Life history responses of Daphnia magna feeding on toxic Microcystis aeruginosa alone and mixed with a mixotrophic Poterioochromonas species

The toxic effects of a mixotrophic golden alga (Poterioochromonas sp. strain ZX1) and a cyanobacterium Microcystis aeruginosa grazed by Poterioochromonas to a cladoceran were investigated through life history experiments using Daphnia magna. Poterioochromonas cultured in two ways (fed M. aeruginosa in an inorganic medium or grown in an organic medium) both induced starvation-like effects on D. magna, indicating that Poterioochromonas is neither acutely toxic nor a good food for D. magna. Despite a microcystin-LR content of 10⁻⁸ μg cell⁻¹ in M. aeruginosa, no toxins were accumulated in Poterioochromonas fed the cyanobacterium. The toxic effect of M. aeruginosa to D. magna was significantly reduced in the presence of Poterioochromonas, which may be performed in two ways: decrease M. aeruginosa cells ingestion of D. magna by grazing on M. aeruginosa; and decrease the toxicity of the medium by degrading the toxins released by M. aeruginosa. This study provides new information on the interactions between a cyanobacterium and its grazer under laboratory conditions and may increase our understanding of the ecological significance of such interactions in the aquatic food webs.     

Surface-retained organic matter of Microcystis aeruginosa inhibiting coagulation with polyaluminum chloride in drinking water treatment

Algogenic organic matter produced by the excess growth of cyanobacteria in semi-closed water areas causes coagulation inhibition in drinking water production. In this study, hydrophilic substances of Microcystis aeruginosa, which were mainly composed of lipopolysaccharide (LPS) and RNA, were prepared, and the involvement of these cyanobacterial hydrophilic substances in coagulation inhibition was investigated. As a result, it was found that the negatively charged hydrophilic substances with a molecular weight higher than 10 kDa have a significant role in coagulation inhibition. Further fractionation of cyanobacterial hydrophilic substances revealed that surface-retained organic matter (SOM), including LPS, could exhibit a potent inhibitory effect on the coagulation using polyaluminum chloride (PACl), presumably because of the direct interaction of hydrophilic SOM with cations originated from PACl, which could impede the hydrolysis of the coagulant.     

Impact of sonication at 20 kHz on Microcystis aeruginosa, Anabaena circinalis and Chlorella sp

Blooms of toxic cyanobacteria such as Microcystis aeruginosa periodically occur within wastewater treatment lagoons in the warmer months, and may consequently cause contamination of downstream water and outages of the supply of recycled wastewater. Lab-scale sonication (20 kHz) was conducted on suspensions of M. aeruginosa isolated from a wastewater treatment lagoon, and two other algal strains, Anabaena circinalis and Chlorella sp., to investigate cell reduction, growth inhibition, release of microcystin and sonication efficiency in controlling the growth of the M. aeruginosa. For M. aeruginosa, for all sonication intensities and exposure times trialled, sonication led to an immediate reduction in the population, the highest reduction rate occurring within the initial 5 min. Sonication for 5 min at 0.32 W/mL, or for a longer exposure time (>10 min) at a lower power intensity (0.043 W/mL), led to an immediate increase in microcystin level in the treated suspensions. However, prolonged exposure (>10 min) to sonication at higher power intensities reduced the microcystin concentration significantly. Under the same sonication conditions, the order of decreasing growth inhibition of the three algal species was: A. circinalis > M. aeruginosa > Chlorella sp., demonstrating sonication has the potential to selectively remove/deactivate harmful cyanobacteria from the algal communities in wastewater treatment lagoons.     

Bio-reaction of nitrobenzene with Microcystis aeruginosa: characteristics, kinetics and application

The bio-reaction of nitrobenzene (NB) with Microcystis aeruginosa was investigated at different initial algal densities and NB concentrations by performing static experiments. The results showed that the elimination of NB was enhanced by the bio-reaction, and the reaction rate varied as a function of the reaction time. Moreover, the reaction rate was significantly affected by the algal density and NB concentration. A kinetic analysis showed that the elimination of NB in a solution without algae appeared to be pseudo-first-order with respect to the NB concentration, whereas a first-order model was too oversimplified to describe the elimination of NB in a solution with algae. Assuming that different algal cells have the same effect on the bio-reaction under the same conditions, the bio-reaction can be described as dC_NB = −k₀C_A^mA_NBⁿdt (where k₀ is the reaction rate constant, C_A is the algae density and C_NB is the concentration of NB). When the growth of algae was not considered, the value of k₀, m and n were 8.170 × 10⁻⁴, 0.5887 and 1.692, respectively. Alternatively, when algae were in the exponential growth phase, the value of k₀, m and n were 1.6871 × 10⁻⁵, 0.7248 and 2.5407, respectively, according to a nonlinear fitting analysis. The kinetic model was also used to elucidate the effect of nutritional limitation on the bio-reaction.     

The role of nitrobenzene on the yield of trihalomethane formation potential in aqueous solutions with Microcystis aeruginosa

Algae are one of the most important disinfection by-product (DBP) precursors in aquatic environments. The contents of DBP precursors in algae are influenced by not only environmental factors but also some xenobiotics. Trihalomethane formation potential (THMFP) in both the separate and interactive pollution of Microcystis aeruginosa and Nitrobenzene (NB) was investigated in batch experiment to discover the effects of xenobiotics on the yield of DBP precursors in the algal solution. The results show that in the separate NB solution, NB did not react with Cl₂ to form trihalomethane (THM), whereas in the algae solution, THMFP had a significant positive linear correlation with M. aeruginosa density in both solution and extracellular organic matter (EOM). The correlation coefficients were 0.9845 (p = 3.567 × 10⁻⁴) and 0.9854 (p = 1.406 × 10⁻⁴), respectively. According to regression results, about 77.9% of the total THMFP came from the algal cells, while the rest came from EOM. When the interactive pollution of M. aeruginosa and NB occurred, the growth of algae was inhibited by NB. The density of M. aeruginosa in a high concentration NB solution (280 μg/L) was only 71.1% of that in the solution without NB after 5 days of incubation. However, THMFP in the mixture (algae and NB) and the EOM did not change significantly, and the productivity of THMFP by the algae (THMFP/10⁸cells) increased with the increase in NB concentration. There was a significant linear correlation between THMFP/10⁸cell and NB concentration (r = 0.9117, p < 0.01), which shows the contribution of the algae to THM formation was enhanced by NB. This result might be caused by the increased protein productivity and the biodegradation of NB by M. aeruginosa.     

Biomass and nutrient content of sessile oak (Quercus petraea (Matt.) Liebl.) and beech (Fagus sylvatica L.) stem and branches in a mixed stand in southern Belgium

Accurate estimates of the amounts of nutrients immobilised in the organs and tissues of different tree species are of prime importance to make appropriate tree species selection and determine the harvesting regime that will ensure forest sustainability. Sixteen sessile oaks (Quercus petraea (Matt.) Liebl.) (64-129 years; stem diameters: 17-57 cm) and twelve beeches (Fagus sylvatica L.) (43-86 years; stem diameters: 9-50 cm) were destructively sampled from a mixed stand located on an acid brown soil in southern Belgium. Statistical models were developed to investigate the differences in nutrient concentrations between tree species, between aboveground tree compartments of the same species, and between tissues of the same compartment. For stem tissues, vertical concentration profiles were described using a versatile equation. Allometric equations were used to predict biomass and nutrient content of tree compartments based on tree dimensions. Broadly speaking, nutrient concentrations tended to be somewhat higher for oak compared with beech, but the amplitude and the direction of inter-species differences varied greatly, depending on the nutrient and the tree compartment. For both species, living branch nutrient concentrations tended to decrease with increasing branch diameter, except for Ca (oak) and Mg (beech). Nutrient concentrations were consistently higher in bark than in wood; this difference between tissues was quite pronounced for Ca, particularly in the case of oak. The biomass and nutrient content equations were used to investigate the effects of tree species and harvesting regime on nutrient exports at harvesting. For equivalent harvesting scenarios, beech was found to induce higher Mg exports than oak, and inversely for Ca. Assuming stand clear cutting, complete tree harvesting would increase average nutrient exports from 65% (Ca) to 162% (P) compared with a stem-only harvesting scenario. These results provide valuable information in the current context of the more intensive utilization of forest products.     

铜绿微囊藻衰亡过程中产甲烷动态及关键影响因子

为探究铜绿微囊藻水华自然衰亡和水华堆积区藻快速衰亡对CH_4释放的影响,构建了室内模拟实验,研究了铜绿微囊藻自然衰亡和快速衰亡过程中水环境因子和CH_4、CO_2气体释放通量的变化。结果表明:高密度铜绿微囊藻衰亡中水环境因子和产气的变化遵循相似的时间模式,且衰亡对CH_4和CO_2的产生有显著的促进作用。自然衰亡和快速衰亡的CH_4累积释放量分别为对照组的22.80倍37.72倍,CO_2累积释放量分别为对照组的5.36倍和4.03倍。自然衰亡与快速衰亡的总碳(C)释放量差异不大,但快速衰亡释放的气体组分中CH_4的占比是自然衰亡的1.86倍。相关性分析表明,溶解性有机碳(DOC)是铜绿微囊藻衰亡中影响CH_4和CO_2释放的主导因子,且DOC中的芳香蛋白类物质和溶解性微生物代谢产物是CH_4和CO_2产生的主要底物来源。     

Evaluation of the mechanisms of the effect of ultrasound on Microcystis aeruginosa at different ultrasonic frequencies

Blooms of cyanobacteria are now considered to be a common environmental issue. They are hazardous to both domestic and wild animals and humans. Current treatments are unable to effectively control such blooms as they become tolerant to biocides and it is difficult to degrade cyanobacterial toxins in water. Alternative methods for control are currently under investigation. One potential effective method is ultrasonic irradiation. Ultrasound inactivates algal and cyanobacteria cells through cavitation by generating extreme conditions, resulting in a number of physical, mechanical and chemical effects. The aim of this study was to investigate the effect of ultrasound at different frequencies on Microcystis aeruginosa. Flow cytometry was used to measure cyanobacterial metabolic cell viability in addition to the more commonly used haemocytometry, optical density and fluorimetry. Results indicate low frequency 20 kHz ultrasound with high intensity (0.0403 W cm⁻³) is effective for the inactivation of cyanobacterial cells. Higher frequencies of 580 kHz (0.0041 W cm⁻³) also resulted in an inactivation effect, but 1146 kHz (0.0018 W cm⁻³) showed a declumping effect as evidenced by flow cytometry. Ultrasonic treatment over time under different sonication conditions demonstrates the following:1. Acoustic cavitation via mechanical effects can induce sufficient shear forces to directly rupture cyanobacteria cells.2. At higher ultrasonic frequencies the mechanical energy of cavitation is less but a larger proportion of free radicals are produced from the ultrasonic degradation of water, which chemically attacks and weakens the cyanobacteria cell walls.3. At higher frequencies free radicals also damage chlorophyll a leading to a loss in photosynthetic cell viability.4. At low powers ultrasonic energy results in declumping of cyanobacteria.     

Assessing the ecotoxicity of vinyl chloride using green alga P. subcapitata, nematode C. elegans, and the SOS chromotest in a closed system without headspace

The ecotoxicity of vinyl chloride (VC) was evaluated using green alga, nematode, and the SOS chromotest. The green alga and nematode tested were Pseudokirchneriella subcapitata, and Caenorhabditis elegans, respectively. Because of the tendency of VC to escape from an aqueous exposure medium to the air phase, all tests in the present study were performed in a closed system without headspace to minimize the losses of VC. Previous studies on VC toxicity were performed in an open system or closed system with headspace. VC inhibits the growth of P. subcapitata. The 48-h IC50 value for P. subcapitata exposed to VC was calculated to be 5.15 mg/L. The survival of C. elegans was not influenced at concentrations of up to 60 mg/L; however, VC has an adverse effect on the reproduction of C. elegans. In a stress-related gene expression test using C. elegans, a significant and concentration-dependent expression of heat shock protein 16.2 was observed, indicating that VC induces the stress of C. elegans at the genetic level. The results of the SOS chromotest using Escherichia coli PQ37 showed an IF_max value of 1.11, indicating that VC is not genotoxic. The present study demonstrated that VC has an adverse effect on the algal growth and reproductive and genetic levels of C. elegans. A closed system without headspace is an effective method of testing the aquatic toxicity of volatile organic compounds such as VC.     

Phosphorus addition reverses the positive effect of zebra mussels (Dreissena polymorpha) on the toxic cyanobacterium, Microcystis aeruginosa

We tested the hypothesis that zebra mussels (Dreissena polymorpha) have positive effects on the toxin-producing cyanobacterium, Microcystis aeruginosa, at low phosphorus (P) concentrations, but negative effects on M. aeruginosa at high P, with a large-scale enclosure experiment in an oligotrophic lake. After three weeks, mussels had a significantly positive effect on M. aeruginosa at ambient P (total phosphorus, TP ∼10 μg L⁻¹), and a significantly negative effect at high P (simulating a TP of ∼40 μg L⁻¹ in lakes). Positive and negative effects were strong and very similar in magnitude. Thus, we were able to ameliorate a negative effect of Dreissena invasion on water quality (i.e., promotion of Microcystis) by adding P to water from an oligotrophic lake. Our results are congruent with many field observations of Microcystis response to Dreissena invasion across ecosystems of varying P availability.     

Influence of sampling strategies on the monitoring of cyanobacteria in shallow lakes: lessons from a case study in France

Sampling cyanobacteria in freshwater ecosystems is a crucial aspect of monitoring programs in both basic and applied research. Despite this, few papers have dealt with this aspect, and a high proportion of cyanobacteria monitoring programs are still based on monthly or twice-monthly water sampling, usually performed at a single location. In this study, we conducted high frequency spatial and temporal water sampling in a small eutrophic shallow lake that experiences cyanobacterial blooms every year. We demonstrate that the spatial and temporal aspects of the sampling strategy had a considerable impact on the findings of cyanobacteria monitoring in this lake. In particular, two peaks of Aphanizomenon flos-aquae cell abundances were usually not picked up by the various temporal sampling strategies tested. In contrast, sampling once a month was sufficient to provide a good overall estimation of the population dynamics of Microcystis aeruginosa. The spatial frequency of sampling was also important, and the choice in the location of the sampling points around the lake was very important if only two or three sampling points were used. When four or five sampling points were used, this reduced the impact of the choice of the location of the sampling points, and allowed to obtain fairly similar results than when six sampling points were used. These findings demonstrate the importance of the sampling strategy in cyanobacteria monitoring, and the fact that it is impossible to propose a single universal sampling strategy that is appropriate for all freshwater ecosystems and also for all cyanobacteria.     

Effects of low or medium-pressure UV irradiation on the release of intracellular microcystin

UV irradiation could be an alternative growth inhibition treatment against toxic Microcystis blooms in lakes. This study examined the effect of UV irradiation on the release of toxic intracellular microcystin. Conventional algicidal treatment (e.g. copper sulfate) was known to cause rapid release of intracellular microcystin and additional problems, but no studies have examined the case of UV treatment. A pure culture of Microcystis aeruginosa PCC 7806 was exposed to monochromatic low-pressure or polychromatic medium-pressure UV lamps. Irradiated pure culture suspension was subsequently incubated for 14 days under white light fluorescent lamps. During incubation, profiles of the number of cells, intracellular and extracellular microcystin concentration were determined. After the UV exposure, the number of cells gradually diminished; the net log cell reduction after 6 days of incubation was 1.6 log or 2.0 log for 600 mJ/cm2 of LP or MP UV irradiation, respectively. There were three findings for UV irradiation effects on the release of intracellular microcystin. First, because UV exposure can inhibit increases in the number of cells for about 6 days, it inhibited the increase of microcystin concentrations in water. Second, intracellular microcystin was gradually released into the surrounding water through a gradual reduction in the number of cells, preventing a rapid increase of microcystin concentration in water. Microcystin concentration in water was not significantly higher in UV-irradiated samples than non-UV-irradiated sample. Third, UV irradiation was able to degrade intracellular microcystin; it was reduced from 24.6 to 7.06 or 7.16 fg/cell by 600 mJ/cm2 of LP or MP UV irradiation, respectively. This contributed to reduce increases in microcystin concentrations in water. UV treatment can inhibit Microcystis growth and reduce intracellular microcystin content without rapid release of intracellular microcystin.     

Delayed influence of dam storage and discharge on the determination of seasonal proliferations of Microcystis aeruginosa and Stephanodiscus hantzschii in a regulated river system of the lower Nakdong River (South Korea)

This study presents the relationship between the dam hydrology and phytoplankton proliferations in a regulated river system in East Asia. A long-term ecological study reveals that multi-purpose dam regulation on river flow in the Nakdong River (South Korea) affects proliferating patterns of phytoplankton, especially during summer (June-August) and winter (December-next February) as Microcystis aeruginosa and Stephanodiscus hantzschii, respectively, dominate each season by over 80%. Eutrophication was observed in the lower Nakdong River (South Korea) (mean+/-standard deviation: nitrate-N, 2.8+/-0.9 mg L(-1); phosphate-P, 40.3+/-31.3 microg L(-1); chlorophyll a, 45.2+/-84.9 microg L(-1); n=449), and the limnological characteristics responded to climatic variations such as monsoon and summer typhoons. The river basin experiences concentrated rainfall during the rainy season (June-September, over ca. 60% of total annual rainfall). Correlation and cross-correlation for time-delayed relationship revealed the hydrological environments (i.e., quantity of dam storage and discharge) had significantly negative relationship with the population dynamics of the two bloom-forming species for up to the next 2 years. S. hantzschii had clearer relationship with dam storage and discharge, which might be due to the seasonality exhibited by the species. The results support the necessity of "smart flow control" which may enable destruction of bloom formation by the two species with an adequate pulse of discharge generated by upstream dams in the far-east Asian river systems. This would increase the efficiency of water resource management system.     

EnergyPlus vs. DOE-2.1e: The effect of ground-coupling on energy use of a code house with basement in a hot-humid climate

For low-rise buildings, the heat losses or gains through the ground coupled building envelope can be a significant load component. Studies have shown that current simulation tools give dissimilar results for the ground coupled heat transfer (GCHT) with basements. This paper quantifies and explains the differences between EnergyPlus and DOE-2.1e (DOE-2) basement GCHTs based on their results for an all electric code house in a hot and humid climate zone. The code house was modeled with two basement configurations i.e. a conditioned basement and an unconditioned crawlspace. DOE-2 was used with Winkelmann's basement model and EnergyPlus was used with its GCHT calculator utility, Basement. The results revealed that the ground isolated EnergyPlus houses used 3-23% more cooling, 12-29% less heating and 3-7% lower overall HVAC electricity use when compared to the ground isolated DOE-2 houses. Ground coupling added up to three times more heat loss in EnergyPlus than in DOE-2. This increased the overall energy use difference between these two programs from 3-7% (ground isolated) to 14-25% (ground coupled). These results showed that a truth standard is required for basement heat transfer calculations of low-rise buildings.     

Possible interaction between a rodenticide treatment and a pathogen in common vole (Microtus arvalis) during a population peak

A common vole (Microtus arvalis) population peak in Northern Spain in 2007 was treated with large scale application of chlorophacinone, an anticoagulant rodenticide of the indandione family. Voles found dead and trapped alive were collected in treated and untreated areas. Residues of chlorophacinone were analyzed in liver of voles by HPLC-UV. Also, the presence of the pathogen Francisella tularensis was analyzed by PCR in samples of vole spleen. Chlorophacinone (82-3800 ng/g; wet weight liver) was only detected in voles found dead in treated areas (55.5%). The prevalence of F. tularensis in voles found dead in treated areas was also particularly high (66.7%). Moreover, chlorophacinone levels were lower in voles that were PCR-positive for F. tularensis (geometric mean [95% CI], 418 [143-1219] ng/g) than in those that were PCR-negative (1084 [581-2121] ng/g). Interactions between pathogens and rodenticides might be considered to reduce the doses used in baits or to avoid the use of the more toxic 2nd generation anticoagulant rodenticides.     

Differentiation and identification of Shigella spp. and enteroinvasive Escherichia coli in environmental waters by a molecular method and biochemical test

Both Shigella spp. and enteroinvasive Escherichia coli (EIEC) are important human pathogens that are responsible for the majority of cases of endemic bacillary dysentery. However, they are difficult to identify and differentiate by biochemical tests or molecular methods alone. In this study, we developed a procedure to detect Shigella spp. and EIEC from environmental water samples using membrane filtration followed by nutrient broth enrichment, isolation using selective culture plates, and identification of the invasion plasmid antigen H (ipaH) gene by PCR amplification and DNA sequencing. Finally, we used a biochemical test and a serological assay to differentiate between Shigella and EIEC. Among the 93 water samples from nine reservoirs and one watershed, 76 (81.7%) water samples of culture plates had candidate colonies of Shigella and EIEC and 5 water samples were positive (5.4%) for a Shigella- and EIEC-specific polymerase chain reaction targeting the ipaH gene. Guided by the molecular method, the biochemical test, and the serological assay, 11 ipaH gene-positive isolates from 5 water samples were all identified as EIEC.     

Influence of physicochemical and operational parameters on Nitrobacter and Nitrospira communities in an aerobic activated sludge bioreactor

To understand how to optimize performance of a partially nitrifying plant, the dynamics of Nitrospira and Nitrobacter abundance were studied over a 1 year period using quantitative polymerase chain reaction (qPCR) and their relative contributions to nitrite oxidation assessed including the affects of temperature and dissolved oxygen (DO). Correlation coefficients linking shifts in the community composition of nitrite-oxidizing bacteria (NOB) to operational or environmental variables indicated Nitrospira was significantly and negatively correlated to nitrite concentrations (r = −0.45, P < 0.01) and DO (r = −0.46, P < 0.01), while temperature showed a strong positive correlation (r = 0.59, P < 0.0001). However, the Nitrobacter portion of the total NOB populations showed a positive correlations with DO (r = 0.38, P < 0.01) and hydraulic retention time (HRT) (r = 0.33, P < 0.05), as well as being negatively correlated with temperature (r = −0.49, P < 0.001) suggesting specific niche adaptations within the NOB community. Nitrospira was dominant being better adapted to the low DO and shorter sludge retention times (SRT) of this plant, while Nitrobacter increased in abundance during the winter months, when temperatures were lower and DO concentrations higher. Principal component analysis (PCA) results supported these findings by the close proximity of Nitrospira and temperature biplots of PC1 and PC2 as well as grouping Nitrobacter, NO₂⁻-N, HRT, and DO in the loadings together. The clustering of samples from specific dates also exhibited a strong seasonality.     

Improvement of metal adsorption onto chitosan/Sargassum sp. composite sorbent by an innovative ion-imprint technology

Technology for immobilization of biomass has attracted a great interest due to the high sorption capacity of biomass for sequestration of toxic metals from industrial effluents. However, the currently practiced immobilization methods normally reduce the metal sorption capacities. In this study, an innovative ion-imprint technology was developed to overcome the drawback. Copper ion was first imprinted onto the functional groups of chitosan that formed a pellet-typed sorbent through the granulation with Sargassum sp.; the imprinted copper ion was chemically detached from the sorbent, leading to the formation of a novel copper ion-imprinted chitosan/Sargassum sp. (CICS) composite adsorbent. The copper sorption on CICS was found to be highly pH-dependent and the maximum uptake capacity was achieved at pH 4.7-5.5. The adsorption isotherm study showed the maximum sorption capacity of CICS of 1.08 mmol/g, much higher than the non-imprinted chitosan/Sargassum sp. sorbent (NICS) (0.49 mmol/g). The used sorbent was reusable after being regenerated through desorption. The FTIR and XPS studies revealed that the greater sorption of heavy metal was attributed to the large number of primary amine groups available on the surfaces of the ion-imprinted chitosan and the abundant carboxyl groups on Sargassum sp. Finally, an intraparticle surface diffusion controlled model well described the sorption history of the sorbents.     

Method for rapid and sensitive detection of Enterococcus sp. and Enterococcus faecalis/faecium cells in potable water samples

We have developed a rapid and robust technological solution including a membrane filtration and dissolution method followed by a molecular enrichment and a real-time PCR assay, for detecting the presence of Enterococcus sp. or Enterococcus faecalis/faecium per 100 mL of water in less than 5 h and we compared it to Method 1600 on mEI agar in terms of specificity, sensitivity, and limit of detection. The mEI and the Enterococcus sp.-specific assay detected respectively 73 (64.0%) and 114 (100%) of the 114 enterococcal strains tested. None of the 150 non-enterococcal strains tested was detected by both methods with the exception of Tetragenococcus solitarius for the Enterococcus sp. assay. The multiplexed E. faecalis/faecium assay efficiently amplified DNA from 47 of 47 (100%) E. faecalis and 27 of 27 (100%) E. faecium strains tested respectively, whereas none of the 191 non-E. faecalis/faecium strains tested was detected. By simultaneously detecting the predominant fecal enterococcal species, the E. faecalis/faecium-specific assay allows a better distinction between enterococcal strains of fecal origin and those provided by the environment than Method 1600. Our procedure allows the detection of 4.5 enterococcal colony forming units (CFU) per 100 mL in less than 5 h, whereas the mEI method detected 2.3 CFU/100 mL in 24 h (95% confidence). Thus, our innovative and highly effective method provides a rapid and easy approach to concentrate very low numbers of enterococcal cells present in a 100 mL water sample and allows a better distinction between fecal and environmental enterococcal cells than Method 1600.     

Impact of toxic cyanobacteria on gastropods and microcystin accumulation in a eutrophic lake (Grand-Lieu, France) with special reference to Physa (= Physella) acuta

Hepatotoxic microcystins (MCs) produced by cyanobacteria are known to accumulate in gastropods following grazing of toxic cyanobacteria and/or absorption of MCs dissolved in water, with adverse effects on life history traits demonstrated in the laboratory. In the field, such effects may vary depending on species, according to their relative sensitivity and ecology. The aims of this study were to i) establish how various intensities of MC-producing cyanobacteria proliferations alter the structure of gastropod community and ii) compare MC tissue concentration in gastropods in the field with those obtained in our previous laboratory experiments on the prosobranch Potamopyrgus antipodarum and the pulmonate Lymnaea stagnalis. We explored these questions through a one-year field study at three stations at Grand-Lieu Lake (France) affected by different intensities of cyanobacteria proliferations. A survey of the community structure and MC content of both cyanobacteria and gastropods was associated with a caging experiment involving P. antipodarum and L. stagnalis. In total, 2592 gastropods belonging to 7 prosobranch and 16 pulmonate species were collected. However, distribution among the stations was unequal with 62% vs 2% of gastropods sampled respectively at the stations with the lowest vs highest concentrations of MC. Irrespective of the station, pulmonates were always more diverse, more abundant and occurred at higher frequencies than prosobranchs. Only the pulmonate Physa acuta occurred at all stations, with abundance and MC tissue concentration (≤ 4.32 µg g DW^− 1) depending on the degrees of MC-producing cyanobacteria proliferations in the stations; therefore, P. acuta is proposed as a potential sentinel species. The caging experiment demonstrated a higher MC accumulation in L. stagnalis (≤ 0.36 µg g DW^− 1 for 71% of individuals) than in P. antipodarum (≤ 0.02 µg g DW^− 1 for 12%), corroborating previous laboratory observations. Results are discussed in terms of differential gastropod sensitivity and MC transfer through the food web.