The fifth conference of the European Pond Conservation Network (Luxembourg, June 2012) brought together researchers, environmental managers, and other stakeholders with the aim to share stateof-the-art knowledge on the ecology, management, and conservation of ponds in the context of the many challenges facing the wider water environment. Although well-known ecological patterns apply to most ponds in Europe and elsewhere, recent data highlight that part of the environmental variables governing pond biodiversity remain specific to climatic/biogeographic regions and to elevation ranges, suggesting that, in addition to common practice, management plans should include range-specific measures. Beyond the contribution of individual ponds to the aquatic and terrestrial life, connected networks of ponds are vital in the provision of new climate space as a response to global climate change, by allowing the observed northward and/or upward movements of species. In terms of services, ponds offer sustainable solutions to key issues of water management and climate change such as nutrient retention, rainfall interception, or carbon sequestration. While the ecological role of ponds is now wellestablished, authoritative research-based advice remains needed to inform future direction in the conservation of small water bodies and to further bridge the gap between science and practice.

Campylobacteriosis has increased markedly in Luxembourg during recent years. We sought to determine which Campylobacter genotypes infect humans, where they may originate from, and how they may infect humans. Multilocus sequence typing was performed on 1153 Campylobacter jejuni and 136 C. coli human strains to be attributed to three putative animal reservoirs (poultry, ruminants, pigs) and to environmental water using the asymmetric island model. A nationwide case-control study (2010–2013) for domestic campylobacteriosis was also conducted, including 367 C. jejuni and 48 C. coli cases, and 624 controls. Risk factors were investigated by Campylobacter species, and for strains attributed to different sources using a combined case-control and source attribution analysis. 282 sequence types (STs) were identified: ST-21, ST-48, ST-572, ST-50 and ST-257 were prevailing. Most cases were attributed to poultry (61.2%) and ruminants (33.3%). Consuming chicken outside the home was the dominant risk factor for both Campylobacter species. Newly identified risk factors included contact with garden soil for either species, and consuming beef specifically for C. coli. Poultry-associated campylobacteriosis was linked to poultry consumption in wintertime, and ruminant-associated campylobacteriosis to tap-water provider type. Besides confirming chicken as campylobacteriosis primary source, additional evidence was found for other reservoirs and transmission routes.

Biofilms colonizing surfaces inside drinking water distribution networks may provide a habitat and shelter to pathogenic viruses and parasites. If released from biofilms, these pathogens may disseminate in the water distribution system and cause waterborne diseases. Our study aimed to investigate the interactions of protozoan parasites (Cryptosporidium parvum and Giardia lamblia [oo]cysts) and viruses (vaccinal poliovirus type 1, X174, and MS2) with two contrasting biofilms. First, attachment, persistence, and detachment of the protozoan parasites and the viruses were assessed with a drinking water biofilm. This biofilm was allowed to develop inside a rotating annular reactor fed with tap water for 7 months prior to the inoculation. Our results show that viable parasites and infectious viruses attached to the drinking water biofilm within 1 h and persisted within the biofilm. Indeed, infectious viruses were detected in the drinking water biofilm up to 6 days after the inoculation, while viral genome and viable parasites were still detected at day 34, corresponding to the last day of the monitoring period. Since viral genome was detected much longer than infectious particles, our results raise the question of the significance of detecting viral genomes in biofilms. A transfer of viable parasites and viruses from the biofilm to the water phase was observed after the flow velocity was increased but also with a constant laminar flow rate. Similar results regarding parasite and virus attachment and detachment were obtained using a treated wastewater biofilm, suggesting that our observations might be extrapolated to a wide range of environmental biofilms and confirming that biofilms can be considered a potential secondary source of contamination.

This study presents an original rDNA PCR and microscopic survey of pelagic freshwater fungal communities, and was designed to unveil the diversity of true Fungi (i.e. the kingdom Eumycota) in three contrasting lake ecosystems (Lakes Pavin, Aydat and Vassivière) located in the French Massif Central. Three clone libraries were constructed from samples collected in the euphotic layers of the lakes during spring 2007. Phylogenetic analysis of the combined data from the three lakes clustered our sequences into thee divisions: Chytridiomycota (50% of total sequences), Ascomycota (40%) and Basidiomycota (10% in Pavin and Aydat only). Several sequences were assigned to a novel Chytridiomycota clade first recovered in Lake Pavin in 2005. Most of the sequences retrieved in the investigated lakes were affiliated with known fungal species, most of which were apparently well adapted to thrive in the pelagic realm. Their main functions (i.e. parasitism and saprophytism), putatively inferred from the closest relatives of the retrieved molecular sequences, were confirmed by microscopic approaches and by enrichment experiments with pollen grains. The occurrence of three fungal forms (zoosporic, yeast and mycelial) was associated with different trophic modes, establishing fungi as strong potential competitors for various niches in pelagic ecosystems, primarily in relation to the processing of particulate organic matter and the production of propagule food sources for grazers. For the first time, this study provides insight into the diversity and the associated functions of all members of the Kingdom Eumycota investigated in the whole plankton fraction of aquatic ecosystems.

In the temperate climate of the northern hemisphere, winter survival of woody plants is determined by the ability to acclimate to freezing temperatures and to undergo a period of dormancy. Cold acclimation in many woody plants is initially induced by short photoperiod and low, non-freezing temperatures. These two factors (5°C and short photoperiod) were used to study changes in the proteome of bark tissues of 1-year-old peach trees. Difference in-gel electrophoresis technology, a gel-based approach involving the labeling of proteins with different fluorescent dyes, was used to conduct a quantitative assessment of changes in the peach bark proteome during cold acclimation. Using this approach, we were able to identify differentially expressed proteins and to assign them to a class of either 'temperature-responsive' or 'photoperiodresponsive' proteins. The most significant factor affecting the proteome appeared to be low temperature, while the combination of low temperature and short photoperiod was shown to act either synergistically or additively on the expression of some proteins. Fifty-seven protein spots on gels were identified by mass spectrometry. They included proteins involved in carbohydrate metabolism (e.g., enolase, malate dehydrogenase, etc), defense or protective mechanisms (e.g., dehydrin, HSPs, and PR-proteins), energy production and electron transport (e.g., adenosine triphosphate synthases and lyases), and cytoskeleton organization (e.g., tubulins and actins). The information derived from the analysis of the proteome is discussed as a function of the two treatment factors: low temperature and short photoperiod.

High-resolution vertical sampling and determination of viral and prokaryotic parameters in a deep volcanic lake shows that in the absence of thermal stratification but within light, oxygen, and chlorophyll gradients, host availability empirically is prevalent over the physical and chemical environments and favors lytic over lysogenic "viral life cycles." Viral activity is crucial to microbial mortality, diversity, potential gene transfer, and lysogenic conversion in aquatic systems (see references 18 and 19 for recent reviews). Viruses can affect ecological processes and nutrient cycles (23), but this ultimately depends on whether the virus is lytic or temperate. Studies examining large data sets have revealed that prokaryotes form the major host reservoir for viruses in pelagic systems (2). Protistan grazing and viral lysis are major sources of bacterial mortality in these systems. In general, bacterivory dominates, but losses of bacteria from viral lysis at times are comparable to those from protistan bacterivory (13). In conditions such as anoxia, bacteriolysis generally is much higher than bacterivory based on evidence from theoretical (12) and empirical (2) field investigations. In contrast to bacterivory and lytic viral infection, which have been examined in a variety of marine systems, only two studies have investigated the two viral "life cycles" simultaneously, but not together with bacterivory (11, 21). For freshwater, one study has examined temperate phages in the surface layer of Lake Superior (17).Pelagic viral ecology thus lacks studies in which potential prophage induction, lytic viral infection, and bacterivory are investigated simultaneously. In addition, most of the studies on these processes have ignored the well-known contribution of depth-related gradients as a major forcing factor in aquatic systems. Inclusion of such a contribution is needed to fully assess the potential roles of viruses for microbial food web processes and to address hypotheses that (i) in suboxic and anoxic waters the importance of lytic viral infection increases relative to bacterivory (2, 13, 20) and (ii) lysogeny increases as lytic viruses decrease with their host densities and production (21). Here we address these questions by examining the vertical distribution of the frequencies of lytically and lysogenically infected cells together with the potential bacterivory from heterotrophic nanoflagellates in a deep meromictic lake. Samples were collected in early spring, when the isothermal water column showed gradients of light, oxygen, and chlorophyll, in order to avoid false interpretations of correlations that often result from the covariation of two variables with respect to a third factor, usually temperature.Lake Pavin is a deep (Z max ϭ 92 m) meromictic mountain lake located in the French Massif Central (2°56ЈE, 45°29ЈN). Samples were taken in triplicate on 20 April 2004 at the reference station (located in the middle and the deepest part of the lake), using an 8-liter Van Dorn bottle. A total of 15 depths were sa...

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