Grazing pressure-induced shift in planktonic bacterial communities with the dominance of acIII-A1 actinobacterial lineage in soda pans.

Astatic soda pans of the Pannonian Steppe are unique environments with respect to their multiple extreme physical and chemical characteristics (high daily water temperature fluctuation, high turbidity, alkaline pH, salinity, polyhumic organic carbon concentration, hypertrophic state and special ionic composition). However, little is known about the seasonal dynamics of the bacterial communities inhabiting these lakes and the role of environmental factors that have the main impact on their structure. Therefore, two soda pans were sampled monthly between April 2013 and July 2014 to reveal changes in the planktonic community. By late spring in both years, a sudden shift in the community structure was observed, the previous algae-associated bacterial communities had collapsed, resulting the highest ratio of Actinobacteria within the bacterioplankton (89%, with the dominance of acIII-A1 lineage) ever reported in the literature. Before these peaks, an extremely high abundance (> 10,000 individuum l-1) of microcrustaceans (Moina brachiata and Arctodiaptomus spinosus) was observed. OTU-based statistical approaches showed that in addition to algal blooms and water-level fluctuations, zooplankton densities had the strongest effect on the composition of bacterial communities. In these extreme environments, this implies a surprisingly strong, community-shaping top-down role of microcrustacean grazers.

Parasite transmission in a natural multihost-multiparasite community.

Understanding the transmission and dynamics of infectious diseases in natural communities requires understanding the extent to which the ecology, evolution and epidemiology of those diseases are shaped by alternative hosts. We performed laboratory experiments to test how parasite spillover affected traits associated with transmission in two co-occurring parasites: the bacterium Pasteuria ramosa and the fungus Metschnikowia bicuspidata Both parasites were capable of transmission from the reservoir host (Daphnia dentifera) to the spillover host (Ceriodaphnia dubia), but this occurred at a much higher rate for the fungus than the bacterium. We quantified transmission potential by combining information on parasite transmission and growth rate, and used this to compare parasite fitness in the two host species. For both parasites, transmission potential was lower in the spillover host. For the bacterium, virulence was higher in the spillover host. Transmission back to the original host was high for both parasites, with spillover influencing transmission rate of the fungus but not the bacterium. Thus, while inferior, the spillover host is not a dead-end for either parasite. Overall, our results demonstrate that the presence of multiple hosts in a community can have important consequences for disease transmission, and host and parasite fitness.This article is part of the themed issue 'Opening the black box: re-examining the ecology and evolution of parasite transmission'.

Rapid evolution of tolerance to toxic Microcystis in two cladoceran grazers.

Evolutionary adaptation could assist organisms to cope with environmental changes, yet few experimental systems allow us to directly track evolutionary trajectory. Using experimental evolution, evolutionary tolerance to Microcystis aeruginosa was investigated in two cladocerans (Daphnia pulex and Simocephalus vetulus) to test the hypothesis that cladoceran grazers rapidly adapt to toxic cyanobacteria. After exposure for either three or six months, both grazers evolved a higher tolerance. The intrinsic rate of population increases in S. vetulus feeding on cyanobacteria was negatively correlated with that on green algae, which suggests that evolutionary adaptation in tolerance would carry a cost in the absence of cyanobacteria. However, the cyanobacterial selection resulted in a general increase in D. pulex when fed both cyanobacteria and green algae. Following a three-month relaxation of selection, S. vetulus in the selection line exhibited reverse evolution back to their original state when their diets were switched back to pure green algae. The present experimental evolution, both forwards and reverse, not only demonstrates the evolutionary responses of cladoceran grazers to toxic cyanobacterial cells in the laboratory, but also indicates that the grazer-cyanobacteria interaction would be an effective system to empirically study rapid evolution to environmental changes.

Effects of phytoplankton extracts containing the toxin microcystin-LR on the survival and reproduction of cladocerans.

The use of ecotoxicological techniques for the evaluation of the quality of limnetic waters allows the early detection of toxic agents that pose risks to human health. In this study Moina micrura (two clones), Daphnia laevis (two clones) and Daphnia similis, a temperate species, were used to evaluate the toxicity of six Microcystis extracts from two Colombian reservoirs. Toxin was detected and quantified by HPLC. Microcystin-LR was found in all extracts with the highest concentrations in one sample from each reservoir (434 µg g(-1) and 538 µg g(-1)). The extracts that had the highest toxin concentration also had the highest toxicities to cladocerans. Measurement of 48-h LC50 showed consistent differences between cladoceran species but not clones, Also, reproduction data in two species were consistent with the MC-LR content of one sample tested, with decreased reproduction and disruption of egg production. However, only some growth results of neonates exposed to extracts were consistent with the acute response. In conclusion, Daphnia species are a good model for monitoring cyanotoxins as they respond in a sensitive way to natural phytoplankton samples containing microcystin-LR.

Distribution, diversity and density of wolbachial infections in cladocerans and copepods from Thailand.

Species of the genus Wolbachia comprise a group of Rickettsia-like, maternally-inherited bacteria that cause several reproductive alterations in arthropod hosts. The best known are cytoplasmic incompatibility and feminization. Here, the first systematic surveys of wolbachial infections in cladocerans and copepods from six geographic regions of Thailand, including Northern, Northeastern, Western, Central, Eastern and Southern are reported. Using gene amplification assays with wsp and groE primers, wolbachiae were detected in 239 (4 spp.) of 1885 (57 spp.) copepods and cladocerans from all regions of Thailand surveyed. Screening results obtained with wsp primers or groE primers were similar in all cases. The presence of wolbachiae was only detected in copepods, not in cladocerans. Sex ratio analyses of the progeny of two species of copepods, Mesocyclops aspericornis and Mesocyclops thermocyclopoides, naturally or artificially infected with wolbachiae showed infection causes feminization (female-bias). The relative density if infection in naturally infected populations of three copepod species, M. thermocyclopoides, Heliodiaptomus elegans and Neodiaptomus blachei, were determined using real-time quantitative PCR assay based on the wsp gene. The density of wolbachiae in M. thermocyclopoides was significantly higher than in the other two species. These results suggest that wolbachial infections are distributed throughout Thailand, and that possibly the natural occurrence of these in copepods may be due to their predation on mosquito larvae. This apparent novel biology may have importance as a genetic drive system for control of vector borne diseases in the future.

Clonal variation in growth plasticity within a Bosmina longirostris population: the potential for resistance to toxic cyanobacteria.

Many aquatic organisms respond phenotypically, through morphological, behavioral, and physiological plasticity, to environmental changes. The small-size cladoceran Bosminalongirostris, a dominant zooplankter in eutrophic waters, displayed reduced growth rates in response to the presence of a toxic cyanobacterium, Microcystisaeruginosa, in their diets. The magnitude of growth reduction differed among 15 clones recently isolated from a single population. A significant interaction between clone and food type indicated a genetic basis for the difference in growth plasticity. The variation in phenotypic plasticity was visualized by plotting reaction norms with two diets. The resistance of each clone to dietary cyanobacteria was measured as the relative change in growth rates on the "poor" diet compared with the "good" diet. The enhanced resistance to M. aeruginosa in B. longirostris was derived from both the reduced slope of reaction norms and the increased mean growth rates with two diets. The large clonal variation within a B. longirostris population may contribute to local adaptation to toxic cyanobacteria and influence ecosystem function via clonal succession.

Interaction of Vibrio cholerae non-O1/non-O139 with copepods, cladocerans and competing bacteria in the large alkaline lake Neusiedler See, Austria.

Vibrio cholerae is a human pathogen and natural inhabitant of aquatic environments. Serogroups O1/O139 have been associated with epidemic cholera, while non-O1/non-O139 serogroups usually cause human disease other than classical cholera. V. cholerae non-O1/non-O139 from the Neusiedler See, a large Central European lake, have caused ear and wound infections, including one case of fatal septicaemia. Recent investigations demonstrated rapid planktonic growth of V. cholerae non-O1/non-O139 and correlation with zooplankton biomass. The aim of this study was to elucidate the interaction of autochthonous V. cholerae with two dominant crustacean zooplankton species in the lake and investigate the influence of the natural bacterial community on this interaction. An existing data set was evaluated for statistical relationships between zooplankton species and V. cholerae and co-culture experiments were performed in the laboratory. A new fluorescence in situ hybridisation protocol was applied for quantification of V. cholerae non-O1/non-O139 cells, which significantly reduced analysis time. The experiments clearly demonstrated a significant relationship of autochthonous V. cholerae non-O1/non-O139 with cladocerans by promoting growth of V. cholerae non-O1/non-O139 in the water and on the surfaces of the cladocerans. In contrast, copepods had a negative effect on the growth of V. cholerae non-O1/non-O139 via competing bacteria from their surfaces. Thus, beside other known factors, biofilm formation by V. cholerae on crustacean zooplankton appears to be zooplankton taxon specific and may be controlled by the natural bacterial community.

[Changes in the behaviour of Moina macrocopa (Crustacea: Cladocera) under the influence of Gurleya sp. (Microsporidia: Gurleyidae)].

It is established, that infestation with Gurleya sp. (Microsporidia, Gurleyidae) changes the behaviour of its host Moina macrocopa (Straus, 1820), a crustacean inhabiting pools. Heavily infested host individuals attach to surface film of water before their death by senescence. As a consequence, their dead bodies filled with spores of the parasite adhere to objects dipped in water that probably facilitates the transfer of spores by birds and cattle to other pools. Weakly infested or uninfested individuals drown after the end of their ontogenesis. Distribution of the microsporidian spores by two different ways (with attached dead bodies of the hosts or with bottom sediments) makes invasion of a new host more probable.

Temporal variations in the dynamics of potentially microcystin-producing strains in a bloom-forming Planktothrix agardhii (Cyanobacterium) population.

The concentration of microcystins (MCs) produced during blooms depends on variations in both the proportion of strains containing the genes involved in MC production and the MC cell quota (the ratio between the MC concentration and the density of cells with the mcyA genotype) for toxic strains. In order to assess the dynamics of MC-producing and non-MC-producing strains and to identify the impact of environmental factors on the relative proportions of these two subpopulations, we performed a 2-year survey of a perennial bloom of Planktothrix agardhii (cyanobacteria). Applying quantitative real-time PCR to the mcyA and phycocyanin genes, we found that the proportion of cells with the mcyA genotype varied considerably over time (ranging from 30 to 80% of the population). The changes in the proportion of cells with the mcyA genotype appeared to be inversely correlated to changes in the density of P. agardhii cells and also, to a lesser extent, to the availability of certain nutrients and the abundance of cladocerans. Among toxic cells, the MC cell quota varied throughout the survey. However, a negative correlation between the MC cell quota and the mcyA cell number during two short periods characterized by marked changes in the cyanobacterial biomass was found. Finally, only 54% of the variation in the MC concentrations measured in the lake can be explained by the dynamics of the density of cells with the MC producer genotype, suggesting that this measurement is not a satisfactory method for use in monitoring programs intended to predict the toxic risk associated with cyanobacterial proliferation.

Risks to the aquatic ecosystem from the application of Metarhizium anisopliae for locust control in Australia.

Laboratory tests of Metarhizium anisopliae var acridum Driver & Milner, at a dose of 1.3 x 10(6) conidia ml-1, had no adverse effects on nymphs of mayfly, Ulmerophlebia sp or 8-week-old fry of the rainbow fish, Melanotaenia duboulayi Castelnau. This dose was toxic to the cladoceran, Ceriodaphnia dubia Richard, causing 100% mortality in 48 h. When this test was repeated at doses of up to 6.7 x 10(3) conidia ml-1, there was only 5% mortality after 192 h. Spraying of artificial water sources with a very high dose of the fungus as an aqueous spray resulted in 80-130 conidia ml-1 at 15 cm depth in the first 24 h after spraying. The conidia rapidly settled out and were absent from the top 15 cm layer of water after about 50 h. A similar experiment using the oil formulation as used in field control resulted in a 2- to 20-fold lower level of conidia in the water. Finally, sampling actual water sources in spray areas revealed a very low level of contamination of the water, with a maximum mean level of 29 conidia ml-1 in the first 24 h after treatment. Thus the level of conidia likely to enter water during control campaigns is a small fraction of that required to kill cladocerans, the only sensitive non-target organism tested. It is concluded that the biopesticide is very unlikely to pose any hazard to aquatic organisms.