1

Physical, Biological, and Chemical Indicators of Soil Health and Microorganism

Presence in a Fallow Field versus a Sod Field in Cambridge, Ontario
Maria Legault (ID #20266913)*
November 30, 2009
ENVS 200; Sarah Ashpole

*Department of Environment and Resource Studies, University of Waterloo, email:
mklegaul@uwaterloo.ca

Abstract
Soil health is a critical issue for future environmental sustainability, but soil
degradation can occur on sod farms from excessive tillage and fertilizer applications
(Perez et al., 1995). This degradation is often caused by the loss of important
microorganism populations, which are negatively affected by alterations to the physical,
biological, and chemical components of the soil. In this study, these alterations were
determined through qualitative and quantitative data gathering during two field studies
and one chemical analysis of collected soil samples. Physically, alterations to the soil
horizons and vegetation communities in the sod field could reduce microorganism
diversity and activity levels. Biologically, the sod field has sufficient earthworm
(Lumbricus terrestris) populations to contribute to healthy microorganism functioning,
but the number of rocks in this field suggests that microorganism populations are limited
by poor soil quality. Chemically, regular fertilizer applications in the sod field could limit
microorganism diversity and environmental safety. The results of this study suggest
that the high level of management on sod farms is causing soil degradation and that
less intensive management efforts are needed.
Key words: soil health, soil degradation, tillage, microorganisms, earthworms
2



Table of Contents
1. Introduction ............................................................................................................... 3
2. Objective and Methodology ..................................................................................... 4
2.1 Physical Soil Components ............................................................................. 4
2.2 Biological Soil Components ........................................................................... 4
2.3 Chemical Soil Components....................5
3. Location of Practical Work and Equipment Used.6
4. Results and Discussion7
4.1 Physical Soil Results.....7
4.2. Biological Soil Results.....7
4.3 Chemical Soil Results...8
4.4 Discussion of Results.............................9
4.4.1 Discussion of Physical Soil Results..............................................................9
4.4.2 Discussion of Biological Soil Results..........................................................10
4.4.3 Discussion of Chemical Soil Results...........................................................11
5. Conclusions and Recommendations for Soil Health........12
6. Acknowledgements....13
7. References..................13
8. Appendices...............................................................................................................16



List of Tables
Table 1: Rationale for and type of equipment used in studies..........................................6
Table 2: Physical characteristics of the representative soil sites......................................7
Table 3: Data collected from each soil site during the second field study........................8
Table 4: Chemical soil results from the first set of soil samples collected........................9

List of Figures
Figure 1: Comparison of soil temperature and earthworm populations............................8
Figure 2: pH levels from each soil site..............................................................................9



3

1. Introduction
Soil health and degradation are two key issues for future environmental
sustainability and are greatly influenced by management regimes (Hill et al., 2000; Lal,
1993). Intensive management involving frequent tillage and fertilizer applications can
cause alterations to the physical, biological, and chemical components of the soil
(Doran et al., 1999). This, in turn, can impair microorganism population size, diversity,
and functioning, all of which are critical components of soil formation, nutrient cycling,
and the breakdown of toxic chemicals (Bardgett et al., 1999; Fierer, Schimel, & Holden,
2003; Hackl et al., 2005; Kirk et al., 2004; Zhou et al., 2008).
Bacteria, actinomycetes, and protozoa are microorganisms that experience
differential impacts from anthropogenic disturbance (Meriles et al., 2009). Alterations to
soil horizons and vegetation can harm bacteria and actinomycetes by limiting
rhizosphere functioning and organic matter content (Ashman & Puri, 2002). Earthworms
feed on and support bacteria and protozoa, making them important for the maintenance
of microorganism populations (Stott, Kennedy, & Cambardella, 1999). Each type of
microorganism also responds quickly to changes in the environment, suggesting that
their populations could fluctuate with disturbances such as fertilizer applications.
This study was interested in describing the qualitative and quantitative attributes
of a fallow field, sod field, and non-agricultural control area to estimate microorganism
presence as an indicator of soil health. It was expected that a comparison of each site
would reveal microorganism deficiencies in the sod field; this hypothesis is supported in
the results, suggesting that sod field management should become less intensive.
4

2. Objective and Methodology
Information on the physical, chemical, and biological components of the soil from
each study site was collection during two separate field studies. The objective of these
studies was to determine the differences between each site as indicators of soil health.
2. 1 Physical Soil Components
The first field study on October 24, 2009, was conducted according to the
supporting literature to determine physical soil attributes (see Feng et al., 2003; Hackl et
al., 2005; Rajaniemi & Allison, 2009). Three representative soil sites were randomly
chosen within each area of interest. Soil plots were 30 cm deep, and four soil samples
were collected at varying depths (first 5 cm, 5-10 cm, 10-20 cm, and 20-30 cm). These
samples were then bagged, marked, and transported to a refrigerator where they were
stored at a temperature of roughly 4 Celsius. Information on the slope, vegetation, and
soil of each area was qualitatively described using the B.C. Ministry of Environment,
Lands, and Parks (BCMCELP) Field Manual for Describing Terrestrial Ecosystems.
2.2 Biological Soil Components
As the majority of microbial biomass is located in the first several centimetres of
soil, soil samples from 0-10 cm were tested for microorganism content on October 26,
2009 (Fierer et al., 2003). One cup of soil, two cups lukewarm water, and one cup of
softener salt were regularly stirred together in a pan for one hour; however, this
experiment yielded no living microorganisms and the thesis of this paper had to be
expanded. More advanced scientific methods, such as phospholipid fatty acid (PLFA)
5

tests or fatty acid methyl ester (FAME) profiles, could have accurately determined the
presence of living microorganism communities (Hill et al., 2000; Meriles et al., 2009).
The second field study was conducted on November 1, 2009, to determine the
number of earthworms at each soil site. Three different 30 cm deep plots were dug, the
soil was removed, and soil temperature was taken at 10 cm. Rocks and earthworms
were counted and classified after sifting the soil. One litre of a mixture composed of
mustard (35 grams) and water (3 litres) was poured into each site; however, this failed
to bring aneic (deep-burrowing) earthworms to the surface (Edwards, 2009).
2.3 Chemical Soil Components
Chemical characteristics of the soil samples collected October 24 were
determined on November 5, 2009, using the LaMotte Company (2001) Soil Testing Kit.
Prior to testing, soil samples were mixed, placed on plastic sheets, and left to dry for 24
hours. The soil was prepared using 2.5 grams of soil mixed for one minute with 4mL of
Universal Extracting Solution, after which it was filtered. For the nitrate nitrogen tests,
1mL of soil filtrate was mixed with ten drops of Nitrate Reagent 1 and 0.5 grams of
Nitrate Reagent 2 on a spot plate and results were taken after five minutes.
Phosphorous results were taken immediately after 1mL soil filtrate was mixed with six
drops of Phosphorous Reagent 2 and one tablet of Phosphorous Reagent 3. Potassium
tests required one Potassium Reagent B Tablet to be dissolved in the soil extract along
with Potassium Reagent C; this mixture was slowly transferred to a reading stand using
a pipette. For pH tests, the Duplex and Bromthymol Blue Indicators were added to
unaltered soil on a clean spot plate to determine pH levels to one decimal place.
6

3. Location of Practical Work and Equipment Used
The geographical location for this study was 555 Riverbank Drive, Cambridge,
Ontario and at the adjoining sod field owned by Greenhorizons Group of Farms Limited
(latitude 43.430215; longitude -80.402484). Parent material of this area was
determined to be morainal because the soil appeared to be a heterogeneous mix of
sand, silt, and clay (BCMELP, 1998). The soils in this area were classified as gray-
brown podzols, which possess high humus content and are slightly acidic from leaching
(Davies et al., 1973). This type of soil is very fertile and provides highly productive
farmland throughout southern Ontario, making it ideal for sod production (Ashman &
Puri, 2002). Equipment used in all field and in-lab studies is outlined in Table 1, below.
Table 1: Rationale for and type of equipment used in studies, along with the outcome of each
Type of Equipment Purpose Outcomes
Shovel and three 10.5 litre
buckets
To remove soil and contain
it for analysis
Soil samples were collected
30 cm ruler To measure soil depth Soil samples were removed
at varying depths
12 plastic baggies and 1
insulated lunch box
To contain and cool soil
samples
Soil samples were safely
transported to refrigerator
3 porcelain pans and 1 stir
stick
To remove microorganisms
from water and salt solution
No microorganisms were
found in soil samples
Metal sieve (10 mm hole
size)
To sift out rocks and
earthworms from the soil
Rocks and earthworms
were removed and counted
Thermometer To measure temperature of
soil at 10 cm depth
Soil temperature taken at
each site
4 mL LaMotte Universal
Extraction Solution
To prepare soil for reagent
testing
Soil was mixed and filtered
to produce soil filtrate
LaMotte soil testing
reagents
To analyze soil nutrients Nitrate nitrogen, potassium,
phosphorous levels taken
LaMotte pH testing
chemicals
To determine acidity of the
soil
Soil pH was identified to
one decimal place
Spot plate and test tubes To mix soil filtrate with
LaMotte chemicals
Nutrient and pH levels of
soil were determined
Pipette and 0.5 gram soil
scoop
To transfer soil and liquid
solutions
Liquids and solids were
safely transferred
7

4. Results and Discussion
4.1 Physical Soil Results
Level of use, vegetation present, slope, and soil characteristics were gathered
from the initial field study and are displayed in Table 2. The majority of this information
was derived from the authors observations using BCMELP (1998) and consultation with
the owner of 555 Riverbank Drive. The results presented are comparable because the
samples were taken in similar locations of slope, which limits the potential for soil
degradation from water and wind erosion (Table 2) (Stocking & Murnaghan, 2001).
Table 2: Physical characteristics of the representative soil sites from the fallow field, sod field, and control
area; vegetation was keyed using Dickinson & Royer (1999) as well as Petrides (1958)
Location Level of use Vegetation Present Slope Soil Characteristics
Fallow
Field
40 years since
last ploughing
event and
fertilizer
application
Alfalfa (Medicago
sativa L.); White
Clover (Trifolium
repens L.); Barnyard
Grass (Echinochloa
crusgalli L. Beauv.)
Slight slope;
samples taken
near top of
slope
Sandy loam;
moderate amount of
roots; weak cast; O,
H, A, and beginning
of E layer (past 30
cm)
Sod Field 5-6 sod removals
during season,
fertilizer and
pesticide
application once
per month
Kentucky Blue
Grass (Poa
pratensis L.)
Slight slope;
samples taken
near top of
slope
Sandy loam; grass
roots; weak cast; light
colour of soil
suggests only E layer
present
Control
Area
None White Elm (Ulmus
americana), Black
Walnut (Juglans
nigra L.)
No slope
(level)
Sandy clay loam; no
roots; moderate cast;
deep O and H layer

4.2 Biological Soil Results
The number of rocks and earthworms present in the soil, as well as soil
temperature, was determined in the second field study. There were many rocks of
varying sizes found in the sod field soil (Table 3). The earthworms found were all
8

classified as Lumbricus terrestris, and their populations fluctuated regardless of soil
temperature, which was similar at each site (Figure 1).

Table 3: Data collected from each soil site during the second field study, including the number of
earthworms and rocks counted along with rock size. Small rocks are defined as less than 2 cm; medium
rocks less than 6 cm; and large rocks less than 10 cm in width
Number of
Earthworms
Number of Rocks Size of Rocks
Fallow Field 27 45 Small
Sod Field 25 90 Small, medium,
and large
Control Area 13 33 Small


Figure 1: Comparison of soil temperature and earthworm populations in the fallow field, sod field, and
control area; Maria Legault, 2009
4.3 Chemical Soil Results
Results from the LaMotte Company (2001) Soil Testing Kit are displayed in Table
4 and Figure 2. These results show that the control area had the highest levels of
potassium and nitrate nitrogen, while the sod field was deficient in all nutrients (Table
4). The pH tests indicate that acidity is within the acceptable range of 5-8.5 pH for
vegetation growth and microorganism functioning (Hackl et al., 2005). Additionally, the
alkaline soil of the sod field is likely a result of regular limestone applications meant to
foster vital grass growth (Perez et al., 1995).
0
5
10
15
20
25
30
Fallow Field Sod Field Control Area
Influence of Temperature on Earthworm Populations
Number of
Earthworms
Soil Temperature
(Celsius)
9

Table 4: Chemical soil results from the first set of soil samples collected, including potassium, nitrate
nitrogen, and phosphorous levels
Location Potassium Levels
(Lbs. per acre
Potassium)
Nitrate Nitrogen
Levels (Lbs. per acre
Nitrate Nitrogen)
Phosphorous Levels
(Lbs. per acre
Available
Phosphorous)
Control Area 120 10 Less than 10
Sod Field Less than 100 Less than 10 Less than 10
Farm Field 100 Less than 10 Less than 10


Figure 2: pH levels from each soil site, which suggests that pH is not limiting to vegetation growth or
microorganism populations Maria Legault, 2009

4.4 Discussion of Results
4.4.1 Discussion of Physical Soil Results
Tillage negatively affects microorganism respiration and activity levels by altering
soil horizons through reduced organic matter content (Feng et al., 2003; Lal, 1993;
Meriles et al., 2009). While the fallow field has not been ploughed for 40 years, the sod
field experiences regular topsoil removal events and appears to be composed entirely of
the E horizon. This is shown through the sod fields light brown colour, high sand
content, and grainy feeling (Aswathanarayana, 1999). In contrast, the thick leaf litter
layer in the control area and the plentiful roots in the fallow field contribute to more
organic matter; a rich O, H, and A layer; and high levels of rhizosphere activity
6.6
6.8
7
7.2
7.4
7.6
7.8
Control Area Sod Field Farm Field
pH Levels
10

(Aswathanarayana, 1999). This, in turn, creates more soil organic carbon and
increases the activity levels of microorganism communities (Stott et al., 1999).
Vegetation at the three soil sites also affects microorganism communities.
Herbaceous vegetation in the fallow field, such as alfalfa, decreases erosion and adds
more carbon to the soil, thereby increasing microbial biomass (Faniran & Areola, 1978;
Rajaniemi & Allison, 2009). This vegetation can also create the abiotic conditions
needed for diverse microorganism communities to survive by maintaining soil pH (Hackl
et al., 2005). In contrast, the single grass species present in the sod field will support
less diverse and active microorganisms (Fierer et al., 2003).
4.4.2 Discussion of Biological Soil Results
A similar number of earthworms were found in the sod and fallow fields,
indicating that microorganism populations are being maintained. Earthworms are
closely tied to microorganisms because they help to break down organic matter, transfer
soil nutrients, and rely on bacteria as a source of nutrition (Ashman & Puri, 2002). The
low number of earthworms in the control area could be a result of the thick moss layer
limiting their ability to burrow into the soil (Davies et al., 1973). In contrast, the well-
turned, porous soil in the sod field is easier to penetrate; this suggests that the
relationship between earthworms and microorganisms in this case does not indicate soil
health, but a high level of management.
Additionally, the number of rocks in the sod field suggests that the soil organic
matter needed to support large microorganism populations has been lost (Lal, 1993). At
the end of the growing season, sod farms remove surface vegetation so that the area
11

can be re-seeded in the spring (Perez et al., 1995). This practise, along with regular
topsoil removal throughout the year, results in more rocks appearing at the surface as
the upper levels of soil are stripped away (Stocking & Murnaghan, 2001).
Subsequently, soil becomes vulnerable to compaction, erosion, and nutrient imbalances
which reduce the ability of microorganisms to survive (Doran et al., 1999). In contrast,
smaller and fewer rocks were found at the fallow field and control sites, indicating the
health of the soil in these areas.
4.4.3 Discussion of Chemical Soil Results
The chemical soil results suggest that regular fertilizer applications are needed in
the sod field, which could negatively affect environmental safety (Bardgett et al., 1999;
Zhengfei et al., 2005). Nitrogen is critical for encouraging the fast growth of grasses,
but was found to be available in low amounts in the sod field (Maryland Department of
Agriculture [MDA], 2009). Regular applications of nitrate nitrogen to the soil to combat
this deficiency could cause leaching into adjacent water sources, posing an
environmental threat (Aswathanarayana, 1999).
Other nutrients in the sod field were also found to be limiting to microorganism
communities. Phosphorous contributes to root growth and therefore soil carbon; low
levels of this nutrient in the sod field indicate depressed microbial biomass (Bardgett et
al., 1999; Murdock, n.d.; Zhou et al., 2008). The development of a substantial
rhizosphere is important for microorganism survival and is strongly influenced by
potassium levels (Ashman & Puri, 2002; MDA, 2009). This suggests that the low levels
of potassium in the sod field may decrease the diversity of microorganisms present.
12

5. Conclusions and Recommendations for Soil Health
This study compared the physical, biological, and chemical components of the
soil to determine the differences between a fallow and sod field. Results supported the
belief that intensive management on sod farms is damaging to soil health. Tillage
causes the loss of the upper soil horizons, which can subsequently increase the
proportion of rocks near the surface and decrease microorganism activity levels.
Fertilizer applications can also damage soil health by causing fluctuations to sensitive
microorganism populations. These negative impacts are significant because
microorganisms are critical to the soil resources which provide the basis for human
nourishment and economic activities. Accordingly, it is recommended that sod
companies reduce their management efforts and try to find more sustainable methods
for extracting their product. Their failure to do so could result in the degradation of large
tracts of soil which could otherwise have benefitted agriculture or ecosystems.
This study was limited by a lack of data, but this was overcome with literature
research. Limited quantitative data was gathered because of the lack of
microorganisms found in the initial field study, perhaps due to the reduction in soil
biomass which occurs during the fall (Bardgett et al., 1999). To overcome this
limitation, literature research was used to understand how qualitative factors affect soil
health and microorganism populations. This study was also unable to discuss how
herbicides (e.g. phenoxyacetic acids) and pesticides affect microorganisms (Garcia-
Rivero, Saucedo-Castaneda, & Gutierrex-Rojas, 2007; Landscape Management, 2005;
Thompson et al., 1998). More studies should be done on this topic to better understand
the interactions between microorganisms and chemicals.
13

6. Acknowledgements
This study was made possible by the generous help and support of staff in the
University of Waterloo Ecology Lab, Doug Mader from Compact Sod, and Rieta Fietsch.
7. References
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Bardgett, R.D., Lovell, R.D., Hobbs, P.J., & Jarvis, S.C. (1999). Seasonal changes in
soil microbial communities along a fertility gradient of temperate grasslands. Soil
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Davies, N.D., Stoker, D.G., Windsor, D.E., Ashcroft, M.T., Coburn, M.C., & Andrews,
W.A. (1973). A Guide to the Study of Soil Ecology. Scarborough, Ontario:
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Dickinson, R., & Royer, F. (1999). Weeds of Canada. Vancouver, BC: Lone Pine
Publishing.
Doran, J.W., Jones, A.J., Arshad, M.A., & Gilley, J.E. (1999). Determinants of soil
quality and health. In R. Lal, Soil Quality and Soil Erosion (pp. 17-36). London:
Soil and Water Conservation Society.
Edwards, C.A. (2009). Earthworms. Chapter 8 in The Soil Biology Primer. Retrieved on
October 29, 2009, from
http://soils.usda.gov/sqi/concepts/soil_biology/earthworms.html
Faniran, A. & Areola, O. (1978). Essentials of Soil Study: With Special Reference to
Tropical Areas. London: Heinemann Educational Books Ltd.
Feng, Y, Motta, A.C., Reeves, D.W., Burmester, C.H., Santen, E.V., & Osborne, J.A.
(2003). Soil microbial communities under conventional-till and no-till continuous
cotton systems. Soil Biology and Biochemistry 35 (12), 1693-1703.
Fierer, N., Schimel, J.P., & Holden, P.A. (2003). Variations in microbial community
composition through two soil depth profiles. Soil Biology and Biochemistry 35 (1),
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Garcia-Rivero, M., Saucedo-Castaneda, G., Gutierrex-Rojas, M. (2007). Organic
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J.T. (2004). Methods of studying soil microbial diversity. Journal of
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Lal, R. (1993). Tillage effects on soil degradation, soil resilience, soil quality, and
sustainability. Soil and Tillage Research 27 (1-4), 1-8.
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Management: The Preferred Source for Large Contractors. Retrieved October
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04
Maryland Department of Agriculture. (2009). Nutrient Recommendations for Sod
Production. Retrieved on October 28, 2009, from
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nt_recommendations_for_sod_production.php
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systems: Characterization of microbial population dynamics, soil microbial
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Production. Cooperative Education Service, University of Kentucky College of
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Agriculture. Retrieved October 29, 2009, from
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Perez, A., Harwood, J., Johnson, D., Somwaru, A., & Zepp, G. (1995). Turfgrass sod:
An economic assessment of the feasibility of providing multiple-peril crop
insurance. Economic Research Service, USDA. Retrieved October 28, 2009,
from http://www.rma.usda.gov/pilots/feasible/PDF/turfsod.pdf
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73). London: Soil and Water Conservation Society.
Thompson, I.P, Bailey, M.J., Ellis, R.J., Maguire, N., & Meharg, A.A. (1998). Response
of soil microbial communities to single and multiple doses of an organic pollutant.
Soil Biology and Biochemistry 31 (1), 95-105.
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16

8. Appendices
Appendix 1: Map of Study Location


Appendix 2: Map of Land Uses at Study Location

17

Appendix 3: Vegetation in a) the fallow field, b) sod field, and c) control area
a)
Maria Legault, 2009
b)
Maria Legault, 2009
c)
Maria Legault, 2009

Appendix 4: Example of soil sample depth in the fallow field
Maria Legault, 2009




18

Appendix 5: Experiment conducted to remove microorganisms from soil samples
Maria Legault, 2009

Appendix 6: Images of rocks found in sod field within first 10 cm of sample
Maria Legault, 2009