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Estuarine, Coastal and Shelf Science 76 (2008) 21e28 www.elsevier.com/locate/ecss

Long-term abundance patterns of macroalgae in relation to environmental variables in the Tagus Estuary (Portugal)
Abel Sousa-Dias, Ricardo A. Melo*
Universidade de Lisboa, Faculdade de Cie ncias, Instituto de Oceanograa, Campo Grande, 1749-016 Lisboa, Portugal Received 28 February 2007; accepted 28 May 2007 Available online 23 July 2007

Abstract Seasonal and inter-annual patterns of macroalgal abundance in a Tagus Estuary oyster reef are described. Macroalgal abundance was estimated as canopy percent cover by three permanent point intercept transects over a 7-year period. Four categories were dened, corresponding to bare substrate and three different macroalgal functional-form groups: (1) ULVA, foliose group, included Ulva spp.; (2) GRACIL, terete corticated macrophyte group, included only Gracilaria gracilis; and (3) FILAM, small (<10 cm) lamentous group, including eight species. A canonical correspondence analysis (CCA) showed that: (1) ULVA were associated with long and hot days, being usually dominant during spring and especially summer; (2) FILAM were associated with mild temperatures and relatively long days, abundant in spring but showed frequent peaks in summer; and (3) GRACIL were also favoured by spring season, although associated to lower temperature and less daylight hours than FILAM. GRACIL and FILAM were present throughout the year. On the contrary, ULVA were absent or with low cover during colder periods. A negative correlation between GRACIL and FILAM seems to indicate competition between the two categories. The applied models explained 23.3% of the temporal variance in category abundance. Rainfall negatively affected macroalgal cover, as indicated by the positive correlation between rainfall and bare substrate. Our conclusions are in agreement with previous studies that consider algae as excellent environmental integrators, even on a small scale, due to a strong link between the macroalgal communities and relevant environmental variables. It is also relevant that this study used open-access databases of environmental variables, which open up new possibilities for mining existing data resources in new ways. Due to large inter-annual variability, long-term studies are essential to understand population dynamics in estuarine phytobenthic communities. 2007 Elsevier Ltd. All rights reserved.
Keywords: algae; estuaries; multivariate analysis; oyster reefs; phytobenthos; transitional waters Regional index terms: Portugal; Lisbon; Tagus Estuary; 38 440 N 9 080 W

1. Introduction Macrophytobenthos is of great importance in estuarine and near shore marine ecosystems because of high productivity, key roles in nutrient dynamics, mainly carbon xation and nutrient scrubbing (Flindt et al., 1999), incorporation of pollutants such as heavy metals (Coelho et al., 2005), and

* Corresponding author. E-mail addresses: abelsd@gmail.com (A. Sousa-Dias), rmelo@fc.ul.pt (R.A. Melo). 0272-7714/$ - see front matter 2007 Elsevier Ltd. All rights reserved. doi:10.1016/j.ecss.2007.05.039

contribution to faunal diversity and maintenance (Eriksson et al., 2004). In the Tagus Estuary the macrophytobenthos includes both macroalgae and salt marsh plants, there are no seagrasses (Ferreira et al., 2003), and, as in other mesotidal estuaries, these communities have considerable areal representation and ecological roles (Simas et al., 2001; AlveraAzcarate et al., 2003). Availability of suitable hard substrate or attachment points has been associated with elevated macroalgal abundance in different estuaries (Middelboe et al., 1998; Nedwell et al., 2002), but in the Tagus Estuary these areas, including soft substrates with gravel or rock fragments and shallow oyster reefs, represent only about 5% of the total

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intertidal area (Ferreira and Ramos, 1989). Even so, a recent modelling study estimated that intertidal macroalgae accounted for 21% of the total carbon xed by all primary producers in the Tagus Estuary, being responsible for nutrient uptake equivalent to the annual loading of about 490,000 people (Alvera-Azcarate et al., 2003). However, contrary to other important estuarine primary producers, like the phytoplankton, for which a number of studies have been performed (e.g., Gameiro et al., 2007 and references therein), seaweed production has received a limited amount of attention, mostly through the application of models (e.g., Ferreira and Ramos, 1989; Alvera-Azcarate et al., 2003). Numerous studies have shown that algae are excellent integrators of uctuating abiotic variables (Lowe, 2006), and can be used to indicate shifts in ecological status (Orfanidis et al., 2003). Temperature and solar radiation are generally considered very relevant environmental factors determining macroalgal distribution both at worldwide and local geographical scales (Lu ning, 1990; Santos, 1993). However, macroalgal diversity and abundance in estuaries are very complex, being inuenced by a variety of other abiotic factors such as salinity, turbidity, and nutrient concentration (Middelboe et al., 1998). Rainfall has also been valued as a very important forcing factor in estuaries as it inuences the input of nutrients from freshwater and fertilizer runoff, the depth of light penetration, through transport of particulate matter and sediment resuspension, and river discharge and ow velocity (Gameiro et al., 2007). Increased ow velocity in turn may cause suspended sediment scour, which may result in the removal of phytobenthos (Francoeur and Biggs, 2006). The Portuguese governmental institution in charge of hydric resources administration collects these kinds of environmental variables, and has made them available online on an open-access basis (http:// snirh.pt, English version not yet available). Previous authors have characterized the nutrient status of the Tagus Estuary. Annual nitrogen loading in the Tagus Estuary was estimated to be 26,000 tons in 1997 (Cabrita and Brotas, 2000), and Gameiro et al. (2004) reported average dissolved inorganic nitrogen (DIN) in the range of 22.1e 53.8 mmol L1 for two sites in the upper Tagus Estuary in 1999e2000. According to Ferreira et al. (2003), the Overall Eutrophic Condition (OEC) index classied the estuary into the Moderate Low category and the Overall Human Impact (OHI) index classied the impact of nutrient inputs to the estuary as Low. The nitrate concentration in the water was far below the limit considered in Directive 91/676/EEC (Ferreira et al., 2003), and in an estuarine quality index (EQUATION index) the Tagus Estuary obtained a classication of fair (Ferreira, 2000). Furthermore, according to Gameiro et al. (2004) mean dissolved inorganic nitrogen has been stable for the last 20 years. Here we report on the seasonal and inter-annual variability of macroalgal abundance, measured as percent cover, on an intertidal oyster reef in the Tagus Estuary, Portugal, over a 7-year period. Our main goal was to quantify the relationships between the patterns of macroalgal abundance and the seasonal dependent environmental variables temperature,

daylight hours and rainfall obtained from a public environmental database. 2. Materials and methods 2.1. Site description Covering about 320 km2 the Tagus is one of the largest estuaries in Europe and is located on the central west coast of Portugal (38 440 N, 9 080 W). The climate is mild with a mean air temperature of 16.3  C and a total annual precipitation of 700 mm (Gameiro et al., 2004) but highly variable (Trigo et al., 2004). The intertidal area corresponds to 40% (138 km2) of the total estuarine area in spring tides (Ferreira et al., 2003) and the substrate varies from mudats to ne sands and oyster reefs, which are no longer commercially exploited, and occupy an area of approximately 16 km2 (Ferreira and Ramos, 1989). The present study was carried out at the i oyster reef (Fig. 1), which emerges only on Ponta do Destro spring tides lower than 0.7 m above tidal datum. The River Tagus mean annual discharge showed very distinct values ranging from 52.8 m3 s1 in the very dry 2005 to 735.5 m3 s1 in rainy 2001 (Gameiro et al., 2007). Other water properties mea i from 1999 to 2005 (Gameiro and Brosured at Ponta do Destro tas, personal communication) were the following: mean water temperature 18.4  C (range 8.0e24.0  C), average salinity 24.0 (range 2.1e35), average suspended particulate matter 23.3 mg L1 (range 3.9e48.3 mg L1), and mean light extinction coefcient 1.1 m1 (range 0.6e2.3 m1). Several macroalgal species have been previously recorded from this site (Silva and Melo, unpublished results): Ulva lactuca Linnaeus and other unidentied Ulva species

Fig. 1. Map of the study site area, Tagus Estuary, with location of Ponta do i oyster reef. Grey areas represent intertidal mud ats. Destro

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23

(Chlorophyceae), Gracilaria gracilis (Stackhouse) Steentoft, L.M. Irvine & Farnham (Rhodophyceae), and several small (<10 cm) thin lamentous species: Cladophora prolifera (Roth) Ku tzing (Chlorophyceae), and Aglaothamnion pseudobyssoides (Crouan & Crouan) Halos, Ceramium virgatum Roth, Erythroglossum laciniatum (Lightfoot) Maggs & Hommersand, Erythroglossum sandrianum (Zanardini) Kylin, Polysiphonia denudata (Dillwyn) Greville, Polysiphonia nigra (Hudson) Batters, Polysiphonia sp. (all Rhodophyceae). Nomenclature follows Algaebase (http://www.algaebase.org). Since both Ulva spp. and G. gracilis were easily recognizable in the eld but not the smaller lamentous species, we opted to subdivide algal taxa by functional-form groups sensu Steneck and Dethier (1994). 2.2. Macroalgal sampling Macroalgal abundance was sampled seasonally from winter 1999 to winter 2006, during spring low tides with an amplitude range > 3.4 m, the only periods when the reef emerged. Due to unfavourable weather some sampling dates were missed (Table 1). Abundance was estimated as canopy cover, i.e., the vegetation covering the substrate above the surface, by point intercept (Elzinga et al., 2001), along linear transects, our sampling units, consisting of thin cables with ink marks at 1 m intervals. On each sampling date three transects were sampled perpendicular to the shoreline at 200 m intervals. At every meter mark, the presence of either of four categories was recorded as: (1) ULVA, foliose group, including only Ulva spp.; (2) GRACIL, corticated terete macrophyte group, including only Gracilaria gracilis; (3) FILAM, including all small (<10 cm) lamentous group species; or (4) SUBST, corresponding to bare substrate, i.e., without macroalgal cover. The total number of point intercept hits recorded was dependent on the tidal amplitude but transects were at least 120 m long. Seasonal percent cover for each of the categories was calculated as the average of the three transects sampled in each date. The same process was applied for category SUBST to measure transect and seasonal percent bare substrate. 2.3. Environmental variables Environmental data were obtained from the Sistema Naciodricos-SNIRH (National nal de Informac ~ ao de Recursos H
Table 1 i oyster reef, Tagus Macroalgal abundance sampling dates at Ponta do Destro Estuary Year 1999 2000 2001 2002 2003 2004 2005 2006 Winter 19/2 e 12/3 29/1 20/3 9/3 11/2 1/2 Spring 15/6 6/4 23/6 26/4 14/6 14/6 24/5 e Summer 27/9 3/8 18/9 9/9 29/9 3/8 23/7 e Autumn e 14/11 e 5/12 26/11 13/11 17/10 e

Hydric Resources System), publicly available online (http:// snirh.pt/). Different environmental variables available in the database were tested but only those that showed robust statistical correlations were selected. These were the following: (1) mean daylight hours between sampling dates (D), as a proxy for incident solar radiation, calculated from monthly averages; (2) total accumulated daily rainfall between sampling dates (R) in mm; and (3) average between sampling dates of mean daily air temperatures (T ), as a proxy for water temperature, in  C. Data available in this database for factors such as salinity, turbidity or inorganic nutrients in the Tagus Estuary were generally too few and temporally sparse, and from sites too i to warrant their utilization. distant from Ponta do Destro 2.4. Data analysis To determine whether there were signicant differences between transect percent cover within each sampling date, throughout the study period, Friedmans test (Siegel and Castellan, 1998) was performed for the macroalgal categories, using the 26 sampling dates as replicates. On each sampling date category percent cover was averaged over the three transects performed, since no signicant differences were found between transects within sampling dates ( p > 0.05). To test for signicant trends in patterns of seasonal and annual variation of percent cover during the sampling period, Kendalls coefcient of rank correlation (Sokal and Rohlf, 1995; Zar, 1999) was used. The same test was chosen to test for signicant correlations between all categories. All statistical analyses were performed using the SPSS package 14.0 (Anon., 1997). The relationships between macroalgal abundance and the environmental variables were investigated with canonical milauer, correspondence analysis e CCA (ter Braak and S 2002), again using the average of the three transects per sampling date for each category. A global Monte Carlo permutation test was used to evaluate the signicance ( p < 0.05) of the rst ordination axis and the sum of all canonical axes milauer, 2002). Multivariate analyses were (ter Braak and S milauer, carried out using CANOCO 4.5 (ter Braak and S 2002). Kendalls coefcient of rank correlation (Sokal and Rohlf, 1995) was also used to test the results of CCA showing association between categories and environmental variables. 3. Results 3.1. Macroalgal percent cover Macroalgal percent cover varied largely between the seasons and the years (Fig. 2). The lowest overall macroalgal percent cover value was measured in winter 2001 (14%) and the highest overall value (77%) occurred in spring 2003. Category FILAM, after a peak in summer 2002, declined until winter 2004 and increased again until summer 2005 when the highest percent cover of the entire study period for this category was observed (64%), showing a marked inter-annual variation. FILAM were present throughout the year, although usually more abundant during springesummer. Exceptions occurred

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100 90

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FILAM GRACIL ULVA

Macroalgal cover (%)

80 70 60 50 40 30 20 10 0 W 1999 Su W 2000 Su W 2001 Su W 2002 Su W 2003 Su W 2004 Su W 2005 Su W 2006

Fig. 2. Variation of macroalgal percent cover (FILAM e small (<10 cm) lamentous group, includes eight species; GRACIL e corticated terete macrophyte group, i oyster reef, Tagus includes Gracilaria gracilis; ULVA e foliose group, includes Ulva spp.; and SUBST e bare substrate without algal cover), in Ponta do Destro Estuary, throughout the study period (W winter; Sp spring; Su summer; and A autumn).

in autumn 2000 and 2004, which had the highest values for those years, and spring 2002, the lowest cover of that year. The minimum percent cover occurred in winter 2004 (9.1%). This category dominated throughout 2005, except in autumn, when ULVA was clearly more abundant. In fact, the highest ULVA percent cover of the study period was 35% in autumn 2005, followed by spring 2003 (30%) and summer 2002 and 2003 (22% and 27%, respectively). ULVA was almost absent (less than 1%) during winter 1999, 2001, 2005 (no ULVA present), spring 1999 and autumn 2004. Although ULVA usually peaked in springesummer, in 2000 and 2005 the season with the maximal ULVA percent cover was autumn. FILAM and GRACIL seemed to alternate in dominance. As FILAM, GRACIL were also present throughout the year, but with maxima usually in winterespring, although abundant in some summers. In summer 2000 and spring 2003 maxima for this category (30%) were observed. However, in spring and summer 2005, GRACIL percent cover was only residual (less than 1%). From spring 2003 to spring 2004 GRACIL maintained similar representation, without a seasonal pattern. To emphasize the apparent seasonal pattern the data were clustered into seasons (Fig. 3). Although a seasonal pattern was apparent when considering the whole macroalgal community, autumn had unexpected higher average values that were mainly due to the unusual high cover of ULVA in autumn 2005. Lack of data from autumn 1999, 2001 and 2006 probably also contributed to these high averages. Nevertheless, an

increase of macroalgal cover during springesummer and a decrease in autumnewinter were evident. Contrary to the seasonal pattern described for the whole macroalgal community, when analysed separately, macroalgal categories showed some differences. ULVA peaked during summer and were less abundant during winter. GRACIL peaked during spring and were less abundant during autumn. FILAM were present throughout the year but with higher coverage during summer. Differences between seasons were much more marked for ULVA than for the other two macroalgal categories. Inter-annual variation of substrate occupation by macroalgae was observed (Fig. 4). Annual substrate cover by macroalgae was usually over 50% except on 2001 and 2004, when the annual average was around 40%; FILAM were dominant during the study period, except in 2003, when GRACIL dominated (w24%), followed by ULVA (w19%), the annual maxima for the latter categories. In 2002 and 2005, FILAM reached over 30% and 40%, respectively. No signicant increasing or decreasing trends for each category during the study period were found ( p > 0.05). There were signicant correlations among some categories (Table 2). 3.2. Environmental variables During the study period, the local climate showed a wide range of variation reected in contrasting dry years, 1999 and 2005, and a very rainy 2001. In general, summers were

FILAM 70 60 50 40 30 20 10 0 W

GRACIL

ULVA

All macroalgae

Macroalgal cover (%)

Sp

Su

Fig. 3. Seasonal variation of macroalgal percent cover during 1999e2006 (for terminology see Fig. 2). Bar graph seasonal averages standard error; solid line sum of all categories.

A. Sousa-Dias, R.A. Melo / Estuarine, Coastal and Shelf Science 76 (2008) 21e28
70

25

All macroalgae

FILAM

GRACIL

ULVA

Macroalgal cover (%)

60 50 40 30 20 10 0
1999 2000 2001 2002 2003 2004 2005 2006

Fig. 4. Variation of average mean annual percent cover of macroalgae (for terminology see Fig. 2); bars total macroalgal percent cover (ULVA GRACIL FILAM).

dry, with the exception of the two rst summers of the sampling period (1999 and 2000), when signicant rainfall occurred (Fig. 5). As expected, winters were the rainiest seasons, but in 2001 unusually high decadal rainfall values were recorded. On the contrary, winter 2005 was unusually dry and spring 2000 had rainfall values more typical of winter seasons (w400 mm). Mean daily air temperature showed a pattern of regular seasonal uctuations, with maxima in summer and minima in winter (Fig. 5). Winter 1999 and 2005 were the coldest of the sampling period (average around 10  C). Spring 1999, 2001, 2003 and 2004 were the warmest of the sampling period (w17  C) and the remaining (2000, 2002 and 2005) were the coldest (w14  C). Summers, as expected, were the warmest season during the study period except in 2000 and 2005, when autumn was the warmest season. Signicant correlations between macroalgal categories and environmental variables are presented in Table 3.

lesser daylight hours than FILAM. ULVA were associated with longer, warmer days, and with the cluster of summer samples. 4. Discussion Our results showed that temperature, daylight hours, and rainfall appear to have signicant effects on the seasonal and inter-annual patterns of macroalgal abundance in Ponta do i oyster reef. Over a 7-year period macroalgal cover Destro ranged between 14% and 77%, with an inter-annual average of 50.5%. Nedwell et al. (2002) reported that for the nutrient-enriched River Deben Estuary average intertidal macroalgae reached a maximum of 50% coverage. A seasonal trend was apparent in our samples with the highest percent cover in springesummer and lowest in autumnewinter. A similar trend was reported for the Tagus Estuary by Ferreira and Ramos (1989), as well as other locations (Solidoro et al., 1997, Raffaelli, 1999; Nedwell et al., 2002; Ferreira et al., 2003; Plus et al., 2005). We found a signicant positive correlation between macroalgal cover, and daylight hours and temperature (Table 2). Increased day length, irradiance (Malta et al., 2003; Raven and Taylor, 2003) and temperature (Plus et al., 2005) were previously associated with a spring increase of macroalgal biomass. Categories FILAM and GRACIL were present throughout the year, although the latter were sometimes residual, as in springesummer 2005. This unusual low abundance was probably also due to the unusual high FILAM coverage, which may compete with GRACIL for substrate occupation, as suggested by the signicant negative correlation between them. As category FILAM species composition may change throughout time, its abundance also peaked in other seasons. Category ULVA that showed a signicant positive correlation with temperature and day length had the highest cover during springesummer, except for autumn 2000 and 2005, which were the hottest season for those years. Ferreira and Ramos (1989) found that Ulva lactuca biomass peaked in spring and summer in the Tagus Estuary. The same is true for other locations and other Ulva species (e.g., Lavery et al., 1991; Solidoro et al., 1997; Plus et al., 2005). Peralta et al. (1997) also found higher growth rates of Ulva spp. in summer and autumn in the Palmones Estuary. CCA showed that samples formed clear seasonal clusters. The winter cluster was the more heterogeneous due essentially

3.3. Canonical correspondence analysis (CCA) The rst two CCA ordination axes explained 23.3% of the categories temporal variability. Other ordination results obtained by CCA are presented in Table 4. The global Monte Carlo permutation test showed that relationships between categories percent cover and environmental variables were statistically signicant ( p < 0.05), for the rst canonical axis (F-ratio 4.985), as well as for the sum of all canonical axes (F-ratio 2.228). In general, samples formed dened seasonal clusters, except for spring 2000 that clustered with winter samples, being especially similar to winter 2002 (Fig. 6). FILAM macroalgae seemed to be favoured by mild temperatures and relatively long days. GRACIL were also favoured in spring, although associated to lower temperature and
Table 2 Signicant correlations found between all categories using Kendalls coefcient of rank correlation; FILAM e small (<10 cm) lamentous algae group; GRACIL e corticated terete macrophyte group; ULVA e foliose algae group; and SUBST e bare substrate without algal cover Correlation coefcient FILAM/SUBST ULVA/SUBST GRACIL/FILAM 0.311 0.495 0.434 Signicance (two-tailed) 0.026 0.000 0.002

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25 20 15 10 5 0
Sp99 Su99 Sp00 Su00 Sp01 Su01 Sp02 Su02 A02 Sp03 Su03 Sp04 Su04 A04 Sp05 Su05 W99 W01 W02 W03 W04 W05 A05 W06 A00 A03

Air temperature (C)

Rainfall (mm)

600 500 400 300 200 100 0

Fig. 5. Accumulated rainfall (bars), and mean daily air temperatures (line graph) between sampling dates. First accumulated rainfall value (W99) is the sum of daily rainfall registered during the 3 months before rst sampling date on February 19, 1999. First mean daily air temperatures value is the average of 1 month before the rst sampling date (W winter; Sp spring; Su summer; and A autumn.). Data obtained from SNIRH (http://snirh.inag.pt/).

to the highly variable annual rainfall regimes during the study period. Winter 2001 and 2005 were both extreme in terms of rainfall which is reected in the macroalgal cover, much lower in 2001 the rainiest winter of the study period than in 2005, the driest year. The fact that both GRACIL and FILAM macroalgal categories were not associated with high temperatures (no signicant correlation) as ULVA is probably one of the main factors allowing their presence throughout the year. If, as predicted by current global warming scenarios, average temperature rises but other factors are kept at similar levels (e.g. nutrient input and irradiance), an increased dominance of ULVA would be expected. In addition, GRACIL abundance, being associated with lower temperatures, will probably be the most affected. The positive correlation between bare substrate and rainfall stresses the importance of this parameter in limiting macroalgal abundance in the Tagus Estuary. Moreover, CCA showed a negative association between rainfall and all macroalgal categories, with ULVA the most affected, and GRACIL the least. The negative effect of rainfall events on substrate occupation could be due to lowered salinity, which affects macroalgal growth (Fong et al., 1996; Martins et al., 1999; McAvoy and Klug, 2005), and increased river ow and suspended sediment scour (Francoeur and Biggs, 2006). As macroalgal mats affect macrofaunal abundance (Raffaelli, 1999; Cardoso et al., 2004), changes in macroalgal cover will probably induce
Table 3 Signicant correlations found between macroalgal categories (FILAM e small (<10 cm) lamentous group; GRACIL e corticated terete macrophyte group; ULVA e foliose group; and SUBST e bare substrate without algal cover), and environmental variables (D e mean daylight hours between sampling dates; T e average between sampling dates of mean daily air temperatures; R e total accumulated daily rainfall between sampling dates) using Kendalls coefcient of rank correlation Correlation coefcient D/SUBST D/ULVA T/SUBST T/ULVA R/SUBST 0.428 0.378 0.354 0.428 0.296 Signicance (two-tailed) 0.002 0.007 0.011 0.002 0.034

modications in the macrofaunal assemblages. Coincidentally, Silva et al. (2006) showed that the main source of non-cyclic variations (i.e., not related to seasonal cycles) of invertebrate abundance in the Tagus Estuary was rainfall, either by directly inuencing the macrofaunal community or by inducing changes in the sediments. Our conclusions agree with previous studies that consider algae as excellent environmental integrators (Lowe, 2006), even on a small scale, due to a strong link between the macroalgal communities and relevant environmental variables (Eriksson and Bergstro m, 2005). It is also relevant that this study included the use of open-access environmental databases, which opens up new possibilities for mining existing data in new ways. Because published studies based on eld data of macroalgal abundance and dynamics are scarce, both for the Tagus and estuaries in general (Nedwell et al., 2002; Bettencourt et al., 2004), we hope to contribute to full this lacuna. The present study also showed that long-term data series are an essential tool to assess the structure and dynamics of estuarine macroalgal benthic assemblages, including both perennial and opportunistic species, because they allow the detection of inter- and intra-annual uctuations and their relationships with uctuating environmental parameters. However, more environmental variables, mainly relating to nutrient status, would be needed for a better understanding of macroalgal dynamics in the Tagus Estuary.

Table 4 Ordination results obtained by canonical correspondence analysis (CCA) performed for all macroalgal categories and environmental variables Axis I Eigenvalue Categories-environment correlations Cumulative percentage variance of categories-environment relationships Intraset correlations of variables Day length (D) Mean lagged air temperature (T ) Total lagged rainfall (R) 0.042 0.617 79.3 Axis II 0.011 0.521 99.8

0.5958 0.5265 0.5056

0.0153 0.2717 0.1677

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0.8

27

Sp05

W05 W99

FILAM
Sp04 Sp01 Su05 Su02 Sp99 Sp03 Su00 Su01 Su04

GRACIL
Sp02 W04 W03 Sp00 W02 W06

D T
ULVA

Su99 Su03 A05

A00

SUBST
A03

A04

A02

W01

-1.2

1.2

Fig. 6. Canonical correspondence analysis (CCA) ordination of macroalgal cover in relation to the environmental variables. ULVA, GRACIL, FILAM and SUBST scores (the value of each category on an ordination axis) are indicated by diamonds, samples by circles (W winter; Sp spring; Su summer; and A autumn), and arrows represent environmental variables (T average between samples of mean daily air temperatures; D average daylight hours between samples; and R total accumulated daily rainfall between samples).

Acknowledgements This study was conducted within the framework of a multidisciplinary monitoring program of the Tagus Estuary nanced by Valorsul, SA (http://www.valorsul.pt). The Oceanography Institute e FCUL is funded by Foundation for Science and Technology, Ministry of Science, Technology and Higher Education (FCT-MCTES, POCTI-ISFL-6-199). Thanks to many fellow scientists and students who helped in the eld and to boat master Domingos Chefe who unfailingly carried i reef even through the thickest us to the Ponta do Destro fog. ASD has presented these results as part of his Masters thesis at the University of Lisbon.

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