Experimental Studies on Zooplankton-Toxic Cyanobacteria Interactions: A Review.

Cyanobacterial blooms have been recognized as a problem in fresh water for about 150 years. Over the past 50 years, experimental studies on the subject have gained importance considering the increasing need to control toxic cyanobacterial blooms. This article presents information on the different lines of research that have been undertaken on zooplankton-cyanobacteria interactions over the past 50 years. These include information on filtering/ingestion rates and phytoplankton preferences of small and large rotifers, cladocerans, and copepods; growth rates of zooplankton on cyanobacterial diets; feeding rates of other freshwater invertebrates on cyanobacteria; role of zooplankton in top-down biomanipulation efforts; effect of cyanotoxins on zooplankton; bioaccumulation of cyanotoxins; and physical and chemical control of cyanobacterial blooms. We also highlight measures that have led to successful lake management and improvement of water quality in selected waterbodies.

Effects of the endocrine disruptor 4-nonylphenol on the demography of rotifers Plationus patulus and Brachionus havanaensis: a multigenerational study.

The emerging contaminant 4-nonylphenol is an active ingredient in many surfactants. Though 4-nonylphenol has been reported in Mexican waterbodies, no stringent law is available to establish its permissible limits. Most ecotoxicological assays using zooplankton species are based on previously unexposed populations, and multigenerational experiments are rare. Plationus patulus and Brachionus havanaensis are widely distributed species in Mexico and have been used in ecotoxicological assessments. In this work, the median lethal concentration of 4-nonylphenol (LC50, 24 h) for both rotifer species was derived. Based on our acute toxicity data (24 h LC50 of 4-nonylphenol for P. patulus and B. havanaensis were 500 and 250 µg L-1, respectively), three sublethal concentrations (µg L-1) (7.81, 15.82 and 31.25 for B. havanaensis and 15.62, 31.25 and 62.5 for P. patulus) were used in the life table demography tests for two successive generations (F0 and F1) of either rotifer species. Regardless of the concentrations used, 4-nonylphenol had an adverse effect on both rotifer species over two successive generations. P. patulus was more sensitive than B. havanaensis in chronic toxicity tests. F1 generation of both rotifer species was far more adversely affected than the F0 in all toxicant treatments.

Population responses and fatty acid profiles of Brachionus calyciflorus (Rotifera) in relation to different thermal regimes.

Diurnal temperature fluctuations affect ectothermic species more than endothermic taxa. We tested the effect of three fixed temperatures (20, 25 and 30 °C) and a 24 h variable (20-30 °C) on the population growth and fatty acid profiles of the common rotifer Brachionus calyciflorus. Depending on the temperature treatment, the peak population abundances of B. calyciflorus varied from 65 to 80 ind./ml, the lowest being on variable temperature range. The rate of population increase varied from 0.31 to 0.52 per day, highest being at 30 °C. There was a curvilinear relationship between the population density and the egg ratio (number of eggs/female) in all the tested temperature regimes. The egg ratio was higher (>0.6) for treatments involving fixed temperatures, but for variable temperature regime, the egg ratios were lower (<0.5). Temperature also induced changes in the fatty acid content of B. calyciflorus. While the total saturated fatty acids increased, both mono-unsaturated and poly-unsaturated fatty acids decreased with increasing temperature regime. These results have been interpreted in relation to the role of temperature (both intensity and mode of exposure) on the population growth characteristics of rotifers.

A Long-Term Study on the Effect of Cyanobacterial Crude Extracts from Lake Chapultepec (Mexico City) on Selected Zooplankton Species.

Many urban lakes in Mexico City such as Lake Chapultepec are infested with high densities of cyanobacteria, particularly Microcystis. We tested the effect of cyanotoxins from cyanobacterial crude extracts on the demographic variables of zooplankton. The rotifers Brachionus havanaensis and Brachionus calyciflorus, and the cladocerans Ceriodaphnia dubia and Moina macrocopa were used for the assays. Temperature effects on the response of B. calyciflorus and 2 clones of M. macrocopa were tested. We hypothesized that with an increase in cyanotoxin concentration and temperature there would be an increase in the adverse effect on the test species and that the clone of Moina previously exposed to cyanobacteria from Lake Chapultepec would be more resistant to the cyanotoxins. Demography experiments showed that B. havanaensis was more sensitive than C. dubia. The negative effect of the cyanobacterial crude extract on B. calyciflorus was greater at 30 °C than at 20 °C or 25 °C. The strain of M. macrocopa isolated from Lake Chapultepec was more resistant to the cyanotoxins than the strain that had not been previously exposed to the cyanobacteria. The present study indicated that cyanobacteria in Lake Chapultepec are highly toxic and, considering the recreational use of this lake, should be controlled. Environ Toxicol Chem 2020;39:2409-2419. © 2020 SETAC.

Effect of salinity and temperature on the acute and chronic toxicity of arsenic to the marine rotifers Proales similis and Brachionus ibericus.

Several abiotic factors influence the ecological responses of aquatic invertebrates to metal toxicity. We examined the effect of salinity (10, 20, and 30 psu) and temperature (25 and 32 °C) on acute and chronic arsenic (As) toxicity to the euryhaline rotifers, Proales similis and Brachionus ibericus. In general, higher salinities and low temperature resulted in lower arsenic toxicity. The population growth studies indicated that P. similis was more sensitive than B. ibericus to As. Arsenic toxicity intensified the vulnerability of P. similis to B. ibericus competition. Life table parameters decreased with increasing As levels in the medium. Chronic toxicity bioassays were more sensitive than acute toxicity tests for determining the adverse effect of As to rotifers. Our findings provide useful insights on the effect of arsenic on rotifer populations exposed to different temperature and salinity scenarios. Proales similis could be an important complement to brachionid rotifers for marine toxicity bioassays.

Toxicity of cyanobacterial blooms from the reservoir Valle de Bravo (Mexico): A case study on the rotifer Brachionus calyciflorus.

Proliferating cyanobacterial blooms due eutrophication in reservoirs is a major global problem. The production of cyanotoxins often increases with grazing pressure and temperature while the sensitivity of zooplankton to cyanotoxins is directly related to temperature. Here we evaluate the effect of different concentrations of the crude extract of cyanobacteria from Valle de Bravo reservoir during dry (January) and rainy (September) seasons at 20 and 25 °C on the rotifer Brachionus calyciflorus based on acute and chronic toxicity tests. We filtered 20 or 150 l of lake water, depending on the intensity of the bloom, and estimated the density and diversity of the cyanobacteria. The crude extracts, after 5 cycles of freezing, thawing and sonication at 14 MHz, were filtered and the microcystin concentration quantified based on ELISA. The extracts were used to conduct the acute and chronic toxicity tests, all in quadruplicate. Acute toxicity tests were based on 24 h mortality. Chronic toxicity tests (population growth and life table experiments) were conducted at 5 and 10% of the median lethal concentration. The field samples were dominated by Microcystis sp. (January) or Woronichinia naegeliana (September). The microcystin concentration in lake water was 9.57 µg/l and 0.097 µg/l and the median lethal concentration was 5.34 µg microcystin/L and 0.35 µg microcystin/L in January and September, respectively. Survival and reproduction of B. calyciflorus were lower in the presence of the cyanobacteria crude extract, more so at 20° than at 25 °C. Our results highlight the urgency of regular monitoring based on zooplankton assays for reservoirs in tropical and temperate regions, subject to frequent and dominant cyanobacterial blooms, often as a result of climate change.

Demographic responses of Cladocerans (Cladocera) in relation to different concentrations of humic substances.

Cladocerans are constantly exposed to humic substances in nature, yet the effects of these substances on their survival and reproduction are not well known. Here, the effects of humic substances (20 and 40 mg L-1) (HS) on the life history variables of three common cladocerans, Ceriodaphnia dubia, Moina macrocopa, and Daphnia pulex were evaluated. The results showed that the effect of humic substances on the tested cladocerans is species-specific, affecting either survival, reproduction or both. For M. macrocopa, exposed to HS at a concentration of 40 mg L-1, the average lifespan and the life expectancy at birth were significantly reduced as compared to controls, but for C. dubia and D. pulex these parameters were increased. Gross reproductive rate was unaffected by the HS level for both D. pulex and M. macrocopa, but it was significantly higher for C. dubia. When compared to the corresponding controls, for HS-exposed cladocerans, the rate of population increase was significantly reduced in case of D. pulex while it was stimulated for both C. dubia and M. macrocopa. It appears that humic substances had a slightly stronger influence on survivorship than on reproduction of the tested cladocerans.

Bioaccumulation of microcystins in seston, zooplankton and fish: A case study in Lake Zumpango, Mexico.

Cyanotoxins from toxic blooms in lakes or eutrophic reservoirs are harmful to several organisms including zooplankton, which often act as vectors of these secondary metabolites, because they consume cyanobacteria, bioaccumulate the cyanotoxins and pass them on along the food chain. Microcystins are among the most commonly found cyanotoxins and often cause zooplankton mortality. Although cyanobacterial blooms are common and persistent in Mexican water bodies, information on the bioaccumulation of cyanotoxins is scarce. In this study we present data on the bioaccumulation of cyanotoxins from Planktothrix agardhii, Microcystis sp., Cylindrospermopsis raciborskii and Dolichospermum planctonicum blooms in the seston (suspended particulate matter more than 1.2 µm) by zooplankton and fish (tilapia (Oreochromis niloticus) and mesa silverside (Chirostoma jordani) samples from Lake Zumpango (Mexico City). The cyanotoxins were extracted from the seston, zooplankton and fish tissue by disintegration using mechanical homogenization and 75% methanol. After extraction, microcystins were measured using an ELISA kit (Envirologix). Concentration of microcystins expressed as equivalents, reached a maximum value of 117 µg g-1 on sestonic samples; in zooplankton they were in the range of 0.0070-0.29 µg g-1. The dominant zooplankton taxa included Acanthocyclops americanus copepodites, Daphnia laevis and Bosmina longirostris. Our results indicate twice the permissible limits of microcystins (0.04 µg kg-1 d-1) for consumption of cyanobacterial products in whole fish tissue of Chirostoma jordani. The data have been discussed with emphasis on the importance of regular monitoring of water bodies in Mexico to test the ecotoxicological impacts of cyanobacterial blooms and the risk that consumption of products with microcystins could promote.

Combined effects of temperature and salinity on the demographic response of Proales similis (Beauchamp, 1907) and Brachionus plicatilis (Müller, 1786) (Rotifera) to mercury.

The demographic response of the brackish-water rotifers Proales similis and Brachionus plicatilis to mercury (0.5, 2, 8 and 32 µg L-1 of HgCl2) at different salinity levels (10 and 20‰) and two temperature (25 °C and 32 °C) regimes were evaluated. Median lethal concentration (LC50) for P. similis and B. plicatilis was 10 and 16 µg L-1, respectively, showing that Proales similis was more sensitive to mercury than B. plicatilis. The rate of population increase (r) for both species was greater at 10‰ salinity and 32 °C (ranged from 0.6 to 0.95 d-1). The r-value decreased as the concentration of mercury in the medium increased. Regardless of the temperature, at lower salinity and higher mercury concentration (32 µg L-1), P. similis died within six days. The survivorship of P. similis and B. plicatilis was higher at 25 °C than at 32 °C (ranged from 5 to 8 and 7-13 d, respectively). Fecundity was higher at 32 °C than at 25 °C for both rotifers species. There was a significant effect of the interaction among salinity, temperature, and mercury in both species on the reproductive variables such as net and gross reproductive rates, generation time and the rate of population increase. Considering the sensitivity of P. similis, we suggest that this species be included in the list of marine bioassay organisms.

Multigenerational effects of triclosan on the demography of Plationus patulus and Brachionus havanaensis (ROTIFERA).

Triclosan is a personal care product widely used in North America, Europe and Asia as antimicrobial ingredient in many consumer chemical products. In Mexico concentrations of triclosan have been reported in aquatic systems. However, there is no law regulating the presence of chemicals such as triclosan, in aquatic systems. The scarce data about this chemical has increased concern among ecotoxicologists regarding possible effects on aquatic organisms. Moreover, multigenerational studies are rarely studied and the results vary depending on the contaminant. Rotifers, are a dominant group of zooplankton, and have been used in aquatic risk assessments of personal care products due to their sensitivity and high reproductive rates. Plationus patulus and Brachionus havanaensis are common rotifers distributed in aquatic ecosystems of Mexico and have been used in ecotoxicological bioassays. In this study, the median lethal concentration (LC50, 24h) of P. patulus and B. havanaensis exposed to triclosan was determined. Based on the LC50, we tested three sublethal concentrations of triclosan to quantify the demographic responses of both rotifers for two successive generations (F0, and F1). The 24h LC50 of triclosan for P. patulus and B. havanaensis were 300 and 500µgL-1 respectively. Despite the concentration, triclosan had an adverse effect on both Plationus patulus and Brachionus havanaensis in both generations exposed. Experiments show that P. patulus was more sensitive than B. havanaensis when exposed to triclosan. When exposed to triclosan the parental generation (F0) of P. patulus was far more affected than F1.

Effect of crude extracts from cyanobacterial blooms in Lake Texcoco (Mexico) on the population growth of Brachionus calyciflorus (Rotifera).

Unlike temperate regions, tropical ecosystems are characterized by high temperatures (>18 °C) all year, promoting blooms of cyanobacteria which often produce secondary metabolites toxic to zooplankton. Nabor Carillo and the Recreational Lake are part of the saline, Lake Texcoco, in Central Mexico which is filled nowadays with treated waste water. Both water bodies are dominated by Planktothrix, Anabaenopsis, Spirulina and Microcystis. In this study we present the concentration of microcystins in these waterbodies over an annual cycle. We also evaluated the chronic effects of cyanobacterial crude extracts from both lakes on two clones of the rotifer Brachionus calyciflorus, one from Nabor Carrillo Lake and the other from a canal in the shallow, Lake Xochimilco. The experiments on population growth were performed, beginning with 10 individuals per container for each of the following treatments: control (no crude extract), concentrated crude extract, and diluted crude extract (50:50) with moderately hard water and Chlorella vulgaris in a concentration of 0.5 × 106 cells ml-1. The cyanotoxin levels were measured using an ELISA test and ranged between 0.20 and 2.4 µg L-1 in the lake water. The results showed that the Recreational Lake extracts were more toxic, killing the rotifers in less than five days. The r values ranged from -1.74 to 0.48 in the presence of the crude extracts and 0.16 and 0.24 in the controls. The results have been discussed with emphasis on the importance of conducting regular studies to test ecotoxicological impacts of cyanobacterial blooms in tropical waters.

Four Transgenerational Demographic Performance of Moina macrocopa Exposed to Chronic Levels of Cadmium.

In this study, we quantified intergenerational, demographic variability of Moina macrocopa subjected to cadmium stress. Exposure of M macrocopa to cadmium (0.2, 0.3, and 0.4 mg/L as CdCl2) through 4 consecutive generations revealed changes in demographic responses not only in survivorship variables but also in reproductive parameters. Long-term demographic responses varied differently, depending on the demographic trait and the concentration of heavy metal in the medium. With the exception of generation time, all life history traits were significantly and adversely influenced due to increase in Cd concentrations. The average life span of M macrocopa varied up to 40% depending on Cd level and the generation of exposure. The highest gross reproductive rates were recorded in controls, while the lowest (∼30% less) were recorded at the highest Cd level. Survival-weighted net reproductive rates were reduced by nearly 50% due to Cd toxicity. The rate of population increase per day of M macrocopa was also significantly affected (∼25%) by Cd as compared to controls. This cladoceran showed a dose-response to Cd toxicity with a significance in both magnitude and frequency of offspring production.

Is Aluminum Innocuous to Zooplankton at pH Below 6?

Aluminum (Al) use has increased greatly during the last two decades, yet little information is available on its toxic effects in relation to pH particularly on zooplankton. In this work, we determined the acute toxicity (LC50) and life table responses for Moina micrura exposed to 0.008, 0.016 and 0.08 mg of Al L-1 at pH of 5, 6 and 7. The age-specific survivorship and reproduction showed a steep decline (80% mortality by the second day) at pH 5, independent of Al level. Both gross and net reproductive rates were significantly lower at pH 6 compared to pH 7, regardless of Al concentration. At pH 7 the rate of population increase of M. micrura was not significantly influenced by the Al level, while at pH 6 it was significantly lower (p < 0.05), suggesting that M. micrura is sensitive to changes in Al under slightly acidic conditions.

Temperature and age affect the life history characteristics and fatty acid profiles of Moina macrocopa (Cladocera).

Demographic responses and fatty acid profiles of Moina macrocopa were quantified under different temperature regimes (20°C, 25°C and 30°C and diurnally variable 20-30°C) and at fixed ration (10.65µgDWml(-1)) of Chlorella. Highest constant temperature (30°C) reduced the density of M. macrocopa. The cladocerans under the fluctuating temperature regime too had lower population growth (about 50% lower than that at constant 25°C). The survivorship of M. macrocopa was higher at 20°C than that at 25°C and 30°C or at variable temperature regime. Gross and net reproductive rates were higher at 25°C. At 20°C, neonates had the highest proportion (67%) of myristic, palmitic and stearic acids while the adults had the lowest (26%) proportion. For both adults and neonates, palmitoleic, linoleic and linolenic comprised of 15-35% of the total fatty acids. Higher percentage (19%) of linoleic acid was present in adults than neonates (7%). Adults had linolenic acid level which was 3-times higher than in neonates. Linoleic and linolenic fatty acids decreased with increasing temperature for neonates and adults from 20°C to 30°C. The demographic responses and fatty acid profiles of M. macrocopa were discussed in relation to level and mode of temperature exposure.

Effect of crude extracts of Dolichospermum planctonicum on the demography of Plationus patulus (Rotifera) and Ceriodaphnia cornuta (Cladocera).

In this study we evaluated the toxicological effects of crude extracts of Dolichospermum planctonicum on Plationus patulus (Rotifera) and Ceriodaphnia cornuta (Cladocera). The active compound from the laboratory-cultured cyanobacteria was extracted by lyophilizing it at -70 °C for 48 h, after which the sample was thawed. The cycle was repeated five times, and between each cycle the cells were sonicated to lyse them. Using the extract of Anabaena, experiments of population growth and life table demography of the two zooplankton species were compared in media with and without (controls) the cyanotoxins. In both experiments two toxic concentrations diluted in US EPA medium were used, one in a 5:15 proportion, and the other in a 2.5:17.5. We found that the rotifers were more resistant to the toxicants than the cladocerans. Their population growth rates ranged from 0.11 d(-1) in controls to 0.08 d(-1) in the presence of the crude extracts for P. patulus and from 0.18 in controls to -0.27 in the presence of the crude extracts for C. cornuta. The crude extract also had a significant negative impact on the demographic parameters, adversely affecting reproduction more than survival. The data have been discussed with emphasis on the importance of further research using cyanobacterial crude extracts.

Feeding and filtration rates of zooplankton (rotifers and cladocerans) fed toxic cyanobacterium (Microcystis aeruginosa).

Microcystis aeruginosa is generally dominant in many Mexican freshwater ecosystems interacting with zooplankton species. Hence, feeding and filtration rates were quantified for three cladoceran (Daphnia pulex, Moina micrura and Ceriodaphnia dubia) and three rotifer species (Brachionus calyciflorus, Brachionus rubens and Plationus patulus) using sonicated M. aeruginosa alone or mixed with Scenedesmus acutus in different proportions (25, 50 and 75%, based on cell density), offering a combined initial density of 100,000 cells·ml(-1). All the three cladoceran species ingested M. aeruginosa (100-300 cells ind(-1) min(-1)) when fed exclusively with cyanobacterium. When green alga offered as exclusive diet, the number of cells ingested by the tested cladocerans varied from 80 to 400 cells ind(-1) min(-1). Compared to cladocerans, rotifers in general consumed much lower quantity (< 200 cells ind(-1) min(-1)) of M. aeruginosa and S. acutus. The filtration rate for Daphnia pulex was inversely related to the proportion of green alga in the diet. For other tested cladocerans, no such clear trend was evident. In mixed treatments containing M. aeruginosa, the filtration rate of Daphnia was highest (about 220 µl ind(-1) min(-1)) when the medium contained 75% of S. acutus. Among the rotifer species, P. patulus filtered highest volume (100 µl ind(-1) min(-1) from mixed diets containing higher proportions (50 or 75%) of M. aeruginosa. Thus, there were species-specific differences in the filtration and feeding rates of zooplankton when offered mixed diets of green algae and toxic cyanobacteria. These probably explain the coexistence of different zooplankton species in Microcystis-dominant waterbodies.

Effect of malathion on the demography of Daphnia pulex Leydig and Diaphanosoma birgei Korinek (Cladocera).

Malathion is a common pesticide used to control insects in agricultural, domestic and industrial sectors in different parts of the world. In this work we evaluated the effects of sublethal concentrations of malathion on the survivorship and reproductive variables of two cladoceran species Daphnia pulex and Diaphanosoma birgei using standard life table demography method. Based on preliminary tests, we selected four sublethal concentrations of malathion for each cladoceran species. For D. pulex, the malathion concentrations were 0. 225, 0.45, 0.9 and 1.8ng I(-1) and for D. birgei, these were 0.0281, 0.0562, 0.1125, 0.225ng I(-1). Our results showed that in general, Daphnia pulex was less sensitive than Diaphanosoma birgei to malathion. The average lifespan of Daphnia pulex in controls was about 19 days while under similar conditions, that of D. birgei was about 21 days. For either cladoceran species, increased pesticide concentration resulted in decreased survival; this was more evident in the treatment containing the highest concentration of malathion (0.225 ng l(-1)) for D. birgei. Fecundity of D. pulex and D. birgei also decreased with increase in the concentration of malathion. For a given cladoceran species, compared to controls, the gross reproductive, net reproductive rates and the population growth rate significantly decreased due to malathion. Compared to the survivorship variables (age-specific survival, life expectancy and average lifespan), the reproductive parameters (gross reproductive rate, net reproductive rate and the rate of population increase) of the two cladoceran species were decreased by the pesticide.

Effect of paracetamol and diclofenac on population growth of Plationus patulus and Moina macrocopa.

Two non-steroidal anti-inflammatory commonly used in Mexico are paracetamol and diclofenac. These drugs reach natural waterbodies transport through the hydrological areas of agriculture and effluent treatment plants and can affect plankton. In this study, we quantified the chronic effects of paracetamol and sodium diclofenac on the population dynamics of the rotifer Plationus patulus and the cladoceran Moina macrocopa under laboratory conditions. Both these species of zooplankton are common in freshwaters and are sensitive to chemicals in the medium. Our results showed that densities of both P. patulus and M. macrocopa decreased with increasing levels of both the analgesic drugs. Both zooplankton species did not survive beyond when paracetamol was applied at 32 mg I(-1) in the medium. Diclofenac in general had more adverse effect than paracetamol for either zooplankton species. P. patulus was more sensitive than M. macrocopa to both analgesic drugs. When diclofenac was present in the medium at > or = 12.5 mg I(-1), rotifer reproduction was inhibited, while the tested cladocerans continued to grow but to lower densities compared to control. The rate of population increase (r) per day of P. patulus and M. macrocopa was significantly and inversely related to the concentration of paracetamol and diclofenac in the medium. However, the relationship between r and drug concentration differed depending on the zooplankton species and drug. In controls, the r of P. patulus was 0.18 d(-1), for M. macrocopa under similar conditions, it was slightly lower (0.16 d(-1)). The rvalues of both zooplankton populations became negative (-0.10 to -0.15 d(-1)) when exposed to paracetamol at 32 mg l(-1) or diclofenac at 25 mg I(-1).

Morphological, morphometrical and molecular (CO1 and ITS) analysis of the rotifer Asplanchna brightwellii from selected freshwater bodies in Central Mexico.

We evaluated different strains of the rotifer Asplanchna brightwellii collected from central Mexico using morphology, morphometry and molecular tools (CO1 and ITS). Three distinct clonal populations from each of the 3 regions (Mexico City, State of Mexico and State of Guerrero) were established under laboratory conditions. For a given waterbody, morphometric comparisons within the populations of A. brightwellii showed almost stable measurements of trophi and with no statistically significant differences among them (p > 0.05). However, asplanchnid body length and width as well as the cyst diameter varied significantly depending on the waterbody from which A. brightwellii was collected. The smallest adults (about 700 microm) were from Valerio Trujano lake (Guerrero State) samples while the largest were from Xochimilco lake. Similar tendencies were reflected in the diameter of resting eggs. In addition, morphologically the cysts of A. brightwellii from the three waterbodies showed slightly different pattern. The number of globular structures on the surface of cysts was smaller for Valerio Trujano strain, while these were larger and less numerous for both Xochimilco and Zumpango strains. The ITS region tree displayed two groups Xochimilco and Valerio Trujano -Zumpango, this analysis did not reflect the morphological grouping; on the contrary the CO1 gene tree separated the populations according to morphological clusters and location (Xochimilco, Valerio Trujano and Zumpango lakes). When the tree was built using the combination of both ITS and CO1 sequences, the phylogenetic relationships observed on CO1 gene were consistent; but showed differences with the relationships observed on ITS region tree (only two groups).

Demography of zooplankton (Anuraeopsis fissa, Brachionus rubens and Moina macrocopa) fed Chlorella vulgaris and Scenedesmus acutus cultured on different media.

Generally zooplankton growth is often limited by the quality of their algal diet. A cheaper common practice in aquaculture, is to culture algae with fertilizers; however, the demography of zooplankton when fed these algae has not yet been evaluated. We studied the population growth and life table demography of the rotifers Anuraeopsis fissa and Brachionus rubens, and the cladoceran Moina macrocopa. For this, the algae Scenedesmus acutus or Chlorella vulgaris were cultured on defined (Bold's basal) medium or the commercial liquid fertilizer (Bayfolan). Experiments were conducted at one algal concentration 1.0 x 10(6) cells/mL of C. vulgaris or its equivalent dry weight of 0.5 x 10(6) cells/mL of S. acutus. The population dynamics were tested at 23 +/- 1 degrees C in 100 mL transparent jars, each with 50mL of the test medium, with an initial density of 0.5indiv/mL, for a total of 48 test jars (3 zooplankton 2 algal species x 2 culture media x 4 replicates). For the life table experiments with M. macrocopa, we introduced 10 neonates (<24h old) into each test jar containing the specific algal type and concentration. For the rotifer experiments, we set 5mL tubes with one neonate each and 10 replicates for each algal species and culture medium. We found that the average rotifer life span was not influenced by the diet, but for M. macrocopa fed S. acutus cultured in Bold's medium, the average lifespan was significantly lower than with the other diets. The gross and net reproductive rates of A. fissa (ranging from 18-36 offspring per female) were significantly higher for C vulgaris cultured in Bold medium. Regardless of the culture medium, Chlorella resulted in significantly higher gross and net reproductive rates for B. rubens than S. acutus diets. The reproductive rates of M. macrocopa were significantly higher in all the tested diets except when fed with S. acutus in Bold medium. The population increase rate, derived from growth experiments of A. fissa and B. rubens, ranged from 0.1-0.25/d and were significantly higher on C vulgaris cultured in liquid fertilizer as compared to the other diets. The growth rates of M. macrocopa ranged from 0.1 to 0.38/d, and were highest with diets of C. vulgaris cultured in Bold medium and S. acutus cultured in fertilizer. Thus, regardless of the culture medium used, the growth rates of the evaluated zooplankton species were higher with Chlorella than with Scenedesmus. The peak population density was highest (2 800ind/mL) for A. fissa fed Chlorella that was cultured on liquid fertilizers, while B. rubens and M. macrocopa had peak abundances of 480 and 12ind/mL, respectively under similar conditions.