0.05; 1999: F= 13.1, p<0.01; Table 3.7). In 1999, Daphnia populations
abruptly collapsed at the end of May after a brief peak (Figure 3.9), reappearing as an isolated
population at the end of the growing season.

44
PLANT-ASSOCIATED CLADOCERA DYNAMICS

900

800
Site 1 Site 2 Site 3 Site 4 Site 5
700
cumulative no./ litre

Growing season
600

500
400
300

200
100

0
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46
Date

Figure 3.8. Daphnia cumulative density (ind l-1) during the two years sampled. Underlined are growing
season dates (April to October). The size of areas gives a measure of the changing relative importance of
each site’s populations.

Ceriodaphnia and Bosmina, on the other hand, were found at comparable densities across
sites in the first year (F-test, p>0.05; Table 3.7). In the second year Ceriodaphnia densities in
lily-bed sites were about double those in more open waters (F=35.6, p<0.001; Tables 3.6 &
3.7) with variance in the order of 600 times the mean (Table 3.5). Populations, particularly of
Daphnia and Ceriodaphnia, aggregated more among lilies.

Table 3.5. Variance-to-mean ratios in lilies and open water for the five open-water taxa (Daphnia,
Ceriodaphnia, Bosmina longirostris, Eudiaptomus gracilis, Cyclops sp and total copepod nauplii) from
samples taken during growing seasons ’98 and ’99 (April to October) in Little Mere (Cheshire). To
calculate ratios all data were included.

Variance/Mean 1998 1999


Lilies Open water Lilies Open water
Daphnia spp 118 74 191 91
Ceriodaphnia spp 557 341 94 43
Bosmina longirostris 76 9 658 611
Eudiaptomus gracilis 7 68 7 218
Cyclops spp 49 58 47 77
Total nauplii 198 38 50 43

45
CHAPTER 3

Daphnia spp. Daphnia spp.

Growing season 1998 Growing season 1999


200 200
individuals/litre

individuals/litre
100 100

Type of site Type of site

Open water Open water

017-Sept Lily bed 0 Lily bed


25-Sept

13-Sept
27-Sept
18-Jun
25-Jun

10-Jun
17-Jun
2-Sept
9-Sept
1-Jun

17-Aug
27-Apr

15-Aug
21-Apr
28-Apr
10-Jul
23-Jul
29-Jul

15-Jul
22-Jul
26-Oct

26-Oct
18-May

5-Aug
1-Apr

13-May
19-May
26-May
2-Jul

1-Jul
8-Jul
7-Oct

8-Oct
5-May
Figure 3.9. Daphnia density (ind l-1) across sites in Little Mere (Cheshire), grouped as lily or open water
sites during growing seasons (April to October) '98 (left) and '99 (right).

Eudiaptomus gracilis, Cyclops spp and total copepod nauplii (i.e. both cyclopoid and calanoid
nauplii) were more abundant in the open water (F=31.0 & F=198.7, respectively, p<0.01;
Tables 3.6 & 3.7). Ceriodaphnia and Bosmina, which were scarce during this growing season,
were found at comparably low densities in both lily beds and more open waters (see Table
3.6).

46
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.6. Marginal densities of pelagic species in lily or open water (OW) areas during growing seasons ’98 and ’99 (April to October)
in Little Mere (Cheshire). Means ± standard error are of densities higher than 1 ind l-1. For a Map, see Figure 3.2.

Means ± S.E. Daphnia spp Ceriodaphnia Bosmina Eudiaptomus Cyclops sp Total copepod
(ind l-1) sp longirostris gracilis nauplii

Lilies 78.8 ± 16.4 38.6 ± 11.4 18.3 ± 8.3 5.0 ± 1.4 25.3 ± 8.5 31.5 ± 16.3
1998

Open water 53.4 ± 11.0 37.5 ± 8.2 3.1 ± 1.3 28.1 ± 24.8 33.5 ± 12.1 48.8 ± 7.6

Lilies 32.4 ± 7.8 203.1 ± 48.2 167.8 ± 55.9 6.2 ± 1.3 35.3 ± 5.1 32.5 ± 5.1
1999

Open water 54.0 ± 16.5 74.9 ± 23.5 120.9 ± 45.4 30.3 ± 7.3 61.9 ± 7.9 52.0 ± 5.5

47
CHAPTER 3

Table 3.7. Summary of significance tests for main factor effects (date, site and ‘lily-open’, or grouped lily-sites versus grouped open-
water sites) and first-order interactions in Univariate ANOVA models for open-water taxa densities (ind l-1). Analyses are of results
deriving from samples taken at 5 sites across Little Mere (Cheshire) during growing seasons ’98 and ’99 (April to October). *F-test is
significant at the 0.05 level, **at the 0.01 level; ***at the 0.001 level.

Full factorial Daphnia spp Ceriodaphnia sp Bosmina Eudiaptomus Cyclops sp Copepod nauplii
models longirostris gracilis
(SS-typeIII)
Source SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig.

DATE 17.9 1.19 17.6*** 26.67 1.78 33.9*** 4.54 0.38 9.4** 3.34 0.28 7.3* 9.38 0.63 37.8*** 13.07 0.87 6.4**
1998

SITE 0.97 0.32 4.7* 0.05 0.02 0.33 0.38 0.13 3.2 3.11 1.04 27.1** 5.26 1.75 106.0*** 1.04 0.35 2.5

DATE*SITE 7.8 0.21 3.03* 4.16 0.14 2.64 4.26 0.28 7.1** 4.24 0.24 6.2* 5.95 0.21 12.4*** 6.41 0.15 1.1
* ** ***
LILY-OPEN 0.37 0.37 5.4 0.01 0.01 0.17 0.03 0.03 0.774 1.19 1.19 31.0 3.29 3.29 198.7 2.57 2.57 18.9**

Source SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig. SS MS Fsig.

DATE 21.75 1.45 19.0*** 109.65 6.09 140.3*** 63.73 3.54 35.2*** 5.90 0.33 11.2*** 20.98 1.17 32.9*** 29.25 1.63 36.5***
1999

SITE 1.08 0.36 4.7** 0.67 0.22 5.2** 2.02 0.67 6.7** 16.70 5.57 191.1*** 0.34 0.11 3.2* 0.24 0.08 1.8

DATE*SITE 9.26 0.27 3.6*** 7.03 0.14 3.2*** 10.83 0.27 2.7*** 4.96 0.12 4.3*** 11.01 0.20 5.8*** 9.95 0.18 4.1***

LILY*OPEN 1.0 1.0 13.1** 1.55 1.55 35.6*** 0.24 0.24 2.4 12.63 12.63 433.5*** 1.05 1.05 29.5*** 3.14 3.14 70.6***

48
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Ceriodaphnia followed bimodal dynamics in the lily beds, with peaks separated by about 2-3
weeks but only displayed one population peak in the open water. Bosmina and copepods
(Eudiaptomus gracilis and Cyclops sp) had unimodal dynamics. In Cyclops and copepod
nauplii no time pattern was evident, but populations fluctuated irregularly, though within the
non-overlapping ranges of lily or open sites. Eudiaptomus, on the contrary, slowly built a
population maximum that subsequently declined across two dates.

3.3.2. Plant-associated cladocerans on lilies.

The years sampled were very different with respect to densities of particular plant-associated
species and community composition (Table 3.8).

Table 3.8. Overall comparisons across years of grand means (i.e. mean of all data together), site means
and lily leaf / petiole means for each plant-associated cladoceran in Little Mere (Cheshire). Data are from
growing seasons ’98 & ’99. Results summarize significance of t-tests (0.05-level); +significant; -not
significant.

Between-year Sites were Between-year Between-year


differences in significantly differences in differences in
grand means different across mean densities mean densities
years on lily petioles* on lily leaves*
Acroperus + Cannot + +
compare**
Alona + Site 2 - +
Chydorus + Sites 1, 2 & 5 + +
Eurycercus - Sites 1 & 2 - -
Graptoleberis + Sites 1, 2 & 5 + +
Peracantha + Site 2 + +
Polyphemus - Site 2 + -
Scapholeberis - Site 2 + -
Sida + Sites 1 & 5 + +
Simocephalus - Sites 2 & 5 - -
*for details on the direction of these differences, see Table 3.10. **only trace densities were
detected in 1999 and, therefore, no comparison for sites can be done.

In particular, all periphyton scrapers with the exception of Eurycercus, which was very scarce
both years, had significantly lower densities during the second year sampled. Scapholeberis
mucronata and Polyphemus pediculus had comparable densities both years. The main plant-
associated filter-feeders, i.e. Sida crystallina and Simocephalus vetulus, were more abundant
the second year, although this difference was only statistically significant in the case of Sida
(Table 3.9).

49
CHAPTER 3

Table 3.9. Grand means (ind g-1DW of lily), standard errors and conventional confidence bounds for the
mean of the 10 species recorded during growing seasons ’98 and ’99 in Little Mere (Cheshire). All data
are included for the calculation of means (see section 3.2.5.1 for details on methods).

GRAND MEANS 1998 1999


(ind g-1 DW of lily)
95 % confidence limits 95 % confidence limits
Food habit Species Mean Std. Lower Upper Mean Std. Lower Upper
error bound bound error bound bound
Periphyton Acroperus harpae 6.6 1.8 2.9 10.3 Trace densities recorded
scrapers Alona sp 9.5 2.4 4.7 14.4 2.8 6.0 2.2 3.4
Chydorus spp 19.6 2.3 15.0 24.3 0.8 0.1 0.6 1.1
Eurycercus lamellatus 1.1 0.15 0.8 1.4 0.2 0.03 0.1 0.2
Graptoleberis testudinaria 14.2 1.0 12.2 16.3 0.7 0.1 0.4 0.9
Peracantha truncata 11.6 1.6 8.3 15.0 3.4 0.4 2.5 4.2
Raptorial Polyphemus pediculus 15.6 8.5 0 32.8 23.8 4.9 14.1 33.6
Filter- Scapholeberis mucronata 4.2 0.7 2.6 5.7 8.3 1.0 6.2 10.4
feeders Sida crystallina 0.8 0.2 0.4 1.2 22.1 3.8 14.6 29.7
Simocephalus vetulus 7.1 0.5 6 8.2 9.4 0.8 7.9 10.9

A) Periphyton scrapers: chydorids.

More animals were generally found on leaves than on petioles in both years (Table 3.10).
Table 3.10. Mean densities (ind.g-1 DW of lily) comparing leaves and petioles in each year separately, for
the main periphyton scraper species. In bold significantly larger values (F-test, significance level: *0.05;
**0.01; ***0.001). All data have been used for the estimation of means. Standard errors indicate variation
in the mean estimate, but cannot be used to compare means statistically (see section 3.2.5.1 for details). See
Table 3.11 for the filter-feeder species’ means ± SE.

Mean ± Std. error 1998 1999


(ind.g-1 DW lily)
Leaves Petioles Leaves Petioles
Acroperus harpae 3.8±0.8 9.5±3.0 Trace densities recorded
Alona sp 9.6±2.7** 9.5±3.0 2.8±0.3 2.8±0.5
Chydorus spp 21.0±2.2** 18.2±3.4 1.0±0.2 0.7±0.2
Eurycercus lamellatus 0.8±0.1 1.4±0.3 0.1±0.03 0.3±0.06
Graptoleberis testudinaria 18.5±1.4*** 10.0±1.5 0.4±0.1 0.9±0.2
Peracantha truncata 17.4±3.4*** 5.9±1.3 5.7±0.7*** 1.1±0.3

B) Filter-feeders.

Plant-associated filter-feeders were found in higher densities near or on lily leaves than on
petioles (Table 3.11), although differences were only statistically significant in the second
year, for both Simocephalus vetulus and Sida crystallina (Table 3.11).

The behaviour of Sida populations in the two years sampled was quite unpredictable. In 1998,
an important population peak was detected in spring (i.e. 250+ ind g-1DW leaf of Sida; Fig.
3.10). Although much larger densities were found on leaves than on petioles in this first year
of sampling, these differences could not be separated from sampling error (F=2.36, p>0.05).

50
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Differences in densities across sites were swamped by the very large standard errors when
estimating means.

50 Sida crystallina , 1998 Sida crystallina , 1999


500
45
450
40
400
35
350
leaves
ind g-1 DW

30

ind g DW
300 petioles
25 leaves

-1
250
20 petioles
200
15
150
10
100
5
50
0
0
06-abr

27-abr

18-may

01-jun

18-jun

25-jun

02-jul

10-jul

29-jul

17-ago

02-sep

17-sep

01-abr

21-abr

28-abr

05-may

13-may

19-may

26-may

10-jun

17-jun

01-jul

08-jul

15-jul

22-jul

05-ago

15-ago

13-sep

27-sep

08-oct
Figure 3.10. Sida crystallina mean densities (ind.g-1DW) with standard error bars (N=5) in leaf and petiole
samples collected across the three lily beds sampled in Little Mere (Cheshire) (April-October). Left: 1998;
right: 1999.

In 1999, leaf samples were significantly richer (by a factor of two) in Sida crystallina than
petiole samples (F=30.96, p<0.001).

Table 3.11. Mean densities (ind.g-1 DW lily) comparing leaves and petioles in each year separately, for the
main plant-associated filter-feeding species in Little Mere during growing seasons ’98 and ‘99. In bold
significantly larger values (F-test, significance level: *0.05; **0.01;***0.001, as in Table 3.14). All data
were used to estimate means (1998: 170 leaves and 150 petioles; 1999: 270 leaves and 270 petioles).
Standard errors indicate variation in the mean estimate, but cannot be used to infer differences between
means (see section 3.2.5.1 for details).

Mean ± S.E. 1998 1999


(ind g-1DW lily)
Leaves Petioles Leaves Petioles
Scapholeberis mucronata 5.0±1.4 3.3±0.9 8.1±1.5 8.5±1.7
Sida crystallina 0.5±0.09 1.1±0.4 23.8±4.6*** 20.5±7.2
Simocephalus vetulus 9.2±0.9** 5.1±0.8 12.6±1.1*** 6.1±0.8
Polyphemus pediculus 23.8±17.1 7.4±1.8 18.3±5.3 29.4±7.7

Numbers of Simocephalus on leaves were 50-100 % larger than those on petioles in both
years (1998: F=4.571, p<0.01; 1999: F=17.87, p<0.001).

51
CHAPTER 3

Simocephalus vetulus , 1998 Simocephalus vetulus , 1999


90
70
80
60
leaves 70
50 petioles
60 leaves
ind g-1 DW

ind g-1 DW
40 petioles
50
30 40

20 30

10 20

10
0
06-abr

27-abr

18-may

01-jun

18-jun

25-jun

02-jul

10-jul

29-jul

17-ago

02-sep

17-sep
0

01-abr

21-abr

28-abr

05-may

13-may

19-may

26-may

10-jun

17-jun

01-jul

08-jul

15-jul

22-jul

05-ago

15-ago

13-sep

27-sep

08-oct
Figure 3.11. Simocephalus vetulus mean densities (ind.g-1DW) with one standard error at either side of the
mean (N=5) on lily leaves and petioles across the three lily beds sampled in Little Mere (Cheshire) (April-
October). Left: 1998, right: 1999.

Densities achieved were higher in 1999 than in 1998. While maxima were reached in mid-
summer in growing season ’98, peaks were reached by the end of the summer in 1999.

52
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.12. Summary of three factor mixed General Linear Models (ANOVA) for periphyton scraper species in Little Mere (Cheshire) during growing seasons
(April to October) ’98 and ‘99. Results are for main factor effects (site, leaf or petiole and date) and interactions. Date is the grouping variable. See section 3.2.5.1
for details.

1998 1999
SITE TYPE OF SITE* TYPE OF SITE* SITE* SITE TYPE OF SITE* TYPE OF SITE* SITE*
Source SAMPLE DATE SAMPLE* TYPE OF TYPE OF SAMPLE DATE SAMPLE* TYPE OF TYPE OF
(Leaf or petiole) DATE SAMPLE SAMPLE* (Leaf or petiole) DATE SAMPLE SAMPLE*
DATE DATE
Acroperus harpae SS1 3.525 0.119 14.031 1.81 0.061 2.716 Trace densities recorded
MS 1.763 0.119 0.702 0.181 0.0308 0.136
F 24.65*** 1.623 9.811*** 0.463* 0.411 1.813*
Alona sp SS 2.753 1.73 8.64 2.11 0.0894 5.77 0.793 0.416 1.655 4.384 0.477 1.822
MS 1.372 1.73 0.54 0.264 0.0447 0.361 0.396 0.416 0.138 0.731 0.238 0.152
F 15.913*** 9.092** 6.242*** 1.387 0.52 4.192*** 3.774* 1.868 1.315 3.278* 1.818 1.158
Chydorus spp SS 14.229 0.885 11.296 2.132 0.638 1.206 0.202 0.346 2.228 0.714 0.525 4.51
MS 7.115 0.885 0.807 0.305 0.319 0.0861 0.101 0.346 0.0795 0.0509 0.263 0.161
F 50.909*** 9.964** 5.774*** 3.337* 1.827 0.424 1.169 4.799* 0.92 0.708 3.641* 2.233**
Eurycercus SS 0.612 0.0182 3.709 1.001 0.388 0.661 0.462 0.0433 0.919 0.146 0.011 0.348
lamellatus
MS 0.306 0.0182 0.232 0.125 0.194 0.0413 0.231 0.0433 0.919 0.0292 0.005 0.0348
F 5.484** 0.267 4.154*** 1.833 2.468 0.526 13.909*** 1.070 5.529** 0.722 0.124 0.759
Graptoleberis SS 13.493 4.092 14.224 0.607 0.915 1.803 1.875 0.0757 5.066 0.37 0.014 0.916
testudinaria
MS 6.747 4.092 1.423 0.121 0.457 0.18 0.938 0.0757 0.181 0.0264 0.008 0.0327
F 48.484*** 35.304*** 10.226*** 1.048 3.325* 1.311 18.448*** 1.592 3.56*** 0.556 0.223 0.987
Peracantha truncata SS 3.475 5.237 5.919 1.4 0.529 2.259 0.409 4.337 2.625 1.077 0.31 0.497
MS 1.737 5.237 0.592 0.28 0.264 0.226 0.204 4.337 0.336 0.269 0.155 0.062
F 11.149*** 23.336*** 3.798** 1.248 2.174 1.857 2.635 35.095*** 4.329** 2.179 1.16 0.465
1
Type III; *0.05 significance level; **0.01; ***0.001

53
CHAPTER 3

Table 3.13. Summary of three factor mixed General Linear Models (ANOVA) for filter-feeding species and Polyphemus pediculus in Little Mere (Cheshire) during
growing seasons ’98 and ‘99. Results are for main factor effects (site, leaf or petiole and date) and interactions. Date is the grouping variable. See section 3.2.5.1 for
details.

1998 1999
SITE TYPE OF SITE* TYPE OF SITE* SITE* SITE TYPE OF TYPE OF LEAFPET* SITE* SITE*
Source SAMPLE DATE SAMPLE* TYPE OF TYPE OF SAMPLE SAMPLE* DATE TYPE OF TYPE OF
(Leaf or DATE SAMPLE SAMPLE* (Leaf or petiole) DATE SAMPLE SAMPLE*
petiole) DATE DATE
Scapholeberis SS1 0.693 0.557 6.757 0.172 0.676 1.664 2.232 0.972 12.764 0.506 0.209 2.038
mucronata
MS 0.347 0.557 0.845 0.0429 0.338 0.208 1.116 0.972 0.912 0.0722 0.105 0.146
F 3.677* 4.3 8.963*** 0.332 3.836* 2.362 4.326* 4.191 3.534** 0.311 0.549 0.763
Sida crystallina SS 0.0861 0.148 1.424 0.149 0.0398 0.584 4.95 3.131 9.467 0.982 0.0613 1.063
MS 0.043 0.148 0.102 0.0212 0.0199 0.0417 2.475 3.131 0.947 0.196 0.0307 0.106
F 0.602 2.365 1.422 0.339 0.248 0.52 16.204*** 30.955*** 6.198*** 1.941 0.253 0.878
Simocephalus vetulus SS 0.953 6.856 2.294 0.565 0.366 2.211 9.354 2.021 5.313 2.175 0.106 1.646
MS 0.477 0.571 2.294 0.0941 0.183 0.184 4.677 2.021 0.266 0.217 0.0528 0.0823
F 3.814* 4.571** 18.599** 0.763 1.038 1.046 30.36*** 17.869*** 1.72 1.923 0.559 0.871

Polyphemus pediculus SS 0.628 0.0782 4.944 0.502 0.302 1.87 3.041 0.332 23.538 1.602 0.652 2.902
MS 0.314 0.0782 0.494 0.100 0.151 0.187 1.52 0.332 1.962 0.267 0.326 0.242
F 1.030 0.229 1.623 0.293 0.670 0.83 4.841* 1.058 6.246*** 0.851 1.329 0.986
1
Type III; *0.05 significance level; **0.01; ***0.001

54
PLANT-ASSOCIATED CLADOCERA DYNAMICS

3.3.3. Submerged vegetation and plant-associated Cladocera.

There were appreciable differences in densities of the plant-associated Cladocera sampled


from submerged vegetation between both years (see Table 3.14). These differences, though,
were generally not statistically significant due to large replicate variability.

Table 3.14. Mean density (ind.g-1 DW of plant) of all plant-associated Cladocera species recorded in
submerged plant samples (N=5) collected at surface (‘top’) and at arm’s length (‘bottom’) in Little Mere
(Cheshire) during two growing seasons (April to October; ’98 and ’99). Significantly larger densities in a
top to bottom comparison using a t-test (0.05 level) are indicated with an asterisk and in bold.

Top vs. Bottom 1998 1999


Statistical comparison
t-tests (0.05 level) Top Bottom Top Bottom
Acroperus harpae 69±20 384*+±108 29.2±6.6 8.9±5.4
Alona sp 21.0±5.8 19.6±5.0 183±94 67.4±22
Chydorus spp 81±58 74±34 538±172 179±38
Eurycercus lamellatus 318±206 236±56 48±36 111±32
Graptoleberis testudinaria 116+±22 103±58 38.7*±15.2 2.6±1.6
Peracantha truncata 47±22 30±26 308±152 45.9±15.2
Polyphemus pediculus 43*±40 0±0 None recorded
Scapholeberis mucronata 23.4±10.6 7.6±6.4 8.2±2.8 7.1±2.4
Sida crystallina 6.8±3.4 7.7±7.2 604±334 248+±60
Simocephalus vetulus 637±1001 673±921 594±354 832±334
*significantly larger in a top-to-bottom comparison (t-test, at the 0.05 level); +significantly larger in a
between-year comparison.

Sida crystallina.

In the second year, Sida was found at high densities in vegetation close to the surface when
populations in lilies had either sharply decreased (sites 1 and 5) or were falling rapidly (site
2). Compare Figures 3.10 and 3.12. Conversely, only small populations were found when the
species was abundant in the lily beds (Sept.13th, 1999).

16 1000
Sida crystallina Top
14 Sida crystallina 900
Top Site 4 Bottom
SITE 4,
number / gram plant

800
number / gram plant

12 SUBMERGED Submerged vegetation


Bottom 700
10 VEGETATION, 1998 1999
600
8 500
6 400
300
4
200
2
100
0 0
18-Jun 25-Jun 29-Jul 17-Aug 02-Sep 17-Sep 17-Jun 13-Sep 27-Sep 8-Oct

Figure 3.12. Mean densities (ind g-1 DW plant) of Sida crystallina with standard error bars (N=5) in
submerged vegetation samples (‘top’ and ‘bottom’) taken in Little Mere (Cheshire) during two growing
seasons (April to October). Left:1998; Right:1999.

55
CHAPTER 3

In the first year, Sida maintained small populations associated with submerged plants when
the species had totally disappeared from lily beds (see Fig. 3.12).

Simocephalus vetulus.

Dense patches of Simocephalus (i.e. around 1,000 ind g-1 DW plant, or around 20,000 ind m-2
lake bottom), were sporadically found at the height of the summer in 1998, and at the end of
the season in 1999 (see Fig. 3.13).

1800 1400
Simocephalus vetulus
Simocephalus vetulus Top
1600 SITE 4, 1200 Site 4 Bottom

number / gram plant


number / gram plant

1400 SUBMERGED
1000 Submerged vegetation
1200
VEGETATION, 1998
1999
1000 800

800 600
Top
600
400
400 Bottom
200
200
* *
0 0
18-Jun 25-Jun 29-Jul 17-Aug 02-Sep 17-Sep 17-Jun 13-Sep 27-Sep 8-Oct

Figure 3.13. Mean densities of Simocephalus vetulus (ind g-1 DW plant) with standard error bars (N=5) in
submerged plant samples taken in Little Mere (Cheshire) during two growing seasons (April to October).
Left: 1998; Right: 1999. Asterisks indicate bottom samples were not taken those dates (see Discussion for
details).

3.3.4. Population size-structure changes: Daphnia spp (mainly Daphnia hyalina-


longispina) and Simocephalus vetulus.

Size-structure changes of Daphnia populations were generally periodic and clearly


identifiable (see, for example, Fig. 3.14). Simocephalus vetulus, in contrast, presented either
apparently random changes in size-structure (i.e. growing season ’98), or few significant
changes (i.e. growing season ’99) (see Figs. 3.16 & 3.17).

56
PLANT-ASSOCIATED CLADOCERA DYNAMICS

50
1-6-98 45 10-7-98
45
40
40
35 35
lily beds
30 lily beds
30 open waters
open waters
25
%

25

%
20 20
15 15
10 10
5 5
0
0
0.5 0.7 0.9 1.1 1.3 1.5 1.7
0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm) size (mm)

45 45
18-6-98 29-7-98
40 40
35 35
30 lily beds
30
open waters lily beds
25 25 open waters
%

%
20 20
15 15
10 10
5 5
0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm) size (mm)

50
25-6-98 90 17-8-98
45
80
40
70
35 lily beds
lily beds 60
30 open waters
open waters
50
%

25
%

20 40

15 30
10 20
5 10
0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm) size (mm)

35 2-7-98
30

25 lily beds
open waters
20
%

15

10

0
0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm)

Figure 3.14. Size-structure of Daphnia populations in lily beds and more open waters in Little Mere
(Cheshire) during growing season ’98 (April to October).

57
CHAPTER 3

60 5-5-99 50 26-5-99
45
50 40
35 lily beds
40
lily beds 30 open waters
open waters

%
%

30 25
20
20
15
10
10
5
0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm) size (mm)

13-5-99 50 13-9-99
60
45
50 40
35
40
30 lily beds
open waters

%
lily beds 25
%

30
open waters 20
20
15

10 10
5
0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm) size (mm)

60 19-5-99
50

40
%

30 lily beds
open waters
20

10

0
0.5 0.7 0.9 1.1 1.3 1.5 1.7
size (mm)

Figure 3.15. Size-structure of Daphnia populations in lily beds and more open waters in Little Mere
(Cheshire) during growing season ’99 (April to October).

Daphnia spp (mainly Daphnia hyalina-longispina).

Populations of Daphnia inhabiting lily beds and those sampled from open waters had clearly
different size-structures (see Figs 3.14 & 3.15). Changes in size-structure of populations in
lily beds and open waters paralleled those of density in both habitats. In 1998, there were
proportionally more of the larger individuals in lily beds than in open waters during most of
June and then again in August. During July, in contrast, the larger individuals were found in
the open water. Most animals were 0.7-1.1 mm in size.

In 1999, there were large changes in size-structure. Open waters had populations of smaller
individuals than those in lily beds. This difference intensified as the month progressed. By
late May, Daphnia populations in lily beds were largely made up of animals of 1.5 mm in size

58
PLANT-ASSOCIATED CLADOCERA DYNAMICS

or bigger. In stark contrast, populations in open waters on this date included 80 % of


individuals smaller than 0.7 mm. Populations declined after May.

Populations recovered in September (Figure 3.15). Size-structure after recovery was similar in
lily beds and open waters, with a predominance of very small animals.

Simocephalus vetulus.

Large animals (i.e. >1.1 mm) were sampled throughout summer ‘98. However, animals were
generally smaller in June than in July. Small animals were again proportionally abundant in
August. In September, a high percentage of the population was made up of animals larger
than 1.0 mm (see Fig. 3.16).

40 Simocephalus vetulus , 1st June, 1998 Simocephalus vetulus , 25th June, 1998
60
35
50
30

25 40
%

20
%

30
15
20
10

5 10

0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1
Body length (mm)
Body length (mm)

Simocephalus vetulus , 10th July, 1998


45

40

35

30

25
%

20

15

10

0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1
Body length (mm)

Figure 3.16. Changes in size-structure of Simocephalus vetulus populations in Little Mere (Cheshire) mid-
summer ’98.

59
CHAPTER 3

Simocephalus vetulus , 8th July, 1999 Simocephalus vetulus , 5th August, 1999
60 45

40
50
35
40 30

25
%

%
30
20

20 15

10
10
5

0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1 0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1
Body length (mm) Body length (mm)

Simocephalus vetulus , 15th July, 1999 Simocephalus vetulus , 15th August, 1999
45 45
40 40
35 35
30 30
25 25
%

20 %20

15 15

10 10

5 5

0 0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1 0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1
Body length (mm) Body length (mm)

Simocephalus vetulus, 13th September, 1999


45

40

35

30

25
%

20

15

10

0
0.5 0.7 0.9 1.1 1.3 1.5 1.7 1.9 2.1
Body length (mm)

Figure 3.17. Size-structure changes of Simocephalus vetulus populations in Little Mere (Cheshire) during
growing season ’99.

Populations of Simocephalus vetulus were late developing in 1999. Significant numbers were
only sampled in July. Size-structure was, compared with 1998, remarkably consistent across
lily beds and very stable (Fig. 3.17). In general, animals were small, with a predominance of
the 0.7 mm size-class in all dates. A larger proportion of big animals was observed in
populations sampled in August and September. At the end of the growing season (i.e. 8th of
October) most animals were small and few were larger than about one millimetre.

60
PLANT-ASSOCIATED CLADOCERA DYNAMICS

3.3.5. Egg-ratio models: Simocephalus vetulus and Daphnia.


Daphnia

Birth rates were higher in lily beds than in open waters, although there was much variation in
any given year.

Large differences in birth rate between lily beds and more open waters were particularly
evident in 1999 (Fig. 3.18, plots on the right). In open waters, growth rates were generally
negative. With few exceptions, lily-bed populations had large birth rates, but also much larger
death rates, as estimated from the difference between apparent growth and recruitment (i.e.
births).

In 1998, death rates were fairly constant in lily beds throughout the growing season, while in
more open waters, births and population growth diminished into the summer (black bars in
Fig. 3.18).

01-jun 18-jun 25-jun 02-jul 10-jul 29-jul 17-ago 25-sep 05-may 13-may 19-may 26-may 13-sep 27-sep 08-oct
0.300 200 0.300 180

Daphnia , lily beds, 1998 Daphnia , lilies, 1999


180 160
0.250
0.200
160 140
0.200
140
0.100 120

Density (ind l-1)


Density (ind l-1)

0.150
120
b and r

100
b and r

0.100 100 0.000


80

80
0.050
-0.100 60
60
0.000 40
40 -0.200

-0.050 20
20
Birth rate (b) Growth rate (r) Density
-0.300 0
-0.100 0 Birth rate (b) Growth rate (r) Density

01-jun 18-jun 25-jun 02-jul 10-jul 29-jul 17-ago


05-may 13-may 19-may 26-may
0.250 120
0.300 25

Daphnia , open water, 1998 Daphnia , open water, 1999


0.200
100 0.200
20
0.150
0.100
80
Density (ind l-1)

0.100
Density (ind l-1)

15
0.000
b and r
b and r

0.050 60

-0.100
10
0.000
40
-0.200
-0.050
5
20 -0.300
-0.100

Birth rate (b) Growth rate (r) Density


-0.400 Birth rate (b) Growth rate (r) Density 0
-0.150 0

Figure 3.18. Birth and growth rates of Daphnia populations in Little Mere (Cheshire) during growing
seasons ’98 (left plots) and ’99 (right plots) among lilies (top plots) and in the open water (bottom plots).
Death rates are represented by the difference between birth rate and growth rate lines. White bars
indicate periods when population growth exceeds apparent birth rates; black bars indicate periods when
birth rate is lower than growth of the population. This could happen as a result of deaths, but also if birth
rate had been underestimated (see Discussion in Chapter 3 for details).

61
CHAPTER 3

Simocephalus vetulus.

The fertility of Simocephalus vetulus (i.e. proportion of ovigerous individuals) was low
throughout the growing season of 1998 (Fig. 3.19). Periods with larger individuals also had a
larger proportion of ovigerous individuals. Site differences in egg-ratios were not apparent
from these data. In 1999, birth rates greatly exceeded growth rates on many occasions (Fig.
3.19).

Comparisons between lily beds as regards growth dynamics are hampered by the scarcity of
data. There were very large differences in the percentage of ovigerous in different lily-bed
sites in any given date (Tables 3.15 & 3.16).

1st June 18th June 25th June 2nd July 10th July 29th July 17th Aug. 2nd Sept. 17th Sept. 8th July 15th July 22nd July 5th Aug. 15th Aug. 13th Sept. 27th Sept. 8th Oct.
0.350 50 0.200 70

Simocephalus vetulus Simocephalus vetulus


45
Lily leaves, 1998 Lily leaves, 1999 60
0.250 0.150
40

Density (ind g-1 DW leaf)


50
35
Density (ind g DW leaf)

0.150 0.100

30
40
b and r
b and r

-1

0.050 25 0.050

30
20

-0.050 0.000
15 20

10
-0.150 -0.050
10

Birth rate (b) Growth rate (r) Density -0.100 Birth rate (b) Growth rate (r) Density 0
-0.250 0

Figure 3.19. Birth and growth rates of Simocephalus vetulus populations in Little Mere (Cheshire) during
growing seasons ’98 (left) and ’99 (right) (April to October). Death rates are represented by the difference
between these two lines. White bars indicate periods when population growth exceeds apparent birth
rates; black bars indicate periods when birth rate is lower than growth of the population. This could
happen as a result of deaths, but also if birth rate had been underestimated (see Discussion for details).

Table 3.15. Sample size and egg-ratio variables (number of ovigerous individuals in each sample, %
ovigerous, total number of eggs and egg-ratio; see 3.2.5.5 for details on calculations) for Simocephalus
vetulus sampled on floating lily leaves, estimated for each date in three lily beds in Little Mere (Cheshire)
during growing season ’98.

Sample Ovigerous % Eggs Egg-ratio


size individuals ovigerous (total)
1st June 192 0 0 0 0.000
18th June 16 0 0 0 0.000
25th June 446 13 2.9 75 0.168
2nd July 684 10 1.5 36 0.053
10th July 346 39 11.3 142 0.410
29th July 59 2 3.4 7 0.119
17th Aug. 16 1 6.3 5 0.313
2nd Sept. 65 15 23.1 51 0.785
17th Sept. 82 0 0.0 0 0.000

62
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.16. Sample size and egg-ratio variables (number of ovigerous individuals in sample, % ovigerous,
total number of eggs and egg-ratio) for Simocephalus vetulus sampled on floating lily leaves, estimated for
each date in three lily beds in Little Mere (Cheshire) during growing season ’99.

Sample Ovigerous % Eggs Egg-ratio


size individuals ovigerous (total)
8th July 442 14 3.2 108 0.244
15th July 247 37 15.0 184 0.745
22nd July 76 1 1.3 8 0.105
5th Aug. 676 62 9.2 417 0.617
15th Aug. 380 71 18.7 424 1.116
13th Sept. 743 128 17.2 599 0.806
27th Sept. 267 17 6.4 48 0.180
8th Oct. 295 3 1.0 8 0.027

3.3.6. Small-scale distribution of species.

No clear relationship between crowding levels and patchiness, as indicated by Lloyd’s index,
was apparent across species. However, some species with large seasonal changes in numbers
(e.g. Graptoleberis testudinaria and Eurycercus lamellatus) showed highest patchiness when
populations were scarce. Plant-associated filterers often showed very regular distributions (i.e.
patchiness index close to 1.0), despite large variations in mean crowding. Sida crystallina was
very regularly distributed at its population peaks in both years (1998 & 1999).

Simocephalus vetulus showed the strongest consistency in patchiness across dates and sites.
Mean crowding was large at times of population growth (max. 134 ind ind-1 in 1998; 162 in
1999).

A general classification of “habitats” for microcrustacean species in Little Mere is


summarized in Tables 3.17-3.19. Filter-feeders were generally more abundant on lily leaves
than on petioles, while the pattern was not as clear for chydorid species (Tables 3.17 & 3.18).
Chydorid species were generally abundant on submerged plant parts (Table 3.17). Daphnia
and Ceriodaphnia were generally more abundant in lily beds. Bosmina and copepods were
found at higher densities in the open water (Table 3.19). In all cases there was large
variability in numbers both across years and across sampling sites.

63
CHAPTER 3

Table 3.17. Variables summarizing the ‘habitat’ association of the 6 chydorid species (Acroperus harpae,
Alona sp, Chydorus spp, Eurycercus lamellatus, Graptoleberis testudinaria and Peracantha truncata)
sampled from Little Mere over the two-year sampling programme. Tabled are the lily part (leaf or petiole)
where denser, and the maximum densities (in individuals sample-1) found in both parts in the two-year
period, the part of submerged plant (top or bottom, see section 3.2.1 for details) where densities were
frequently more abundant and the max. density found, the across-year variability in densities, judged
from the differences in grand means (see Table 3.9) and size of population peaks in each year, the
percentage decrease or increase in average density between years (in brackets, year when maximum), and
the month when the maximum density of each species was reached (i.e. ‘seasonality).

Concept Acroperus Alona sp Chydorus Eurycercus Graptoleberis Peracantha


harpae spp lamellatus testudinaria truncata
Lily leaf- petiole ‘preference’ * Petioles Leaves-sites Leaves Petioles Leaves Leaves
(n.s.) 1,5 (n.s.)
Petioles-site 2
Max. density on leaves 209 450 418 33 551 380
(ind. sample-1)
Max. density on petioles 173 128 193 12 64 37
(ind. sample-1)
Part of submerged plant where Bottom Top No pref. Bottom Top-Bottom Top
more frequently abundant * (n.s.) (n.s.) (n.s. 1998) (n.s.)
Max. density on submerged 55 15 15 76 67 27
plant (ind. sample-1) Abundant Very scarce Very scarce Very abundant Abundant Scarce
Across-year variability Extremely var. Variable Variable Variable Extremely var. Variable
% of each year grand mean ± 600 % ± 300 % ± 2000 % ± 2000 % ± 500 % (1998) ± 300 %
(1998) (1998) (1998) (1998) (1998)
Seasonality (month of max.) September September July ’98 June & Sept. July ’98 September
September ’98 June ‘99
‘99 June ‘99

Table 3.18. Variables summarizing the ‘habitat’ association of the main plant-associated filter-feeders
(Scapholeberis mucronata, Sida crystallina and Simocephalus vetulus) and of the raptorial species
Polyphemus pediculus, sampled from Little Mere over the two-year sampling programme. Tabled are the
lily part (leaf or petiole) where denser, the maximum densities (in individuals sample-1) found in both
parts in the two-year period, the part of submerged plant (top or bottom, see section 3.2.1 for details)
where densities were frequently more abundant and the max. density found, the across-year variability in
densities, judged from the differences in grand means (see Table 3.9) and size of population peaks in each
year, the percentage decrease or increase in average density between years (in brackets, year when
maximum), and the month when the maximum density of each species was reached (i.e. ‘seasonality).

Concept Polyphemus Scapholeberis Sida crystallina Simocephalus


pediculus mucronata vetulus
Lily leaf- petiole ‘preference’ * Leaves Leaves Leaves Leaves
(n.s.) (n.s.)
Max. density on leaves 2700 270 1731 386
(ind. sample-1)
Max. density on petioles 259 87 876 133
(ind. sample-1)
Part of submerged plant where more No pref. Top Top No pref.
frequently abundant (n.s.)
Max. density on submerged plant 10 5 171 353
(ind. sample-1) Very scarce Very scarce Abundant Very abundant
Across-year variability Variable Variable Extremely var. Not variable
% of each year grand mean ±150 % ±200 % (1999) ±2700 % (1999) ±30 % (1999)
(1999)
Seasonality (month of max.) June-July ‘98 July April ’98 June-July ’98
July-Aug. ’99 Sept.Oct. ‘99 Aug. ‘99
‘n.s.’ indicates the differences in densities between leaves and petioles are not significant (see Table 3.14,
for details).

64
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Table 3.19. Variables summarizing the ‘habitat’ association and seasonality of the main ‘open-water’
species in Little Mere (Bosmina longirostris, Ceriodaphnia spp, Daphnia spp, Cyclops spp and Eudiaptomus
gracilis) over the two-year sampling programme. Tabled are the lily-bed or open-water ‘preference’, the
site where maximum densities were generally reached, the maximum density (in individuals l-1) in each
year, the across-year variability in densities, as judged from the increase/decrease in densities in both lily
beds and open waters, the percentage increase or decrease in density in either habitat (lily beds, LB, and
open waters, OW) and the month when population peaks were reached (i.e. ‘seasonality’).

Concept Bosmina Ceriodaphnia Daphnia spp Cyclops spp Eudiaptomus


longirostris spp gracilis
Lily bed or open water* Open water Lily beds Lily beds Open water Open water
(n.s.) (occasionally more
in lily beds)
Site ‘preference’ 3, 4 > 1, 2, 5 1, 2, 5 > 3, 4 5 in 1998 3, 4 > 1, 2, 5 3, 4 > 1, 2, 5
1 in 1999
Max. density (ind. l-1) 1998: 135 1998: 204 1998: 409 1998: 146 1998: 127
1999: 1264 1999: 1358 1999: 456 1999: 264 1999: 610
Across-year variability Extremely var. Extremely var. Variable Variable Extremely var.
% more in lily beds (LB) or 1998: 350 % OW 1998: 20 % LB 1998: 37 % LB 1998: 100 % OW 1998: 600 % OW
in open waters (OW) ** 1999: 2 % OW 1999: 250 % LB 1999: 61 % LB 1999: 60 % OW 1999: 1400 % OW
Seasonality (month of max.) 1998: October 1998: Sept.Oct. 1998: July 1998: Sept.Oct. 1998: November
1999: July-Aug. 1999: July-Aug. 1999: May & Sept. 1999: July-Aug. 1999: April
‘n.s.’ indicates densities in lily beds or open waters are not significantly different (see Table 3.7, for
details); ** percentages have been calculated from the marginal means for lily beds and open waters (see
Table 3.6).

65
CHAPTER 3

3.4. Discussion.

A few very clear patterns in spatial distribution of species were observed. Daphnia
populations were significantly larger in lily beds in Little Mere, lending support to the idea
that lilies provide a refuge against fish predation for the zooplankton. ‘Open waters’ (i.e.
outside lily beds) had little submerged vegetation that could act as refuge during most of both
growing seasons sampled. Differences in the size of populations between lilies and open
waters increased towards mid-summer, perhaps coinciding with a late recruitment of
planktivorous fish.

Gliwicz & Rykowska (1992) interpreted the onshore-offshore gradient in pelagic species’
densities as the result of predator-induced behaviour of the zooplankton. However, the
smaller, not the larger, populations were found in the littoral areas, dominated by a belt of
common reed (Phragmites australis). Lauridsen & Buenk (1996) found that nighttime
densities of Daphnia increased in the open water, whilst in the littoral zone, vegetated with
the fine-leaved submerged Potamogeton pectinatus, densities were much lower. The highest
daytime densities were at the edge of the plant beds, and not inside them. Lauridsen et al.
(1996) compared the dynamics of zooplankton in and out of different-diameter plant beds.
Beds with large edge-to-size ratios were the most effective daytime refugia against fish
predation.

Refuge effectiveness has been found to be dependent on a balance between predator pressure
(as measured by areal fish density) and submerged plant density (Schriver et al. 1995). Little
is known about the differential refuge effectiveness for zooplankton of floating and
submerged plants. There has been controversy concerning the potential refuge role of
nymphaeids as opposed to completely submerged vegetation (Venugopal & Winfield 1993;
Scheffer 1998, Blindow et al. 2000). Venugopal & Winfield (1993) found higher perch
densities in large yellow lily stands (Nuphar lutea) in Priest Pot, English Lake district.
However, these differences were not significantly different (F-test, p>0.05), perhaps because
results were based on a small sample size (i.e. 61 perch over five weeks). Lower light-levels
under the floating leaf canopy may provide a relative refuge for zooplankton in lily beds
(Scheffer 1998). However, the foraging behaviour of most planktivorous fish is hardly
impaired by the reduction in ambient light due to shading in lily beds (Wright & Shapiro
1990). Timms & Moss (1984) proposed it is the direct ‘physical interference’ with the feeding

66
PLANT-ASSOCIATED CLADOCERA DYNAMICS

behaviour of planktivores that may explain the apparent refuge role of nymphaeids (see also
Moss, Kórnijow & Measey 1998). However, Winfield (1986) found that the foraging
performances of rudd (Scardinius erythropththalmus) and perch (Perca fluviatilis) were
actually increased in a dense stand of Nuphar spp (albeit an artificial stand) relative to those
shown in open water, perhaps owing to a reduction in ‘perceived’ predation risk from
piscivores. Moss et al. (1998) have shown that lilies (Nuphar lutea) in Little Mere (England)
reduced predation by perch (Perca fluviatilis) on Daphnia spp at high plant densities.

The clearest evidence for the refuge role of lily beds was obtained here in the second year of
sampling. Daphnia was only dominant in the first two months of this growing season (i.e.
April & May). By the beginning of June only residual populations could be found. The
severity of the impact, as compared to this species’ population dynamics the previous year,
and the fact it concerned the whole lake, suggest it could have been the result of heavy
predation by young-of-the-year (YOY) fish once they reached the size to have any significant
effect on Daphnia populations. These yearly variations in predation pressure from
planktivorous fish on zooplankton are common in shallow lakes, especially those of high
trophy (see Scheffer et al. 1997). Only residual populations were maintained until significant
numbers were detected at a single sampling station by the end of the growing season (i.e.
September ’99; see Figure 3.8).

This observation has implications for the estimation of zooplankton grazing impacts as
calculated from single measurements of density, generally from open-water sampling stations
(e.g. Blindow et al. 2000). Also, Daphnia have been shown to migrate vertically even in such
shallow depths as those in Little Mere (F-test; p<0.05), albeit only in enclosure studies (see
Chapter 5). It is unlikely Daphnia gains any refuge from visually-predating fish by migrating
vertically in this lake and the persistence of this behaviour in Little Mere may simply be a
relict trait of populations with strong vertical migrations and that migrated in from the deep
Mere Mere, upstream (see Chapter 2 for details; Fig. 2.3).

Two small cladocerans, Bosmina longirostris and Ceriodaphnia spp dominated the
zooplankton community in the absence of Daphnia species in July and part of August in the
second year of sampling (Figure 3.8). Daphnia is a more efficient filter-feeder than smaller
cladocerans (Brooks & Dodson 1965; Chow-Fraser & Knoechel 1985; Mourelatos & Lacroix
1990). In the presence of intense planktivory by visually-predating fish, smaller cladocerans

67
CHAPTER 3

would come to predominate over the larger, and thus more vulnerable, Daphnia species. This
could be because the fish would deplete invertebrate predators feeding preferentially on
smaller zooplankton, or, more directly, because fish would predate preferentially on larger
zooplankton. In Little Mere, the apparent shift in community composition lends further
support to the idea intense fish predation was responsible for the temporary disappearance of
Daphnia from the lake in growing season ’99.

Larger-bodied Daphnia were consistently found in lily beds, too (Figures 3.14 & 3.15). Very
few individuals larger than 1 mm were found in the two open-water sites in 1998, with the
exception of two dates in July, when very large densities were found throughout the lake. This
would be expected if individuals of different cohorts had a chance to grow in the face of fish
predation. The magnitude of this apparent refuge effect in 1998, was variable. In July, larger
Daphnia could be found in open waters, and densities were high in both lilies and open
waters. By contrast, in August 17th virtually all Daphnia in the lake were found among the
lilies. The few Daphnia sampled from outside lily beds were smaller than 0.7 mm in size,
while about half the populations in lily beds were larger than 0.9 mm (see Fig. 3.14). Again,
all these facts are consistent with the idea lily beds offer a very powerful refuge for Daphnia
in the face of fish predation.

Changes in size-structure of Daphnia populations in 1999 give further support to the idea lily
beds act as refuge. Before hatching of young-of-the-year in May, size-structure across sites
was comparable across sites, both in lily beds and out. In May, most animals in open waters
were in small size-classes. The potential for population growth at the end of May was very
large. Most of the populations were in larger size classes and 55-84 % of the populations in
lily beds (but not in open waters) were made up of large ovigerous individuals (see section
3.3.5). This could be explained by larger Daphnia migrating less out of relatively protective
lily beds in response to a very high predation risk outside the beds or that size-selective fish
predation was more intense outside lily beds. Densities then were substantially lower in the
two open-water sites (see Fig. 3.9). Further support for this hypothesis would need direct
evidence that the overall decline of Daphnia populations was related to fish moving into the
lily beds. Large numbers of small perch (Perca fluviatilis) were found in surveys with seine
nets, conducted in Little Mere in November ’99 (see Appendix A).
Evidently, lily beds only delayed the demise of populations, but a threshold fish density can
be imagined below which this ‘refuge effect’ would be sufficient to maintain relatively stable

68
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Daphnia populations, other things being equal (Schriver et al. 1995; see, for example, Moss,
Kornijow & Measey 1998; see also Discussion in Chapter 5).

Large-scale distribution of plant-associated cladocerans in Little Mere.

The plant-associated filter-feeders were never very abundant in the open-water (see Table
3.18), although, occasionally, Simocephalus vetulus was found in considerable numbers on
Ceratophyllum demersum. Large numbers were found when Daphnia populations were
strongly increasing in the waters above the submerged plant beds. Daphnia were seldom
found in submerged plant parts (a max. of 451 ind sample-1 were present in a sample on July
29th, 1998; 21 individuals were counted in one sample on October 8th, 1999). However, these
Daphnia could have been inadvertently introduced from the above waters during collection of
the sample. In any case large numbers of Simocephalus were never found along with large
numbers of Daphnia in submerged plant beds, suggesting these two species favour different
niches.

In lily beds, Simocephalus was associated with leaves, and few individuals were caught in
water samples taken among the lilies. High Daphnia densities were found in lily beds together
with considerable numbers of Simocephalus (e.g. compare Figures 3.9 & 3.11). Again, this
seems to point towards both species partitioning resources available in adjacent ‘habitats’, i.e.
lily leaves for Simocephalus, and water among the lily structure for Daphnia spp.

Simocephalus is an extremely poor swimmer (Sharma & Pant 1984). Movements of


populations out of lily beds have not been observed (Lauridsen et al. 1996). In submerged
plant beds in Little Mere, dense patches of Simocephalus (i.e. around 1,000 ind g-1DW, or
around 20,000 ind m-2) were sporadically found, at the height of summer ‘98, and at the end
of growing season ‘99 (Fig. 3.13). Populations in submerged plant beds are probably quite
independent of those in lily beds.

Size-structure changes of Simocephalus were more conspicuous in the first year. There was
little pattern to these changes, though. During the second year, size-structure did not change
visibly even during dates with maximal fish presence (as implied from Daphnia
disappearance; i.e. July mainly). This would be expected if Simocephalus was less vulnerable
to fish, as predation pressures were apparently stronger the second year. The species in the
plant-associated cladoceran community do not seem to be subject to the same predation

69
CHAPTER 3

pressures of ‘open-water’ grazers (Beklioglu & Moss 1995, 1996), perhaps owing to their
lower visibility while remaining still (Fairchild 1981, but see Fairchild 1982). However, it is
difficult to imagine what factors may drive size-structure dynamics of Simocephalus in Little
Mere. Fish predation, as with Daphnia, is probably not a negligible influence, even if at lower
intensities than over the faster-moving, and therefore perhaps more visible, Daphnia. In
support of this, more of the larger Simocephalus were found in late summer ’98. In 1999,
Simocephalus individuals slowly grew from August onwards, and most of the September
peaks were made up of small animals (i.e. <0.7 mm), offspring of the larger August animals.

Sida crystallina, on the other hand, seldom co-occurred with high Daphnia densities in lily
beds. The extreme seasonality of Sida (Table 3.18; Figs. 3.10 & 3.12) may be an alternative
pathway to co-existence with competing filter-feeders such as Daphnia. Thus, this species
may increase before Daphnia’s spring peak in April-May (e.g. in 1998), or after the demise of
Daphnia populations, at the end of the growing season (e.g. in 1999). It may be a pioneer
species. It thrives when it has no competition from other species, suggesting it takes
advantage from occupying competitor-free space (Fairchild 1981). Daphnia and Sida did
occasionally co-occur, however, albeit in localized areas of the lake (e.g. site 5 at the end of
growing season ’99). Indeed, very high densities of Sida occurred occasionally in some lily
beds, but not others. In addition, large populations developed in submerged plant beds in the
second growing season. Lily beds seem to be the preferred ‘habitat’, however, as large
numbers first developed there. Only when populations reached densities well in excess of
30,000 ind m-2 (e.g. Sept. 27th, 1999), was Sida also observed in more open waters (e.g. Oct.
8th, 1999).

Unfortunately, little can be said on top-bottom differences in submerged plant beds because
samples could only be taken of both fractions on two occasions. In addition, it would have
been interesting to extend sampling to other sites with submerged plants in order to gauge an
estimate on horizontal variability in Sida’s density estimates. This may be crucial when
extrapolating point figures to whole areas, for example when evaluating grazing impacts of
whole populations, (see Chapter 4). Important plant-associated filter-feeders such as Sida may
find submerged plant surfaces an attractive option, particularly when ‘sources’ (i.e. lily beds)
become food-depleted.

70
PLANT-ASSOCIATED CLADOCERA DYNAMICS

Sida crystallina is largely confined to the littoral zone as it is a sessile phytoplankton filter-
feeder typically attached to plant surfaces. Availability of attachment areas is not thought to
limit Sida’s distribution as both fish predation (Fairchild 1983) and food conditions (Fairchild
1981) generally regulate population size below a size at which attachment would be scarce.
However, in this study Sida was found at very high densities on some dates in 1999 (i.e. >800
ind g-1DW, or 40,000+ ind m-2; see Fig. 3.10). Attachment space could then have been scarce,
to the extent of animals having to swim despite their known preference for living directly
attached to surfaces (Szlauer 1973). Attachment to sub-optimal surfaces such as the underside
of lily leaves (Frodge et al. 1990) may also have been forced by crowding. Fairchild (1981)
found Sida crystallina in stands of Nymphaea to swim more in search for reattachment in
areas with fewer lily leaves than in denser parts of the same lily bed. A higher density of Sida
was found on isolated plants or sparsely distributed ones, particularly those on margins of
stands of aquatic vegetation. This implies density (as expressed per m2) was uniform across
areas and not denser where plant biomass was higher. Indeed from my data, Sida density can
be seen to increase with decreasing plant availability. It can be shown density (in ind g-1DW)
increased as lilies died down at the end of the growing season, particularly at sites where
populations were not very crowded.

Fairchild (1981) also found density of this species was higher on younger plants. Significant
differences in densities across depths were observed in immature individuals but not in adults.
Juvenile Sida were significantly more abundant near the surface. The behaviour of Sida was
not observed to change at night (Fairchild 1981). However, it is not known what determines
the distribution of this species at a larger scale, tens of metres (e.g. between sampling stations
in this study). The only published detailed study on the spatial distribution on this species is
that of Fairchild (1981) and was based on a restricted number of samples of mainly emergent
plant species (Scirpus). Few references have been made to population movements of this
species, for instance between plant beds, except a passing reference to 30 metres being a good
approximation to the range covered by any given population (Fairchild 1983). Confusingly,
Lauridsen et al. (1996) found density differences between lily beds and more open waters
were significant during the day but not at night. Fairchild (1981), however, did not observe
diel behaviour changes of this species.

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CHAPTER 3

Population dynamics.

Standard error bars are large, particularly when densities are highest, and therefore it could be
we are actually comparing different ‘patches’ in egg-ratio models (see section 3.3.6 for details
(see Keen & Nasaar 1981). Occasionally, apparent birth rates exceeded actual population
growth rates. These inconsistencies indicate the necessity of drawing error bars for parameters
used in egg-ratio models. On the other hand, the predictably higher yield of young of
Simocephalus after the 10th of July could not be detected, as perhaps 3 or 4 generations of
young would have succeeded at the temperatures found those dates. This highlights the need
for sampling at periods close to the egg developmental period, which is shorter at higher
temperatures, in order to detect all significant dynamics of populations.

There was a tendency in both years towards a larger proportion of big Simocephalus towards
late summer. On many occasions actual growth rates of Simocephalus populations cannot be
justified by new births suggesting a source of mortality. Dynamics was more irregular in 1999
than in the previous year, perhaps as a result of larger fish predation pressure.

In 1998, both Simocephalus and Daphnia presented relatively high birth rates at the end of
June (see Figs. 3.18 & 3.19), particularly in lily beds. Simocephalus maintained the highest
birth rates of the two species, perhaps gaining advantage of the added food source of
periphyton, generally unavailable to Daphnia. Birth rates of Daphnia were larger within lily
beds than in more open waters. A reduction in perceived predation risk among lilies could
release time and effort to feeding, increasing population growth.

Small-scale distribution of species.

Crowding levels were generally correlated with population size. However, crowded
populations were found to be very regularly distributed. Lloyd (1967) thought patchiness
should be lower at higher densities, as crowding pressures lead to homogenisation. However,
Whiteside (1974) found that increased densities of chydorid populations at Elk Lake
(Minnesota) did not lead to more uniform arrangements.

There are two main contesting hypotheses for patchiness generation:

72
PLANT-ASSOCIATED CLADOCERA DYNAMICS

1) Patches are the result of availability and search for best food patches. Crowded patches
will be increasingly unattractive to foraging animals and relatively empty spaces will be at
a premium (Lloyd 1967).
2) Patchiness is the consequence of low-intensity and unspecific fish predation (Whiteside
1974).

Fish pressures mid-summer ’99 were likely so intense that even chydorid populations were
affected. Patchiness of these populations was extremely variable. In contrast, the larger
populations developing by late summer ’99 were regularly distributed, in agreement with
Whiteside’s hypothesis of predation-driven patchiness patterns.

The limited mobility of chydorids, as they generally crawl rather than swim (see Fryer 1968),
would make the strategy of searching and then staying in ‘best food patches’ even more
advantageous than for swimmers. Moreover, delays in how spatial arrangement would reflect
underlying resource competition would be expected in chydorids, especially when food
conditions changed rapidly. Data from 1999, however, seem to indicate chydorids’ patchiness
was in direct relation to fish predation.

In contrast with chydorids, distribution of plant-associated filterers was independent of fish


predation pressure, and crowded populations were very regularly distributed. Extraordinarily
crowded conditions were experienced locally by Sida populations at the end of growing
season ’99. The dense aggregations were not patchily distributed. Again this observation is in
support to the idea it is intraspecific, and perhaps interspecific (e.g. Daphnia &
Simocephalus), competition that mainly regulated the small-scale distribution of cladocerans
in Little Mere during the two years sampled, rather than fish.

The comparison between microcrustacean populations in lily beds and more open waters in
this chapter will be extended to a comparison of their respective trophic role in the following
chapter. The main hypothesis tested is that plant-associated microcrustacea have an important
role to play in the maintenance of water transparency in Little Mere.

73
CHAPTER 3

Conclusions.

• There were significantly more Daphnia in lily beds than in more open waters. The
individuals in lily-bed populations were also larger. Both observations lend support to the
hypothesis lily beds constitute an effective refuge for large-bodied Cladocera in the face
of fish predation risk.

• Sida crystallina developed large but short-lived populations at the beginning of growing
season ’98 (April) and at the end of the season in 1999 (September-October), maintaining
residual populations during the rest of both summers (Fig. 3.10).

• Sida crystallina was occasionally very abundant in Little Mere, reaching maximum
densities of 1205 ind g-1DW lily leaf (1731 Sida in one leaf sample; see Table 3.18).

• Simocephalus vetulus reached maximum densities in July and August in both years, even
when high fish densities were apparent (as inferred from Daphnia’s low numbers). This
suggests this species is not as exposed to fish predation as more pelagial species such as
Daphnia.

• Large populations of most chydorid species developed in the first growing season. During
growing season ’99, by contrast, numbers were between 3 and 20 times lower (see Table
3.17).

• Lowest patchiness values were found at periods of maximum crowding in all species
particularly during the first growing season, suggesting food limitation and intra-specific
competition were the main drivers of the spatial arrangement of populations at the small
scale (Lloyd 1967).

• During growing season ’99 (April to October), spatial distributions of chydorid species
seemed to be driven by fish predation pressure. Patchiness was extremely variable during
periods of high fish predation. By contrast, the generally much larger populations in late
summer ’99 were very regularly distributed (i.e. not very patchy). This is in agreement
with Whiteside’s hypothesis of predation-driven patchiness patterns (Whiteside 1974).

74
CHAPTER 4

Grazing rates of the plant-associated and open-water


cladoceran communities in Little Mere.
CHAPTER 4

4.1. Introduction.

Water transparency is influenced by multiple processes (Frodge et al. 1990; Barko & James
1991; Hansson 1992; Schriver et al. 1995; Karjalainen 1998; Gross 1999; see Chapter 1 for
details). Many of these processes are particularly intense within plant beds in shallow lakes,
and are thought to buffer the whole lake from a “switch” towards a turbid, phytoplankton-
dominated state (Leah et al. 1980, Timms & Moss 1984, Scheffer 1989, Blindow et al. 1993).
The relative importance of grazing and nutrient limitation may be some function of lake depth
(Moss et al. 1997). Moss et al. (1994) found in the West Midland Meres (Cheshire) that in the
deep meres (i.e. > 3 metres) it was nitrogen which limited phytoplankton growth. In the
shallow meres, inverse correlations between peak chlorophyll-a concentrations and
cladoceran density (number of individuals per litre), particularly of Daphnia species, were
taken as indicative of top-down regulation of algae. Elser et al. (1990) suggested algal
biomass in oligotrophic lakes is regulated by nutrient availability rather than by zooplankton
grazing, and Carpenter et al. (1997) proposed a nutrient loading gradient to organize the
strength of the main regulatory forces of primary and secondary production in shallow lakes.
In this gradient, grazing is considered a negligible control of algae under high nutrient
loadings. In Little Mere, a shallow fertile lake (see Chapter 2 for details), top-down control by
Daphnia seems to have been the most powerful influence on phytoplankton (Stephen, Moss &
Phillips 1998). During the 1991-1999 period, Daphnia density was inversely related to
chlorophyll-a values. However, in particular years and within periods of growing seasons (i.e.
April to October), low chlorophyll-a concentration and high water transparency cannot be
accounted for by Daphnia grazing. What is not known is what are the relative roles of
Daphnia and plant-associated grazers. Very little attention has been paid to the role of
microcrustacean grazers other than Daphnia species, particularly plant-associated genera such
as Sida and Simocephalus, may have in the maintenance of clear water in shallow lakes.

Also, information is needed on the relative contributions of 'direct' mechanisms mediated by


plants, for example the nutrient uptake by plants in competition with planktonic algae, and
'indirect' ones, for instance the grazing of phytoplankton by the plant-associated genera (e.g.
Sida and Simocephalus).

Planktonic zooplankton and even more so their plant-associated counterparts, exist in a


bewildering array of forms and sizes in shallow lakes (Fryer 1968). This diversity may mean

76
GRAZING RATES

we cannot easily identify what fractions of the biota are the main grazers of phytoplankton
(Frost 1984). Furthermore, accurate estimation of grazing rates in natural communities is
difficult, and several ways of estimating these have been devised (see, for example, Gliwicz
1968, Enright 1969, Haney 1971, Porter 1973, Gulati et al. 1982). In broad terms, methods
can be grouped into two classes.

The first group of methods estimates grazing rates directly, from field death rates of algae,
estimated from repeated sampling of phytoplankton populations (e.g. Allen 1922), or through
the in situ incubation of phytoplankton and zooplankton assemblages (Porter 1973). These
methods are subject to large sampling variability and algae cell division rates applicable to
natural environments must be known with a degree of accuracy. The second type of estimates
involves estimating herbivory indirectly, from field abundance and clearance rates of grazers
(e.g. Gulati et al. 1982). The main disadvantage of the latter type of estimates derives from
the large number of variables normally involved in calculations (e.g. field abundance of
grazers, calculated volumes of water and/or areal plant biomass estimates, clearance rates).
Thus, errors are multiplied and interpretation of results can be complicated (see Discussion
for details).

Indeed, the feeding rates of microcrustacea are influenced by many interacting environmental
factors. Species compete for resources (Horton et al. 1978) and predation influences the
spatial distribution of grazing pressures (Kerfoot 1987). Many of these influences and factors
change spatially, seasonally and even daily, quantitative and qualitatively. Therefore it was
hardly surprising that results obtained in feeding experiments under controlled conditions in
the laboratory were generally poorly representative of natural conditions (see Peters &
Downing 1984).

The first in situ attempt at measuring feeding rates of particular species involved the
comparison of algae, bacteria and detritus food in a plankton trap submerged in the lake water
with an anaesthetising agent to stop grazing activity at a given time (Gliwicz 1968). The
feeding rate was then estimated from the difference in cell counts generally in a simple
medium, before and after the timed period during which the animals had been left to feed (e.g.
Fuller & Clarke 1936).

Methodological problems with this technique (e.g. resuspension of faeces, varying food
concentrations) prompted a series of seminal papers in which a technique was developed that

77
CHAPTER 4

was less labour-intensive than the cell-count method. This method involved uptake of
radioactively-labelled food items in short-term experiments (Marshall & Orr 1955; Nauwerck
1959 cited in Rigler 1971; Malovitskaya & Sorokin 1961; Monakov & Sorokin 1961, cited in
Burns & Rigler 1967). Relatively accurate instantaneous feeding rates could be obtained
quickly and easily.

Schindler (1971) stressed the necessity of using more than one food type with respect to both
size and shape, or “taste”, as ingestion rates seemed to vary with food species presented to the
animals. The issue of particle size feeding selection was first approached by Lampert (1974)
and Gophen et al. (1974). Lampert used populations of Daphnia pulex, offering tritium-
labelled bacteria and carbon-14 labelled algae as food organisms, then calculating an index of
food preference from the relative proportions of these two isotopes in the animal compared
with the feeding medium.

Haney (1971, 1973) designed an in situ method improving on Gliwicz’s 1968 apparatus,
bringing together the advantages of in situ experimentation and the short-term radioactive
technique. This innovative approach relied on using a plankton trap, or other means of
enclosing a volume of lake water with its natural biota (i.e. a “grazing chamber”), with a
piston that could rapidly introduce a small amount of highly radioactive food once the
chamber had been lowered at a certain depth into the lake. A typical experiment would last 5
minutes to an hour (depending on the gut passage time of the animal species concerned),
during which the zooplankton were allowed to feed. A critical step in this procedure is to kill
the animals before food has had time to complete gut passage. Moreover, the short feeding
period reduces the chance of relevant biological and chemical processes having a significant
effect on rates. On the other hand, the precision and ease of use of radioactivity counting
apparatus (e.g. liquid scintillation counters) greatly reduced the amount of time previously
needed to estimate rates from cell-count differences. Used with in situ experiments, this
technique allowed study of variables thought to play a role in the clearance rates on a spatial
and temporal scale under natural conditions (e.g. food concentration, temperature, vertical
migration of feeding animals).

Downing and Peters (1980) modified Haney’s method by enclosing plants in the chamber.
Thus, they were able to study the effect of body size and food concentration on the in situ
feeding of a plant-associated cladoceran, Sida crystallina. Littoral cladocerans apparently can
feed on food in suspended algae or scrape periphyton (Fryer 1968). In a later paper, Downing

78
GRAZING RATES

(1981) investigated whether littoral cladoceran species (i.e. Alona affinis, Chydorus
sphaericus and Sida crystallina) feed on these different food items on the basis of availability
or if other mechanisms are in play. Both food compartments were labelled with different
isotopes as in Lampert’s (1974) technique.

In this chapter, data on field abundance of grazers (reported in Chapter 3), plant biomass
estimates, and clearance rate estimates from in situ experiments carried out in Little Mere are
brought together in order to calculate measures of grazing rate (% of lake volume filtered per
day, i.e. % day-1). Clearance rates were estimated from in situ short-term feeding experiments
using a modified radio-labelling technique in which the animals were presented with both
periphyton and planktonic algae labelled with different isotopes (H-3 and C-14). The aim of
the differential labelling was to pick up differences in uptake related to food selectivity of the
animals (see Fryer 1968). Periphyton ingestion rates (in mass of periphyton ingested per day)
were also estimated from the carbon-14 activity taken up by animals.

The main aim of this chapter is to compare the grazing rates (% day-1) of the ‘open-water’
microcrustacean grazer community with those of the Cladocera associated with plants, in
order to establishing the relative roles of Daphnia and of hitherto often ignored plant-
associated filter-feeders in Little Mere. Multiple and interacting mechanisms may be
responsible for variations in phytoplankton density and water transparency (see Figure 1.1).
Because of this complexity, I am directly addressing the question of what is the absolute
contribution to these variations of cladoceran grazing. The validity of grazing rate estimates is
discussed in relation to sources of error (i.e. estimates of field abundance of main grazers,
clearance rates and areal plant biomass estimates).

79
CHAPTER 4

4.2. Materials and Methods.

4.2.1. In situ short-term feeding experiments.

A series of nine short-term experiments was conducted between the 26th of April 1999 and
29th of October 1999 in Little Mere (Cheshire, England), see Table 4.1 for details of each
experiment.

Table 4.1. Date, weather and number of samples of the 9 in situ experiments.

Experiment Date Weather Total number of


number samples per experiment
1 26th April Sunny spells, slight breeze. 51
2 12th May Showery, mild, no wind. 65
3 1st June Sunny spells, no wind. 69
4 7th July Rainy, windy. 43
5 28th July Sunny, no wind. 68
6 13th Aug. Showery, mild, no wind. 61
7 23rd Sept. Overcast, breezee. 100
8 5th Oct. Sunny, no wind. 93
9 29th Oct. Variable, slight breeze. 100

In a typical in situ grazing experiment using radio-labelled algal food, a plankton trap or other
means of enclosing a volume of lake water with their natural biota (i.e. a “grazing chamber”)
is submerged in the lake. The radioactively labelled food is injected rapidly and left for a short
period (i.e. 5 minutes to an hour, depending on animal’s gut passage time) during which the
animals are allowed to feed. I used a 7 mm thick perspex chamber of capacity 6 litres (see
Figure 4.1), based on an original design by Downing & Peters (1980).

Figure 4.1. Photo and schematic picture of the grazing chamber used in experiments. Dimensions are 10 x
20 x 30 cm. The chamber was constructed by sealing perspex pieces. The closure was made water-tight by
using strips of neoprene and silicone around the inside. The chamber can open allowing a thorough
cleaning before a new experiment is initiated. The sampling port allows the retrieval of the strips with
periphyton before the sampling of the suspended algae and the animals at the end of each experiment.
Adapted from an original design by Downing & Peters (1980).

80
GRAZING RATES

To approach the issue of periphyton versus suspended algae feeding selectivity of animals,
radioactive periphyton and planktonic algae, both of which had been previously labelled in
the laboratory with different isotopes, were introduced into the grazing chamber along with
the animals.

1. Preparation of periphyton food.


- Carbon-14 labelled periphyton.

An artificial, plastic, substrate with a rough surface was chosen for periphyton colonization
for a series of reasons. In the first place, it facilitates enormously the labelling process. The
natural substrate for the algae, a plant, would be expected to compete for the uptake of label.
Many aquatic plant species are known to use bicarbonate (i.e. the carbon-14 carrier used in
these experiments) as a carbon source. It is also easier when using plastic strips to standardise
rates to unit area. A smooth and bright polythene plastic was an option discarded because it is
thought to be more “unnatural” from the point of view of periphyton growth (Eminson &
Moss 1980).

The periphytic community colonizing plastic substrates resembles that on more natural
surfaces (Eminson & Moss 1980, Hansson 1992, but see Young 1945, Tippett 1970). Also,
the potential problem of periphyton availability to feeding animals within the box can be
minimized by increasing the number of strips with periphyton presented to them. The problem
remains, however, of how natural the strips may be to potential microcrustacean scrapers.

Four small strips (i.e. 10 x 2 cm; see Fig. 4.3) were used. A set of strips was left for at least
three weeks prior to each experiment, dangling from wires attached to a raft anchored in the
experimental lake (see Fig. 4.2). To label the periphyton, generally 50 µCi of carbon-14
activity (54 mCi mmol-1, aqueous solution; ICN Pharmaceuticals, Inc.) was added to each of
four 250 ml flasks in 200 ml of GF/C-filtered lake water collected from the experimental lake.
The flasks were then left under continuous tungsten bulb light for at least 72 hours prior to
each in situ experiment. Details on final strip activities achieved for each in situ experiment
are shown in Table 4.2.

81
CHAPTER 4

Figure 4.2. Moment when the raft for periphyton in situ colonization is placed into the lake.

Strips were transported within 2 hours to Little Mere in a plastic bottle with GF/C-filtered
lake water, to be used in the in situ experiment.

Table 4.2. Carbon-14 activities, incubation periods (weeks) and labelling times (hours) for strips used in
each in situ experiment. Radioactivity of strips is shown as both total C-14 average activity (N=4) per strip
(MBq) and C-14 average activity (Bq) per µg of periphytic chlorophyll-a.

Experiment Incubation Labelling Mean activity Mean activity per strip


(date) period time (hours) per strip (Bq µg-1 periphytic chl-a)
(weeks) (MBq)
1. 26th April 3 4 0.258 8.5
2. 12th May 2 5 0.054 1.6
3. 1st June 3 5 0.081 0.9
4. 7th July 2 5 0.040 2.4
5. 28th July 2.5 5 0.017 1.5
6. 13th August 2.5 5 0.099 12.7
7. 23rd Sept. 5 5 0.033 2.7
8. 5th October 2.5 5 0.036 2.4
9. 29th October 4 2 0.004 0.1

- Estimation of periphyton concentrations.

In order to compare feeding rates of animals on periphyton across experiments we need a


specific measure of carbon-14 activity (i.e. radioactive counts per minute CPM µg-1 periphytic
chlorophyll-a, or CPM µg-1 dry weight of periphyton). To obtain these measures, each strip
was cut in half lengthwise (see Fig. 4.3 for a flow chart of procedures). A set of halves was

82
GRAZING RATES

processed for the estimation of chlorophyll-a content (phaeopigment acid-correction method


on SFS 5772 Finnish standard, based on ISO 10260, 1992) and dry weight. The periphyton
was scraped off the plastic strip using a steel wire brush into a known volume of distilled
water and sieved through a 140 µm mesh to remove larger elements, such as filamentous
algae (see Figure 4.4).

RADIO- To lake In situ SAMPLE


LABELLING EXPERIMENT PREPARATION
(2-4 days in the lab.) AND COUNTING
In situ
INCUBATION Strips: 2 x (10 x 21 cm.)
cm.)
(3-4 weeks)

Strips:
4 x (10 x 42cm.)
cm.)
CHLOROPHYLL-a
ESTIMATION
Strips: 2 x (10 x 21cm.)
cm.)

Figure 4.3. Sequence of steps to prepare radio-labelled strips and take samples at the end of each in situ
experiment. See text for details.

To estimate the chlorophyll-a content, a sample of the resulting periphyton suspended algae
was collected onto a GF/C filter and extracted using ethanol 90 % in a hot bath (T=75 °C) for
5 minutes (SFS 5772 Finnish standard). Absorbance was read at 750 and 665 nanometres.
Another sample collected onto a tared GF/C filter was oven-dried for 24 hours at 105 °C and
weighed on an electronic balance to estimate dry weight of the periphytic algae. This was
calculated by subtraction from the final weight of the tared filter. Other contributions to the
final weight of the filter, such as inorganic solids, were considered negligible and therefore
not included in dry weight calculations.

- Periphyton activity.

At the end of the timed feeding period, the sampling port on the grazing box was opened and
the strips retrieved with forceps and placed in four separate bottles with distilled water. Once
on shore the radioactive periphyton is thoroughly scraped off using a steel wire brush, as was
used in the laboratory for the estimation of concentration parameters (see previous section).
Again, the resulting periphyton suspended algae were sieved through a 140 µm mesh to
remove larger particles and the whole volume was collected onto GF/C filters with the aid of
a hand pump.

83
CHAPTER 4

Carbon-14 radioactivity measured from periphyton on strips,


detached by shaking or by whole-strip assays
6000 Strip assay (activity per strip)
Shaking and filter (activity per strip)

Counts per minute (103)


5000

4000

3000

2000

1000

0
6 24 48 72 168
Labelling period (hours)
Figure 4.4. Carbon-14 activity of periphyton colonizing plastic strips, measured in a time-course uptake
experiment at times 6, 24, 48, 72 and 168 hours (1 week). Plotted are means (N=3 strips) and one standard
error at either side of the mean. Two methods for measuring carbon-14 on the periphyton were compared
in this test. In the first, the plastic strip was cut into small pieces which were then introduced into
scintillation vials and radioactivity measured. In the second, periphyton was detached from strips by
shaking in water during one minute. The periphyton was then collected onto GF/C filters and these placed
into vials for activity measurement. Direct assay of strips showed generally higher activities than assays
with GF/C filters. Because of large variability, however, differences between these two treatments were
not statistically significant.

Each strip was cut into small pieces and both the filter and its corresponding scraped strip (see
Fig. 4.3) were introduced into marked scintillation glass vials with 1 ml of tissue solubilizer
(soluene-350 and toluene, quaternary ammonium hydroxide mixture, Canberra-Packard).
These vials were left for digestion to take place for around 24 hours at 40 °C, before about 10
ml of scintillation cocktail were added (toluene-based) with 50 µl of glacial acetic acid as a
buffer. Swings in pH have been shown to interfere with radioactivity counting (Downing &
Peters 1980).

Average strip carbon-14 activities


600 in in situ experiments

500
CPM (103) microg-1 chl-a

400

300

200

100

0
26-abr 12-may 01-jun 07-jul 28-jul 13-ago 23-sep 05-oct 29-oct

Figure 4.5. Average strip activity (CPM (103) µg-1 chlorophyll-a), bars are of one standard error at either
side of the mean (N=4 strips).

84
GRAZING RATES

Differences in uptake cannot be related to labelling period or input activity, but are date-
specific. The suggestion is that different components of the periphytic community are more
efficient at using bicarbonate, the carbon-14 carrier, as a carbon source.

2. Planktonic algae food.

The natural seston in the chamber was labelled using as “tracer” a dense culture of
Scenedesmus spp. This species was chosen as a “tracer” of phytoplankton ingestion because
of the ease with which it can be cultured and maintained without significant contamination.
Experiments were short (i.e. around 15 minutes; see Table 4.4) and significant losses due to
sinking within the chamber were thus not expected. It was assumed the “tracer species” was
representative of the range of sizes ingested by the animals under natural conditions. This
assumption is appropriate and avoids the problem of unequal label uptake by different algae
species in the natural assemblage (see, for example, Lampert 1988). In addition, the small
volumes injected into the 6-l chamber are comparatively small, thus not changing the natural
food conditions in the lake.

- Preparation of the H-3 labelled Scenedesmus “tracer”.

The minimum volume of Scenedesmus culture to be injected in the chamber for ingestion to
be detectable by the scintillation counter depends on culture density. Downing (1981) using
the yeast species Rhodotorula as food organism for Sida crystallina judged 1000 cells ml-1 to
be a minimum. Considering the capacity of the chamber (6 litres), the much smaller size of
the Scenedesmus cell and the small volume needed for fast injection and mixing (40 ml), a
density would be required of about 2 x 105 cells ml-1-tracer. Trial tests using 500 ml flasks
and Daphnia spp showed an increased chance of detectable ingestion using around 10,000
cells ml-1 in the flask, as opposed to the 1000 cells ml-1 used by Downing (1981).

85
CHAPTER 4

Figure 4.6. Flow chart showing the sequence of steps followed to prepare the H-3 labelled Scenedesmus
‘tracer’ and take samples at the end of each in situ experiment. See text for details.

Another important parameter of the labelled culture is the specific activity of the label at the
start of the experiment. For the uptake to be measurable after the time spans employed in
short-term feeding experiments (5 minutes to an hour) the in-chamber activity should be at
least 1,500 CPM ml-1, or around 1 CPM cell-1 if densities are 1000 cells ml-1 (Downing &
Peters 1980). At least 5000 CPM ml-1 were shown to be necessary for ingestion by lake
Daphnia spp to be detectable in preliminary tests in the laboratory. However, and given the
higher “tracer” concentrations used in the in situ experiments here, a higher per millilitre
activity was achieved (around 15,000 CPM ml-1, see Fig. 4.7).

86
GRAZING RATES

Suspension radioactivities sampled in the grazing


chamber at the end of each feeding experiment
25000
GF/C means
GF/F means
20000
CPM ml-1

15000

10000

5000

0
26-abr

12-may

01-jun

07-jul

28-jul

13-ago

23-sep

05-oct

29-oct
Figure 4.7. Final in-box activities of suspended material achieved in experiments (in CPM ml-1). Bars
plotted are the averages of three 50 ml aliquots, collected on either GF/C (1.2 µm pore diameter) or GF/F
filters (0.7 µm). Error bars are of one standard deviation either side of the mean. Details in text.

To obtain a fast estimate of culture density, a curve relating density (i.e. cells ml-1) to direct
spectrophotometric readings can be used (see Fig. 4.8).

Prediction of approximate cell density (cells ml-1) from


direct spectrophotometric readings (abs. 665 nm)
12000
1.2
10000
cells ml-1 (x103)

8000
0.55
6000

4000 0.39
0.27 Density = 8048.9 * Abs665 + 970.48
2000 0.19 r2 = 0.9945, p<0.001
0
0
0 0.2 0.4 0.6 0.8 1 1.2 1.4
Absorbance (665 nm)

Figure 4.8. Curve relating cell density (cells ml-1) with absorbance at 665 nm for a Scenedesmus culture.

The culture of known density (1 or 2 x 106 cells ml-1) was prepared by dilution with an
inorganic nutrient-enriched commercially available solution (7 % vol. total N; 1.3 % P, as
water-soluble pentaoxide; 6 % potassium oxide; 50 mgKg-1 Zn; 30 mgKg-1 B; 20mgKg-1 Mo;
10mgKg-1 Mn; ASB-Greenworld Ltd., Lincolnshire, UK). About 40 ml of this culture were
then introduced into a 50 ml. flask and 59.2 MBq (1.6 mCi) of tritiated sodium acetate added

87
CHAPTER 4

(ICN Pharm. Inc., sodium salt in ethanol). The culture was left to label under continuous light
on a fly rotator for 48-72 hours (see Table 4.3).

Table 4.3. Labelling periods and input activity for the Scenedesmus ‘tracer’ in the 9 experiments. Also
shown are measures of the residual activity (expressed as counts per minute, CPM and as a percentage of
the total initial activity) left at the end of the labelling period in the medium used for labelling the algae.
This residual activity is calculated as an average of 3 x 100 µl aliquot samples of the supernatant, after
centrifugation of the medium to isolate the algae. See Fig. 4.7 and text for details.

Experiment Labelling Initial input H-3 activity left in % input activity


period (hours) activity labelling medium remaining at the end
(MBq) (100 µl) of the labelling
period
1 48 59.2 445748 8.1
2 48 59.2 169495 3.1
3 48 59.2 193481 3.5
4 72 59.2 220422 4.0
5 96 59.2 250728 4.6
6 72 59.2 295092 5.4
7 96 59.2 236071 4.3
8 72 59.2 147402 2.7
9 96 59.2 141636 2.6

This culture was shown to take up a maximum of 53 % of the initial activity after 48 hours
(see Fig. 4.9). However, after around 72 hours the label was apparently lost from the cells,
suggesting either the uptake concerns mainly labile compartments of cell function, rather than
structural parts, or that once cell lysis starts, label is released to the medium.

%INITIAL H-3 TAKEN UP BY Scenedesmus


60

X
50

40 X
% initial H-3 activ ity

30 X

20 X

10
X X
X
0X

-10
0 20 40 60 80 100 120 140 160 180
Labelling time (hrs.)

Figure 4.9. Tritium uptake curve for a mixed Scenedemus culture during a time-course experiment with a
mixed Scenedesmus culture. Uptake is expressed as percentage of total tritium activity introduced at the
beginning of the labelling period measured in the algae. Plotted are means and standard deviations of
three aliquot samples taken from the algae culture at times 0, 1, 3, 6, 24, 48 and 72 hours.

88
GRAZING RATES

TRITIATED LABELLING MEDIUM TIME-COURSE

100 X

90
X
80 X
X
70
X X
X X
60
103 CPM/mL

50
40
30
20

10
0
0 20 40 60 80 100 120 140 160 180
Labelling time (hrs.)

Figure 4.10. Tritium activity remaining in the labelling medium during a time-course experiment with a
mixed Scenedesmus culture. Plotted are means (in 103 of CPM ml-1 of labelling medium) with standard
deviations of three aliquot samples taken from the supernatant after centrifugation to isolate the algae (see
also Fig. 4.9). Samples were taken at times 0, 1, 3, 6, 24, 48 and 72 hours after introduction of H-3-sodium
acetate. Most of the tritium is taken up by the algae in the first 24 hours. Note that there was little loss of
activity in the labelling medium due to the exchange with the air and water during the 7-day time-course.

At the end of the labelling period the volume of labelled culture was double-centrifuged at
1,500 rpm for 10 minutes each time, resuspending once in fresh, unlabelled, medium. The
centrifugation was repeated to eliminate any remaining unincorporated label. Samples of the
first supernatant always presented very low activities, suggesting most of the label was within
the cell. The labelled culture was kept in a 50 ml closed-cap centrifuge tube until the moment
it was introduced into the 40 ml syringe for the experiment in situ. It was shown specific
activity did not decrease significantly after two hours from the time the labelling medium was
removed from the labelled algae.

- Lake suspended algae food measures prior to each in situ experiment.

Clearance rates have been shown to be strongly dependent on body size of the filter-feeder
(Chow-Fraser & Knoechel 1985; Knoechel & Holtby 1986a & b). Effects of temperature
(Lampert & Taylor 1985; Bogdan & Gilbert 1982; Mourelatos & Lacroix 1990), and food
concentration (e.g. Downing & Peters 1980), are equivocal in in situ experiments, although
the influence of food conditions on rates in laboratory experiments is well documented. These
effects, however, rarely account for more than 30 % of variation in feeding rates (Peters &
Downing 1984).

89
CHAPTER 4

I assumed that the highly radioactive Scenedesmus culture injected in the chamber as “tracer”
did not change significantly the within-chamber concentrations of algae. Thus, samples were
taken directly from the lake to estimate chlorophyll-a and dry weight of suspended matter
prior to each experiment.

Two one-litre samples of unfiltered water and two of water passed through a 25 µm mesh
were taken at the experimental site using 1-l Pyrex bottles. Cladocerans can probably only
ingest particles smaller than 100 µm, depending on shape, and more likely smaller than 25 µm
(Burns 1968). The fraction sampled represents most of the “more edible” algae available to
these grazers. In the laboratory, three 0.5 litre aliquots of filtered and three of unfiltered water
were passed through pre-combusted and tared GF/C filters and the filters dried (T=105° C, 24
hours) and weighed to a precision of 10 µg in an electronic balance. They were then
combusted at 400° C for 0.5 hours and re-weighed for ashfree weight estimations. Preliminary
tests showed all organic matter combusted in the first half hour. A measure of chlorophyll-a
concentration was also taken for the filtered and unfiltered water by collecting two 300-ml
aliquots onto GF/C filters and extracting the pigment with hot 90 % ethanol in a water bath
(T=75°C for 5 minutes).

-Within-chamber suspended algae sampling after each in situ experiment (i.e. planktonic
algae activity).

At the end of each experiment three 50 ml aliquot samples of the suspended algae were taken
from within the box for radioactivity counting. The assumption is that this activity does not
change significantly because of the grazer activity, or for any other reason, in the short time of
the experiment (generally about 15 minutes, see Table 4.6).

The samples were taken by collecting 50 millilitres of the suspended algae onto glass fibre
filters in the field by means of a hand pump, then placing the filter into a glass vial with tissue
solubilizer. As with periphyton samples, filters were introduced into marked glass vials with
one millilitre of tissue solubilizer and left to digest overnight at 40°C before scintillation
cocktail and buffer were added, prior to counting. The filtering of the suspended algae
samples onto glass fibre filters and the processing of the periphyton samples was done in the
field. This reduced the time during which the highly labile tritium could exchange with the
water and other living fractions in the water, and minimized the slow detachment of the
carbon-14 labelled periphyton.

90
GRAZING RATES

3. Field procedures.

All experiments were conducted at the same site, located in more open and deeper waters (i.e.
outside the lily beds) to facilitate the submersion of the box.

Before the experiment began, samples of the lake water were taken at this site (see site
location on Map; ‘site 3’ in Fig. 4.14). The animals to be used in each experiment were
collected from neighbouring plants using both the sampler especially designed for lily leaves
(see section 3.2.3 for details) and a “tube sampler” for taking depth-integrated samples of the
water within the lily bed. The aim was to maximize the representation of species present in
the lake at the experimental date (see Table 4.6). See Fig. 4.11 for a flow chart of field
procedures.

The box was filled with lake water by submersion. The previously sampled animals and the
carbon-14 labelled strips with periphyton were then introduced into the box through the
sampling port (see Fig. 4.1), the port was tightly closed and the box lowered into the lake
attached to the side of the boat with a rope. The beginning of each experiment was marked by
the injection into the box of the radioactive tracer algae. The injection was effected by means
of a small syringe connected through a plastic tube to an inlet on the top of the chamber. The
water in the chamber was then mixed using the same syringe for around 30 seconds (i.e. 6 or
7 syringe-fulls). Mixing tests in the laboratory using acrydine as a dye have demonstrated
effective mixing after this time.

Given that periphyton ingestion rates are much lower than suspended algae feeding rates (e.g.
Downing 1981), the time before experiment initiation, when animals were left with the
carbon-14 labelled strips, is considered negligible compared with the feeding period. In
addition, some time was assumed for food location by periphyton scrapers. Although an inlet
was prepared for introduction of the experimental animals, it was subsequently preferred to
use the sampling port directly to minimize potential disturbance to their feeding behaviour
through injection.

Strips were retrieved and a suspended algae sample taken before the experiment was
terminated, with about two minutes allowed for this (see Table 4.6 for experimental periods).
The animal sample was then taken (see section 4.2.1) and submerged in liquid nitrogen for

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CHAPTER 4

transport back to the laboratory. Back on the shore, samples of periphyton and suspended
algae were collected on glass fibre filters, as described in the appropriate sections.

• Carbon-14 labelled strips


Collection of animals from
To open-water site
plant bed • Animals

Lowered into water Injection of the H-3 labelled Scenedesmus


culture + mixing (30 seconds)

FEEDING PERIOD
(12-15 minutes)

• Periphyton strips removed.


• Suspension activity sample.
• Collection of feeding animals on mesh + N2 freezing.

Figure 4.11. Field procedures during each in situ experiment. Each experiment started with the collection
of animals from neighbouring lily beds and ended with the collection of samples of periphyton, suspended
algae and the feeding animals from the grazing chamber. See text for details.

4. Animal samples.

In the laboratory, the mesh was defrosted and the animals, algae and other material, washed
into a bottle with distilled water (Burns & Rigler 1967). Losses of radioactivity into the
sorting fluid (i.e. distilled water) are not thought to be large because isotonical concentrations
can be reached rapidly.

Individual animals were measured, under a stereomicroscope with an ocular micrometer


(maximum precision 10 µm) and placed in marked glass vials into which 0.5 ml of distilled

92
GRAZING RATES

water had been added. Animals were sorted for at least two hours, but often only a fraction of
those collected on the mesh were radio-assayed. A record was kept of size and species and
other features of potential interest, for example presence of ephippia or asexual eggs in the
brood pouch. One mililitre of tissue solubilizer was added and the samples incubated for 24
hours at 40°C in an oven, as were periphyton and suspended algae samples collected during
the experiment, as described in previous sections.

Counts were made on both C-14 and H-3 channels simultaneously. Radioactivity was
measured on 650 animals of 5 cladoceran species (see Table 4.4).

-Radioactivity counting: quench and spillover corrections.

When using radioactively labelled food we are essentially comparing the differences between
the activity levels derived from the animals with those in the feeding medium, then attributing
these differences to the ingestion of radioactive food by the animals during the experimental
period (Rigler 1971). The comparison is made using simple arithmetic relationships:

C.R. = Ra * 24 / Rm * t ‘C.R.’ is the clearance rate, in ml ind-1day-1

P = (Ra’ – residual) * 24 / (Rm’ * t) ‘P’ is the periphyton ingestion rate, in µg


periphyton ind-1day-1.

where Ra and Ra’ are the H-3 and carbon-14 radioactivities of one animal (cpm or
counts per minute per individual).

Rm and Rm’ are the H-3 and carbon-14 radioactivities of the feeding media
(cpm per ml, and cpm per µg of periphytic chlorophyll-a, respectively).

‘t’ is the experimental period (hours), and

Residual = (F*resid.*t/24) (in cpm per individual); ‘resid.’ is the residual


carbon-14 activity in the suspended algae (i.e. detached carbon-14 labelled
periphyton). The residual calculated by the expression represents the detached
periphyton filtered, not ingested by the animals after scraping off a surface
such as that of the plastic strips used in the experiments here. Carbon-14
activities in animals may increase through filtering this naturally detached
periphyton, a process different from scraping or ‘browsing’ (see Horton et al.
1978). Thus, periphyton ingestion was corrected for this “accidental” uptake
(see Downing 1981).

In dual isotope experiments both carbon-14 and H-3 contribute to ‘Ra’ in one single sample.
Activities from both can be counted in a liquid scintillation analyzer simultaneously because

93
CHAPTER 4

the energy profiles of the particles emitted hardly overlap. Likewise, residual carbon-14
activity may be found in ‘Rm’, from detached labelled periphytic cells.

Before clearance rates can be calculated, values for ‘Ra’ and ‘Rm’ should be corrected for
their overlapping energy profiles and for a measure of interference with mainly the
scintillation liquids, called ‘quench’.

Efficiency of detection by the counter is reduced by interference of the scintillation cocktail


and by absorption of particles by the tissues in animal samples (i.e. “self-absorption”, see, for
example, Wang & Williams 1965; Wang, Willis & Loveland 1975). Quench may be also due
to plastic in some of the periphyton samples, or partially digested glass fibre in suspended
material samples.

The energy profiles of both isotopes used overlap slightly (around 7-14 % in an unquenched
sample). Moreover, quench influences the degree of overlap. A curve relating “quench level”
(as determined by the liquid scintillation analyser) to efficiency was drawn (Fig. 4.12). From
the quench level given for each sample an approximate correction was made.

70 tSIE and % of unquenched

60
% of unquenched CPM

50

40

30
R2 = 0.9195
20

10

0
908 748 559 479 351 270 204 165 131 104
tSIE

Figure 4.12. Curve relating quench index (i.e. tSIE, or Transformed Spectral Index of the internal
standard spectrum, an index generated by the counter and measuring ‘quench’; high values indicate little
quench) to efficiency (percentage of theoretical activity detected in spite of interference or ‘quench’).

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GRAZING RATES

4.2.2. Plant surveys.

To estimate total population sizes of the main plant-associated grazers (densities in ind g-1DW
plant), areal measures of plant biomass (in gm-2DW) were needed. Thus, two surveys were
conducted in each of two growing seasons (1998 and 1999). Each year, the first survey was in
mid-summer, when the plant communities were at maximal growth, and the second in late
summer, when senescence had set in. Each survey generally lasted two or three days.

About 50 plant samples were collected in each survey, by removing all the plant biomass
enclosed by a cylinder of known base area (0.105 m2; see Figure 4.13). Samples were sorted
into species, and the lily parts further classified into petiole and leaf parts. Plants were then
dried at 60°C. Plant biomass was then expressed as dry weight per m2 of lake bottom (i.e. g
m-2).

Sampling locations were randomized by drawing 10-12 transects across the lake with a rope
that was marked with tape every 10 metres (for transect maps see Appendix C). Every mark
was assigned a number and a subset of about 50 samples of all possible numbers along
transects was then selected at random.

Figure 4.13. Sampler used to isolate a known area


from within which all plant biomass is removed
using a rake (see text for details). The base area is
0.105 m2 (diameter = 36 cm).

Diameter = 36 cm

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GRAZING RATES

Plant cover.

Coverage of the different plant species in Little Mere was visually assessed in the first survey
by noting the vegetation along the transects. In subsequent surveys gross changes in the
vegetation communities and lily bed coverage were assessed in large sectors of the lake.

Depth.

The depth of the water column at each sampling point was determined using a marked pole.
Two lines of stakes were set at 10-m intervals in the lake. Stakes were numbered and with the
aid of conspicuous landmarks on the shoreline, a reference was kept of the approximate
position while taking measurements. A detailed depth map was then drawn by connecting
sites of similar depth.

96
GRAZING RATES

[Here "fused" Map]

Map on paper (A4 landscape) with lily bed outline, extrapolation sectors and sampling site location +
overlaid A4 transparency with bathymetry.

Figure 4.14. Map of Little Mere (Cheshire) showing the lily beds’ outline (black interior line), equal depth
lines, sampling site location and extrapolation sectors for each sampling site. Note that the sector
represented by site 5 is made up of three separate areas. The boundaries of each sector approximate the
actual lily bed outline.

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GRAZING RATES

98
GRAZING RATES

4.2.3. Estimation of grazing rates.

Grazing rates for the main filter-feeding species in Little Mere were separately estimated for
the two years sampled (i.e. 1998 and 1999). Large spatial variation in grazing rates has for
long been acknowledged (e.g. Gulati, Siewertsen & Postema 1982). For Daphnia spp, sites
were grouped into either lily or “open water” sites, while for Simocephalus vetulus and Sida
crystallina data for the three lily bed sites were grouped.

Table 4.4. Approximate parameters used for extrapolation of density data and grazing rate calculation
(see text for details). Values are given for the area (m2), volume (m3), mean depth (m) of each sector of the
lake used in extrapolations and represented by each sampling point.

TYPE OF SITE* Area Volume Mean depth Surface


SITE (m2) (m3) (m) (m2)
Lily bed 1 1,100 770 0.7
sites
2 2,500 1,750 0.7 10,500 (38 %)
5 6,900 4,300 0.6
‘Open 3 7,900 6,500 0.8 17,500 (62 %)
water’
sites 4 9,600 7,600 0.8
*see Fig. 4.14 for a map of sampling site locations.

The following equation gives the grazing rate for any given species (G, in % lake volume
filtered per day, i.e. % day-1), for each sector separately (see Figure 4.14 for a Map of
sectors):

×

T F
Equation 1.
G
V × 1000

ind. × ml day ind.


-1 -1
-1
% day = l × ml l
-1

See Table 4.5 for the meaning of variables used in equation 1.

Variability in clearance rates with body size of the animal was taken into account when
calculating grazing rates by applying a different rate for each size class. A ‘weighted’
clearance rate is calculated for each date and species. The weights are on the basis of size-
class proportions in the populations sampled from lily leaves in the case of Simocephalus
vetulus, and from tube samples in the case of Daphnia spp. Three size classes were used,
small individuals (0.5-0.9 mm), medium-sized (0.91-1.5 mm) and larger individuals (> 1.51
mm). When no size-structure information was available (e.g. when densities were too small to

99
GRAZING RATES

obtain reliable information), sizes were assumed to be those of the closest date or/and site. For
Sida, an average clearance rate for a 1.5 mm individual was used.

Table 4.5. Variables used in the calculation of grazing rates (% day-1).

VARIABLE UNITS DESCRIPTION ESTIMATION


T individuals Total number of individuals of a given species T = D * P * C
(ind) (plant-associated) in a lake sector. ind ind g-1 g m-2 m2
T’ ind Total number of individuals of a given species T’ = D’ * V
(‘open-water’) in a lake sector. ind ind l-1 l
-1 -1
F ml day ind Clearance rate. See section 4.2.1 for details.
D ind g-1 Number of individuals per gram of lily See Chapter 3 for details.
(leaf + petiole).
D’ ind l-1 Number of individuals per litre of lake water. ditto.
P g m-2 Grams of lily (leaf + petiole) per unit area of See section 4.2.2 for details.
lake bottom.
C m2 Surface area of a sector of the lake around ditto.
each sampling section (see Fig. 4.15 for a
Map).
V l Approximate volume of lake water in a given V = C * z * 1000
sector of the lake. l m2 m l m-3
z m Average depth of the water column in any See section 4.2.2 for details.
given sector of the lake.

For the estimation of total population sizes ('T' in equation 1), data from lily leaf and petiole
samples were added together to calculate a ‘lily-integrated’ density for each animal species.
Likewise, errors in density estimates were calculated from the pooled data (i.e. leaf + petiole
samples). When calculating whole population sizes from densities (i.e. ind g-1DW) two values
of plant biomass were used per season, corresponding to the two surveys conducted (section
4.2.2). The first factor is used for April to the end of July density data, while the second value
helped extrapolate the rest of densities.

I will restrict the analysis to a comparison of filter-feeders of both ‘communities’ (i.e. open-
water and plant-associated). The estimation of periphyton ingestion rates and the total
removal by whole populations involves much larger variability and, therefore, results are
difficult to interpret in terms of potential effects on the ecosystem.

4.3. Results.

Clearance rates.

The data consist of 530 individual animal clearance and periphyton ingestion rates (Table
4.10). Highly significant (F-ratio test, p<0.001, except Scapholeberis mucronata; p=0.014)

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GRAZING RATES

regression models for clearance rates were obtained for five of the six species (Table 4.9).
For periphyton ingestion rates highly significant models were found for only four of the six
species, Eurycercus lamellatus, Simocephalus vetulus, Sida crystallina, and Daphnia spp. Ten
species were collected in experiments (Table 4.6), but breadth of response data only allowed
analysis on six (Daphnia, Simocephalus vetulus, Sida crystallina, Eurycercus lamellatus and
Ceriodaphnia sp.)

Table 4.6. Experimental periods and composition of animal samples of each in situ experiment. The period
to freezing the filter is taken as the feeding period.
Experiment Feeding period (mins.) Number Species’ names*
To box To filter- Animals Species
retrieval freezing
1 14 18 51 4 D/Ch/B/Sim
2 14 19 65 6 Sim/D/B/C/P/A
3 - 15 69 6 Sim/D/C/E/P/A
4 - 14 43 4 D/C/B/Sim
5 - 15 68 4 Sim/Sc/C
6 - 15 61 3 Sim/Sc/C
7 - 15 100 4 Sim/S/D/E
8 9 12 93 5 Sim/S/D/G/E
9 9 13 100 4 Sim/S/D/E
*Species: A-Alona spp; B- Bosmina longirostris; C- Ceriodaphnia spp; Ch-Chydorus spp; D-Daphnia spp;
E-Eurycercus lamellatus; P-Peracantha truncata; Sc-Scapholeberis mucronata; Sim. Simocephalus vetulus;
S-Sida crystallina.

Table 4.7. Ranges for clearance rate (CR), periphyton ingestion rate (P) and the main independent
variables in models for each species in in situ dual-isotope feeding experiments in Little Mere (Cheshire,
UK).

Variable Simocephalus Daphnia Sida Eurycercus Scapholeberis


vetulus* spp* crystallina* lamellatus mucronata
Observations 264 130 78 35 23
Number of dates 7 8 3 5 2
CR (ml ind-1day-1) 0.3-26.3 0.4-29.4 0.59-260.0 0.0-1.6 1.2-9.3
P (µg ind-1day-1) 0.1-429 0.1-252 0.2-656 0.1-768 0.4-23
Size (mm) 0.44-2.00 0.25-2.94 0.75-3.44 0.56-2.25 0.38-0.69
Temperature (°C) 9.9-24.4 8.7-23.9 10.8-14.7 10.8-14.7 20.8-24.4
Periphyton chl-a (µg cm-2) 0.8-8.9 1.2-8.9 1.2-4.8 1.2-8.9 0.8-1.1
<25 µm planktonic chl-a (µg l-1) 6.9-45 6.9-45 6.9-45 6.9-45 40.1-44
Total planktonic chl-a (µg l-1) 11.3-93.6 11.3-93.6 13.7-57.7 13.7-57.7 58.3-93.6
Total suspended solids (mg l-1) 2.5-14.7 2.5-13 2.5-5.6 2.5-7.4 13-14.7
*after exclusion of outliers. See text for details.

See Tables 4.9 & 4.10 for the variation accounted for by each model. All accounted for 21.7-
69.7 % of variance in clearance rate measurements and 11.4-73.2 % of periphyton ingestion
estimates. The most important of the variables tested was body size, accounting alone for 22-
62 % of the total variance observed in clearance rates. Results for variables selected for
periphyton ingestion rates were quite variable (Table 4.10).

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GRAZING RATES

Table 4.8. Transformation of response data (clearance rate and periphyton ingestion rate estimates) from
in situ dual-isotope experiments conducted in Little Mere (Cheshire) during growing season ‘99 (April to
October) for each species and linearizing effect of transformations, as expressed by the coefficient of
determination (r2) when regressed against body size (mm). See text for selection criteria for type of
transformation.

Species Type of Periphyton Effect of Effect of


transformation ingestion rate transformation transformation
for Clearance data (P) (CR) (P)
rate response r2 r2
data (CR)
Before After Before After
Daphnia spp1 Log Log 0.46** 0.499** 0.0005 0.047*
Scapholeberis mucronata Log Log 0.238* 0.26* 0.018 0.038
Sida crystallina1 Log Log 0.46** 0.57** 0.007 0.125**
Simocephalus vetulus1 Log Sqrt 0.51** 0.54** 0.17** 0.231**
Eurycercus lamellatus Sqrt Untransformed 0.6** 0.62** 0.55** -
1
Outliers suppressed (see text for details). * p<0.05; ** p<0.01.

Food conditions for the nine experiments were spread across quite a wide range, allowing the
relationship of clearance rate to food concentration to be investigated (see Fig. 4.15).

2
Periphyton (micrograms DW cm )
100 Total chlorophyll-a 16
<25 micrometer chl-a
90 14
TSS

Suspended solids (mg l -1)


80 <25 micrometer SS
chl-a (micrograms l-1)

12
70
60 8.9
10

50 8
40 6
30 3.3
3.0 4
20 4.8
1.6
10 1.1 1.2 2
0.8
1.5
0 0
26-Apr 12-May 01-Jun 07-Jul 28-Jul 13-Aug 23-Sep 05-Oct 29-Oct

Figure 4.15. Food conditions during feeding experiments at the experimental site. Plotted are total and less
than 25 micrometer fractions of both chlorophyll-a (µg l-1) and suspended solids (mg l-1). Periphyton
concentrations (in µg dry weight cm-2) within the grazing chamber have been plotted as histograms (see
text for details).

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GRAZING RATES

Table 4.9. Clearance rate best linear regression models relating clearance rate to size and other variables in experiments carried out in Little Mere (Cheshire). Basic
statistics and ANOVA tables for the species analysed are given. The top table shows the multiple linear regression models obtained for Simocephalus vetulus and
Daphnia spp. The best linear model for the other four species includes only one predictor (size), for details on the transformation chosen for the response data, see
Table 4.8. See text for details.

Simocephalus vetulus Daphnia spp


2 2
Predictor variable Coeff. SE Part. F Adj.r Signif. F- Predictor variable Coeff. SE Part. F Adj.r Signif. F-
increase increase
MODEL Size (mm) 0.691 0.035 305.2 0.538 <0.001 Size 0.248 0.031 120.5 0.481 <0.001
-2
Strip chl-a (µg cm ) -0.008 0.002 19.1 0.568 <0.001 Temperature (°C) -0.048 0.006 58.8 0.642 <0.001
-1
Temperature (°C) 0.015 0.003 12.1 0.586 0.001 <25µm chl-a (µg l ) -0.006 0.002 18.05 0.685 <0.001

<25 µm ashfree SS
-2
-0.032 0.01 9.4 0.599 0.002 Strip chl-a (µg cm ) 0.09 0.037 6.25 0.697 0.014
-1
(mgl )
constant -0.146 0.057 constant 0.08 0.09

df SS MS F p df SS MS F p

ANOVA Total 261 23.48 Total 129 14.06

Regression 4 14.21 3.55 98.5 <0.001 Regression 4 9.93 2.48 75.28 <0.001

Residual 257 9.27 0.036 Residual 125 4.12 0.033

Sida crystallina Eurycercus lamellatus Scapholeberis mucronata


2 2 2
Var. Coeff. SE Part. F Adj.r p Coeff. SE F Adj.r p Coeff. SE F Adj.r p

Size 0.511 0.05 105.14 0.569 <0.001 2.5 0.33 56.2 0.62 <0.001 3.18 1.19 7.11 0.217 0.014

constant 0.619 0.098 -0.03 0.37 0.28 0.65

ANOVA df SS MS F p df SS MS F p df SS MS F p

Total 79 11.81 34 47.6 22 6.16

Regression 1 11.7 11.7 105.14 <0.001 1 30 30 56.2 <0.001 1 1.56 1.56 7.11 0.014

Residual 78 0.11 0.0014 33 17.6 0.53 21 4.6 0.22

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GRAZING RATES

Table 4.10. Best periphyton ingestion linear regression models relating periphyton ingestion rates to size and other variables in experiments carried out in Little
Mere (Cheshire). Basic statistics and ANOVA tables for the species analysed are given. The top table shows the multiple regression models. The second table shows
the simple models found for two species.

Simocephalus vetulus Daphnia spp Eurycercus lamellatus


2 2 2
Predictor Coeff. SE Part. F Adj.r p Predictor Coeff. SE Part. F Adj.r p Predictor Coeff. SE Part. F Adj.r p
variable variable variable
MODEL Temperature -0.456 0.03 86.85 0.248 <0.001 Periphyton 0.091 0.067 436.04 0.771 <0.001 Size 303.4 36.36 40.62 0.54 <0.001
-2
(°C) (mg cm ) (mm)
<25µm chl-a 0.141 0.011 202.92 0.576 <0.001 <25µm chl-a 0.024 0.003 86.9 0.863 <0.001 <25µm 83.68 16.6 18.58 0.7 <0.001
-1 -1
(µg l ) (µg l ) ashfree SS
Size (mm) 3.06 0.426 41.15 0.637 <0.001 constant -0.17 0.029 Temperature 23.13 10.33 5.01 0.732 0.032
(°C)
Periphyton -0.124 0.029 17.99 0.656 <0.001 constant -754.04 177.05
-2
(mg cm )
Constant 4.585 0.683

Source df SS MS F p Source df SS MS F p Source df SS MS F p

ANOVA Total 261 3962 Total 129 25.59 Total 34 719573

Regression 4 2619.9 654.97 125.4 <0.001 Regression 2 22.124 11.07 407.75 <0.001 Regression 3 543915 181305 31.99 <0.001

Residual 257 1342.3 5.22 Residual 127 3.45 0.027 Residual 31 175659 5666

Scapholeberis mucronata Sida crystallina


2 2
Predictor Coeff. SE Part. F Adj.r p Predictor Coeff. SE Part. F Adj.r p

Size * * 0.841 -0.007 n.s. Size -0.357 0.107 11.116 0.114 0.001
(mm) (mm)
constant constant 1.873 0.211

ANOVA

df SS MS F p df SS MS F p

Total 22 1.975 Total 79 45.61

Regression 1 0.076 0.076 0.841 n.s. Regression 1 5.69 5.69 11.116 0.001

Residual 21 1.899 0.009 Residual 78 39.92 0.512

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GRAZING RATES

Simocephalus vetulus yielded the most complex models. Size accounted for 53.8 % of
clearance rate variation (Table 4.9). An additional 3 % was explained by the depressive effect
of periphyton concentration on rates. With respect to the range of values predicted, values are
about 3 times larger than those predicted by Ivanova & Klekowsky (1972), but about half
those estimated by Sharma & Pant (1982) using cell-count methods (see Table 4.11).

Periphyton ingestion of this species could be explained in 65.6 % of cases, with four variables
(see Table 4.10). Temperature had a strong depressive effect on periphyton ingestion.
Periphyton concentration (mg cm-2 dry weight) was also inversely correlated with ingestion
rate. Larger animals tended to ingest larger amounts of periphyton.

Table 4.11. Clearance rate estimates (ml ind-1day-1) for the main filter-feeders in Little Mere (Cheshire).
Tabled are own estimates (CR) and values gleaned from the literature (CR', CR''). The ratios between
own estimates and literature values are also shown.

Clearance rate comparisons Size CR CR' CR'' CR/CR' CR/CR''


(ml ind-1day-1) and ratios (mm)
Daphnia spp 0.7 3.0 4.0 3.1 0.8 0.97
1.2 5.7 11.6 14.6 0.5 0.4
1.8 9.7 23.2 40.3 0.4 0.2
2.1 12.1 - 86.1 - 0.1
Sida crystallina 1.2 5.0 29.5 19.1 0.17 0.26
1.8 18.2 84.3 45.5 0.22 0.4
2.1 30.5 130.5 61.0 0.23 0.5
Simocephalus vetulus 1.2 5.7 10.3 1.8 0.6 3.2
1.8 12.6 36.6 6 0.3 2.1
2.1 17.4 - 10.8 - 1.6
CR'. Daphnia spp: Mourelatos & Lacroix (1990); Sida crystallina: Downing & Peters (1980); Simocephalus
vetulus: Brendelberger (1991). CR''. Daphnia spp: Burns & Rigler (1967); Sida crystallina: Downing
(1981); Simocephalus vetulus: Ivanova & Klekowski (1972).

Following are the equations drawn by back-logging regressions between body length (mm)
and clearance rate estimates (ml ind-1day-1), from in situ feeding experiments in Little Mere:

Clearance rate (CR: ml ind-1day-1) vs. Length (L: mm):

Simocephalus vetulus: CR = (102.96* Log (L+1) - 0.19) - 1


Daphnia spp: CR = (101.97* Log (L+1) + 0.15) - 1
Sida crystallina: CR = (104.84* Log (L+1) - 0.88) -1

In Little Mere, Daphnia apparently filtered less efficiently than Simocephalus and Sida,
particularly larger-bodied animals (see Table 4.11 & Fig. 4.16).

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GRAZING RATES

Periphyton ingestion rates of this species were positively correlated with the amount of
periphyton presented to the animals in the box. On the other hand, suspended algae food (<25
µm chlorophyll-a) also had a positive effect on ingestion of periphyton.

Sida crystallina filtered about double the volume of similar-sized Daphnia, particularly the
larger individuals (Table 4.11). Strikingly, the larger Sida ingested less periphyton than small
individuals (see Table 4.10).

Eurycercus lamellatus. Clearance rates (CR) were only significantly correlated with size of
the animal (Table 4.9). This species is thought to be mainly a detritivore and periphyton
scraper (Smirnov 1962). However, a surprising 62 % of CR variation could be explained by
carapace length, in a similar fashion to filter-feeders. For periphyton ingestion it was possible
to build a multiple regression model with two additional variables representing suspended
algae food concentration (<25 µm ashfree weight of suspended solids) and temperature,
accounting for 73.2 % of total variation.

Only a small amount of variation in clearance rates of Scapholeberis mucronata could be


explained by models (27 %), and no valid model was found for periphyton ingestion, given
the small sample size (N=23). Size was the only variable with sufficient range of values for
meaningful analysis, but its effects on ingestion were not significant. No significant clearance
rate or periphyton ingestion models for Ceriodaphnia spp could be found, as very few
countings were made on this species (Table 4.7).

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GRAZING RATES

1.6 Daphnia spp : clearance rate vs. body size


log(CR+1) = 1,97* log (L+1) + 0,15
2
1.4 r = 0,499

1.2

log(ml day ind +1)


-1
-1 1.0

0.8

0.6

0.4

0.2

0.0
0.0 0.1 0.2 0.3 0.4 0.5 0.6
log (mm + 1)

Simocephalus vetulus : clearance rate vs. body size


1.6

1.4 log(CR+1) = 2.9583 * log (L+1) + 0.1864


2
R = 0.5569
1.2

1.0

0.8

0.6

0.4

0.2

0.0
0.0 0.1 0.2 0.3 0.4 0.5 0.6
log(mm + 1)

3.0 Sida crystallina : clearance rate vs. body size

log (CR+1) = 4.8441*log(L+1) - 0.8769


2
2.5 r = 0.56

2.0

1.5

1.0

0.5

0.0
0.2 0.3 0.4 0.5 0.6 0.7
log (L+1)

Figure 4.16. Clearance rate (in ml ind-1day-1; log-transformed) vs. animal body length (L(mm); log-transf.)
for Daphnia spp (top), Simocephalus vetulus (middle) and Sida crystallina (bottom). Lines represent the
linear regression models shown on each plot (see Table 4.8).

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GRAZING RATES

Least-squares multiple regression analyses were performed to establish the relationship


between feeding (i.e. clearance rates or periphyton ingestion rates; CR or P, respectively), and
the individual body length, algal concentration, periphyton concentration and temperature (see
Table 4.12). Any response, however complex, may be more than adequately described by a
polynomial if most of the relevant variables are known. The question, then, is really what
order polynomial are we prepared to accept as workable, given, of course, that the model
describes a statistically significant effect. Linear models were preferred to those involving
power or higher order polynomials, because of their generally better fit to response data. All
species’ data had to be fitted to a same-shape regression model too, i.e. linear, so comparisons
between species could be easily made.

Table 4.12. Candidate predictor variables in stepwise multiple regression analysis.

Suspended algae Suspended solids Periphyton density Temperature Size


chlorophyll-a
• Total • Total • Per cm2 periphytic
• <25 µm • <25 µm fraction chlorophyll-a
fraction • Total ashfree • Per cm2 periphyton
• <25 µm ashfree dry weight

Equations were fitted for the main species found in experiments, Simocephalus vetulus,
Daphnia spp, Ceriodaphnia spp, Sida crystallina, Scapholeberis mucronata and Eurycercus
lamellatus. The simplest equation was found for each species by sequentially adding terms
with significant coefficients (F-ratio test at the 0.05 significance level). Further refinement of
models involved the consideration of the explanatory value of each term, as expressed by the
additional variation in feeding rates accounted for in the model (i.e. the r2).

Initially, all variables were included in the dataset. Sometimes the forward stepwise procedure
added variables that were highly correlated or biologically related (e.g. total suspended solids
and total chlorophyll-a). The analysis was then re-run with the first variable entering in the
stepwise procedure as representative of all other related variables (e.g. total suspended solids
would now represent total and small chlorophyll, <25 µm suspended solids, total suspended
solids ashfree weight and <25 µm suspended solids ashfree weight). The model selected was
that one having the maximum significant (F-ratio test, alfa=0.05) predictive power (i.e. max.
r2). However, very small step increases were not added to the model, even if the final model
was highly significant (maximum simplicity principle).

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GRAZING RATES

In addition, variables were only considered candidate predictors in each model if more than
three values were possible for any given species (i.e. three or more dates). Thus, only ‘size’
was considered a potential predictor for Scapholeberis mucronata (2 dates) and Sida
crystallina (3 dates). Analogously, no periphyton ingestion model was drawn for
Ceriodaphnia spp because of insufficient breadth of response data.

Unstable variance of filtering rate measurements


20 (Simocephalus vetulus )
15

10
Residuals

0
0 0.5 1 1.5 2
-5

-10
Size (mm.)
-15

Figure 4.17. Plot of residuals of the linear regression model of clearance rate (ml ind-1day-1) of
Simocephalus vetulus on size (mm) before transformation of data. Variance of response data was found to
follow a pattern of increasing variance with increasing means. Details on the type of transformation
chosen for each model and species are given in Table 4.8.

For the full interpretation of regression models, two basic conditions of data must be met. In
the first place, response for given values of the predictors must be about normally distributed.
Secondly, the variance of these responses across the range of values adopted by the predictors
must be similar (i.e. variance is said to be ‘stable’). Variance in feeding rates was shown to be
unstable, with a tendency to a larger scatter of response in larger animals, which is a common
pattern (Samuels 1991; see Fig. 4.17). To make the response linear and alleviate error
unstability, both periphyton ingestion and clearance rates were transformed. For details on the
type of transformation see Table 4.8. The logarithmic transformation was often chosen over
the commonly used power-transformation (see, for example, Downing & Peters 1980). The
type of transformation was chosen based on three criteria. First, it maximized the amount of
response accounted by the linear model (i.e. max. r2). It also stabilized variance and normality
of response, while at the same time lowering the value of residuals (i.e. minimum mean sum
of residuals for the transformations tested).

Regression models can sometimes describe data well, but it is perhaps their predictive
function which is more often highlighted (Bailey 1992). Thus, extreme values (with respect to
the regression model, i.e. ‘outliers’) weighing heavily on predictive power were identified by

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GRAZING RATES

visual inspection of residual plots and removed from subsequent analysis (see Table 4.8).
Outliers were always found to be the maximal values of clearance rate, perhaps explained by
the inadvertent inclusion of labelled algae material into animal vials. Therefore their
suppression is justified because they do not belong to the distribution but respond to a
methodological error (Samuels 1991).

Plant cover and biomass.

Lily growth the first year was about half that in the second year (Fig. 4.18). However, mean
estimates are not accurate enough to estimate growth rates in the interval between the first and
second surveys. Years were very different with respect to average changes between surveys,
though. Thus, in the second year, the cycle of lily growth was shifted later in the season, and
peaks of biomass were generally larger in the second survey. However, differences between
years could not be tested statistically due to large variation in biomass.

Leaf average weight across sites in 1998 Petiole average weight across sites in 1998
80 50
July '98
70 45
September'98 July'98
40
60 September'98
35
DW (gr.m-2)

DW (gr/m-2)

50 30
40 25

30 20
15
20
10
10 5
0 0
site 1 site 2 site 5 site 1 site 2 site 5

Leaf average weight across sites in 1999 Petiole average weight across sites in 1999
160 90
June'99
80 June'99
140 September'99
September'99
120 70
60
DW (gr.m-2)

DW (gr.m-2)

100
50
80
40
60
30
40
20
20 10
0 0
site 1 site 2 site 5 site 1 site 2 site 5

Figure 4.18. Lily leaf and petiole average weights (gm-2DW) in plant surveys conducted in Little Mere
during two growing seasons (1998 & 1999). Top and bottom plots are 1998 and 1999 surveys, respectively.
First and second ‘replicate’ surveys are left and right, respectively. Bars are of one standard error each
side of the mean. See text and Appendix B for details.

As regards plant cover, in the ‘open water’ (i.e. outside the lily-covered littoral area; for a
Map see Fig. 4.14), both years were characterized by an increasing dominance of
Ceratophyllum demersum as the growing season proceeded. Maximum biomass of this
species was comparable in both years.

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GRAZING RATES

Callitriche hermaphroditica was found in late summer the first year, in very dense patches
(i.e. up to 265 g m-2DW). In the second year, in contrast, it was not found at all among the 67
samples taken in late summer, but was present in June, albeit at lower densities than in the
previous year.

Potamogeton berchtoldii was present in the first year, although no biomass estimates are
available. The second growing season it was only occasionally spotted. On the other hand,
Elodea canadensis was hardly seen the first year, but was relatively abundant by September in
the second year (i.e. 40-80 gm-2DW; Appendix C) even if patchily distributed.

Grazing.

The grazing role of the plant-associated filter-feeders in Little Mere, represented by


Simocephalus vetulus and Sida crystallina, was very small compared with that of Daphnia
(Figs. 4.19 to 4.21).

Daphnia spp grazing rates were much higher and persistent in 1998 than in 1999 (Fig. 4.19).
In the second growing season, grazing rates of this species were virtually zero from the
beginning of June to mid-August. A high peak was measured in spring within the lily beds
this year.

Daphnia grazing rates during growing season 1998 Daphnia grazing during growing season 1999
250
90
Grazing in lily beds
80 Grazing in lily beds
200
Grazing in open Grazing in open waters
70 waters
Grazing rate (% day )
-1
Grazing rate (% day )
-1

60
150
50

40 100
30

20 50

10 *
0 0
1/4
8/4
15/4
22/4
29/4
6/5
13/5
20/5
27/5
3/6
10/6
17/6
24/6
1/7
8/7
15/7
22/7
29/7
5/8
12/8
19/8
26/8
2/9
9/9
16/9
23/9
30/9
7/10
14/10
21/10
1/6

2/7

2/9

9/9
27/4

18/5

18/6

25/6

10/7

23/7

29/7

17/8

17/9

25/9

7/10

26/10

Figure 4.19. Daphnia spp grazing rates (% day –1; i.e. percentage site-area volume (litres) filtered per day)
during the two growing seasons (left: 1998, right: 1999).

Simocephalus vetulus had a maximum grazing rate of 8-10 % day-1 at certain times of the
growing season, with a localized peak of 15-30 % day-1 (Fig. 4.20). There were important

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GRAZING RATES

differences in the timing of maxima between years, and in the first growing season maximum
grazing impact occurred mid-summer while in the second year it was restricted to the end of
the season.

4 12
Simocephalus vetulus , 1998
Simocephalus vetulus , 1999
3,5
10
3 Min. plant biomass

Grazing rate (% day-1)


Grazing rate (% day-1)

Max. plant biomass 8 Min. plant biomass


2,5 Max. plant biomass

2 6

1,5
4
1

2
0,5

0 0
6/4

27/4

18/5

1/6

18/6

25/6

2/7

10/7

29/7

17/8

2/9

17/9

1/4
21/4
28/4

5/5
13/5
19/5

26/5
10/6
17/6

1/7
8/7
15/7

22/7
5/8
15/8
13/9
27/9
8/10
Figure 4.20. Simocephalus vetulus grazing rates (in % day-1, i.e. percentage volume filtered per day) across
lily beds in Little Mere (Cheshire) (Left: 1998; right: 1999). Different-colour bars indicate rates estimated
from the min. and max. plant biomass. Total populations of the species are calculated using the min. and
max. values found during plant surveys. Bars are one standard error at either side of the mean, calculated
from density estimate errors (for details see section 4.2.3).

In both years, Simocephalus density differences detected across lily beds paralleled those of
grazing rates. Rates were about twice larger in 1999 than in 1998 (Fig. 4.20). In 1998 the
largest grazing rates occurred in June and July, while in 1999 rates were larger in late
summer.

30 180 70
Sida crystallina , 1998 Sida crystallina, 1999
160
25 60
140 Min. plant biomass
Grazing rate (% day-1)

Max. plant biomass


Grazing rate (% day-1)

50
20 120 Chlorophyll-a
Chl-a (microg l-1)

Min. plant biomass


Max. plant biomass 100 40
15
80 30

10 60
20
40
5 10
20

0 0
0
1/4
21/4
28/4
5/5
13/5
19/5
26/5
10/6
17/6
1/7
8/7
15/7
22/7
5/8
15/8
13/9
27/9
8/10
6/4

27/4

18/5

1/6

18/6

25/6

2/7

10/7

29/7

17/8

2/9

17/9

Figure 4.21. Sida crystallina grazing rates (% day-1; i.e. percentage volume filtered per day) across lily
beds in Little Mere (Cheshire) (Left: 1998; right: 1999). Different-colour bars indicate rates estimated
from the min. and max. plant biomass at each site. Total populations of the species are calculated using
the min. and max. values found during plant surveys. Bars are one standard error at either side of the
mean, calculated from density estimate errors (for details see section 4.2.3).

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GRAZING RATES

Sida crystallina showed important differences in grazing rate both between sites and between
years. Highest grazing rates in the first year occurred at the beginning of the growing season
(i.e. April 1998), while during the second year only small peaks were detected in April. In

contrast, very high grazing rates were measured at the end of this growing season (i.e.
September-October ‘99) (Fig. 4.21). Maximum grazing rates in the second year were an order
of magnitude higher than in 1998 in some sites, too. Higher rates were located around site 1 in
both years (for a Map see Fig. 4.14). Significant reductions in algal biomass concomitant with
high grazing rates of this species were apparent in September-October 1999 (Fig. 4.21).
Unfortunately, no chlorophyll data are available for the beginning of growing season ‘98,
when relatively high rates were measured for this species. During approximately two months
at the end of this growing season, very low chlorophyll-a values were measured in Little
Mere, particularly within the lily beds (see Table 4.14). Implications for a top-down
regulation of algae by this grazer are discussed below.

4.4. Discussion.

Clearance and periphyton ingestion estimates.

Estimation of accurate feeding rates is fraught with difficulty. For instance, complete tracer
mixing in the grazing chamber is necessary for uptake to be representative. Very small species
and small individuals of larger species may encounter labelled cells at random in the short
feeding period, reflecting, rather than filtering rate, cell concentration in the box and/or
medium heterogeneity (e.g. Chow-Fraser & Knoechel 1984; Mourelatos & Lacroix 1990).
However, tests with a dye in the laboratory showed 30 seconds to be enough for complete
mixing. On the other hand, to obtain the minimum specific activity for in situ experiments I
had to be sure the algal density was neither too high compared with natural lake algal
densities, nor so low that homogeneous mixing in the box might become more of an issue,
given experiments are short-term. During the labelling period (48-72 hours) the algae
increased by at least an order of magnitude, making the culture denser than originally
intended. Thus, relationships between clearance rate and food conditions should be regarded
with caution. Periphyton activity in the box during each experiment had relatively narrow
bounds (Fig. 4.5).

It is still matter of controversy whether cladocerans feed continuously under natural


conditions. The short spans of time employed by necessity in radio-tracer feeding studies may
compound this effect but are difficult to detect. Haney (1973) brought attention to the fact diel

113
GRAZING RATES

rate depressions are overlooked in short-term experiments. Changes in animal behaviour


related to diel migratory rhythms are not taken into account either (see Peters & Downing
1984). Albeit important, these types of error are thought to be smaller than those controlled in
the experiment (e.g. tracer mixing, experiment fast termination).

Crowding is thought to explain a lot of the across-author variability in clearance rate estimates
(Peters & Downing 1984; Szlauer 1973). To avoid crowding, few animals were introduced
into the box (see section 4.2.1 for details).

Clearance rate models.

Slopes of relationships between clearance rates of species and body size are within the range
reported in the literature (see Table 4.11 & Fig. 4.16). Detailed cross-study comparisons are
not easily made on the basis of predictive regressions (Chow-Fraser & Knoechel 1984),
however they can point towards significant trends (see, for example, Peters & Downing
1984). Solutions of equations are subject to large uncertainty (see r2 values in Table 4.8) and,
thus, their primary use is heuristic (see Peters & Downing 1984).

Simocephalus vetulus emerged as a relatively unefficient filter-feeder, particularly in the


larger size range (i.e. > 1.2 mm; see Fig. 4.16 & Table 4.11). There is very large across-
worker variability in clearance rates estimated for this species. The trend, though, is for a
relatively low filtering efficiency as compared with other cladoceran filter-feeders.
Simocephalus vetulus is a slow-growing species (Bottrell 1975) and phytoplankton ingestion
at the rates measured may be sufficient to provide for this growth. In addition, one would
expect this species, given its strong preference for living in close proximity to plants (see, for
example, Lauridsen et al. 1996), to apportion a considerable part of its diet to alternative food
sources, such as periphyton, abundant within plant beds.

Sida crystallina, on the other hand, was found to be quite an efficient filterer, although rates
obtained were considerably lower than those presented by Downing & Peters (1980) and
Downing (1981). Values obtained by these authors for this species exceed by an order of
magnitude common values of clearance rate estimated for Daphnia (Table 4.11). In this study,
Sida crystallina was found to filter over twice as fast as similar-sized Daphnia. Because the
relationship between length and biomass of animals is similar for both species (Dumont 1975;
Downing & Peters 1980), Sida is considered to have a superior filtering capacity than

114
GRAZING RATES

Daphnia. However, predominance, or even abundance, of Sida has been seldom reported (see
Discussion in Chapter 3 for details).

In Little Mere, Sida clearance rates were higher with lower suspension and periphyton
concentrations (e.g. October 5th, 1999; see Fig. 4.10). Downing & Peters (1980) suggested
periphyton had some regulatory effect on the nutrition of Sida, as clearance rates decreased
with increasing periphyton concentration. Furthermore, Downing (1981) found this
'inhibitory' effect of periphyton on clearance rates to be enhanced at high suspension
concentrations. Clearance rates would be expected to decrease with increasing particle
availability anyway, irrespective of periphyton concentration (McMahon & Rigler 1965). This
highlights the difficulty of separating the influences on clearance rate of periphyton and
planktonic algae.

Clearance rates for Daphnia are conservative estimates. The small number of individuals
larger than 1.2 mm in experiments (see Fig. 4.16, top) may explain in part the relatively low
estimates for this species (see Table 4.13). Periphyton had a slightly 'negative' effect on
Simocephalus filtering activity, while the reverse was true for Daphnia. The latter result
should be regarded with caution, though, as 55 % of Daphnia samples were collected in the
first two experiments, biasing correlations towards the values of periphyton presented on the
first two dates (see Figure 4.10 and Table 4.15).

Periphyton ingestion models.

In Little Mere, larger Simocephalus ingested more periphyton than smaller individuals.
Whether these age-differences can be related or not to feeding habit can only be guessed at.
On the other hand, Simocephalus vetulus was inhibited in its periphyton ingestion activity by
the presence of the strips. Simocephalus may either actively swim away from plastic strips
colonized by periphyton (e.g. Lauridsen & Lodge 1996) or choose to filter-feed when
periphyton concentrations increase. Perhaps only the larger individuals were repelled by the
periphyton-covered strips, thereby filter-feeding on more carbon-14-free algae, while smaller
individuals not avoiding so much the proximity of the carbon-14 labelled algae, filtered more
of the detached algae in the process. However, this could also come about through more
swimming, in which case a more pelagic habit could be attributed to the younger animals,

115
GRAZING RATES

similarly to Sida crystallina (Fairchild 1981). To test these ideas feeding experiments are
needed which present both foods while they are closely monitored.

With regards to multivariate periphyton ingestion models (see Table 4.10), there is hardly any
consistency in variables selected across species. One would expect larger Daphnia individuals
to switch to browsing more frequently than smaller animals (Horton et al. 1979), collecting in
the process periphyton from strips. However, no significant correlation with size was detected
for periphyton ingestion rates. Periphyton concentration actually increased ingestion rate, as
would be expected if Daphnia utilized this source normally, which adds to the inconsistency.

Large Sida fed less on periphyton than smaller animals. Again this is an unexpected result, as
juveniles have been reported to swim faster and more frequently than older individuals
(Fairchild 1981), thereby probably associating less with periphyton-burdened surfaces. The
relationship is significant, albeit with a very flat slope, but very difficult to interpret
biologically. On the other hand, the most common methodological error involves not washing
away labelled cells attached to body surfaces. Larger animals would be expected to be more
prone to produce this error, giving a positive slope, rather than the one observed.

Clearance rates are the basis for the estimation of measures of grazing “impact” (% day-1; i.e.
% lake volume filtered per day). However, these measures are subject to further error, as new
variables come into play during calculations (see section 4.2.3 for details on methods). Errors
associated with each variable are multiplied when we calculate grazing rates. Potential
sources of error or natural variability when estimating grazing rates are listed in Table 4.13.

116
GRAZING RATES

Table 4.13. Potential sources of error in grazing rate estimation (see Equation 1).

Source of error* Units Description


Animal density (D & D’) ind g-1 Extrapolation errors: point-samples density in areas.
ind l-1 Large variability across replicates.
Natural spatial variation.
Sampling error.
Potential disturbance during sampling operations,
producing differences between replicates.
producing differences between species.
producing differences between samplers.
Areal plant biomass (P) g m-2 Unaccounted spatial heterogeneity.
Seasonal changes.
Sampling procedures and estimate accuracy.
Extrapolation errors: point-samples whole-bed
averages.
Total population size of ind Different sampling frequency of plant biomass and
animal species (T & T’) animal density.
In-bed plant biomass differences and extrapolated total
bed animal population sizes.
Error implicit in the linear regression between plant
biomass and density of associated animals.
Cover and Depth (C & z) m2 Seasonality of plant cover, particularly of submerged
m plants.
Averaged depth for each lake sector.
Season and inter-annual differences in water level (i.e.
depth, ‘z’).
Clearance rate (CR) ml day-1 ind-1 See section 4.2.1 for details.
*see meaning of variable codes in Table 4.5.

Estimates of field abundance of cladoceran species.

Plankton sampling techniques are bound to underestimate densities of microcrustacea living


attached or in close association to plant surfaces, particularly when these are low (Fig. 4.22).
Also, there is large variability in densities across replicate samples (Figure 4.23). Plant-
associated species densities are especially variable (see section 3.3.6 for details).

Plankton sampling technique and lily leaf sampler


6,0 Simocephalus vetulus , site 2 (1999)

5,0

plankton technique
4,0 leaf sampler
Nm-2 (x10-4)

3,0

2,0

1,0

0,0
01-jul 08-jul 15-jul 22-jul 05-ago 15-ago 13-sep 27-sep 08-oct

Figure 4.22. Comparison of the effectiveness of plant-associated Cladocera sampling using conventional
plankton techniques and a specifically designed lily leaf sampler. Densities are in ind.m-2. Plotted are
results for a population peak of Simocephalus vetulus sampled from a very shallow lily bed (site 2) during
growing season ’99. For details on extrapolations, see text.

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GRAZING RATES

Sampling procedures are bound to affect how representative samples are of real populations.
It is important, though, that errors due to sampling procedures are not systematic errors. No
apparent pattern of disturbance was detected in my dataset (see Fig. 4.23).

Extrapolation of point-samples to estimate densities in whole areas is an important source of


error (De Bernardi 1984). In littoral areas and in shallow lakes in general, swarming and
contagious distribution of populations, are likely to be more important than in pelagial
environments (Folt 1987; Folt et al. 1993; see Discussion in Chapter 3).

450

400

350

300
% of mean

250

200

150
2
r = 0,026
100

50

0
0 1 2 3 4 5
Replicate order

Figure 4.23. Replicate order and % of mean density sampled with each replicate during a population peak
of Simocephalus vetulus (site 1, growing season 1999). No trend for a disproportionate amount of animals
in the first sample suggests that there was no systematic effect of disturbance during sampling operations.

Areal plant biomass and cover estimates .

During the height of the growing season, errors overall are smaller, simply because there is
generally less time between plant survey and sampling of the animals. This pattern of error
may apply to whole plant beds, but within beds it is likely that growth and/or senescence
proceeds at different rates in different areas (Spence 1982).

A main assumption in the extrapolation of point-measures of animal density (ind g-1DW) to


whole areas is that plant biomass is linearly related to the density of associated animals. No
significant relationship exists between leaf weight and number of Simocephalus vetulus (or
Sida crystallina) per leaf. The unit of relevance is probably surface area, but estimating such a
parameter routinely is difficult. However, because leaf weight in samples is a perfectly
random variable (i.e. it fits the Poisson distribution very well), the large number of leaves
sampled allow us to consider the variable ‘leaf weight’ as normally distributed (Bailey 1992).
Thus, leaf weight, when thought of in terms of populations of leaves, is an excellent predictor

118
GRAZING RATES

of total number of associated animals (see Fig. 4.24). Analogous considerations apply to lily
petiole weight.

Extrapolation errors: cumulative numbers of Simocephalus


vetulus (both years) and cum. lily leaf weight against number of
10000 lily leaves 500
9000 cum. Simocephalus 450
8000 400

cum. lily leaf weight


7000 cum. leaf weight 350
Simocephalus
cum. number

6000 300

(gr.)
5000 250
4000 200
3000 150
2000 100
1000 50
0 0
0 50 100 150 200
cumulative number of lily leaves

Figure 4.24. Prediction of total numbers of Simocephalus vetulus on lily leaves from the weight (grams) of
a large population of lily leaves. See text for details.

Grazing rates and water clarity in Little Mere.

Daphnia was generally the main grazer in Little Mere (Figs. 4.19 to 4.21). Variations in
chlorophyll-a are apparently in an inverse relation to Daphnia density during most of the
growing season. On numerous occasions, however, high water transparency (i.e. low chl-a
concentration) coincided with small Daphnia populations, suggesting other processes aside
from strict Daphnia top-down control may have been in operation during these periods (see
Chapter 1 for details). Sida crystallina was the most important plant-associated filter-feeder
during the two years sampled. Simocephalus vetulus, by contrast, was generally not an
important grazer in Little Mere, although, in combination with Sida, could account for
changes in water clarity (see Fig. 4.21).

No conclusions can be drawn on when and where precisely is the large grazing impact of Sida
going to occur. However, population peaks and, therefore, high grazing rates, seem to occur
every growing season. This species appeared at the very beginning and end of each growing
season, perhaps exploiting the window of opportunity oferred by Daphnia´s generally low
numbers during these periods. The suggestion is these two species compete, either through
direct interference or through competition for common resources (i.e. planktonic algae).
Sida's strategy seems to be to rapidly outnumber other potentially competing species,

exhausting resources and, thus, disappearing after episodes of extreme population crowding
(see section 3.3.6 for details).

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GRAZING RATES

The question remains, however, of what actual effects on water clarity the plant-associated
filter-feeder community are having during these periods of calculated high grazing rate. The
discussion following will be centred around Sida crystallina and the clear-water phase
occurring at the end of growing season ‘99 (see Table 4.14 for details).

It is difficult to predict how relatively localized impacts will affect whole lake water
transparency, particularly as exchange rates between water masses are not known (see James
& Barko 1991). Indeed on particular dates with high plant-associated herbivory, mainly of
Sida crystallina (see Figure 4.21), a clear-cut difference in transparency and chlorophyll-a
levels was apparent between open-water sites and lily bed sites (see Table 4.14). For example,
at the end of growing season ‘99, and during about two months, very low chlorophyll-a
values were measured (i.e. less than 20 µg l-1) concomitantly with low Daphnia densities (i.e.
less than 10 ind l-1 in lily beds; less than 20 ind l-1 in open-water sites; see Fig. 3.8). On the
other hand, grazing rates of the plant-associated filter-feeders (i.e. Simocephalus vetulus and
Sida crystallina) were sufficient to account for the low chlorophyll-a values, while those of
Daphnia, be it in plants beds or in more open-waters, were not (see Fig. 4.21 for Sida
crystallina, Fig. 4.19 for Daphnia).

There is much evidence to support the idea that Sida was the main regulator of water
transparency during these dates. For instance, during the first two dates of clear-water (i.e.
September 13th and Sept. 27th, 1999; see Table 4.14) relatively low Sida densities were found
in the open-water sites, dominated by submerged plant beds (i.e. sites 3 and 4, with no lilies;
for a Map see Fig. 4.14). Chlorophyll-a measured in these sites these dates was twice to four
times higher than in the lily beds.

Very high Sida densities (17,000-40,000 ind m-2) were found on October 8th in submerged
vegetation, coinciding with low chlorophyll-a concentrations (< 10 µg l-1), suggesting Sida
was indeed the main grazer. Negligible grazing rates of Daphnia were measured in the open
water during the clear-water phase (see Fig. 4.19).

The clear-water phase is difficult to explain by other processes aside from herbivory. Direct
plant competition with the algae for nutrients in the water is one possibility (see, for example,
Howard-Williams 1981). Ceratophyllum demersum, a completely submerged plant species,
increased in cover and biomass towards the end of the growing season (see section 4.3). This

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GRAZING RATES

mechanism, however, cannot account for the fact water was clearer in lily beds than in more
open waters. In addition, nutrients (both nitrogen and phosphorus) were not limiting during
the clear-water phase. For instance, 234 µg l-1 of NH4+ and 129 µg l-1 total phosphorus were
measured on October 5th, with values of less than 10 µg l-1 of chlorophyll-a (see Table 4.14).
Without an estimation of rates of exchange of water masses between the quieter lily areas and
more open waters and resulting nutrient exchange, the importance of plant uptake cannot be
properly assessed, though.

Ceratophyllum demersum is thought to have strong allelopathic effects (Gross 1999; see
Chapter 1 for details). The increase in biomass of this species coincided with the clear water
(see Table 2.2), but again, plant beds were largely confined to outside the lily-covered areas,
while the clearer water was found in the lily beds (see Table 4.14).

Wash-out of algae cells was most likely not an important factor in maintaining water
transparency during the observed clear-water phase, with relatively low outflows and sinking
rates within and outwith lily beds probably comparable throughout the growing season (see
Table 4.14). Nevertheless, sinking may explain differences across sites to a certain extent,
particularly during some dates. For instance, proportionally more small algae (i.e. < 25 µm)
could be found at the sheltered site 2 (for a Map see Fig. 4.14), than in other lily areas, and
clearly smaller ratios were consistently found in deeper areas, without lilies.

Table 4.14. Clear-water phase at the end of the 1999 growing season. Small to total chlorophyll-a (%) and
total chlorophyll-a values (µg l-1) across sampling sites in Little Mere (Cheshire).

Clear- % small/large chlorophyll-a Total chlorophyll-a


water
Lily bed Open water Lily bed Open water
phase
Site 1 2 5 3 4 1 2 5 3 4
13th Sept. 91.6 100 92.5 55.5 38.3 9.5 6.3 10.7 28 90
27th Sept. 84.0 100 62.5 68.8 42.9 26.2 15.5 61.5 33.7 83.7
8th Oct. 77.6 71.1 77.4 62.1 82.0 10.7 9.7 11.5 8.7 13.3

In conclusion, top-down effects of Daphnia are important in Little Mere, and, in combination
with plant-associated herbivory, could explain most changes in water clarity in this lake.
Plant-associated grazers such as Sida crystallina are a by no means negligible factor in
controlling algal biomass, at least during certain periods of the growing season (April to
October). Experiments would be needed, however, in order to assess the absolute

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GRAZING RATES

contributions of these important grazers, isolating any confounding effects of, for instance,
nutrient uptake by plants, in competition with algae.

Herbivory as a regulating factor is under a set of varying nutrient regimes and fish predation
pressures in natural conditions. Next chapter presents the results of two mesocosms
experiments looking into how herbivory, and thereby water transparency, is regulated by
nutrients and fish, and how these factors may interact in Little Mere.

4.5. Conclusions.

Clearance rates (ml ind-1day-1) and periphyton ingestion rates (µg ind-1day-1):

• Significant clearance rate (CR) regression models for five of the six cladoceran species in
experiments were drawn, i.e. Daphnia spp, Simocephalus vetulus, Sida crystallina,
Eurycercus lamellatus and Scapholeberis mucronata.

• All models accounted for 21.7-69.7 % of variance in CR measurements and 11.4-73.2 %


of periphyton ingestion estimates.

• Simocephalus vetulus: body length accounted for 53.8 % of CR variation. An additional 3


% was explained by the deppresive effect of periphyton concentration (mg cm-2 periphytic
chl-a). Temperature had a strong deppresive effect on periphyton ingestion. Periphyton
concentration was also inversely correlated with periphyton ingestion. Larger animals
ingested more periphyton.

• Daphnia spp: this species filtered less efficiently than Simocephalus and Sida, particularly
larger-bodied animals. Periphyton ingestion was positively correlated with periphyton
concentrations in the grazing chamber. Suspended algae food also had a positve effect on
periphyton ingestion.

• Sida crystallina filtered about double the volume of similar-sized Daphnia, particularly
the larger individuals. Both high suspension and periphyton concentrations depressed
clearance rates. Larger Sida ingested less periphyton than small individuals.

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GRAZING RATES

Grazing rates (% day-1):

• Plant-associated filter-feeders (‘alternative grazers’) are a generally minor grazer


component in Little Mere as compared to open-water filterers such as Daphnia spp.

• At the beginning and end of the growing season alternative grazers may have had a
significant impact on phytoplankton, or even been the main grazers during short periods.

• The magnitude of these short-term effects is very variable across years.

• Timing of maximum grazing impact of alternative grazers is also very variable across sites
within any given year.

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CHAPTER 5

The relative role of nutrients and fish predation on


zooplankton dynamics in Little Mere
CHAPTER 5

5.1. Introduction.

The relative weight of physico-chemical ('bottom-up') and biological ('top-down') factors


influencing communities and ecosystem function remains a fundamental issue in aquatic
ecology (e.g. Kairesalo et al. 1999; Blindow et al. 2000). Efforts, however, have often by-
passed the basic questions in search of direct, fast applications (Kitchell et al. 1988; Gophen
1990; Kitchell & Carpenter 1993).

For instance, biomanipulation, or "the deliberate exploitation of the interactions between the
components of the aquatic ecosystem in order to reduce the algal biomass" (Shapiro 1990),
has had a controversial history (see De Melo et al. 1992 for comment), with periods of an
abundance of success cases being reported (e.g. Briand & McCauley 1978; Shapiro & Wright
1984; Carpenter et al. 1987; Post & McQueen 1987; Threlkeld 1988; McQueen et al. 1990),
soon followed by case-studies acknowledging its multiple limitations (for a review see Gulati
1995).

Some of these limitations come from its short-lived effects (e.g. Vijverberg 1984; Mills et al.
1987; Shapiro 1990; McQueen et al. 1990), from unknown complexities of natural
ecosystems interfering with restoration (e.g. invertebrate predation, e.g. Edmondson & Abella
1988; Benndorf 1990; benthivorous fish and sediment resuspension, Meijer et al. 1994; see
Lammens et al. 1990 and references therein), or from blue-green algae, which are thought to
be relatively unedible (Porter 1973, Gliwicz 1990, but see DeBernardi & Giussani 1995)
coming to predominate as more favoured algae are grazed down by zooplankton (e.g. Gliwicz
& Siedlar 1980; Moss et al. 1991). Moreover, it is now recognized, algae may quickly
develop morphological defenses in response to infochemicals exuded by zooplankton (Lurling
1999).

However, biomanipulation seems to be very effective in shallow lakes (Lammens et al. 1990;
Jeppesen et al. 1990; Kasprzak 1995). The success of this restoration technique relies on the
strength of top-down interactions in the lake (e.g. piscivory, herbivory by microcrustacean
grazers). Generally, the aim is to sustain populations of large herbivores (Lampert 1988).
Indeed, smaller zooplankton are thought to be less efficient at controlling algal biomass
(Dawidowicz 1990; Crisman & Beaver 1990).

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

By contrast, nutrient control alone has seldom worked as a restoration tool in shallow lakes
(see, for example, Jeppesen et al. 1990; Hosper & Jagtman 1990; Jeppesen et al. 1991) and
one of the reasons for this could be the increased internal loading from organic-rich sediments
as external loads are reduced (Moss et al. 1991; Phillips et al. 1994; Stephen et al. 1997). In
addition, bioturbation by benthivorous fish (Meijer et al. 1990; Meijer et al. 1994) or
chironomids (Phillips et al. 1994) particularly in already eutrophicated waters (see, for
example, Vanni et al. 1990), may hamper restoration efforts.

Nutrient control, although often insufficient for complete restoration, is always necessary
(see, for example, Moss et al. 1996). Trophy of the lake strongly affects outcomes of
biomanipulation, although it is still not well known to what extent (Carpenter et al. 1997).
Elser et al. (1990), for example, proposed lakes of ‘intermediate productivity’ would be the
best candidates for successful biomanipulation because links between zooplankton and algal
biomass are stronger. For the same nutrient concentration, different herbivore species have
been shown to have a different "pattern of nutrient recycling" (Sterner 1989; Urabe &
Watanabe 1992; Urabe 1993), in part explaining the observed shifts of community structure
with eutrophication toward small, relatively unefficient, herbivores (Pace 1984). On the other
hand, the response to nutrient enrichment depends on trophy too, and the more nutrient-rich,
the less susceptible to nutrient effects (Allan 1980). The need for combining nutrient control
measures with manipulation of the trophic structure of shallow lakes was emphasized at the
Biomanipulation Conference held in Amsterdam in 1989 (Lammens et al. 1990; see, for
example, Benndorf 1988, 1990; Hosper & Jagtman 1990; Sondergaard et al. 1990).

McQueen (1990), for instance, found that a surprisingly low 64 % of manipulations (in a set
of 36 analysed) reported significant effects of zooplankton on algae, while 100 % reported
significant effects of piscivores on fish. He explained this as implying that the trophic cascade
was "damped". Smaller-sized species are less efficient at utilizing available food particles
than could be, for example, piscivorous fish feeding on planktivores. Thus, eutrophication
would determine a wider gap between primary production and algae biomass utilization by
the herbivorous zooplankton (see De Bernardi & Giussani 1995).

De Melo et al.'s (1992) cross-analysis of 50 papers shows that in those cases where
biomanipulation was reported to have been successful, conclusive evidence was not put
forward as to what were the main causal mechanisms for clarification of the water, whether

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CHAPTER 5

zooplankton herbivory alone or grazing in combination with other processes. Moreover, in


their analysed set, 56 % of enclosure studies, and 50 % of lake or pond studies were qualified
as 'undecided' as regards zooplankton grazing being the main causal agent of increased
transparency following manipulation. In fact, examples exist of successful restoration plans
based on biomanipulation where the algae community biomass and structure was shown to
depend mainly on nutrients (Kitchell & Carpenter 1992; Gophen 1995).

Table 5.1. Some problems and indirect effects encountered in the literature during restoration attempts of
shallow lakes using biomanipulation (modified from Kasprzak 1995 & Gulati 1995).

Examples DIFFICULTIES IN Examples INDIRECT AND


APPLYING UNEXPECTED EFFECTS
BIOMANIPULATION
Vijverberg 1984 Planktivorous fish tend to re- Meijer et al. 1994 Nutrient recycling due to
establish high standing stocks. increased sediment
resuspension by benthivorous
fish.
See DeMelo et al. The response is sometimes Edmondson & Daphnia is released from
1992 for criticisms weak (‘damped’). Abella Neomysis predation because of
increase of longfin smelt
(Spirinchus thaleichthys)
Mills et al. 1987 Nutrient limitation of Leah et al. 1980 Increase in macrophyte
zooplankton through ‘over’- biomass and cover with higher
biomanipulation. water transparency and indirect
macrophyte effects, i.e.:
Shading of phytoplankton
Zooplankton refugia
Plant competition for nutrients
in the water
Allelopathy
Faster sinking, …
See Chapter 1 for details.
Gliwicz & Siedlar Phytoplankton develops
1980 morphological defence
mechanisms.
Benndorf 1990 Some planktivorous fish are
more resilient to
biomanipulation in part owing
to different feeding behaviour
and time-scheduling of this.
Vanni et al. 1990 Effectiveness depends on
nutrient status of lake. Nutrient
control is often necessary.

Control of indirect mechanisms playing a role in algal growth may be key to successful
biomanipulation (see, for example, Kairesalo 1999; see Table 5.1). In shallow lakes, many of
these mechanisms are mediated by plants (Jeppesen et al. 1997; see Chapter 1 for details).
Indeed, as water becomes clearer and plant coverage increases, a series of 'buffering
mechanisms' of the plant-dominated state are thought to set in (Ozimek et al. 1990; Moss
1990; Grimm & Backx 1990).

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Because field enclosures ('mesocosms') can be easily controlled and replicated (see Table
5.11) they are a powerful tool for investigating some of these mechanisms and ecosystem
function (Carpenter & Kitchell 1992, but see Frost et al. 1988). Issues that have been
profitably addressed in a mesocosms approach are:

• The relative importance of top-down and bottom-up processes on water clarity


maintenance. The interactions between these types of processes are perhaps more
interesting than a simplistic assessment of their relative importance (see, for example,
Carpenter et al. 1987; Vanni 1987).
• The role of macrophytes in the maintenance of water clarity (e.g. Beklioglu & Moss 1995;
Stephen, Moss & Phillips 1998; Moss, Kornijow & Measey 1998).
• Selected interactions between zooplankton and their predators, and between herbivorous
zooplankton and the algae they graze (see Carpenter & Kitchell 1993).
• Effectiveness of the biomanipulation technique in a particular lake (e.g. Van Donk et al.
1994; Faafeng et al. 1990).

Enclosure studies were conducted in Little Mere (Cheshire) at the height of two summers with
the main objective of understanding how fish presence and different nutrient regimes may
affect, a. the biomass build-up of different zooplankton species and, b. water transparency,
potentially regulated by herbivory, in this lake. A graded series of nutrient loadings was
applied at varying levels of fish predation (i.e. fish density) and the changes in various
variables monitored (see detailed Methods in section 5.2). A further aim was to look at how
the system would respond to increased nutrient loads once plants had been removed. In this
case, the hypothesis was that the “switch” towards an ‘algae-dominated state’ would occur
more easily when plants were removed (see Chapter 1).

Unfortunately, there was not much plant growth inside our enclosures, or even in the lake
itself during the experiments (see Discussion for details). The focus in this chapter is therefore
on the population dynamics of planktonic Crustacea (Cladocera), as opposed to plant-
associated cladocerans, and on water transparency, under the conditions imposed by
treatments (nutrient loadings and applied fish densities) .

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CHAPTER 5

Also, the experiments conducted here were part of a larger project at the European scale
(SWALE, namely, "Shallow Wetland and Lake Function and Restoration in a Changing
European Environment"). Effectively the same experiment was reproduced six times in five
different countries across a latitudinal gradient, i.e. Finland, Sweden, The Netherlands, United
Kingdom and Spain, at the heights of two growing seasons (’98 and ’99). In Spain, two
experiments were performed, one located in the northwestern region, and the other on the
central Mediterranean coast.

The main hypothesis tested in these experiments was that the stability of a shallow lake
ecosystem, defined as its resilience in the face of increasing nutrient loads, would deteriorate
as the current warming trend reflected in water temperatures. Specifically, higher average
temperatures would facilitate a ‘switch’ (see Chapter 1) to the algae-dominated state in many
shallow lakes. This could happen with:

a) increasing average water temperatures (across experiments).


b) increased fish activity at higher temperatures, removing the ‘top-down control’ of algae
exerted by the zooplankton grazers (within each experiment).

Thus, to test this two-pronged hypothesis, experiments were ‘replicated’ across a latitudinal
gradient. The average temperature differences experienced by shallow lakes in very different
latitudes mimicked the changes that could be experienced by these systems as a consequence
of global warming.

5.2. Materials and Methods.

Two experiments using mesocosms were conducted mid-summer 1998 and 1999 in Little
Mere (Cheshire, England; see Chapter 2). The main design differences between them are
summarized in Table 5.2.

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Table 5.2. Experimental designs and treatments of both experiments.

1998 1999
Dates (total number of weeks). June 8th-August 3rd (9) June 28th-August 9th (7)
Replicates (per treatment combination). 3 2
Number of treatment combinations. 12 18
Applied nutrient loadings (Nitrogen/Phosphorus) 0, 8.3, 41.5 & 83.1 mgl-1 NO3 0, 0.13, 0.25, 0.39, 0.55 & 1.32 mgl-1
(in mgl-1 of enclosure water*) 0, 0.44, 2.2 & 4.4 mgl-1 PO4 0, 2.53, 5.05, 7.58. 12.54 & 25.28 mgl-1
Target nutrient loadings (Nitrogen/Phosphorus). 0, 1, 5 and 10 mg1-1 NO3 0, 0.5, 1.2, 1.8, 2.4 and 3 mgl-1 NO3
0, 100, 500 and 1000 µg1-1 PO4 0, 50, 120, 180, 240 and 300 µg1-1 PO4
Number of fish added per enclosure in the 0, 3 & 15 fish 0, 3 & 15 fish
different fish treatment levels*
Target fish density treatment levels. 0, 4 & 20 gm-2 0, 4 & 20 gm-2
*to calculate these weekly additions, we assumed an average depth of 0.95 metres and a total volume of
water enclosed by each enclosure of 750 litres; fish are assumed to have an average wet weight of 1 gram.

The plastic used for the enclosures was soft polyethylene of 125 µm thickness. Each end of
these bags of soft plastic was held onto 1-m diameter plastic rings using cable ties. The
enclosures were then set in place holding them by the frames (see Fig. 5.2), and weighted to
the bottom with bricks (Fig. 5.3). A plastic, coarse mesh was attached covering the whole
outer surface of each block of enclosures, to minimize disturbance from fish and birds (see
Fig. 5.1). See details of the construction of frames and enclosures in Figure 5.2.

Figure 5.1. Materials needed for the experimental setup in the lake. At the front of the picture, the netting
use to cover the outside of the frames can be seen. Frames, stakes and enclosures are behind.

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CHAPTER 5

Figure 5.2. Construction of each frame for four enclosures. Tbe frame was made of PVC tubing. Also
shown are the attachments of each frame to stakes, which were used to fix the frame into position in the
lake. Attachment of the tubing to stakes used cable ties, as shown in the diagram.

Both experiments followed a randomised block design. Each block of enclosures contained
one replicate of each treatment combination, in a random position within the block, so
“position effects” could be ironed out in averaging across treatments. The 1998 experiment
consisted of 3 blocks of 12 enclosures, with 3 levels of fish density applied (one no-fish
control), and 4 nutrient loadings (one no-nutrient addition control). In 1999 the number of
replicates was only two, as 5 different nutrient loads were applied (see Table 5.2 & Figure
5.4).

Figure 5.3. Arrangement of the in situ enclosures into blocks. Two blocks of 12 mesocosms can be seen in
the picture, as the plastic enclosures are set in place.

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Figure 5.4. Arrangement of enclosures in groups of four and in blocks. Each treatment combination was
represented by one enclosure in each block. The position of each treatment combination (e.g. no nutrients,
high fish density) was randomized within each block to ‘iron out’ potential unwanted effects of enclosure
position on a variable sediment. In 1998 there were 3 blocks and 12 treatment combinations. In 1999, 2
blocks, with 18 treatment combinations each.

The fish used were small sticklebacks (Gasterosteus aculeatus) 4-5 cm long, captured with a
benthos net (1 mm mesh) from streams in Ince marshes (Cheshire). Some fish were kept in a
continuous flow aquaria to replenish enclosures where mortality was observed, in order to
maintain desired treatment densities. Fish were added to obtain approximate densities of 0, 4
and 20 gm-2 (0, 3 and 15 one-gram fish, approximately). Phosphorus and nitrogen loadings
were applied as KH2PO4 and Ca(NO3)2.4H2O. Treatments were applied weekly, following the
first, ‘pre-treatment’ week, to increase PO43--P and NO3--N concentrations by 0, 0.1, 0.5 & 1
mgl-1 and 0, 1, 5 & 10 mgl-1, respectively. Treatment differences between experiments are
shown in Table 5.4. The loadings in the 1999 experiment were lower, as results from the first
year suggested the ecologically meaningful range for our lakes was well below 5 mg l-1 for
NO3--N and 0.5 mgl-1 for PO43--P. No significant differences were generally detected across
treatments for the zooplankton groups analysed in this week (see Table 5.5) and, therefore,

133
CHAPTER 5

subsequent analyses were performed only on ‘treatment weeks’, thus assuming a common
baseline for all enclosures.

Zooplankton data were organized either directly as species or in 7 large groups classified
according to size and habit of species (see Table 5.3).

Table 5.3. Pre-treatment week: significance of differences in means (GLM-ANOVA, Sum of Squares type
III) across enclosures grouped as either ‘to-be-treatment-combinations’ or as ‘experimental blocks’.
Blocks are included as a ‘random factor’. Results are for the main seven zooplankton groups analysed in
both years (in µg-C l-1). These groups are: 1- Small Cladocera (<500 µm); 2- Large Cladocera (>500 µm);
3- Cyclopoid copepods; 4- Calanoid copepods; 5- Total copepod nauplii; 5- Raptorial Cladocera &
Copepoda; 7- Filter-feeders. For the species composition of each group refer to Table 5.3. Number of cases
analysed when zero-cases were excluded is shown. There were not enough valid cases for analysis in group
2 (1999) once zero-values were excluded (see text for details).

Significance Group N 1998 1999


(*p<0.05; **p<0.01; ***p<0.001) (1998/1999)
1 15/35 n.s. n.s.
2 15/* n.s. cannot compute
3 13/35 n.s. n.s.
Grouped by treatment combination 4 35/35 n.s. n.s.
(1998: 12; 1999: 18) 5 35/35 n.s. n.s.
5 35/35 n.s. n.s.
7 35/35 n.s. n.s.
All 35/35 n.s. n.s.

1 15/35 n.s. n.s.


2 15/* n.s. cannot compute
3 13/35 n.s. *
Experimental blocks 4 35/35 ** n.s.
(1998: 3; 1999: 2) 5 35/35 n.s. n.s.
5 35/35 ** *
7 35/35 * n.s.
All 35/35 n.s. n.s.

A 10-litre weekly zooplankton sample was taken per enclosure using a tube sampler (see Fig.
3.6). Seven of these 10 litres were strained through a 50 µm mesh to collect the cladoceran
zooplankton. The remaining three litres were separately strained through a 25 µm mesh to
estimate the density of rotifers. Cladocera were counted under a stereomicroscope at 30x
magnification, and rotifers under an inverted microscope at 100x.

Data are abundances (ind l-1) or biomass (in µg-C l-1) for each species and biomass (µg-C l-1)
for groups. The percentages of total zooplankton biomass represented by each group were
also calculated and analysed. Biomass was estimated using regression equations relating body
size to biomass (in µg-C l-1) provided by Luokkanen (1995), Lehtovaara (unpubl.) and
Kankaala et al. (1990). Average size estimates for each species in each experiment were

134
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

drawn from a lumped sample bringing together extreme treatment combinations and
enclosures with largest numbers of any given species. Size-structure of populations and
changes in average size were not taken into account in these estimates. However, differences
due to size changes were expected to be smaller than, for example, sampling errors. Thus, the
extra measuring effort for more precise size estimates was considered unnecessary.

Table 5.4. Species composition of zooplankton groups, classified on the basis of size and feeding habit.

GROUP SPECIES REPRESENTED


1-Small Cladocera (< 500 µm) Bosmina longirostris, Ceriodaphnia spp,
Polyphemus pediculus and Scapholeberis
mucronata.
2-Large Cladocera (> 500 µm) Daphnia spp and Simocephalus vetulus.
3-Cyclopoid copepods mainly Cyclops spp
4-Calanoid copepods mainly Eudiaptomus gracilis.
5- Total copepod nauplii Both cyclopoid and calanoid nauplii.
5-Raptorial Cladocera + Copepoda Cyclops spp and Polyphemus pediculus.
7- Filter-feeders Bosmina longirostris, Ceriodaphnia spp,
Daphnia spp, Eudiaptomus gracilis,
Scapholeberis mucronata, Simocephalus vetulus
and copepod nauplii.

Scarce plant growth had implications on the data finally included in analyses. The data
collected after plant removal in the first year were analysed separately from weeks before
cutting. Thus, results of analyses concerning abundances and biomass before the plant cutting
are tabled separately from those after the cutting. The intensity of perturbation represented by
the plant cutting was deemed to be stronger than any direct effect of plant removal, given the
low plant biomass present in enclosures (see Appendix D). Therefore, in the second year no
plant removal was attempted and, thus, data of all experimental weeks are analysed together.

Statistical analyses.

Nutrient load and fish predation effects on zooplankton density and biomass.

Reliability of means increases as the experiment ‘matures’. The outcome of treatment


application, at the end of the experimental period, is of more value than changes occurring
throughout the experiment. Thus, data were weighted on the basis of the number of weeks
passed from the start. The time-variability of treatment effects was examined separately for
this time-averaged analysis, by means of plots of the three-way interaction
nutrient*fish*week.

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CHAPTER 5

Analyses were conducted using three-way mixed-effect analyses of variance on each


species/group, to identify significant treatment effects (i.e. nutrient loadings and/or fish
densities). Effects of position of treatments within experimental blocks were accounted for by
including ‘block’ as a ‘random variable’ (see Lindman 1974). The power of the test is
considerably increased with respect to totally randomized designs (i.e. ignoring block effects)
(Lindman 1974; Sokal & Rohlf 1981). Furthermore, if significant differences between blocks
with respect to fixed treatments are not taken into account, conclusions can be radically
different. In general, treatment effects are overestimated when blocks are not included in the
design. When a significant effect was detected by the F-test, Tukey’s honestly significant
difference tests were conducted to identify which levels of the treatment were mainly
responsible for the significant effect.

Normality and stability of variance of the dependent variables were visually checked. The
normality assumption was checked by inspection of a plot of the residuals of data against the
cumulative predicted normal line. On the other hand, any patterns in variance across the data
were checked by plotting means versus their standard deviations. Biomass and count data
were generally log-transformed to ‘pull in the long tails’, and percentage data were
arcsin(square-root)-transformed, particularly when patterns in variance were detected.

Abundance of values close to zero may lead to computational problems. This is because the
estimated variance will be unduly low on dates with many zeros, generating a pattern of
unstable variance, with variance much larger for high means than for means close to zero (i.e.
from means estimated from data with many zeros). Transformation is often insufficient to
correct this pattern of unstable variance (Samuels 1991). Because variance cannot be assumed
to be equal across different mean estimates, the F-test has no power to discern differences
between means from those occurring in variances (Fowler & Cohen 1990). Therefore, when
more than half the data were made up of zeros, only non-zero data were analyzed. By this
procedure we re-scale data and assume nil values do not respond to treatment effects. This is
not an unreasonable assumption given treatment effects are assessed in terms of differences in
numbers, and not as producing the presence or absence of any given species. Indeed,
differences in the zooplankton community composition may be more influenced by
differential recruitment from different sediment patches covered by enclosures (DeStasio Jr.
1990), than by any treatment applied. Data from the pre-treatment week were used to assess
whether there were significant differences between enclosures and/or blocks as regards any of

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

the variables of interest (i.e. zooplankton density, chlorophyll-a,…). As no significant


differences were detected (see Table 5.3), the pre-treatment week data was excluded from
subsequent analyses.

Effect of a provoked perturbation on zooplankton dynamics.

The effect of plant-cutting on zooplankton dynamics in the 1998 enclosures was also looked
at. To analyse this effect an ANOVA of biomass of each group (in µg-C l-1) was performed
separately on data before and after the cutting. Significances of effects before and after the
perturbation were then compared for each group. All enclosures were included in these
analyses, i.e. both enclosures where plants were removed (i.e. disturbed) and enclosures
where no plant growth was apparent and therefore had not been disturbed.

5.3. Results.

Experiments in the separate years were characterised by very different zooplankton


communities (see Table 5.5).

Table 5.5. Ranges of biomass (in µg-C l-1) and relative importance (in % of total zooplankton biomass) of
main groups of zooplankton in both experiments. The minimum and maximum in each range refer to the
extremes in any given enclosure.

Group 1998 1999


µg-C l-1 %(over total µg-C l-1 % (over total
zooplankton zooplankton
biomass) biomass)
Small Cladocera 0-298 0-57 0-2,539 0-95
Large Cladocera 0-885 0-98 0-314 0-59
Cyclopoid copepods 0-135 0-55 0-1,013 0-85
Calanoid copepods 0-443 0-97 0-53 0-9
Nauplii 0-221 0-58 0-495 0-53
Raptorial feeders 0-135 0-55 0-1,013 0-85
Open-water filterers 5-1,159 44-100 9-10,025 14-100
Rotifers 0-933 0-100 0-9,983 0-99.8
Total biomass 7-1,288 - 34-10,055 -

The experiment in 1998 was dominated by large Cladocera, mainly Daphnia spp, and among
copepods, calanoids were more frequent and abundant than cyclopoids. In the second year
experiment, small Cladocera (i.e. Bosmina longirostris and Ceriodaphnia spp) and rotifers
were prevalent and populations of larger species only started developing late in the
experiment. Calanoid copepods were scarce in this second experiment, while cyclopoid
copepods were frequently the dominant group of copepods. Rotifer biomass (in µg-C l-1) was
one order of magnitude larger overall in the second year experiment (see Table 5.5). In

137
CHAPTER 5

general, the second year experiment yielded much larger biomass for most groups,
particularly small Cladocera, cyclopoid copepods and rotifers. The latter were by far the
dominant zooplankton group in the 1999 experiment (Table 5.5).

Differences in community composition across experiments and a discussion of possible


factors explaining these differences are given in section 5.3.2 and in the Discussion section.
Results for each experiment are separately described in the following two sections.

5.3.1. The 1998 experiment.

Fish decreased the biomass of large and small Cladocera in significant amounts, both before
and after the plant-cutting (see Table 5.7). In this experiment, large Cladocera (>500 µm)
were largely represented by Daphnia spp. Fish effects on Daphnia biomass were generally
very conspicuous (F=125.0, p<0.001 & F=218.8, p<0.001 in the pre- and post-cutting periods,
respectively; see Table 5.9). Indeed, at the highest fish density, Daphnia populations were
close to being wiped out. At the low fish density treatment, on the other hand, Daphnia
populations were stable at about two thirds of the biomass of those in enclosures with no fish
(see Fig. 5.5). The nutrient treatment interacted with this strong fish predation on Daphnia,
particularly during the post-cutting period (see Table 5.9). The dynamics of this interaction
are examined in detail in the Discussion section.

Contrary to the cladoceran zooplankton groups, rotifer biomass was significantly greater in
the fish treatments than in enclosures without fish (F=2171, p<0.001 and F=25.4, p<0.01, in
the pre- and post-cutting periods, respectively; see Fig. 5.6).

Nutrient effects on the biomass of most zooplankton groups were not significant (see Table
5.9). The general tendency, though, was for enclosures with a higher nutrient loading to have
a larger maximum zooplankton biomass. However, these effects were not significant against
the background of fish predation effects. Nutrient effects were only significant on the biomass
of the smallest zooplankton, i.e. copepod nauplii and rotifers, but not on the relative
proportions (i.e. % of total zooplankton mass) of these groups (see Table 5.9).

138
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Daphnia hyalina Daphnia hyalina


1998, Pre-cutting period 1998, Post-cutting period
300 400

300
micrograms-C/litre

micrograms-C/litre
200

200

100
No fish 100 No fish

Fish- medium Fish- medium

0 Fish- high 0 Fish- high


0 1 2 3 0 1 2 3

Nutrient loadings Nutrient loadings

Figure 5.5. Treatment (nutrient and fish) interaction plot for Daphnia spp in the 1998 experiment (pre-
and post-cutting periods, left and right, respectively). Plotted are mean biomass values (in µg-C l-1) across
the nutrient treatments (control: no addition; level 1: 8.3 mgl-1 N and 0.44 mgl-1 P; level 2: 41.5 mgl-1 N
and 2.2 mgl-1 P and level 3: 83.1 mgl-1 N and 4.4 mgl-1 P; all values are in mg-added per litre of enclosure
water). Each bar shows the results for one fish density (control: no fish; level 1: 4 grams m-2 or ~3 fish per
enclosure, and level 2: 20 grams m-2, or ~15 fish per enclosure).

Total rotifer biomass Total rotifer biomass


1998, Pre-cutting period 1998, Post-cutting period
300 300
micrograms-C/litre

micrograms-C/litre

200 200

100 No fish 100 No fish

Fish-medium Fish- medium

0 Fish- high 0 Fish- high


0 1 2 3 0 1 2 3

Nutrient loadings Nutrient loadings

Figure 5.6. Effect of treatments (nutrient loadings and applied fish densities) on total rotifer biomass (in
µg-C l-1) in the pre- and post-cutting periods. Plotted are means (N=3) grouped into the three different fish
treatments (controls, 4 gm-2 of fish (~3 fish per encl.) or ‘medium density’, and 20 gm-2 or ‘high density’
(~15 fish per enclosure). On the x-axis are nutrient loadings (see Table 5.1 for details).

The last 4 weeks of the experiment (i.e. after plant cutting) show a similar pattern of response
to that before the plant cutting, with fish effects being more important in the determination of
the biomass of most zooplankton groups than nutrients (see, for example, Fig. 5.7). Fish
effects were stronger with respect to the pre-cutting period. However, nutrients were more
important during this period in determining many groups’ biomass than they appeared to be in
the first phase of the experiment (see Table 5.9). Especially during the post-cutting period, the
effects of nutrients on zooplankton biomass were apparently dependent on the fish treatment
considered. Generally, increased nutrient loads lead to higher zooplankton biomass up to a
level, above which, additions were reflected in a decrease in biomass of most groups. This
level seemed to be lower in enclosures with fish (see, for example, Fig. 5.5). In addition, this

139
CHAPTER 5

interaction between the effect of fish and nutrients on zooplankton biomass was stronger in
the post-cutting period (e.g. compare, for example, left and right plots in Fig. 5.5). Because of
this, fish effects are not detected when both periods are lumped together (see Table 5.8),
warranting the separation of analyses on the periods before and after plant cutting. These
time-averaged effects are discussed in detail in the appropriate section.

Table 5.6. Significance of main treatments and interactions on the biomass and the percentage of total
biomass of zooplankton groups (see Table 5.5) when all weeks (i.e. before and after the plant cutting) are
lumped together (1998). All are results of ANOVA tests with balanced design except the group ‘Small
Cladocera’, which was analysed using a GLM-ANOVA for unbalanced designs. Significance of block
effects and of the interaction block * treatment (nutrients or fish) are also indicated.

ZOOPLANKTON GROUP All weeks Stability of variance1


µg-C l-1 & % total mass Block N F N*F Block
interactions
AND TREATMENTS
Small Cladocera Biomass n.s. n.s. n.s. n.s. n.s. None
(zeros excluded)
% n.s. n.s. * n.s. n.s. None

Large Cladocera Biomass n.s. n.s. *** n.s. n.s. 2


% n.s. n.s. *** n.s. n.s. 2
Cyclopoid Biomass n.s. * ** n.s. F: **. 2
copepods % n.s. n.s. ** * n.s. 2

Calanoid copepods Biomass n.s. n.s. n.s. ** F: **. 2


% n.s. * * ** F: * 2

Copepod nauplii Biomass n.s. n.s. * n.s. n.s. 2


% n.s. n.s. ** * n.s. 2
Raptorial feeders Biomass n.s. * ** n.s. F: *. 2
(Cladocera) % n.s. n.s. ** n.s. n.s. 2

Open-water Biomass n.s. n.s. n.s. * n.s. Both


filterers % n.s. n.s. ** n.s. n.s. 2

Rotifers n.s. * ** n.s. n.s. Both


Total zooplankton n.s. n.s. n.s. n.s. n.s. Both
1
’n.s’. not significant; *p<0.05; **p<0.01; ***p<0.001. The two criteria for assessing stability of variance
of the data are: 1- Levene’s test of the hypothesis variances across groups are equal. Rejection of this
hypothesis implies the variance is NOT stable, and 2- Means are regressed against variances. Significance
of this relationship is stated, for each group. Fulfillment of these criteria are indicated as 'both’, ‘none’ or
1 or 2, whichever the case.

140
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Table 5.7. Significance of main treatments and interactions on the biomass and relative biomass proportions (in %) of zooplankton groups (for a definition of these
groups see Table 5.5) before and after the cutting of plants (1998). All are results of ANOVA tests with balanced design (i.e. same number of data points for each
column and row) except the group ‘Small Cladocera’, which was analysed using a GLM-ANOVA for unbalanced designs (type III Sum of Squares). Significance of
block effects and of the interaction block * treatment (nutrients or fish) are also indicated.

ZOOPLANKTON GROUP µg-C l-1 & Pre-cutting Post-cutting Stability of variance1


% of total zooplankton mass and Block N F N*F Block Block N F N*F Block PRE- POST-
& & treatment
TREATMENTS treatment
CUTTING CUTTING
Small Cladocera Biomass n.s. n.s. * n.s. n.s. n.s n.s. * n.s. n.s. Both 2
(zeros
excluded)
% * n.s. * n.s. n.s. * n.s. ** n.s. n.s. None 2
Large Cladocera Biomass n.s. n.s. *** n.s. n.s. n.s. n.s *** n.s n.s. Both 2
% n.s. n.s. * n.s. n.s. n.s. n.s. *** n.s. n.s. 2 2
Cyclopoid copepods Biomass n.s. n.s. *** n.s. n.s. n.s. n.s * * N:*; F:* 2 None
% n.s. n.s. ** n.s. n.s. n.s. n.s. ** ** n.s. Both None
Calanoid copepods Biomass n.s. n.s. ** * N: *; F: * n.s. n.s. n.s. * F:* OK OK
% n.s. n.s. ** ** n.s. n.s. n.s. n.s. ** F:* 2 None
Copepod nauplii Biomass n.s. * *** n.s. n.s. n.s n.s. n.s. n.s n.s. None None
% n.s. n.s. ** n.s. n.s. n.s. n.s. n.s. n.s. n.s. Both Both
Raptorials Biomass n.s. n.s. *** n.s. n.s. n.s * * n.s F: * None None
% n.s. n.s. ** n.s. n.s. n.s. n.s. * * n.s. 2 2
Open-water filterers Biomass n.s. n.s. ** n.s. n.s. n.s. n.s. n.s. * n.s. Both Both
% n.s. n.s. ** n.s. n.s. n.s. n.s. * * n.s. 2 2
Rotifers n.s. * *** n.s. n.s. n.s. n.s. ** n.s. n.s. Both 2
Total zooplankton n.s. n.s. ** n.s. n.s. n.s. n.s. n.s * n.s. Both 2
’n.s’. not significant; *p<0.05; **p<0.01; ***p<0.001.
1
The two criteria for assessing stability of variance of the data are: 1- Levene’s test of the hypothesis variances across groups are equal. Rejection of this hypothesis
implies the variance is NOT stable, and 2- Means are regressed against variances. Significance of this relationship is stated, for each group. Fulfillment of these
criteria are indicated as 'both’, ‘none’ or 1 or 2, whichever the case.

141
CHAPTER 5

Table 5.8. ANOVA table for significance of main treatment effects (N: nutrient additions; F: fish densities) and their interactions on species’ biomass (in µg-C l-1) in
the 1998 experiment. For unbalanced designs (i.e. when zeros are excluded), a GLM-ANOVA with type III Sum of Squares was run (see text for details). Some
statistical comparisons cannot be computed because the exclusion of zeros may eliminate from analysis all values of one or more terms of the statistical comparison.
Significance of block effects and of the interaction block * treatment (nutrients or fish) are also indicated.

SPECIES’ BIOMASS Pre-cutting period Post-cutting period Stability of


(µg-C l-1) AND TREATMENTS variance1
Block N F N*F Block interactions Block N F N*F Block PRE- POST-
i.nteractions
Bosmina longirostris n.s. n.s. n.s. Cannot Cannot compute n.s. n.s. n.s. n.s. n.s. Cannot Both
compute
(zeros excluded) compute
Ceriodaphnia spp n.s. n.s. n.s. Cannot Cannot compute n.s. n.s. n.s. n.s. n.s. 2 Both
(zeros excluded) compute
Cyclops spp n.s. n.s. *** n.s. n.s. n.s. n.s. * * N: *; 2 2
F: *
Daphnia spp n.s. n.s. *** n.s. n.s. n.s. n.s. *** * n.s. 2 2
Eudiaptomus gracilis n.s. n.s. ** *** N: ** n.s. n.s. n.s. * F: * Both 2
F: **
Polyphemus pediculus n.s. * * n.s. N: cannot compute; n.s. n.s. n.s. None 2
(zeros excluded) F: n.s.
Total copepods n.s. * ** * n.s. n.s. n.s. n.s. * F: * 2 2

Rotifers n.s. * *** n.s. n.s. n.s. n.s. ** n.s. n.s. Both 2
Total zooplankton n.s. n.s. ** n.s. n.s. n.s. n.s. n.s. * n.s. Both 2

’n.s’. not significant; *p<0.05; **p<0.01; ***p<0.001. 1The two criteria for assessing stability of variance of the data are: 1- Levene’s test of the hypothesis variances
across groups are equal. Rejection of this hypothesis implies the variance is NOT stable, and 2- Means are regressed against variances. Significance of this
relationship is stated, for each group. Fulfillment of these criteria are indicated as 'both’, ‘none’ or 1 or 2, whichever the case.

142
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

5.3.2. The 1999 experiment.

In stark contrast with the previous experiment, few effects of treatments were detected across
the zooplankton groups analysed. Effects of nutrients and fish were only significant on open-
water filterers’ biomass (see Table 5.9 & Fig. 5.7).

Open water filter-feeders


1999
3000
micrograms-C/litre

2000

1000
No fish

Fish- medium

0 Fish- high
0 1 2 3 4 5

Nutrient loadings

Figure 5.7. Biomass (in µg-C l-1) of the group ‘open water filter-feeders’ under the different treatment
combinations of fish and nutrients in the 1999 experiment. Mean biomass (N=2) estimates are plotted
separately for the three fish treatments (no fish, medium and high density of fish) as different shaded
bars. The x-axis gives increasing nutrient loadings.

When all species were grouped (i.e. total zooplankton biomass) only nutrient effects were
statistically significant (Table 5.9). No significant effects of the treatments were detected on
the biomass of zooplankton species taken separately, this year (see Table 5.10).

Bosmina longirostris Bosmina longirostris


1999. Experimental Block One 1999. Experimental Block Two.
1000 700

600
800
cumulative micrograms-C/l

cumulative micrograms-C/l

500

600
400

300
400

Fish- high 200 Fish- high


200
Fish- medium 100 Fish- medium

0 No fish 0 No fish
0 1 2 3 4 5 0 1 2 3 4 5

Nutrient loadings Nutrient loadings

Figure 5.8. Effect of treatments (nutrient loads and applied fish densities) on Bosmina longirostris biomass
µg-C l-1) in the two experimental blocks during the 1999 experiment. The size of shaded sectors at any
given nutrient loading, on the x-axis, shows the differential effect of the three fish treatments (no fish,
medium and high densities) on this species' biomass (in cumulative µg-C l-1). There is considerable
variability across enclosures, though not related directly to block (see ‘block’ effect in Table 5.10).

143
CHAPTER 5

Treatment effects varied significantly across the two experimental blocks (see Tables 5.9-
5.10). Thus, when a completely randomized design (i.e. ignoring block effects) is analysed
with a two-way ANOVA, both nutrient and, especially, fish effects are generally highly
significant. For example, small cladocerans (e.g. Bosmina and Ceriodaphnia), which were the
dominant species during this experiment, were particularly abundant in enclosures with fish
(Figure 5.8). There were considerable differences between the two experimental blocks in this
respect, though. For example, the highest biomass of Bosmina longirostris was reached in the
high fish density enclosures in one block, but in medium-density treatments in the other (see
Fig. 5.8).

The fish treatment increased the percentage of total biomass of both raptorials and open water
filter-feeders (see Table 5.9). Enclosures with high densities of fish also had the highest
biomass proportions of open water filter-feeders (mainly rotifers, but also Bosmina and
Ceriodaphnia). Raptorials, on the contrary, seemed to be favoured by the low density fish
treatment, but were found in proportions similar to those in controls in enclosures with high
fish density (see Figure 5.9).

Open water filter-feeders Rotifer biomass


1999. Percentage of total biomass 1998, Post-cutting period
100 60

90 50
% of total mass

% of total mass

80 40

70 30

60 No fish 20 No fish

50 Fish- medium 10 Fish- medium

40 Fish- high 0 Fish- high


0 1 2 3 4 5 0 1 2 3 4 5

Nutrient loadings Nutrient loadings

Figure 5.9. Effect of treatments (nutrient loads and applied fish densities) on the percentage of total
zooplankton biomass (in µg-C l-1) of open-water filter-feeders and raptorials (Cladocera and Copepoda) in
the 1999 experiment. Fish effects were not very clear in either case (Table 5.9) and the direction of these
effects differs among the two groups.

Overall, significance of treatment effects was considerably muted with respect to the 1998
experiment. When fish effects were detected, generally these were of increasing biomass.
This effect of increasing zooplankton biomass was particularly evident at the highest fish
density (see, for example, Figure 5.10). Some possible reasons for this anomaly are discussed
in next section.

144
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Table 5.9. ANOVA table for significance of main treatment effects (N: nutrient additions; F: fish densities) and their interactions on each groups’ biomass (in µg-C
l-1) and percentage of total zooplankton mass in the 1999 experiment. Significance of block effects and of the interaction block * treatment (nutrients or fish) are
also indicated. Some statistical comparisons and tests cannot be computed because the exclusion of zeros may eliminate from analysis all values of one or more
terms of the statistical comparison.

ZOOPLANKTON GROUP µg-C l-1 & Block N F N*F Block Stability of


% of total zooplankton mass and TREATMENTS interactions variance1
Small Cladocera Biomass n.s. n.s. n.s. n.s. N: ***; F: *** 2

% n.s. n.s. n.s. n.s. N:**; F:*** None


Large Cladocera with zeros Biomass ** n.s. n.s. n.s. n.s. 2

% * n.s. n.s. n.s. n.s. None


without zeros Biomass n.s. n.s. n.s. n.s. n.s. Both
(N=75)
% n.s. n.s. n.s. n.s. Cannot compute Cannot compute
Cyclopoid copepods Biomass n.s. n.s. n.s. n.s. n.s. Both
% n.s. n.s. * n.s. n.s. Both
Calanoid copepods Biomass n.s. n.s. n.s. n.s. F: * Both
% n.s. n.s. n.s. n.s. F:* 2
Copepod nauplii Biomass n.s. n.s. n.s. n.s. n.s. Both
% n.s. n.s. n.s. n.s. n.s. Both
Raptorial feeders (Cladocera + Copepoda) Biomass n.s. n.s. n.s. n.s. n.s. Both
% n.s. n.s. * n.s. n.s. Both
Open water filter-feeders Biomass n.s. * * n.s. n.s. Both
% n.s. n.s. * n.s. n.s. Both
Total zooplankton n.s. * n.s. n.s. n.s. Both
1
’n.s’. not significant; *p<0.05; **p<0.01; ***p<0.001. The two criteria for assessing stability of variance of the data are: 1- Levene’s test of the hypothesis variances
across groups are equal. Rejection of this hypothesis implies the variance is NOT stable, and 2- Means are regressed against variances. Significance of this
relationship is stated, for each group. Fulfillment of these criteria are indicated as 'both’, ‘none’ or 1 or 2, whichever the case.

145
CHAPTER 5

Table 5.10. ANOVA table for significance of main treatment effects (N: nutrient additions; F: fish densities) and their interactions on each species’ biomass (in µg-
C l-1) in the 1999 experiment. A General Linear Model-ANOVA with type III Sum of Squares was run, allowing for the analysis of datasets with missing cells as a
result of the exclusion of zeros. Effects and interactions with insufficient degrees of freedom cannot be computed and are thus indicated.

SPECIES’ BIOMASS Block N F N*F Block Stability of


µg-C l-1 interactions variance1

Bosmina longirostris n.s. n.s. n.s. n.s. N: ***; F: *** 2


Ceriodaphnia spp ** n.s. n.s. n.s. n.s. 2
Daphnia spp with zeros (N=215) ** n.s. n.s. n.s. n.s. Cannot compute
without zeros (N=55) n.s. n.s. n.s. n.s. n.s. None
Scapholeberis mucronata with zeros (N=215) n.s. n.s. n.s. * N: ** None
without zeros (N=71) n.s. n.s. n.s. n.s. n.s. None
Simocephalus vetulus (N=17) cannot n.s. n.s. cannot cannot compute None
compute compute
Eudiaptomus gracilis n.s. n.s. n.s. n.s. F: *. Both
Cyclops spp n.s. n.s. n.s. n.s. n.s. Both
Nauplii n.s. n.s. n.s. n.s. n.s. Both
Total copepods n.s. n.s. n.s. n.s. N: * 2
Rotifers n.s. n.s. n.s. n.s. n.s. Both
’n.s’. not significant; *p<0.05; **p<0.01; ***p<0.001.1The two criteria for assessing stability of variance of the data are: 1- Levene’s test of the hypothesis variances
across groups are equal. Rejection of this hypothesis implies the variance is NOT stable, and 2- Means are regressed against variances. Significance of this
relationship is stated, for each group. Fulfillment of these criteria are indicated as 'both’, ‘none’ or 1 or 2, whichever the case.

146
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Differences between the 1998 and the 1999 experiment.

The main difference between both experiments concerns the degree to which zooplankton
biomass was controlled by fish predation. In the 1998 experiment, strong effects were
apparent, with much larger total zooplankton biomass in enclosures without fish than with
fish (see Fig. 5.10). Effects were strong throughout this experiment, but particularly
pronounced during the post-cutting period. In the 1999 experiment, there were no appreciable
fish effects (see Table 5.9 & Fig. 5.10). Nutrient effects on zooplankton biomass were not
significant in both experiments. The significance of differences between experiments is
discussed below.

Total zooplankton Total zooplankton


1998, Pre-cutting period 1998, Post-cutting period
400 500

400
micrograms-C/litre

micrograms-C/litre

300
300

200
200 No fish No fish

Fish- medium 100 Fish- medium

100 Fish- high 0 Fish- high


0 1 2 3 0 1 2 3

Nutrient loadings Nutrient loadings

Total zooplankton
1999
3000
micrograms-C/litre

2000

1000 No fish

Fish- medium

0 Fish- high
0 1 2 3 4 5

Nutrient loadings

Figure 5.10. Total zooplankton biomass (in µg-C l-1) and treatments (nutrient loads and applied fish
densities) in both experiments (top left: 1998, pre-cutting period; top right: 1999, post-cutting period;
bottom: 1999). Fish effects are apparent in the 1998 experiment but are not so clear in the 1999
experiment.

Clear top-down effects of fish on algal density, as measured by chlorophyll-a, were apparent
in the 1998 experiment. By contrast, in the 1999 experiment there were no effects of fish on
chlorophyll-a (see Fig. 5.11, bottom). In the 1998 experiment, fish effects were particularly
clear in the post-cutting period (see Fig. 5.10, top). Nutrient effects on chlorophyll-a were not

147
CHAPTER 5

statistically significant (marginally significant in the post-cutting period; see Table 5.12),
although there was a trend for increasing algal biomass with higher nutrient loadings. This
trend seemed to be interrupted at the highest nutrient loading in both periods. This truncated
trend or ‘saturation effect’ was more evident in the post-cutting period. Possible causes for
this are discussed below.

Chlorophyll-a Chlorophyll-a
Pre-cutting period, 1998 Post-cutting, 1998
120 200

100
150
micrograms/litre

micrograms/litre
80

60 100

40
No fish 50 No fish

20
Fish- medium Fish- medium

0 Fish- high 0 Fish- high


No load 1 2 3 No load 1 2 3

Nutrient loadings Nutrient loadings

Figure 5.11. Response of the phytoplankton (i.e. chlorophyll-a, in µgl ) to treatments (nutrient loads and -1

applied fish densities) in the 1998 experiment. Plotted are means of N=3 enclosures, left: before plant
cutting and right: after plant-cutting, for each fish treatment (no fish controls, 4 gm-2 of fish and 20 gm-2).
The x-axis gives nutrient loadings (see Table 5.1 for details).

Chlorophyll-a
1999
400

300
micrograms/litre

200

100 No fish

Fish- medium

0 Fish- high
No load 1 2 3 4 5

Nutrient loadings

Figure 5.12. Response of the phytoplankton (i.e. chlorophyll-a, in µgl-1) to treatments (nutrient loads and
applied fish densities) in the 1999 experiment. Plotted are means of N=2 enclosures for each fish treatment
(no fish controls, 4 gm-2 of fish and 20 gm-2). The x-axis gives nutrient loadings (see Table 5.2 for details).

148
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

5.4. Discussion.

These experiments, with all their limitations (see Table 5.11), have been frequently used to
assess the potential for biomanipulation, i.e. the removal/reduction of predation pressure on
grazer populations with an aim to increasing grazing on phytoplankton. (see, for example,
Lampert 1988; the term ‘biomanipulation’, however, was originally coined with a much wider
meaning, see Shapiro 1990). The main conclusion that can be drawn from experiments here,
is that fish strongly regulate the maximum biomass of large-bodied grazers (Cladocera) in
Little Mere (1998 experiment, pre-cutting F=125.2, p<0.001; post-cutting F=459.9, p<0.001).
Moreover, zooplankton populations cannot survive in the face of fish predation pressure in
excess of about 4 g m-2 (see, for example, Fig. 5.10), which is equivalent to about 2 or 3 small
(1 gram) sticklebacks, at least within enclosures in our experiments (see Discussion below).

In addition, the effects of predation on grazer populations translated immediately into larger
algal biomass (see Table 5.12). More generally, this has obvious implications on the idea
algal biomass can be controlled through ‘biomanipulation’. Results from experiments
conducted in Little Mere in growing seasons ’98 & ‘99 will be discussed in favour and against
biomanipulation as a restoration tool in shallow lakes in general.

Data and theory should be assessed together (see Lehman 1985), and mesocosms experiments
conducted in Little Mere and reported here provided an opportunity to compare the dynamics
of zooplankton in enclosures and in the natural situation (i.e. in the lake), under the treatments
applied (fish densities, and nutrient loadings).

Experiments with enclosures are vulnerable to the cumulative effects of treatments resulting
in effects amplified by time and application of treatments, rather than direct effects of the
experimental manipulation (see Table 5.11 for details). On the other hand, high nutrient
concentrations achieved in enclosures give a unique opportunity to look at the interaction of
nutrient limitation (e.g. Urabe & Watanabe 1992) and fish predation on zooplankton. The
dynamics of this interaction is discussed in detail, below.

Plant beds are thought to act as a ‘refuge’ for zooplankton against fish predation (e.g. Leah et
al. 1980). Open-sediment mesocosm allow plant growth to develop, and cutting of plants mid-
way through the experiment was aimed at testing the effect of submerged plant growth
removal on zooplankton and water transparency, again under the treatments applied.

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CHAPTER 5

Unfortunately, few enclosures developed significant plant biomass. On the other hand, this
apparently random behaviour of enclosures as regards plant growth (see, for example, Fig.
5.14) provides an opportunity to compare the response of enclosures with and without plants,
before and after the perturbation constituted by the plant removal.

Table 5.11. Pros and intrinsic shortcomings of open-sediment mesocosms. See text for details.

ADVANTAGES SHORTCOMINGS
1) They allow the isolation of effects of controlled 1) Problem of representativeness (see Frost et al. 1988)
experimental treatments (see Kitchell & Carpenter Shallow bottom lakes are often spatially very
1993) heterogeneous…
2) Small size permits easy replication which can be 2) Problem of extrapolation.
used, with appropriate statistical techniques, to tease Are mesocosms a good ‘mimic’ of lakes, or are
out spurious effects (e.g. sampling error and initial important factors left out of designs?.
conditions differences).

3) Within the context of their appropriate aims (see 3) Problem of generality.


Introduction for details), they are a powerful method to Shallow lakes are very heterogeneous and varied
obtain reliable answers to some specific research environments. Results from one lake are often not
problems. applicable to other shallow lakes. Both data and theory
must be assessed together. To be aware of particular
features affecting experimental results should be a
prerrogative of any attempt to apply results to other
lakes.
4) Algae sedimentation rates may be significantly
greater inside mesocosms than in more natural
conditions, where turbulence and waves continually
resuspend cells.
5) Artificially high zooplankton densities and
unnaturally high grazing rates (e.g. Faafeng et al.
1990). Therefore top-down effects of herbivores are,
potentially, overestimated.
5) Seasonality of particular species population
dynamics is often not in phase with that of the lake.
7) Higher water temperatures with respect to lake and
altered light climate, especially further down the water
column.
8) Magnified effect of periphytic uptake of nutrients.
Inner enclosure walls provide a large colonizing
surface for periphyton.
9) Variable sediment ephippial recruitment and
“pioneer colonizer” effects. A water column and a
possibly atypical patch of lake bottom are isolated by
each enclosure. Plankton communities in enclosures,
both phytoplanktonic and zooplanktonic, may be
atypical of the lake.
10) Limited to one or two different fish species.
Higher-order trophic interactions and competition
between species, as those existing in nature, cannot be
easily reproduced in an in situ mesocosms approach.

The comparison between the two years’ experiments has provided some information on the
differential response to treatments of very different zooplankton communities. Some of the

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

potential causes of these two very different outcomes as regards zooplankton community
composition in different years are discussed.

Finally, lessons learnt from these experiments can be used to frame an understanding of algal
biomass regulation in Little Mere, and the maximum levels of nutrient load and fish density
that can be expected to support this regulation in the more complex environment of the lake.

Top-down control of zooplankton by fish predation in Little Mere.

The effect of fish predation on zooplankton biomass was clearly stronger in the 1998
experiment than in the 1999 experiment (see Fig. 5.10, top plots), as shown by the generally
lower biomass at high fish density. The composition of the zooplankton community in both
years probably had a strong influence on this outcome. During the 1999 experiment, over 90
% of the total zooplankton biomass was rotifer biomass. Rotifers are generally small species,
probably not under the same predation pressure cladocerans are generally exposed to, under
the assumption fish forage preferentially, or better, on larger zooplankton (Brooks & Dodson
1955; Lynch 1979; Kerfoot & DeAngelis 1989). Indeed no fish predation effect was found on
rotifer biomass in the 1998 experiment. In this experiment, populations of Daphnia spp
developed in all enclosures in the week before fish were added (the ‘pre-treatment week’; see
Fig. 5.13). As fish predation pressure was introduced, differences in Daphnia biomass became
evident between enclosures with fish and without (Fig. 5.13). This had practically immediate
consequences on phytoplankton biomass, as measured by chlorophyll-a, and enclosures with
fewer Daphnia, i.e. those with fish, had significantly more chlorophyll-a (see Table 5.12).
This suggests grazing by this species has indeed a strong control over algal biomass in Little
Mere. Results reported in Chapter 4 concerning algal biomass (as measured by chlorophyll-a)
and Daphnia grazing across areas in Little Mere are consistent with this suggestion.

The coupling between phytoplankton and zooplankton is not always as close as in Little
Mere. Negative feedback effects may dampen the interaction considerably in some lakes (e.g.
Cryer et al. 1985; see McQueen 1990). For instance, increased grazing pressure may reduce
the fast nutrient cycles (i.e. sedimentation of algae cells, phosphorus release from sediments
as dead algae cells promote reducing conditions at the microzone; see, for example, Phillips et
al. 1994). Grazing will also increase light penetration and potentially algal growth, and
grazers may become food-limited in crowded conditions (e.g. Urabe & Watanabe 1992). The
short spans of our experiments (7-9 weeks; see Table 5.3), though, perhaps precluded the

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CHAPTER 5

possibility of these feedback mechanisms operating, or being strong, in enclosures. Faafeng et


al. (1990), for instance, found in bag experiments in the pelagial zone, that grazing was more
important during summer than in spring or autumn, and attributed this to seasonal changes in
the proportion of edible algae in the phytoplankton community. Moss et al. (1991) also
propose phytoplankton seasonal succession should be considered when assessing the potential
for successful biomanipulation.

In the experiments reported here the influence of changes in the phytoplankton assemblage
over zooplankton production cannot be adequately assessed because the effects of large algal
biomass (see Discussion below) are confounded with high cyanophyte densities in enclosures,
particularly during the 1999 experiment. Nevertheless, few cyanophytes developed in the
1998 experiment (an occasional peak of 17 % of total numbers in week 4 in one enclosure,
but generally less than 7 %) and, thus, cannot be responsible for the changes in secondary
production occurring over time. In the 1999 experiment, zooplankton abundances, mainly of
small cladocerans (see Table 5.6), increased irrespective of treatment and it is unlikely the
phytoplankton community composition had any effect on zooplankton abundance. Overall,
seasonal changes and the fact experiments are short-term, are not thought to have affected the
validity of conclusions. Indeed, field observations support the contention that top-down
control of zooplankton in Little Mere is strong, at least under the nutrient concentrations that
were found in the lake’s waters.

Table 5.12. Summary of ANOVA results on treatment effects (applied fish densities and nutrient loadings)
on chlorophyll-a concentration (µg l-1) in the pre- and post-cutting periods of the 1998 experiment.

Chlorophyll-a vs. PRE-CUTTING PERIOD POST-CUTTING PERIOD


Treatments SS MS F SS MS F
Block 0.529 0.255 1.408 0.153 0.077 0.585
FISH 4.209 2.104 15.401* 9.297 4.548 18.04*
NUTRIENT 0.895 0.299 1.301 0.820 0.273 4.14+
FISH*NUTRIENT 1.003 0.157 0.938 0.845 0.141 0.555
+
0.07<p<0.05; *p<0.05

The interaction of nutrient and fish effects, or the relative contribution of top-down and
bottom-up control of zooplankton and phytoplankton in Little Mere.

The effect of nutrients on zooplankton biomass, and particularly on Daphnia biomass, during
the 1998 experiment seemed to be modulated by fish density. The overall pattern of
dependency between Daphnia spp biomass (in µg-C l-1) and treatments (i.e. nutrient additions
and different fish applied densities) in the 1998 experiment is summarized in Figure 5.13.

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Following, the effects of nutrients on Daphnia biomass (experiment 1998) are separately
examined at the three levels of fish density (no fish, medium and high density). Potential
nutrient effects in the 1999 experiment will be examined when discussing the differences in
zooplankton community structure between the two experiments.

• In the absence of fish, nutrient levels apparently increased the ceiling of maximum
zooplankton production as the experiment progressed (Figure 5.13), although this effect
was not statistically significant (see Table 5.7). However, above ‘level 2’ nutrient loadings
(see Table 5.2 for details), biomass reached a maximum and further additions had little
effect on both the phytoplankton, as measured by chlorophyll-a readings (see Fig. 5.11),
and Daphnia biomass. Moreover, further additions may have had deleterious effects on
zooplankton. Three mechanisms can be thought of, accounting for this levelling off and
posterior decline:

a) Lower assimilation efficiency at high nutrient concentrations. Nitrogen and phosphorus


may become limiting to zooplankton growth as N:C and P:C ratios in the algae cells
decrease as carbon availability increases (i.e. at higher nutrient loadings). Although
zooplankton has considerable homeostasis capacities, ratios in body tissues also become
lower at higher nutrient concentrations (Sterner 1990; Urabe & Watanabe 1992) and this
is suggested to be caused by lower assimilation efficiencies at high food concentrations
(Urabe & Watanabe 1991). This could explain the maximum reached at high loadings in
fishless enclosures. No direct evidence of a decrease in C/P ratios is available to directly
test this idea. In any case, declines in Daphnia production occurring in some fishless
enclosures above level 2 loadings (see Fig. 5.13) cannot be explained by limiting factors.
Deleterious effects must be invoked.

b) Deleterious effects on Daphnia, either limiting survival and/or preventing growth. The
decline of Daphnia populations over time parallels that of the populations of this species
in the lake itself. What differs is the rapidity of this decline, which seems to depend on the
nutrient treatment. This could be related to the grazing capacity of the zooplankton
populations. The highest nutrient loads in the 1998 experiment were probably unrealistic,
promoting levels of algal biomass that could have interferred with Daphnia’s filtering
machinery. This effect, though, is not related to filamentous algae as these were generally
scarce during experiments, but to high green algae densities. Lower nutrient loads were

153
CHAPTER 5

tested in the 1999 experiment, also examining a more detailed range of nutrient
applications (see Table 5.2). Unfortunately, this effort was obscured by initial conditions
(compare Figs. 5.11 & 5.12), of high algal densities. ‘Baseline conditions’ in mesocosm
experiments may be of paramount importance. Once the system is isolated from the rest of
the lake, its capacity for change may be linked, and treatment effects may be swamped if
the system is already heavily eutrophicated.

c) Self-shading of algae. The algae community may depend on the nutrient regime (Sommer
1989). Differences in the phytoplankton assemblage and the different chlorophyll-a
content of algal species (Reynolds 1983) may explain some of the chlorophyll-a variation,
but it is hard to believe that the very large differences can be accounted for by only
species composition of the algae community. A likely possibility is that nutrients were not
limiting in many enclosures with the highest nutrient load, exceeding the growth capacity
of the algae. Thus, comparable algal biomass (i.e. chlorophyll-a) was estimated in the high
nutrient load enclosures, irrespective of fish and grazer effects.

Analogous experiments conducted further south in the latitudinal gradient (see section 5.3 for
details) did not report such effects. The southernmost experiment, in Valencia, Spanish central
mediterranean coast, is a case in point. There, even the highest nutrient additions reflected in
higher chlorophyll-a values. This could happen under the higher light levels of the more
southern latitudes. Furthermore, in León, northwest Spain, water temperatures during the
experiment were comparable to those in England. However, no "saturation effect" was
apparent either. Apparently, light may have been the limiting factor at the very high nutrient
loadings in our experiment (see Carpenter et al. 1997).

On the other hand, ‘time-effects’ may have been important. As nutrient loads were
progressively incorporated into the system, lower and lower additions were capable of
decreasing Daphnia biomass (see Figure 5.7). A possible mechanism for these deleterious
effects could be the mechanical interference of the feeding apparati of animals, particularly at
the algal densities promoted by the highest nutrient loadings (Burns 1958; Porter 1973;
Gliwicz & Siedlar 1980; Gliwicz 1990). Significant herbivory effects of Cladocera are, thus,
only expected when grazer populations are able to reach a certain threshold biomass. Below
this, phytoplankton is not grazed down fast enough to compensate for algal growth. Turbidity
due to the algae, then, gradually increases.

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NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

-1
Daphnia biomass (µg-C
Daphnia spp. ug-C/litre
l )
500

Control
No fish (no fish)
Fish-medium density
400
Level 1
Fish- high density
Level 2
Daphnia ug-C/litre

300

200

100

0
Control N2 Control N2 Control N2 Control N2 Control N2 Control N2 Control N2 Control N2 Control N2
N1 N3 N1 N3 N1 N3 N1 N3 N1 N3 N1 N3 N1 N3 N1 N3 N1 N3
Pre-treatment Week 1 2 3 4 5 6 7 8

Figure 5.13. Daphnia spp mean biomass (in µg-C l-1; N=3) across weeks in all treatment combinations.
Different lines are of biomass in the three fish density treatments (no fish, 4gm-2 and 20 gm-2). Each line
represents mean Daphnia biomass across the nutrient levels (control, N1, N2 and N3; see Table 5.2 for
details).

Cyanobacteria are not good food for daphnids. Laboratory growth experiments by De Mott
(1998) have conclusively shown reduced growth and reproduction of 5 daphnid species when
fed on pure diets of the cyanobacterium Synechococcus elongatus. The issue is of import, as
attempts at controlling algae populations through biomanipulation fail if zooplankton cannot
utilize cyanobacteria as food (see De Bernardi & Giussani 1990). Toxic effects of these algae
may be overcome, though, as various authors have documented top-down effects of filter-
feeders on blue-green algae (e.g. De Bernardi et al. 1992). Gliwicz (1990) suggested grazer
control of blue-green algae would be limited to those instances when a large grazing potential
(i.e. large zooplankton biomass) coexisted with small populations of blue-green algae species.

Results from the 1998 experiment suggest large phytoplankton densities (as measured by
chlorophyll-a) occurring in high loading enclosures are sufficient to inhibit feeding and
growth of Daphnia (point b., above). Toxic effects are difficult to argue, as most enclosures
had few cyanobacteria. Thus, direct mechanical interference seems the most likely mechanism
(see Burns 1958; Gliwicz & Siedlar 1980). The occurrence of colonial forms of algal species

155
CHAPTER 5

at higher nutrient concentrations (Holtmann & Hegewald 1985) could enhance this effect.
Recently, colony formation in Scenedesmus, a green algae species, has been related to grazer
presence, cued by infochemicals exuded by the grazers themselves (Lurling & Van Donk
1997), giving support to the idea that mechanical interference may indeed offer some degree
of protection to the algal cell from herbivory.

• In the presence of relatively low fish densities (i.e. 4 grams of fish per m2). More nutrient
reflected in larger Daphnia biomass. This effect, though, was slighter than in fish controls.
Furthermore, at the greater nutrient loads, Daphnia populations died out sooner and never
reached the biomass seen in fishless enclosures. Fish predation on zooplankton resulted in
drastic swings in population size across weeks. At the lower nutrient loads, populations
seemed to be more stable across weeks (see Fig. 5.13), even in the face of comparable fish
predation pressures, although large error bars across enclosures with same treatment
combinations do not allow refinement of this tentative conclusion.

This result can be interpreted as lending support to the idea it is the grazing potential (as given
by zooplankton biomass; see Lampert 1988) which sets a limit to the potentially deleterious
effects of phytoplankton on the grazers (Gliwicz 1990). Thus, in the presence of fish, the
ensuing smaller Daphnia populations would not be able to keep algal populations in check,
and at high nutrient loads, phytoplankton would be so dense as to seriously interfere with
feeding mechanisms. This is also consistent with the cumulative effect of nutrient loadings in
progressively lowering the threshold addition capable of decreasing Daphnia biomass.
However, it is also likely Daphnia populations were slowly reduced by fish predation
pressure along weeks. This would suggest the balance of fish predation and Daphnia biomass
is struck somewhere below the 4 g m-2 threshold (i.e. equivalent to about 3 one-gram fish per
enclosure), rather than above it.

Benndorf (1988) emphasizes the necessity of reaching an optimum zooplanktivorous fish


density, in order to maximize the ‘filtration capacity’ of herbivorous zooplankton. Lampert
(1988) calculated the biomass of zooplankton necessary to attain a given algal biomass
reduction, but gave no suggestion as to what fish pressure would allow such a grazing
capacity to be attained. Indeed lakes may differ with respect to zooplankton community
structure, seasonality of this, and other biotic factors (e.g. mysid invertebrate predators on
zooplankton, in Benndorf et al. 1990) and generalizations are therefore difficult to reach. In

156
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

addition, plant beds are thought to act as refuge for zooplankton in the face of fish predation
(Leah et al. 1980; Timms & Moss 1984). As far as I am aware, the most detailed study of the
interaction fish-zooplankton-phytoplankton in relation to plant refuge availability is that of
Schriver and colleagues (1995). In their large-scale enclosure experiment in Lake Stigsholm
(Denmark), a very shallow lake (mean depth 0.8 m), they estimated ~2 fry m-2 to be the
maximum fish density that large-bodied cladoceran populations could withstand, as long as
the percentage volume infested of aquatic vegetation (PVI) exceeded 15-20 %. This figure of
2 fry m-2 approximates well that found in Little Mere, particularly in the 1998 experiment, of
close to, but less than, 4 gm-2 (approx. 3 fry m-2). Perhaps had our experiment continued,
predation effects on Daphnia would have become more obvious, even at the medium fish
treatment (i.e. 4 g m-2).

• At the highest fish density (i.e. 20 g m-2). Daphnia populations were only briefly sustained
at the lowest nutrient treatments (see pre-treatment week and weeks 1 & 2 in Fig. 5.13). It
could be that the nutrient level at which grazer populations disappear is dependent on the
presence of fish and, perhaps, on fish density. Again, the smaller grazer populations that
can survive in the face of low levels of fish predation are those capable of keeping in
check the algae community. This can only happen, perhaps, when the phytoplankton
reproductive capacity, as given by nutrient loads, is below a threshold (Gliwicz 1990).

Overall, the interaction between nutrient loadings and the fish treatment on Daphnia
production (µg-C l-1) was not statistically significant (F-test, p>0.05). However, visual trends
in Daphnia production in relation to treatments and its changes (see, for example, Fig. 5.13)
are easily interpreted in terms of the grazing capacity of Daphnia populations in relation to
fish predation pressure and the ‘grazability’ of phytoplankton (given by its density), as
discussed above.
Thus, algal biomass in the 1998 experiment was mainly regulated by Daphnia herbivory.
Nutrients had only a small, modulating, effect on zooplankton biomass during the 1998
experiment. However, an understanding of population biomass dynamics cannot be reached if
nutrient concentrations and algal densities experienced by these populations, are not taken
into account. No apparent effect of nutrient loadings was observed in the 1999 experiment.
Differences between the zooplankton community structure in both experiments are discussed
below.

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CHAPTER 5

Plant refugia and fish predation on zooplankton in Little Mere.

The refuge role of plant beds (see Leah et al. 1980; Timms & Moss 1984) might vary with
plant cover (Lauridsen & Buenk 1995; Lauridsen et al. 1995), plant density (Schriver et al.
1995), plant types dominating the lake (i.e. floating-leaved, stemmed emergent, totally
submerged; Carpenter et al. 1997; Scheffer 1998), or bathymetry, although these variations in
refuge role have not, so far, been specifically addressed in an experimental way.

Few plants were present in enclosures in both experiments (i.e. generally less than 1g m-2DW
in the 1998 experiment; see Fig. 5.14, too). The threshold value of 3 fry m-2, suggested as a
threshold-maximum fish density that can sustain large Daphnia populations, could be higher
in the lake, where refugia are available. There is strong evidence from field observations in
Little Mere that lily beds do act as refugia against predation, mainly by perch (Perca
fluviatilis) (see Chapter 3 for details).

Plant biomass (gm-2) and chlorophyll-a (micrograms l-1) in the


three fish treatments (1999 experiment)
10 800
No fish treatment
9 3 fry per enclosure (4gm-2) 700
8 16 fry per enclosure (20gm-2)
Plant biomass (gDWm-2)

chlorophyll-a 600

micrograms l-1 chl-a


7
6 500

5 400
4 300
3
200
2
1 100

0 0

Figure 5.14. Plant biomass (gm-2DW) and chlorophyll-a (µg l-1) in the 36 enclosures, measured in the last
week of the 1999 experiment (Aug. 13th). Enclosures have been grouped into the three fish treatments (no
fish, medium density and high density) and are arranged so from left to right on the x-axis. Bars
indicating plant biomass are shaded accordingly.

No refuge effect, however, was apparent in enclosures in both experiments. In the 1998
experiment hardly any plants grew at all, with yields larger than 1 g m-2DW in only two
enclosures. In the 1999 experiment, plant biomass was generally larger (see Fig. 5.14).
However, few grazer effects were apparent anyway, regardless of plant biomass in the
enclosures, as discussed above, and thus few refuge effects can be expected. Interestingly,
though, the relationship between plant biomass and chlorophyll-a was statistically significant
this year (r2=0.12, p<0.05), albeit with little explanatory value. The apparent effect of plants

158
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

on phytoplankton was especially clear in the enclosures with relatively dense macrophyte
growth, and these had sensibly less chlorophyll-a, irrespective of whether the enclosure
contained fish or not (see different-shaded bars in Fig. 5.14).

Macrophytes may have some relevance in the regulation of algal biomass in this experiment,
although it is unlikely the mechanism was through the harbouring of the small cladoceran
grazers in the presence of fish. More likely, plant competition for nutrients may have
contributed to limiting growth of phytoplankton (see, for example, Howard-Williams 1981).
Allelopathy cannot be discarded as an hypothesis, either. Both these effects, nutrient
competition and allelopathy, are confounded and specific experiments testing different levels
of plant biomass and different species with known allelopathic activity (e.g. Chara; see
Wium-Andersen et al. 1982; but see Forsberg et al. 1990) would be needed to separate their
relative influence on phytoplankton biomass.

Differences in zooplankton community structure between the two experiments.

In the 1998 experiment, the highest rotifer biomass was found in the high fish density
treatment, where few Daphnia could be found. This could be interpreted as Daphnia
generating unfavourable conditions for the development of large rotifer populations, perhaps
competing for resources (Lynch 1979; Vanni 1985, 1987). This suggestion is consistent with
results from the 1999 experiment, when generally high rotifer biomass were found coinciding
with few or no Daphnia. Again this could be interpreted as small herbivores (Ceriodaphnia,
Bosmina) being normally outcompeted by the more efficient filter-feeder Daphnia (Dodson
1974; Lynch 1978, 1979). Low interspecific competition following elimination of the large
Daphnia would allow the smaller-bodied Cladocera to develop (Vanni 1985).

The question remains, though, why did Daphnia not develop in the 1999 experiment as it did
in the 1998 experiment. No block or treatment effects were appreciable at the outset of both
experiments (see Table 5.4). However, the second experiment was initiated when the lake
phytoplankton community measured up to 80 µg l-1 of chlorophyll-a, about four times as
much as that at the outset of the 1998 experiment (compare Figs. 5.11 & 5.12). Lake
phytoplankton densities sharply decreased soon after the 1999 experiment was set up.
However, dense algae communities were sustained within the enclosures. Nutrient additions
in subsequent treatment weeks added to this abnormally-high-for-the-lake algal density.

159
CHAPTER 5

Fish effects, for example on small cladocerans such as Bosmina longirostris, were very
variable and significantly different in both blocks (see Table 5.10 and Fig. 5.8). Block effects
may have obscured main treatment effects, but the general trend was for enclosures to behave
‘independently’ of applied treatments. One reason for these anomalies could have been a
higher fish mortality induced by the dense algal populations developing in many enclosures,
sometimes with considerable densities of cyanobacteria, as discussed in detail above. Large
Cladocera populations (e.g. Daphnia spp) only started developing late in the experiment, in
some enclosures and not consistently across treatment combinations. Indeed, the zooplankton
community developing in the 1999 experiment was a reflection of that developing in the lake
itself. Large-bodied zooplankton in the lake was probably subject to intense predation. In
addition, this may have freed up food resources for the smaller cladocerans as a consequence
of the bigger mortality inflicted by visually-predating fish on the larger and thus more
vulnerable (Brooks & Dodson 1955; Hall et al. 1975), though superior resource-competitor
Daphnia (Vanni 1985, 1987).

Although considerable densities of small cladocerans developed in enclosures (e.g. 7000


Bosmina and 3000 Ceriodaphnia per litre in some enclosures) no strong effects of grazing on
algae were apparent as measured by decreases in chlorophyll-a (small cladoceran
log(biomass) vs. log(chlorophyll-a) regression: r2 = 0.30; p<0.001). Weak top-down effects of
small Cladocera have been observed in numerous occasions, particularly in lowland temperate
lakes of low latitudes but also in more northern lakes (e.g. Schoenberg & Carlson 1984;
Crisman & Beaver 1990).

Biomanipulation in shallow lakes: what do these experiments tell us?.

From a management perspective, results from these experiments suggest maintenance of


water clarity in an environment effectively devoid of aquatic plant cover by herbivory of
zooplankton in the presence of fish is only feasible within a range of nutrient loads and under
a relatively low fish pressure. Only large cladocerans seem to be capable of exerting any
control on phytoplankton populations, under the set of high nutrient regimes imposed in these
experiments.
With regard to biomanipulation as an effective restoration technique in shallow lakes, two
restrictions can be made, supported by the data reported here and extensively covered by
recent scientific literature. The first is that nutrient concentrations at the time of
biomanipulation are important (Lammens et al. 1990 and references therein; Benndorf 1990;

160
NUTRIENTS, FISH AND ZOOPLANKTON DYNAMICS

Jeppesen et al. 1991; Perrow et al. 1994; DeBernardi & Giussani 1995; Carpenter et al. 1997;
Kairesalo et al. 1999). Indeed the highest loading treatment in the 1998 experiment supported
a relatively low zooplankton production in the presence of fish. The second is that populations
of large cladocerans can only coexist with fish under relatively low predation regimes (e.g.
Brooks & Dodson 1955; Halls et al. 1975; Lynch 1979; Vanni 1987, etc.), at least in the
conditions provided by mesocosms. Sustainable herbivory could perhaps be maintained below
a maximum fish biomass of 4 g m-2 (fresh weight).

161
CHAPTER 6

Summarizing discussion
CHAPTER 6

The resilience of the plant-dominated state (see CHAPTER 1) is considerable, and much research
effort has gone into elucidating the specific mechanisms contributing to the maintenance of clear
water. Comparatively little is known, however, about the abundance of plant-associated grazers,
although their potential role as important buffers of the plant-dominated state has been
hypothesized for long. In addition, few investigations into the role of grazing have taken into
account, or even acknowledged, the implications of aggregation of grazers, either into ‘clumps’
(i.e. horizontal distribution) or ‘layers’ (i.e. vertical migration).

Seasonal cycles of both chydorids and other plant-associated cladocerans (e.g. Simocephalus and
Sida) as well as important horizontal differences were identified in this study (CHAPTER 3).
Information on features of species' populations, such as size-structure, and clearance rate
(CHAPTER 4), in turn strongly dependent on animal body size, complement data provided in
CHAPTER 3 on animal densities. Grazing rates (in % lake volume filtered per day) were then

estimated (CHAPTER 5), providing insight into the trophic role of different cladoceran species and
communities.

Daphnia was generally the main grazer in Little Mere. However, water clarity may be maintained
in its absence in this lake. A very large biomass of plant-associated filter-feeders was also found,
at the beginning and end of the two growing seasons sampled (1998 & 1999), always coinciding
with low chlorophyll-a. High localized grazing rates were estimated for these species during
these periods. On the other hand, the fit of regressions between Daphnia and chlorophyll-a was
often poor, albeit inverse correlations were apparent from visual inspection of data (CHAPTER 4).
Lags could not always explain this poor fit.

Studies with a similar approach to that used in this study have reached very different conclusions
as regards the importance of herbivory in their systems. However, this highlights the usefulness
of grazing rates as measures of trophic role of particular grazer components in a lake.

The role of Daphnia in the control of algal crops was also investigated in relation to fish
predation pressure on populations and the potentially modulating effects of nutrient regimes
using data from two field mesocosm experiments conducted in two separate summers (1998-

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SUMMARIZING DISCUSSION

2000) (CHAPTER 5). Top-down effects were apparent in the 1998 experiment, stronger than
effects of nutrients on algal biomass (i.e. chl-a concentration).

There is much scope for investigation of the relative contribution to water transparency of plant-
bed processes, for example through parallel experiments isolating the effect of each mechanism
as much as possible, within the same lake. The grazing on phytoplankton of plant-associated
Cladocera, for one, is a clearly important buffer of the plant-dominated state (see CHAPTER 4), but
also grazing of periphyton by chydorids and other scrapers in Little Mere may have important
feedback links.

Nutrients captured in periphyton and then scraped from surfaces by these species will reach the
sediment with their death. In addition, plant growth may be favoured by the cleaning of plant
surfaces. Thus, chydorids may be an important link that helps sustain plant beds in the face of
increasing nutrient loads and in-lake concentrations, and therefore perpetuate the plant-dominated
state (see CHAPTER 1 for details). Very scarce reliable data are presently available about the
ingestion rates of chydorids (e.g. Alona), estimation of periphyton is difficult and extremely
variable across short distances. Animal abundances can be considerable (i.e. 500-5000 Chydorus
m-2). Despite their generally small size (< 1 mm, but frequently less than 0.7 mm) and probably
low ingestion rates, they are worth investigating in as much as they may be important buffers of
the clear-water state in lakes where plants are widespread and abundant. Predictions have never,
as far as I am aware, been made on the potential trophic role in plant beds and in shallow lakes,
of these periphyton scrapers.

Shallow lake management will greatly benefit from understanding what processes are important
in the maintenance of clear water. This study has shown another mechanism by which plant beds
can indirectly perpetuate the conditions under which they may continue to grow (i.e. mainly high
water transparency), through grazing on phytoplankton by harboured filter-feeders. Periphyton
grazing by chydorids, and other scrapers (e.g. snails) has not been approached at an ecosystem
level and in a quantitative way yet, and their abundance and distribution suggest this line of
research may be worth pursuing in the future.

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CHAPTER 6

On the other hand, results from this study may help to understand better some cases in which
biomanipulation in shallow lakes has not been successful. Plant-associated grazers may
contribute to the sustenance of plant growth and thereby to the buffering of the plant-dominated
state in the face of eutrophication, at least during limited periods of the growing season (April to
October). This highlights, again, the importance of taking into account plant beds and their
structuring role when designing a restoration strategy for a given shallow lake. Plant beds may
act as refugia for zooplankton against fish predation and indeed the potential of providing the
refugia artificially has been explored in the past, albeit with mixed success. However, other
processes mediated by the living surfaces provided by aquatic plants, and the physico-chemical
conditions within natural beds, are probably very important. This study has shown that grazers
making their habitat in aquatic plant beds can be a by no means negligible buffer mechanism of
the plant-dominated state of shallow lakes.

166

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