Freshwater Biology (2002) 47, 14631473

Correlations between nutrient concentrations and zooplankton populations in a mesotrophic reservoir

REZ-MARTI NEZ* J. M. CONDE-PORCUNA,* E. RAMOS-RODR I G U E Z and C . P E *Institute of Water Research, University of Granada, Granada, Spain Department of Animal Biology and Ecology, Faculty of Sciences, University of Granada, Granada, Spain

SUMMARY 1. A year-round study was conducted in a mesotrophic reservoir to determine the dynamics of zooplankton populations as a function of food availability (edible phytoplankton), nutrient concentration, temperature and hydraulic regime. 2. Rotifer biomass was correlated with soluble reactive phosphorus (SRP) concentration. The abundance of the rotifers Keratella cochlearis and Anuraeopsis ssa were not correlated with food availability (measured by chlorophyll and cell counts) but showed a strong dependence on P availability. Another rotifer, Synchaeta oblonga, and crustacean species were not related to nutrient availability but seemed to be dependent on food concentrations, especially of some phytoplankton taxa. 3. In this eld study, rotifers seemed more susceptible than Daphnia or copepods to P-limitation. Among rotifer species, Keratella seemed to be more susceptible than Anuraeopsis to P limitation. Different susceptibilities of zooplankton species to nutrient limitation may be important in explaining the dynamics of these organisms in natural situations. Further analyses are warranted to clarify the interactions between nutrient limitation and energy limitation among zooplankton.
Keywords: Anuraeopsis, Daphnia, eld studies, Keratella, nutrient limitation, P-limitation, phytoplankton, Rotifer

Introduction
Nitrogen and phosphorus concentrations are known to be highly dynamic and to potentially limit the production of aquatic ecosystems (Sterner & Hessen, 1994). Algae have relatively high ratios of C : P and C : N, and the C : N : P ratio of phytoplankton varies considerably under N or P limitation (Goldman, McCarthy & Peavy, 1979; Sterner et al., 1993). These biochemical responses of algae to resource limitation may affect their quality as food for upper trophic levels. Several laboratory studies have shown that the limitation of N and especially P reduces the quality of
Correspondence: J. M. Conde-Porcuna, Department of Animal Biology and Ecology, Faculty of Sciences, University of Granada, Fuentenueva s/n, E-18071 Granada, Spain. E-mail: jmconde@goliat.ugr.es 2002 Blackwell Science Ltd

algae as food for Daphnia (Groeger, Schram & Marzolf, 1991; Sterner et al., 1993; Van Donk et al., 1997; Weers & Gulati, 1997; DeMott, 1998; DeMott, Gulati & Siewertsen, 1998; Schulz & Sterner, 1999). The decrease in food quality of P-limited algae could be a direct effect of P limitation, as well as an indirect effect caused by changes in biochemistry and cell wall morphology (Van Donk & Hessen, 1993; Weers & Gulati, 1997). Urabe, Clasen & Sterner (1997) observed, under experimental conditions of very highly soluble reactive phosphorus (SRP), that Daphnia uptake of inorganic P reverses growth limitation. Boersma (2000) observed that both the mineral and biochemical limitations of phytoplankton play a role in the growth and population dynamics of Daphnia, but suggested that the mineral requirements need to be met rst. Other recent experimental laboratory studies have also shown that P-limitation signicantly reduces the growth rates of the rotifers Brachionus and Anuraeopsis
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and Daphnia abundance in some lakes, which suggested that high seston C : P ratios constrained Daphnia abundance in these lakes. In contrast, they observed little relationship between seston C : P ratios and other crustacean species. However, no eld study has yet analysed the susceptibility of different rotifer taxa to P limitation. The present study investigated the impact of nutrient limitation, especially SRP availability (SRP concentration and DIN : SRP ratio), on the zooplankton community structure in a mesotrophic reservoir during a year-round study. Food concentration, phytoplankton taxonomy, temperature and hydraulic regime were also considered.

(Rothhaupt, 1995; Conde-Porcuna, 2000). Moreover, Conde-Porcuna (2000) showed that A. ssa was more sensitive than Daphnia to P-limitation and that the effect of nutrient limitation on this rotifer was stronger than that of competition from Daphnia in a laboratory experiment. These results suggest that low food quality may reduce the potential for competition between zooplankton species. Because the ratio of dissolved inorganic N to SRP (DIN : SRP), a ratio of nutrient supply, has been used as an indicator of N or P limitation for phytoplankton (Rhee, 1978; Morris & Lewis, 1988; Morales-Baquero et al., 1999), a negative relationship between this ratio and zooplankton biomass or density could be expected in natural systems. Furthermore, this negative relationship should be stronger for some rotifer species than for Daphnia because, as previously commented, some rotifers seem to be more susceptible than Daphnia to P limitation. Numerous eld studies have analysed the effects of nutrient limitation and N : P ratios on phytoplankton and bacterioplankton populations in natural situa rez-Mart nez & Cruz-Pizarro, tions (Elser et al., 1995; Pe 1995; Morales-Baquero et al., 1999; Vrede et al., 1999; Yurkovskis, Kostrichkina & Ikauniece, 1999; Gonzalez 2000). Furthermore, the inuence of the N : P ratio re-supplied by zooplankton on the cycling of N and P in aquatic systems has also been investigated (Sterner, 1990; Urabe, Nakanishi & Kawabata, 1995; Carrillo, Reche & Cruz-Pizarro, 1996). However, studies on the impact of nutrient limitation on zooplankton community structure in natural situations are very scarce (Hessen, 1992; DeMott & Gulati, 1999). In eld studies, zooplankton growth and abundance have been traditionally analysed as a function of food concentration, taxonomic composition of the phytoplankton and abiotic factors such as temperature (McCauley & Kalff, 1981; Pace, 1984; Hewitt & George, 1987; Lampert & Rothhaupt, 1991; CondePorcuna, Morales-Baquero & Cruz-Pizarro, 1994; Morales-Baquero, Conde-Porcuna & Cruz-Pizarro, 1994; Fussmann, 1996; Conde-Porcuna & Declerck, 1998). However, eld data on zooplankton taxa would be expected to show differences in their susceptibility to P limitation. Morales-Baquero & Conde-Porcuna (2000) observed that rotifers were the only zooplankton group in some oligotrophic lakes that seemed to be susceptible to P-limitation. DeMott & Gulati (1999) found negative correlation between seston C : P ratios

Methods Study site

Bermejales reservoir is a medium-size (562 ha) reservoir lying on the limestone located in an open valley area of the South of Spain. The reservoir is mesotrophic, with Secchi disc transparencies of 13 m and mean total phosphorus concentration of 18.3 lg L1. rez-Mart nez & Cruz-Pizarro (1995), According to Pe Bermejales reservoir is a P-limited system with a high N concentration throughout the year. Other limnological characteristics of the Bermejales reservoir are described in Morales-Baquero et al. rez-Mart nez & Sa nchez-Castillo (2002). (1994) and Pe

Sampling regime
Samples for measurements of chlorophyll a concentration (Chla) and nutrient concentration and density of Phytoplankton and Zooplankton were collected monthly from April 1988 to March 1989. These samples were taken from ve depths at the deepest point of the reservoir (Maximum depth: 19 M), using an 8-L Van Dorn Sampler. Water column mean values were used for analyses. Temperature was measured on each sampling day using a YSI probe (Hydrolab 4041) (Yellow Springs Instruments, Yellow Springs, OH, USA). Dissolved nutrient concentrations were analysed after ltering water through a Whatmann GF/C lter (Whatman International Ltd., Maidstone, UK). We used the following analytical methods: Murphy & Riley (1962) for SRP, the nesslerization method for ammonium
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Nutrient limitation of zooplankton in a reservoir

(APHA, 1985), Shinn (1941) for nitrite, and the brucine method for nitrate (APHA, 1985). In the present paper, the sum of the ammonium, nitrite and nitrate concentrations will be considered as DIN. Total suspended solids were measured according to APHA (1985). Food availability for zooplankton was estimated as the Chla concentration of the edible algae fraction (< 40 lm). Chlorophyll-a samples were obtained by ltering water through a 40-lm mesh screen. They were analysed spectrophotometrically on methanolic extracts, and Chla was estimated following Jones (1979). Zooplankton samples (8 L of water ltered onto a 40-lm mesh screen) were preserved in 4% formaldehyde. Zooplankton species were identied and counted under an inverted microscope. Measurements of individuals in each sample (whenever possible, 20 animals per species) were made using a calibrated ocular micrometer. Zooplankton abundances were converted to biomass using the values and formulas given in Bottrell et al. (1976). Phytoplankton samples (100 mL) were immediately xed in acetic Lugols. These samples were also counted and analysed for species identication using an inverted microscope. Thirty cells of the most common phytoplankton species and between one and thirty cells of the rest of the species were measured with an ocular micrometer for every sampling day. Biomass of each individual taxon was determined by multiplying mean cell biomass (considering 1 lm3 = 106 lg fresh weight) by cell population density. The water residence time (hydraulic regime) was estimated monthly as the ratio of water inow to mean water volume in the reservoir. The absolute population changes of each zooplankton species, temperature changes and concentration changes of Chla and SRP were calculated as the difference between each sampling interval. Statistical analyses were performed using the STATISTICA program (Statsoft, Inc. 1998). Pearson correlations were used and probability values were adjusted for the number of simultaneous tests using the sequential Bonferroni method (Rice, 1989). Multiple regression analyses were performed to compensate for covariance, using the forward stepwise selection procedure to select those variables with a signicant F-value that signicantly increase the regression sum of the squares.
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Results
The zooplankton community of the Bermejales reservoir was mainly dominated by cladocerans in terms of biomass, although rotifers were relatively important during autumn and winter (Morales-Baquero et al., 1994). The main zooplankton species were the rotifer Keratella cochlearis Gosse, the cladocerans Daphnia longispina O.F. Mu ller and Ceriodaphnia spp., and the copepod Tropocyclops prasinus Fisher. The biomass of these species was high during autumn and winter, when water residence time was low (Fig. 1). Other important zooplankton species were the rotifers A. ssa Gosse (mean biomass: 2.13 lg L1) and Synchaeta oblonga Ehrenberg (mean biomass: 1.90 lg L1). The phytoplankton community was dominated by dinoagellates and diatoms, especially Ceratium hirundinella O.F. Mu ller and Cyclotella ocellata Panto rez-Mart nez & Cruz-Pizarro, 1995). csek (Pe Regression analyses were performed using the zooplankton populations as dependent variables. Because the effects of food availability, SRP, DIN, temperature and water residence time on zooplankton biomass may be not independent, a multiple forward stepwise regression was undertaken to compensate for covariance. The result showed that SRP was the only signicant variable related to total rotifer biomass (Table 1). In relation to this, Fig. 2 depicts the temporal pattern of total rotifer biomass, SRP and Chla. Total rotifer biomass showed the same temporal pattern as SRP, whereas Chla showed a different pattern. Cladoceran and copepod biomasses were not signicantly correlated with SRP concentration or Chla (P > 0.05). Table 1 also shows that SRP was the most signicant variable (highest partial correlation) related to the biomass of Keratella and was the only signicant variable related to that of Anuraeopsis. No signicant relationships were obtained between SRP and densities of S. oblonga. For crustacean species, neither the biomass of D. longispina nor the biomass of Tropocyclops prasinus was correlated with SRP, whereas Ceriodaphnia biomass was positively correlated with SRP (Table 1). The Chla was signicantly related to the biomass of Synchaeta and Daphnia. Water residence time showed high and signicant partial correlations with Keratella and Daphnia. When all these analyses were repeated using total edible phytoplankton biomass (< 40 lm) instead of Chla as food source for zooplankton, no

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Fig. 1 Population biomass of the main zooplankton species and water residence time (Tw) during 198889 in Bermejales reservoir. Table 1 Results of forward stepwise regression analyses (n = 12). Dependent variables: total rotifer biomass, biomass of rotifer species and biomass of crustacean species. Independent variables: chlorophyll-a content (< 40 lm), SRP and DIN concentrations, water residence time (Tw), and temperature. Only those independent variables with a signicant F-value were selected in the regressions. When two variables were included in the regression, the partial correlation of each variable is shown Rotifers SRP DIN Chla Tw Temperature Adj. R2 0.90*** Keratella 0.80** Anuraeopsis 0.69* 0.75** 0.66* 0.81 0.77 0.47 0.56 0.87*** 0.93*** 0.88 Synchaeta Daphnia Ceriodaphnia 0.67* 0.64* Tropocyclops

0.58

ns = not signicant; *P < 0.05; **P < 0.01; ***P < 0.001. R2 = square multiple correlation coefcient.

signicant effects of phytoplankton biomass were observed. In order to analyse species-specic phytoplankton effects on zooplankton taxa, we repeated the previous regression analyses using the specic biomass of the dominant edible algae species (Table 2). For Keratella and Anuraeopsis, the results were similar to those obtained in Table 1. For Synchaeta and Daphnia, Oocystis spp. was the only phytoplankton taxon included in the regression. Oocystis spp. was the only

signicant variable related to Synchaeta, whereas Oocystis and water residence time were related to Daphnia (Table 2). No signicant relationships were obtained between cladoceran and rotifer species and the biomass of other edible algae, which are normally good food sources for zooplankton (e.g. Dictyosphaerium, Chrysochromulina, Rhodomonas and Cryptomonas). Chrysochromulina and Oocystis were related to T. prasinus biomass but no other variable was signicantly related to this copepod (Table 2). The alga
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Fig. 2 Total rotifer biomass, chlorophyll-a concentration (Chla) and soluble reactive phosphorus (SRP) during 198889 in Bermejales reservoir. Table 2 Results of forward stepwise regression analyses (n = 12). Dependent variables: total rotifer biomass, biomass of rotifer species and biomass of crustacean species. Independent variables: species-specic phytoplankton biomass (< 40 lm), SRP and DIN concentrations, water residence time (Tw) and temperature. Only those independent variables with a signicant F-value were selected in the regressions. When two variables were included in the regression, the partial correlation of each variable is shown Rotifers SRP DIN Cyclotella Oocystis Dictyosphaerium Cryptomonas Rhodomonas Chrysochromulina Tw Temperature Adj. R2 0.90*** Keratella 0.80** Anuraeopsis 0.69* Synchaeta Daphnia Ceriodaphnia Tropocylops

0.77**

0.72*

0.69*

0.74** 0.66* 0.81 0.77 0.47 0.59 0.87*** 0.76 0.75

ns = not signicant; *P < 0.05; **P < 0.01; ***P < 0.001. R2 = square multiple correlation coefcient.

Ceratium hirundinella, which is not edible to zooplankton, was signicantly correlated with zooplankton biomass (r = 0.89; P < 0.001). We also analysed the relationships between the DIN : SRP ratio and the biomass of the main zooplankton groups (rotifers, cladocerans and copepods).
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Because SRP concentration was below detectable levels from June to August, we considered the SRP concentration during these months as 0.1 lg L1 for the calculation of the DIN : SRP ratio. The results revealed that rotifers had a highly signicant and negative correlation with the DIN : SRP ratio (Fig. 3).

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P > 0.05). Multiple regression analyses were also performed between the number of eggs per female of rotifer species (K. cochlearis and A. ssa) and food availability, DIN : SRP ratios, temperature and water residence time. These results showed that DIN : SRP ratios and water residence time were now signicantly correlated with the fecundity of K. cochlearis (partial correlation for DIN : SRP: r = 0.77, P < 0.01; partial correlation for water residence time r = 0.85, P < 0.01). The DIN : SRP was the only signicant variable related to the fecundity of A. ssa. The fecundity of S. oblonga was not analysed because Synchaeta releases its eggs directly into the water and they sink to deeper waters where there are different conditions (low concentrations of food and low temperature) and copepod predation can produce egg mortality, decreasing population fecundity (Stemberger & Gilbert, 1987; Roche, 1990). Pearson correlations showed that population changes of Keratella and Anuraeopsis were correlated with changes in concentration of SRP (Table 3). In contrast, Synchaeta population change was correlated with changes in chlorophyll-a content (Table 3). Because Daphnia may suppress rotifers through competition in Bermejales reservoir (Conde-Porcuna et al., 1994), Daphnia population change was included in these correlations. However, no correlation was observed between Daphnia and any rotifer taxa. Figure 5 shows that the relationship between Keratella population change and SRP concentration change exhibited a higher slope than that between Anuraeopsis and SRP. This could mean that for similar changes in the SRP content, the increase in Keratella density would be higher than that in Anuraeopsis.

Fig. 3 Relationship between total rotifer biomass and the ratio DIN : SRP in Bermejales reservoir (r = 0.88; P < 0.001). Both variables were logarithmic transformed.

Neither cladocerans nor copepods were signicantly correlated with this ratio (r = 0.43 and r = 0.47 for cladocerans and copepods, respectively, P > 0.05). Similar results were obtained when these regression analyses were run excluding JuneAugust data, because of the undetectable SRP concentrations (rotifers: r = 0.78, P < 0.05; Cladocerans: r = 0.37, P > 0.05; copepods: r = 0.48, P > 0.05). In addition to these analyses, to test whether the fecundity (number of eggs per female) of rotifers could be related with P limitation, correlation analyses were performed. In the Pearson correlation analyses, fecundity of A. ssa was signicantly and inversely related to the DIN : SRP ratio (Fig. 4), whereas fecundity of K. cochlearis was not (r = 0.34;

Table 3 Correlations of rotifer species population changes with changes in temperature, SRP, edible algae and Daphnia biomass between sampling intervals (D). Edible algae (<40 lm) were correlated using biomass estimates of algae (bio) and chlorophyll-a concentrations (Chla) D Keratella D Anuraeopsis D Synchaeta D D D D D Temperature SRP edible algae (bio) edible algae (Chla) Daphnia 0.38ns 0.86*** 0.61(*) 0.28ns 0.72* 0.36ns 0.75** 0.55ns 0.15ns 0.59ns 0.13ns 0.39ns 0.28ns 0.80** 0.04ns

Fig. 4 Relationship between the fecundity of Anuraeopsis (number of eggs per female) and the ratio DIN : SRP in Bermejales reservoir (r = 0.67; P < 0.05). Both variables were logarithmic transformed. Analysis was restricted to occasions when Anuraeopsis density was > 1 ind L1.

ns = not signicant; *P < 0.05; **P < 0.01; ***P < 0.001; (*) = not signicant differences using the Bonferroni correction (only one estimate of edible algae, bio or Chla, was considered). 2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473

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Fig. 5 Relationships of rotifer species population changes with changes in SRP concentration between sampling intervals in Bermejales reservoir.

Discussion
The correlation analysis suggests that rotifers were affected by P limitation in Bermejales reservoir, whereas food availability had a minor effect. This P limitation had no apparent effect on cladocerans and crustaceans. DeMott & Gulati (1999) suggest that Daphnia abundance in several lakes was strongly constrained by the seston C : P ratio, whereas other cladoceran species were less sensitive to P limitation, as predicted by stoichiometry theory. However, no previous studies have shown the impact of nutrient limitation on rotifer populations in natural ecosystems. Morales-Baquero & Conde-Porcuna (2000) presented some evidence to support a greater susceptibility of rotifers to P limitation in oligotrophic lakes compared with crustaceans, but rotifer taxa, reproductive rates, food availability and seasonal changes were not considered. Several laboratory studies have shown that P-limited algae are poor food for Daphnia (Sommer, 1992; Sterner & Smith, 1993; Sterner et al., 1993; Van Donk & Hessen, 1993; Weers & Gulati, 1997) and for some rotifer species (Rothhaupt, 1995; Conde-Porcuna, 2000). Changes of the cell wall in P-limited algae have been related to low digestibility of algae by zooplankton (Van Donk & Hessen, 1993; Van Donk et al., 1997). Furthermore, nutrient limitation of algae has been related to fatty acid (Harrison, Thompson & Calderwood, 1990) and mineral limitations (Boersma, 2000). However, P limitation and fatty acid limitation are not mutually exclusive alternatives (Gulati & DeMott, 1997; Boersma, 2000). Direct mineral nutrient limita 2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473

tion of the rotifers caused by unfavourable cell stoichiometries of the food algae cannot be ruled out. rez-Mart nez & Cruz-Pizarro (1995) observed in Pe eld experiments that P is the limiting nutrient for phytoplankton in Bermejales reservoir. In the present study, two rotifer species seemed to be more susceptible than crustaceans to P limitation. Our results show that the fecundity of rotifer species may be affected by P limitation. These ndings are consistent with a previous laboratory study (Conde-Porcuna, 2000). The absence of a clear effect of food availability on rotifers suggests that nutrient limitation may have a greater effect than food concentration itself has on rotifer dynamics in natural systems, although rotifers may specialise in feeding on specic food classes (Edmonson, 1965). However, eld studies on rotifer dynamics have not taken into account nutrient concentrations or N : P ratios. Rotifer population dynamics are usually explained by exploitative competition, interference competition and predation (Gilbert, 1988; May & Jones, 1989; Lampert & Rothhaupt, 1991; Conde-Porcuna et al., 1994; Fussmann, 1996; CondePorcuna, 1998). Conde-Porcuna (2000) showed that food quality may inuence the strength, impact and outcome of competition and that low food quality reduces the effect of Daphnia competition. In Bermejales reservoir, there was no detectable negative relationship between Daphnia and rotifers during the present study, although other studies have shown that Daphnia suppress rotifer populations in this reservoir (Conde-Porcuna et al., 1994). However, nutrient conditions in the latter study were not considered and may have differed from those observed in the present study. High P concentration in zooplankton tissues is associated with high growth through high concentrations of RNA (Elser et al., 1996; Main, Dobberfuhl & Elser, 1997). Elser et al. (1996) showed that there can be strong ontogenetic shifts in body stoichiometry. Daphnids appear to maintain high concentrations of RNA throughout ontogeny, resulting in generally low N : P ratios at all life stages. In rotifers, the N : P ratios are not well known, although Rothhaupt (1995) showed that Brachionus has high N : P ratios. Rotifers, in contrast to crustacean zooplankton, have a xed number of nuclei after embryogenesis and this number is not subsequently altered (eutely). Morales-Baquero & Conde-Porcuna (2000) suggested that P limitation in the environment could be mainly

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bacterial food sources for these rotifer species. Nevertheless, a previous study indicated that phytoplankton are an important food source for rotifers in Bermejales reservoir (Conde-Porcuna et al., 1994). In the present study, edible algae did not increase when SRP concentrations were high. This could be the result of high grazing pressure by zooplankton that suppressed the edible phytoplankton biomass. However, the lack of clear negative relationships between zooplankton and phytoplankton in the edible size fraction suggests that phytoplankton were not strongly or consistently affected by grazing. Some algae species have very high rates of cell division (Sommer & Kilham, 1985) that may explain the lack of a signicant response despite grazing pressure, as observed by rez-Mart nez & Cruz-Pizarro (1995) in this reservoir. Pe Ceratium hirundinella was positively and signicantly correlated with zooplankton biomass. However, this positive relationship is unlikely to be the result of a positive effect of this alga on zooplankton, because it rez-Mart nez & Sa nchezis not edible. Previously, Pe Castillo (2002) observed that Ceratium density was also related to the hydraulic regime of the reservoir. Our results might be explained by changes in the hydraulic regime that caused unmeasured changes in resources that favour rotifers. As high SRP values and low DIN : SRP ratios were reached after the water residence time dropped at the end of the summer, P availability may have been related to the hydraulic regime in the reservoir. Greater water inows could increase the total suspended solids in the water of reservoirs and we found a signicant positive correlation between SRP concentration and total suspended solids in Bermejales reservoir (r = 0.58; P < 0.05). This could mean that greater water inows (short water residence time) could favour an increased SRP concentration during autumn and winter in Bermejales reservoir. However, water residence time and/or water inow in the reservoir were not signicantly associated with SRP concentration or DIN : SRP ratio (P > 0.1). Moreover, the fecundity of K. cochlearis was signicantly related to the DIN : SRP ratio regardless of the hydraulic regime (measured as the water residence time in the reservoir) according to the multiple regression analysis. In summary, P limitation may play an important role in rotifer dynamics, in addition to phytoplankton availability or possible interactions with other zoo 2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473

reected in the numbers of eutelic organisms (rotifers) because they largely allocate resources to reproduction. Our results also suggest that Keratella is more susceptible than Anuraeopsis to P limitation (Fig. 5). Moreover, Synchaeta did not show any signicant relationship with either SRP concentration or DIN : SRP ratio. Schulz & Sterner (1999) showed that the body of Bosmina has a lower percentage of P than does that of Daphnia and suggested that Bosmina has lower P requirements than Daphnia and is better able to grow with a low P diet. Indeed, the different susceptibility that rotifer species seem to show to P limitation in our study could be related to differences in body P content. The outcome of competition between zooplankton populations may shift in favour of zooplankton species with low P requirements under conditions of seston P mineral limitation. However, the susceptibility of various rotifers to P limitation needs to be tested. As an alternative to P limitation, it can be suggested that high SRP favours specic resources that are better used by rotifers. This may be related to changes in the phytoplankton community and bacteria abundance. However, C. hirundinella was the only algae species that showed a positive relationship with SRP (r = 0.69; P < 0.05), and C. hirundinella is not edible to zooplankton. Oocystis and Crysochromulina were the only edible phytoplankton taxa that showed a positive and signicant effect on some zooplankton species. Keratella and Anuraeopsis showed no relationship with any edible phytoplankton species in multiple regression analyses. Several surveys have shown that K. cochlearis and A. ssa also feed on bacteria and detritus (Pourriot, 1977; Starkweather & Bogdan, 1980; Ooms-Wilms, 1991). This could partly explain the lack of signicant relationships between phytoplankton and Keratella and Anuraeopsis in our study. Moreover, some studies have shown that bacterial production increases in response to nutrient availability (Pace & Cole, 1996) and that bacteria are one of the major regenerators of dissolved P in plankton in lakes (Hudson & Taylor, 1996). Bacteria could have higher P contents than algae in Bermejales reservoir and, because some rotifers may also feed on bacteria, they would be less affected by P limitation. Moreover, in our study, high SRP availability could also be related to high bacterial production and abundance and consequently to increased

Nutrient limitation of zooplankton in a reservoir

plankton species. Correlation analyses do not provide direct evidence for the mechanisms involved in complex eld conditions. Nevertheless, our data support the hypothesis that P limitation should be taken into account when studying the dynamics of zooplankton and interactions between populations in natural systems. Further testing is needed to verify the importance of P limitation on grazers.

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Acknowledgments
n Financial support was provided by Confederacio ca del Guadalquivir and CICYT Project Hidrogra guez. NAT91/570, and a FPI Grant to E. Ramos-Rodr We thank W.R. DeMott and an anonymous referee for substantially improving the manuscript. We also thank Mr Richard Davies for assistance with the English.

References
American public health association, American water works association and water pollution control federation. (APHA) (1985) Standard methods for the examination of water and wastewater. APHA, New york. Boersma M. (2000) The nutritional quality of P-limited algae for Daphnia. Limnology and Oceanography, 45, 11571161. Bottrell H.H., Duncan A., Gliwicz Z.M., Grygierik E., Herzig A., Hillbricht-Ilkowska A., Kurasawa H., Larsson P. & Weglenska T. (1976) A review of some problems in zooplankton production studies. Norwegian Journal of Zoology, 24, 419456. Carrillo P., Reche I. & Cruz-Pizarro L. (1996) Intraspecic stoichiometric variability and the ratio of nitrogen to phosphorus resupplied by zooplankton. Freshwater Biology, 36, 363374. Conde-Porcuna J.M. (1998) Chemical interference by Daphnia on Keratella: a life table experiment. Journal of Plankton Research, 20, 16371644. Conde-Porcuna J.M. (2000) Relative importance of competition with Daphnia (Cladocera) and nutrient limitation on Anuraeopsis (Rotifera) population dynamics in a laboratory study. Freshwater Biology, 44, 423430. Conde-Porcuna J.M. & Declerck S. (1998) Regulation of rotifer species by invertebrate predators in a hypertrophic lake: selective predation on egg-bearing females and induction of morphological defences. Journal of Plankton Research, 20, 605618. Conde-Porcuna J.M., Morales-Baquero R. & Cruz-Pizarro L. (1994) Effects of Daphnia longispina on rotifer populations in a natural environment: relative impor 2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473

tance of food limitation and interference competition. Journal of Plankton Research, 16, 691706. DeMott W.R. (1998) Utilization of a cyanobacterium and a phosphorus-decient green alga as complementary resources by daphnids. Ecology, 79, 24632481. DeMott W.R. & Gulati R.D. (1999) Phosphorus limitation in Daphnia: evidence from a long term study of three hypereutrophic Dutch lakes. Limnology and Oceanography, 44, 15571564. DeMott W.R., Gulati R.D. & Siewertsen K. (1998) Effects of phosphorus-decient diets on the carbon and phosphorus balance of Daphnia magna. Limnology and Oceanography, 43, 11471161. Edmonson W.T. (1965) Reproductive rate of planktonic rotifers as related to food and temperature in nature. Ecological Monographs, 35, 61111. Elser J.J., Dobberfuhl D.R., Mackay N.A. & Schampel J.H. (1996) Organism size, life history, and N:P stoichiometry. Bioscience, 46, 674684. Elser J.J., Lubnow F.S., Marzolf E.R., Bret M.T., Dion G. & Goldman C.R. (1995) Factors associated with interannual and intrannual variation in nutrient limitation of phytoplankton growth in Castle Lake, California. Canadian Journal of Fisheries and Aquatic Sciences, 52, 93104. Fussmann G. (1996) The importance of crustacean zooplankton in structuring rotifer and phytoplankton communities: an enclosure study. Journal of Plankton Research, 18, 18971915. Gilbert J.J. (1988) Suppression of rotifers by Daphnia: a review of the evidence, the mechanisms and the effects on zooplankton community structure. Limnology and Oceanography, 33, 12861303. Goldman J.C., McCarthy J.J. & Peavy D.G. (1979) Growth rate inuence on the chemical composition of phytoplankton in oceanic waters. Nature, 279, 210215. Gonzalez E.J. (2000) Nutrient enrichment and zooplankton effects on the phytoplankton community in microcosms from El Andino reservoir. Hydrobiologia, 434, 8196. Groeger A.W., Schram M.D. & Richard G. (1991) Inuence of food quality on growth and reproduction in Daphnia. Freshwater Biology, 26, 1119. Gulati R.D. & DeMott W.R. (1997) The role of food quality for zooplankton: remarks on the state-of-art, perspectives and priorities. Freshwater Biology, 38, 753768. Harrison P.J., Thompson P.A. & Calderwood G.S. (1990) Effects of nutrient and light limitation on the biochemical composition of phytoplankton. Journal of Applied Phycology, 2, 4556. Hessen D.O. (1992) Nutrient element limitation of zooplankton production. American Naturalist, 140, 799814.

1472

J.M. Conde-Porcuna et al.

relationship. Canadian Journal of Fisheries and Aquatic Sciences, 41, 10891096. Pace M.L. & Cole J.J. (1996) Regulation of bacteria by resources and predation tested in whole lake experiments. Limnology and Oceanography, 41, 14481460. rez-Mart nez C. & Cruz-Pizarro L. (1995) SpeciesPe specic phytoplankton responses to nutrients and zooplankton manipulations in enclosure experiments. Freshwater Biology, 33, 193203. rez-Mart nez C. & Sa nchez-Castillo P. (2002) Winter Pe population of Ceratium hirundinella in a south-temperate reservoir. Journal of Plankton Research, 24, 8996. Pourriot R. (1977) Food and feeding habits of rotifera. r Hydrobiologie Beihefte Ergebnisse der LimnolArchive fu ogie, 8, 243260. Rhee G.Y. (1978) Effects of N:P atomic ratios and nitrate limitation on algal growth, cell composition, and nitrate uptake. Limnology and Oceanography, 23, 1025. Rice W.R. (1989) Analyzing tables od statistical tests. Evolution, 43, 223225. Roche K. (1990) Some aspects of vulnerability to cyclopoid predation of zooplankton prey individuals. Hydrobiologia, 198, 153162. Rothhaupt K.O. (1995) Algal nutrient limitation affects rotifer growth rate but not ingestion rate. Limnology and Oceanography, 40, 12011208. Schulz K.L. & Sterner R.W. (1999) Phytoplankton phosphorus limitation and food quality for Bosmina. Limnology and Oceanography, 44, 15491556. Shinn J.A. (1941) A Practical Handbook of Seawater Analyses, 167 pp. Fisheries Research Board Canadian Bulletin, Canada. Sommer U. (1992) Phosphorus-limited Daphnia: intraspecic facilitation instead of competition. Limnology and Oceanography, 37, 966973. Sommer U. & Kilham S.S. (1985) Phytoplankton natural community competition experiments. A reinterpretation. Limnology and Oceanography, 30, 436440. Starkweather P.L. & Bogdan K.G. (1980) Detrital feeding in natural zooplankton communities: Discrimination between live and dead algal foods. Hydrobiologia, 73, 8385. Stemberger R.S. & Gilbert J.J. (1987) Defenses of planktonic rotifers against predators. In: Predation: Direct and Indirect Impacts on Aquatic Communities (Eds W.C. Kerfoot & A. Sih), pp. 227239. University Press of New England, Hanover and London. Sterner R.W. (1990) The ratio of nitrogen to phosphorus resupplied by herbivores: zooplankton and the algal competitive arena. The American Naturalist, 136, 209229. Sterner R.W., Hagemeier D.D., Smith W.L. & Smith R.F. (1993) Phytoplankton nutrient limitation and food quality for Daphnia. Limnology and Oceanography, 38, 857871.
2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473

Hewitt D.P. & George D.G. (1987) The population dynamics of Keratella cochlearis in a hypereutrophic tarn and the possible impact of predation by young roach. Hydrobiologia, 147, 221227. Hudson J.J. & Taylor W.D. (1996) Measuring regeneration of dissolved phosphorus in planktonic communities. Limnology and Oceanography, 41, 15601565. Jones J.G. (1979) A Guide to Methods for Estimating Microbial Numbers and Biomass in Freshwater. FBA Scientic Publication, 39, 1112. Lampert W. & Rothhaupt K.O. (1991) Alternating dynamics of rotifers and Daphnia magna in a shallow r Hydrobiologie, 121, 343353. lake. Archive fu Main T.M., Dobberfuhl D.L. & Elser J.J. (1997) N:P stoichiometry and ontogeny of crustacean zooplankton: a test of the growth rate hypothesis. Limnology and Oceanography, 42, 14741478. May L. & Jones D.H. (1989) Does interference competition from Daphnia affect populations of Keratella cochlearis in Loch Leven, Scotland? Journal of Plankton Research, 11, 445461. McCauley E. & Kalff J. (1981) Empirical relationship between phytoplankton and zooplankton biomass in lakes. Canadian Journal of Fisheries and Aquatic Sciences, 38, 458463. Morales-Baquero R., Carrillo P., Reche I. & nchez-Castillo P. (1999) Nitrogen-phosporus relaSa tionship in high mountain lakes: effects of the size of catchment basins. Canadian Journal of Fisheries and Aquatic Sciences, 56, 18091817. Morales-Baquero R., Conde-Porcuna J.M. & Cruz-Pizarro L. (1994) The zooplankton biomass and food availability in four reservoirs of contrasting trophic status. r Hydrobiologie Beihefte Ergebnisse der LimnolArchive fu ogie, 40, 161173. Morales-Baquero R. & Conde-Porcuna J.M. (2000) Effect of the catchment areas on the abundance of zooplankton in high mountain lakes of Sierra Nevada (Spain). r theoretVerhandlungen der Internationale Vereinigung fu ische und angewandte Limnologie, 27, 18041808. Morris D.P. & Lewis W.M. (1988) Phytoplankton nutrient limitation in Colorado mountain lakes. Freshwater Biology, 20, 315327. Murphy J. & Riley J.P. (1962) A modied single solution method for the determination of phosphate in natural waters. Analytica Chimica Acta, 27, 3136. Ooms-Wilms A.L. (1991) Ingestion of uorescently labelled bacteria by rotifers and cladocerans in lake Loosdrecht as measures of bacterivory: preliminary results. Memorie dellIstituto Italiano di Idrobiologia, 48, 269278. Pace M.L. (1984) Zooplankton community structure, but not biomass, inuences the phosphoruschlorophyll-a

Nutrient limitation of zooplankton in a reservoir

Sterner R.W. & Hessen D.O. (1994) Algal nutrient limitation and the nutrition of aquatic herbivores. Annual Review of Ecology and Systematics, 25, 129. Sterner R.W. & Smith R.F. (1993) Clearance, ingestion and release of N and P by Daphnia pulex feeding on Scenedesmus acutus of varying nutritional quality. Bulletin of Marine Science, 53, 228239. Urabe J., Clasen J. & Sterner R.W. (1997) Phosphorus limitation of Daphnia growth: is it real? Limnology and Oceanography, 42, 14361443. Urabe J., Nakanishi M. & Kawabata K. (1995) Contribution of metazoan plankton to the cycling of nitrogen and phosphorus in Lake Biwa. Limnology and Oceanography, 40, 232241. Van Donk E. & Hessen D.O. (1993) Grazing resistance in nutrient-stressed phytoplankton. Oecologia, 93, 508511. Van Donk E., Lu rling M., Hessen D.O. & Lokhorst G.M. (1997) Altered cell wall morphology in nutrient-de-

1473

cient phytoplankton and its impact on grazers. Limnology and Oceanography, 42, 357364. Vrede K., Vrede T., Isaksson A. & Karlsson A. (1999) Effects of nutrients (phosphorus, nitrogen, carbon) and zooplankton on bacterioplankton and phytoplankton: a seasonal study. Limnology and Oceanography, 44, 16161624. Weers P.M.M. & Gulati R.D. (1997) Growth and reproduction of Daphnia galeata in response to changes in fatty acids, phosphorus, and nitrogen in Chlamydomonas Reinhardtii. Limnology and Oceanography, 42, 15841589. Yurkovskis A., Kostrichkina E. & Ikauniece A. (1999) Seasonal succession and growth in the plankton communities of the Gulf of Riga in relation to long term nutrient dynamics. Hydrobiologia, 393, 8394. (Manuscript accepted 30 November 2001)

2002 Blackwell Science Ltd, Freshwater Biology, 47, 14631473