The aim of this study was to investigate the relationship between antibiotics and antibiotic-resistant fecal bacteria (E. coli) in water along a medical center-wastewater treatment plant-river continuum (4 km). A multiresidue chemical analysis methodology, using solid phase extraction coupled with liquid chromatography tandem mass spectrometry, was performed to detect whether low levels of contamination by 34 antibiotics were related to antibiotic resistance of E. coli and antibiotic use. The contamination of water by antibiotics and antibiotic-resistant E. coli decreased along the continuum. Although amoxicillin was predominantly prescribed, only ofloxacin (1 ng·L(-1)) and sulfamethoxazole (4 ng·L(-1)) persisted in the river. At the retirement home, in the medical center, even though no tetracycline and sulfamethoxazole were consumed, the highest occurrences of antibiotic resistance were in classes of quinolones (42.0%), sulfonamides (24.0%), tetracyclines (38.0%), and penicillins (38.0%), mainly due to the presence of multiple antibiotic-resistance genes on class 1 integrons. Along the continuum, the occurrence of E. coli resistant to antibiotics and those carrying class 1 integrons decreased in water samples (p-value <0.001). Interestingly, in the river, only persistent antibiotic compounds (ofloxacin, sulfamethoxazole) were found, but they did not correspond to the major resistances (tetracycline, amoxicillin) of E. coli.

e To determine if hospital effluent input has an ecological impact on downstream aquatic environment, antibiotic resistance in Enterococcus spp. along a medical center-retirement home-wastewater treatment plant-river continuum in France was determined using a culture-based method. Data on antibiotic consumption among hospitalized and general populations and levels of water contamination by antibiotics were collected. All isolated enterococci were genotypically identified to the species level, tested for in vitro antibiotic susceptibility, and typed by multilocus sequence typing. The erm(B) and mef(A) (macrolide resistance) and tet(M) (tetracycline resistance) genes were detected by PCR. Along the continuum, from 89 to 98% of enterococci, according to the sampled site, were identified as Enterococcus faecium. All E. faecium isolates from hospital and retirement home effluents were multiply resistant to antibiotics, contained erm(B) and mef(A) genes, and belonged to hospital-adapted clonal complex 17 (CC17). Even though this species remained dominant in the downstream continuum, the relative proportion of CC17 isolates progressively decreased in favor of other subpopulations of E. faecium that were more diverse, less resistant to antibiotics, and devoid of the classical macrolide resistance genes and that belonged to various sequence types. Antibiotic concentrations in waters were far below the MICs for susceptible isolates. CC17 E. faecium was probably selected in the gastrointestinal tract of patients under the pressure of administered antibiotics and then excreted together with the resistance genes in waters to progressively decrease along the 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.

14Bovine digital dermatitis (DD), is a highly prevalent disease among dairy cattle 15 characterized by ulcerative and painful lesions. While multiple risk factors are involved 16 in the disease, its precise etiology remains unclear and the effectiveness of the current 17 control strategies still highly variable. However, and even if the main role of Treponema 18 spp. in the development of the disease is consistently recognized, the importance of 19 other bacterial communities and the factors affecting the skin environment remains 20 uncertain. The objective of this study was to investigate the dynamics of microbiotas 21 from feet affected by DD, before and after the implementation of disinfectant footbaths 22 under field conditions. During this investigation, the diversity, structure, and 23 composition of microbiotas from DD lesions over time were explored according to 24 different clinical factors. The findings of this investigation confirmed a particular 25 lesions evidenced a different structure and diversity in comparison to non-proliferative 30 lesions. The composition of microbiotas changed over time revealing the potential main 31 role of Treponema spp., Fusobacterium spp., Mycoplasma spp. and Porphyromonas 32 spp. in the dynamics of DD lesion progression. Further studies are necessaries to 33 confirm if the clinical evolution of DD lesions is driven by a particular microbiota and 34 how this microbiota induces disease. 35 Highlights 39Multiple bacteria have been identified in DD lesions. However, a large part of these 40 microorganisms are inhabitants of the foot skin and the farm environment. For the first 41 time, the microbiotas of DD lesions were followed over 45 days under field conditions 42 to explore their evolution over time and how footbaths practices may affect their 43 dynamics. The results of this investigation confirmed a particular microbiota related to 44 DD lesions dominated by Treponema spp. and highly different from those microbiotas 45 of healthy skin. The microbiotas from DD lesions evolved over the study period and 46 the differential bacteria were identified. Further studies are guaranteed to determine 47 the role of the bacteria composing these microbiotas on the lesion occurrence and its 48 outcome. 49 50 polymicrobial disease spread across the world and characterized by ulcerative and 55 painful lesions which might persist as a chronic condition. DD lesions progress in 56 dynamic ways conditioned by multiple environmental factors that influence the spread 57 of associated-pathogens and the integrity of the foot skin. Currently, the control 58 strategies implemented in dairy farms such as footbaths or individual treatments, have 59 demonstrated variable effectiveness in the healing and prevention of DD lesions 2,3 . 60Although the precise etiology of DD remains unclear, the maceration of the foot skin 61 and the presence of Treponema spp. have been consistently identified as the major 62 etiological components of DD 4-7 . However, multiple other bacteria have also been 63 associa...

Footbaths represent a potentially useful strategy for the prevention of claw infectious diseases by treating a large number of animals concomitantly. Nevertheless, under field conditions, footbath solutions are exposed to increasing number of animal passages and therefore, to different volume losses and concentrations of manure contamination which could alter their presumed bactericidal activity. Across increasing number of cow passages, the organic matter (OM) concentration, the microbial load (ML) and the residual volumes were assessed in six commercial farms. The results indicate that the OM concentration and ML increased linearly with the number of passages of animals, and with the number of defecations in the footbath. The OM concentrations and the ML were not impacted by the farm’s feet hygiene status (clean, fair and dirty), suggesting that probably the increasing number of cow passages and defecations influenced more the contamination of footbaths than the hygiene of the feet. In all the farms the volumes decreased drastically after 200 cow passages (50%). The OM concentrations after 150 and 200 cow passages did not exceed the regulatory concentrations in which disinfectant products should demonstrate to still be effective (20 g/l), and coincide with the often advised renewal rates. The findings of this study suggested that beyond the footbath contamination by OM, the renewal rates must be mainly adapted according to the remaining volume to guarantee that the entire foot is covered and therefore assure the topical action of the solution. This study highlights the importance of footbath designs for the successful implementation of these strategies in practice.

The collective treatment (CT) of an affected herd is commonly advised to control bovine digital dermatitis (DD). Several CT are commercialized, frequently without major evidence supporting their effectiveness. The objective of this systematic review was to evaluate the published evidence that supports CT in the treatment and prevention of DD lesions in dairy herds. Across the evidence, the main limitations in the studies design were identified and the possible sources of inconsistency were investigated. An extensive literature search of publications through electronic databases and gray literature was conducted between July 2015 and January 2016. Studies that did not include an untreated or placebo control group were excluded from the review. The literature search and screening process identified 13 publications with 24 treatment trial comparisons and 18 prevention trial comparisons. The published evidence included studies mostly considered to have a low or unclear risk of bias. Descriptive analyses were performed according to the prevention and treatment outcomes, and case and success definitions were identified for each study and summarized in odds ratios (OR). Pairwise meta-analyses were conducted according to the prevention and treatment outcomes, comparing directly the intervention used in each study, and ignoring any other differences in the intervention characteristics. The results of the meta-analyses indicated a low degree of heterogeneity across the evidence for the prevention outcome [I = 0%, 95% CI: 0 to 37.2%, 95% prediction interval (PI): 0.72 to 1.74)] and a moderate degree for the treatment outcome (I = 25.3%, 95% CI: 0 to 63%, 95% PI: 0.39 to 3.73). Similarly, appraisal of the graphical L'Abbé plot suggested a considerable degree of heterogeneity across the evidence for the treatment outcome. For both outcomes, the frequent small sample sizes of the trials indicate imprecision across the included studies. Additionally, for the treatment and prevention outcomes, an asymmetric funnel plot suggested possible publication bias. The overall quality of the evidence, for both outcomes (prevention and treatment), was therefore considered to be low, indicating that the true effect of CT may be substantially different from that estimated across the included studies. Consequently, this review and meta-analysis does not support an association between the CT considered in the review and a beneficial effect in the prevention and treatment of DD lesions. The effectiveness of CT therefore remains uncertain, and the epidemiological circumstances in which it can be useful must be investigated. These findings highlight the importance of developing high quality, controlled trials to evaluate the effectiveness of CT for DD control.

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