Clostridium botulinum type C, D, C/D, and D/C: An update.

Clostridium botulinum is the main causative agent of botulism, a neurological disease encountered in humans as well as animals. Nine types of botulinum neurotoxins (BoNTs) have been described so far. Amongst these "toxinotypes," the A, the B and E are the most frequently encountered in humans while the C, D, C/D and D/C are mostly affecting domestic and wild birds as well as cattle. In France for instance, many cases and outbreaks are reported in these animal species every year. However, underestimation is very likely at least for avifauna species where the detection of dead animals can be challenging. Knowledge about BoNTs C, D, C/D, and D/C and the diseases they cause in animals and humans is still scarce and unclear. Specifically, the potential role of animal botulism outbreaks in cattle and poultry as a source of human illness needs to be further assessed. In this narrative review, we present the current knowledge about toxinotypes C, D, C/D, and D/C in cattle and poultry with, amongst various other aspects, their epidemiological cycles. We also discuss the zoonotic potential of these toxinotypes and some possible ways of risk mitigation. An adapted and effective management of botulism outbreaks in livestock also requires a better understanding of these less common and known toxinotypes.

Tracing water perturbation using NO3-, doc, particles size determination, and bacteria: A method development for karst aquifer water quality hazard assessment.

Karst systems, as well as springs, are vulnerable to water perturbation brought by infiltration. In this research, sources of water perturbations were examined. The first objective is to provide a method that can determine the origin of the water flowing in the karst outlet. The second objective is to identify the associated water quality hazards caused by the infiltration source. The method relies on these parameters: turbidity, DOC, NO3-, particle size, and bacteria (E. coli, enterococcus and total coliforms). As the method was applied during flood events, measurement of the water flow is also needed to have a basic knowledge on the hydrodynamic of the water resource. The proposed method is based on a high resolution monitoring of physico chemical parameters of the water flowing during flood events. Using this proposed method, (1) the origin of the water can be identified, (2) the type and nature of water perturbation can be described, and (3) the type of water perturbation that accompanies contaminants such as the one with anthropogenic source (e.g. NO3-) and bacterial nature can be determined. In identifying the water origin, this proposed method employed NO3- and DOC data normalization. Values are projected in the NO3-_norm = f(DOC_norm) reference frame. These are aligned to the slope. Depending on the obtained slope (α), water origin can be disclosed. If α > 1, the increase of concentration of DOC weighs more, characterizing water from surface runoff. Whereas, if α < 1, the consideration is more on the increase of NO3- concentration, characterizing water from unsaturated zone. However if α cannot be calculated because there is no evident slope, this characterizes the water already present in the system. Water originating from the surface runoff is prone to inorganic and bacterial contamination adsorbed by the particles. Identifying the type of water perturbation needing water treatment is important in managing the water resource. Hence, the evolution through time of NO3- and DOC with the particle size distribution, anthropogenic nature type of contaminant (i.e. in this study NO3-), and presence or absence of bacteria were examined. This method was applied in the springs of the Toulon, an important drinking water source of the city of Périgueux in France. This site was chosen considering the following factors: (1) its karst nature being vulnerable to infiltrations, having fractures and sinkholes; (2) its land use being influenced by the anthropogenic activities such as agriculture; and (3) its observed pronounced turbidity incidence. The first flood events of two hydrological cycles were assessed. Three water origins of the spring water and the respective water quality hazards were identified: (i) water from saturated zone with minerals, (ii) water from unsaturated zone with nitrate, and (iii) water from surface runoff with the presence of bacteria. The second and third types of water perturbation gave evidence that the Toulon springs can be contaminated. Hence, in terms of resource management, the information obtained can be used as a basis in forecasting and planning the management actions or water quality treatments needed.

Flux estimation, temporal trends and source determination of trace metal contamination in a major tributary of the Seine estuary, France.

Anthropogenic impacts on rivers have increased significantly over the past ~150 years, particularly at the beginning of the industrial revolution. Among other signs, this impact is manifested through the addition of trace metals and metalloid elements to rivers. The Eure River watershed in France covers an area of 6017 km2 and is a major tributary of the Seine estuary. It is not exempt from anthropogenic pressures and has been exposed to significant metal discharges over the last 80 years. The average concentrations of metals (i.e., Cr, Co, Ni, Cu, Zn, Ag, Cd, Sb, and Pb), in suspended particulate matter currently transported by the river are high compared to the local geochemical background. Moreover, the lack of correlation between concentration variations and the hydrosedimentary behaviour of the Eure River suggests that the river is currently under anthropogenic pressure. Analysis of sediment cores indicate strong As contamination during the 1940s, Cr, Co, Ni, Cu, Zn, Ag, and Cd contamination during the 1960s and 1970s, and Sb and Pb contamination during the 1990s and 2000s. The enrichment factors calculation suggests that total anthropogenic pressure within the Eure River watershed since the 1940s was comparable or higher than those in many other French watersheds. An estimation of particulate metal flux in 2017 shows that the Eure River watershed contributed to 7, 8, 9, 10 and 16% of total inputs to the Seine estuary in Cr, Cu, Zn, Cd and Pb respectively. Moreover, the estimation of past theoretical flux indicates that during the 1990s the Eure River watershed was the main contributor of particulate Pb to the estuary. The use of Pb isotopes has revealed that this contamination was primarily of industrial origin.

Author Correction: Global karst springs hydrograph dataset for research and management of the world's fastest-flowing groundwater.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Global karst springs hydrograph dataset for research and management of the world's fastest-flowing groundwater.

Karst aquifers provide drinking water for 10% of the world's population, support agriculture, groundwater-dependent activities, and ecosystems. These aquifers are characterised by complex groundwater-flow systems, hence, they are extremely vulnerable and protecting them requires an in-depth understanding of the systems. Poor data accessibility has limited advances in karst research and realistic representation of karst processes in large-scale hydrological studies. In this study, we present World Karst Spring hydrograph (WoKaS) database, a community-wide effort to improve data accessibility. WoKaS is the first global karst springs discharge database with over 400 spring observations collected from articles, hydrological databases and researchers. The dataset's coverage compares to the global distribution of carbonate rocks with some bias towards the latitudes of more developed countries. WoKaS database will ensure easy access to a large-sample of good quality datasets suitable for a wide range of applications: comparative studies, trend analysis and model evaluation. This database will largely contribute to research advancement in karst hydrology, supports karst groundwater management, and promotes international and interdisciplinary collaborations.

Polycyclic aromatic hydrocarbon transport processes on the scale of a flood event in the rural watershed of Le Bebec, France.

Transport processes of polycyclic aromatic hydrocarbons (PAHs) were investigated in a rural watershed in France, which led to new quantitative results on the scale of a flood event. For that purpose, principal components analysis (PCA) combined with signal decomposition of PAH Breakthrough curves were used. The results obtained by PCA enabled us to distinguish (i) the transport of low molecular weight PAHs in dissolved form and (ii) the transport of high molecular weight PAHs in particulate form. The intermediate molecular weight PAHs are distributed in these two clusters. The application of a signal decomposition method confirms the results of PCA and makes it possible to quantify the mass PAH resulting, respectively, from resuspension (13.3mg, i.e. 77% of the total mass) and from direct transfer (2.8mg=16% of the total mass). For the resuspension phase, desorption of PAH (38% of the total exported mass) could be also distinguished from sediment resuspension (39%).

Assessment of faecal contamination and the relationship between pathogens and faecal bacterial indicators in an estuarine environment (Seine, France).

The Seine estuary, one of the largest estuaries of the European northwest continental shelf, is subjected to numerous anthropogenic influences. Here we present an assessment of the microbial faecal contamination of the estuary water. The most vulnerable areas were defined on the basis of the fluxes of indicator organisms and the occurrence of Salmonella and Cryptosporidium sp. and Giardia sp. (oo)cysts. The microbial quality of the water changes from upstream to downstream: in the upstream area, contamination by faecal-indicator bacteria and Salmonella occurs during periods of high flow; in the urbanized area, mid-way between the uppermost areas of the estuary and its mouth, discharge from a wastewater treatment plant and a tributary degrade water quality; at the estuary mouth, the accumulation of microorganisms attached to particles in the maximum turbidity zone, particularly Clostridium perfringens spores and oocysts of Cryptosporidium, is accompanied by inputs of ThC and Escherichia coli from tributaries. In some areas, significant strong relations are observed between Salmonella, (oo)cysts of protozoan, and levels of faecal indicators.

Change in the Structure of Escherichia coli Population and the Pattern of Virulence Genes along a Rural Aquatic Continuum.

The aim of this study was to investigate the diversity of the Escherichia coli population, focusing on the occurrence of pathogenic E. coli, in surface water draining a rural catchment. Two sampling campaigns were carried out in similar hydrological conditions (wet period, low flow) along a river continuum, characterized by two opposite density gradients of animals (cattle and wild animals) and human populations. While the abundance of E. coli slightly increased along the river continuum, the abundance of both human and ruminant-associated Bacteroidales markers, as well as the number of E. coli multi-resistant to antibiotics, evidenced a fecal contamination originating from animals at upstream rural sites, and from humans at downstream urban sites. A strong spatial modification of the structure of the E. coli population was observed. At the upstream site close to a forest, a higher abundance of the B2 phylogroup and Escherichia clade strains were observed. At the pasture upstream site, a greater proportion of both E and B1 phylogroups was detected, therefore suggesting a fecal contamination of mainly bovine origin. Conversely, in downstream urban sites, A, D, and F phylogroups were more abundant. To assess the occurrence of intestinal pathogenic strains, virulence factors [afaD, stx1, stx2, eltB (LT), estA (ST), ipaH, bfpA, eae, aaiC and aatA] were screened among 651 E. coli isolates. Intestinal pathogenic strains STEC O174:H21 (stx2) and EHEC O26:H11 (eae, stx1) were isolated in water and sediments close to the pasture site. In contrast, in the downstream urban site aEPEC/EAEC and DAEC of human origin, as well as extra-intestinal pathogenic E. coli belonging to clonal group A of D phylogroup, were sampled. Even if the estimated input of STEC (Shiga toxin-producing E. coli) - released in water at the upstream pasture site - at the downstream site was low, we show that STEC could persist in sediment. These results show that, the run-off of small cattle farms contributed, as much as the wastewater effluent, in the dissemination of pathogenic E. coli in both water and sediments, even if the microbiological quality of the water was good or to average quality according to the French water index.

Assessing the Nonlinearity of Karst Response Function under Variable Boundary Conditions.

The hydraulic and transfer response of karst aquifers is complex and often highly nonlinear: due to their high transmissivity and connection with the surface, such systems are very sensitive to modifications of their boundary conditions. The aim of this study was to assess the variation of the response depending on both upstream and downstream parameters, and propose a methodology to simulate the response of the karst system depending on those parameters. The impact of the variations of multiple environmental parameters on the response of a karstic system submitted to tidal variations (Normandy, France) was investigated after a campaign of artificial tracer tests acquired in very different hydrologic conditions (rainfall events, low tide, high tide, low/high piezometric level, and low/high waters). Principal components analysis and hierarchical clustering were applied on both environmental variables and karstic system response variables (parameters of the residence time distribution [RTD] curves). Equations between the RTD parameters and the most relevant variables were established using a symbolic regression algorithm. It appeared that the variations of the RTD parameters depend mainly on the cumulated rainfall preceding the injection, the piezometric level of the aquifer, and on the tide parameters. The hydraulic conditions downstream of the aquifer have a strong influence on the hydraulic and transfer response of the aquifer. The response of the aquifer in various and extreme conditions has been simulated using the equations resulting from the symbolic regression algorithm. Such relationships can be useful for management of water resources in karst media, and support decision making.

Diversity of active microbial communities subjected to long-term exposure to chemical contaminants along a 40-year-old sediment core.

In estuarine ecosystems, metallic and organic contaminants are mainly associated with fine grain sediments which settle on mudflats. Over time, the layers of sediment accumulate and are then transformed by diagenetic processes mainly controlled by microbial activity, recording the history of the estuary's chemical contamination. In an environment of this specific type, we investigated the evolution of the chemical contamination and the structure of both total and active microbial communities, based on PhyloChip analysis of a 4.6-m core corresponding to a 40-year sedimentary record. While the archaeal abundance remained constant along the core, a decrease by one order of magnitude in the bacterial abundance was observed with depth. Both total and active microbial communities were dominated by Proteobacteria, Actinobacteria, and Firmicutes in all sediment samples. Among Proteobacteria, alpha-Proteobacteria dominated both total (from 37 to 60 %) and metabolically active (from 19.7 to 34.6 %) communities, including the Rhizobiales, Rhodobacter, Caulobacterales, and Sphingomonadales orders. Co-inertia analysis revealed a relationship between polycyclic aromatic hydrocarbons, zinc and some polychlorobiphenyls concentrations, and the structure of total and active microbial communities in the oldest and most contaminated sediments (from 1970 to 1975), suggesting that long-term exposure to chemicals shaped the structure of the microbial community.

Antibiotic-resistant Escherichia coli in karstic systems: a biological indicator of the origin of fecal contamination?

Occurrences of antibiotic-resistant Escherichia coli in two springs of a karstic system (NW France) providing drinking water were determined to study the role of aquifers in the dissemination of the resistance genes. Water samples were collected during wet and dry periods and after a heavy rainfall event to investigate E. coli density, antibiotic resistance patterns, and occurrences of class 1, 2, and 3 integrons. By observing patterns of the resistant isolates (i.e. number and type of resistances) and their occurrences, we were able to define two resistant subpopulations, introduced in the aquifer via surface water: (1) R1-2, characterized by one or two resistance(s), essentially to chloramphenicol and/or tetracycline (96.5%), was always found during the heavy rainfall event; (2) R3-10, characterized by three or more resistances, mostly resistant to tetracycline (94.1%) and beta-lactams (86%), was found transiently. Class 1 and 2 integrons were detected, mostly in the R3-10 subpopulation for class 1 integrons. The characteristics of these two subpopulations strongly suggest that the contamination originates from pasture runoff for the R1-2 subpopulation and from wastewater treatment plant effluents for the R3-10 subpopulation. These two subpopulations of E. coli could be used as biological indicators to determine the origin of groundwater contamination.