Research Note : Persistent Salmonella problems in slaughterhouses related to clones linked to poultry companies

Salmonellosis remains one of the main foodborne zoonoses in Europe, with poultry products as the main source of human infections. The slaughterhouse has been identified as a potential source for Salmonella contamination of poultry meat. Despite the mandatory programme of the EU, there are companies with persistent Salmonella that are unable to remove the bacteria from their processing environment, compromising the entire production line. In this context, an intensive sampling study was conducted to investigate a slaughterhouse with persistent Salmonella problems, establishing the genetic relationship among Salmonella strains isolated during the slaughter process. A total of 36 broiler flocks were sampled during processing at the slaughterhouse. Salmonella was identified based on ISO 6579-1:2017 (Annex D), serotyped by Kauffman-White-Le-Minor technique, and the genetic relationship was assessed with ERIC-PCR followed by PFGE. The outcomes showed that 69.4% of the batches sampled carried Salmonella upon arrival at the slaughterhouse and that 46.3% of the different samples from carcasses were contaminated with Salmonella. The two serovars isolated at the different steps in the slaughterhouse were Enteritidis (98.2%) and Kentucky (1.8%). Pulsed-field gel electrophoresis analysis revealed a low genetic diversity, with all S. Enteritidis isolates showing a nearly identical pulsotype (similarity >85%) and S. Kentucky strains showed the same XbaI PFGE profile (95.0% genetic similarity). The results of this study showed a high genetic relationship among isolates recovered from carcasses and environmental samples in the slaughterhouse from both Salmonella -positive and Salmonella -free flocks. Salmonella strains re-circulated across to poultry flocks and re-entered the slaughterhouse to survive on the processing line. Thus, it is necessary to implement molecular diagnosis methods in time at the field level to determine the Salmonella epidemiology of the flock, to make rapid decisions for the control of Salmonella and prevent entry into the slaughterhouse environment

Characterization of Salmonella Frintrop isolated from dromedary camels (Camelus dromedarius)

Different studies have reported the prevalence and antibiotic resistance of Salmonella in dromedary camels and its role in camelid‐associated salmonellosis in humans, but little is known about the epidemiology of Campylobacter in dromedaries. Here, we investigate the prevalence, genetic diversity and antibiotic resistance of Campylobacter and Salmonella in dromedary camels (Camelus dromedarius). A total of 54 individuals were sampled from two different dromedary farms located in Tenerife (Canary Islands, Spain). While all the samples were Campylobacter‐negative, Salmonella prevalence was 5.5% (3/54), and the only serovar isolated was S. Frintrop. Pulsed‐field gel electrophoresis analysis revealed a low genetic diversity, with all isolates showing a nearly identical pulsotype (similarity >95%). Our results indicate that dromedary camels could not be a risk factor for Campylobacter human infection, but seems to be a reservoir for Salmonella transmission. Since camel riding has become one of the main touristic attractions in several countries, and its popularity has increased considerably in recent years, a mandatory control, especially for zoonotic pathogens such as Campylobacter and Salmonella, should be implemented.info:eu-repo/semantics/acceptedVersio

Assessing the role of livestock and sympatric wild ruminants in spreading antimicrobial resistant Campylobacter and Salmonella in alpine ecosystems

Background: Livestock play an important role as reservoir of enteric pathogens and antimicrobial resistance (AMR), a health and economic concern worldwide. However, little is known regarding the transmission and maintenance of these pathogens at the wildlife-livestock interface. In this study, we assessed the occurrence, genetic diversity and AMR of Campylobacter spp. and Salmonella spp. shed by sympatric free-ranging livestock and a wild herbivore in an alpine ecosystem. Results: Campylobacter spp. was isolated from 23.3 % of cattle and 7.7 % of sheep but was not isolated from horses nor Pyrenean chamois (Rupicapra pyrenaica). Campylobacter jejuni was the most frequent species. A high genetic diversity and certain host specificity of C. jejuni isolates was observed. The main AMR detected in Campylobacter isolates was to nalidixic acid (88.2 %), ciprofloxacin (82.4 %) and tetracycline (82.4 %); only 11.7 % of the isolates were pan-susceptible and 17.6 % were multi-resistant. Salmonella ser. Newport was isolated only from one Pyrenean chamois and was pan-susceptible. Conclusions: Results show that free-ranging cattle and sheep are spreaders of Campylobacter as well as their AMR strains in the alpine environment. Therefore, contaminated alpine pastures or streams may constitute a source for the dissemination of AMR enteropathogens. However, apparently, alpine wild ungulates such as Pyrenean chamois play a negligible role in the epidemiology of zoonotic enteropathogens and AMR, and are not potential bioindicators of the burden of alpine environments.info:eu-repo/semantics/publishedVersio

Foraging in the Anthropocene: Feeding plasticity of an opportunistic predator revealed by long term monitoring

For centuries, human activities have altered the population dynamics of wildlife. New anthropogenic food sources provide a predictable and abundant food supply that often induces very significant changes in the size, distribution, and behaviour of many populations, with ultimate consequences on the structure and functioning of natural ecosystems. Here, we combine historical and contemporary feather samples of a population of a su-perabundant, opportunistic predator, the yellow-legged gull Larus michahellis, to assess its trophic ecology and relate it to human activities in the long term. Dietary assessments were based on stable isotope analysis of carbon, nitrogen, and sulphur (δ13C, δ15N and δ34S), and were conducted through three end-point (marine prey, waste from landfills-slaughterhouses, and terrestrial invertebrates) Bayesian mixing models. Our results suggest that gulls’ diet showed a progressive decrease in the consumption of marine prey throughout the most recent period (late 20th century onwards), linked to an increase in the consumption of meat waste and small terrestrial invertebrates. Reported dietary changes over the sampling period correlated positively with the availability of marine resources around the breeding area. We provide evidence suggesting that the ability of gulls to exploit efficiently diverse anthropogenic food subsidies likely resulted in the exponential demographic increase of this population throughout the 20th century. In addition, current regulations affecting the availability of these food resources (e.g., fishing discards and landfill waste) likely reversed this trend over the last decade. Long-term evidence of population trophic plasticity, like the one we present here, is essential to implement and support management and conservation actions that limit the availability of anthropogenic resources, especially when it comes to superabundant, problematic species.info:eu-repo/semantics/publishedVersio

Multi-isotopic assessments of spatio-temporal diet variability: the case of two sympatric gulls in the western Mediterranean

In predator populations, changes in foraging behaviour in response to spatio-temporal variability of prey are expected. Prey depletion might cause trophic niche widening in generalist species, but not in specialists, which are expected to increase their foraging effort without diet shifts. In sympatric species feeding on similar resources, reduced food availability can increase interspecific competition and cause trophic niche segregation. To understand these processes, we studied the spatio-temporal variability in diet and niche width in 2 sympatric gull species, the yellow-legged gull Larus michahellis and Audouin’s gull Ichthyaetus audouinii, which have experienced exponential growth in recent decades due to an increase in anthropogenic food subsidies. We sampled feathers from chicks of both species in several colonies along the western Mediterranean from 2009 to 2011 and performed stable isotope analysis of carbon, nitrogen and sulphur on these feathers. Bayesian modelling shows that both species displayed opportunistic behaviour if different types of resources were available, but could also narrow their trophic niche if 1 resource was abundant. We also provide evidence of trophic segregation between the 2 gull species, suggesting the occurrence of interspecific competition for food. Our meta-population approach provides a comprehensive view of the trophic ecology and the competitive interactions of these gull species. We emphasize the usefulness of 3-dimensional isotope analyses to correctly assess spatio-temporal variability in trophic behaviour of predator species, revealing differences that would remain hidden in single population studies or when using only the isotopic ratios of 2 elements.info:eu-repo/semantics/acceptedVersio

Foraging in the Anthropocene: Feeding plasticity of an opportunistic predator revealed by long term monitoring

For centuries, human activities have altered the population dynamics of wildlife. New anthropogenic food sources provide a predictable and abundant food supply that often induces very significant changes in the size, distribution, and behaviour of many populations, with ultimate consequences on the structure and functioning of natural ecosystems. Here, we combine historical and contemporary feather samples of a population of a superabundant, opportunistic predator, the yellow-legged gull Larus michahellis, to assess its trophic ecology and relate it to human activities in the long term. Dietary assessments were based on stable isotope analysis of carbon, nitrogen, and sulphur (δ13C, δ15N and δ34S), and were conducted through three end-point (marine prey, waste from landfills-slaughterhouses, and terrestrial invertebrates) Bayesian mixing models. Our results suggest that gulls' diet showed a progressive decrease in the consumption of marine prey throughout the most recent period (late 20th century onwards), linked to an increase in the consumption of meat waste and small terrestrial invertebrates. Reported dietary changes over the sampling period correlated positively with the availability of marine resources around the breeding area. We provide evidence suggesting that the ability of gulls to exploit efficiently diverse anthropogenic food subsidies likely resulted in the exponential demographic increase of this population throughout the 20th century. In addition, current regulations affecting the availability of these food resources (e.g., fishing discards and landfill waste) likely reversed this trend over the last decade. Long-term evidence of population trophic plasticity, like the one we present here, is essential to implement and support management and conservation actions that limit the availability of anthropogenic resources, especially when it comes to superabundant, problematic species

Supplementary feeding stations for conservation of vultures could be an important source of monophasic Salmonella typhimurium 1,4,[5],12:i:-

[EN] Vultures are nature's most successful scavengers, feeding on the carcasses of dead animals present in the field. Availability of domestic carrion has been unstable due to rapidly changing agro-grazing economies and increasing sanitary regulations that may require burial or burning of livestock carcasses. Thus, several griffon vulture (Gyps fulvus) recoveries are based on European legislation that guarantees the animals' welfare, avoids intense persecution of the vultures and allows the feeding of threatened wildlife in supplementary feeding stations (SFS). However, in recent years, many studies have speculated on the likelihood that avian scavengers may be infected by feeding on pig carcasses at SFS from intensive livestock. In this context, the present study evaluated whether free-living griffon vultures and pig farms share zoonotic Salmonella strains to test the hypothesis that vulture are infected during consumption of carcasses provided at SFS. Here, the occurrence, serotypes and genomic DNA fingerprinting (phage typing and pulsed-field gel electrophoresis) of isolated strains were carried out in griffon vultures and pig farms authorised to provided carcasses at SFS in Castellón province (eastern Spain). The bacteriological analyses revealed that 21.1% of vultures and 14.5% for pig farms samples tested were Salmonella-positive. Monophasic S. typhimurium 1,4,[5],12:i:- was the most frequently isolated serovar. Comparison of Salmonella strains isolated from vultures and pig farms revealed that monophasic S. typhimurium 1,4,[5],12:i:-, S. Derby and S. Rissen strains were highly genetically homogeneous (similar DNA fingerprint). In conclusion, the current study indicates that free-living griffon vultures and pig farms that provide the carcasses at SFS share several zoonotic Salmonella strains. On this basis, and although transmission could be bidirectional, our result seems to corroborate the pig carcasses-to-vulture transmission and cross-infection at SFS. As an immediate Salmonella control strategy in wild avian scavengers, we suggest the implementation of a programme to guarantee that solely pig carcasses from Salmonella-free farms arrive at SFS.Funding grants from the Ministry of Infrastructures, Territory and Environment (Regional Government/Generalitat Valenciana) and from CEU-UCH (Consolidacion de Indicadores INDI15/16, INDI16/20, INDI17/25) are acknowledged. CERCA Programme from the Generalitat de Catalunya is also acknowledged.Marin-Orenga, C.; Torres, C.; Marco Jiménez, F.; Cerdà-Cuéllar, M.; Sevilla-Navarro, S.; Ayats, T.; Vega-García, S. (2018). Supplementary feeding stations for conservation of vultures could be an important source of monophasic Salmonella typhimurium 1,4,[5],12:i:-. The Science of The Total Environment. 636:449-455. https://doi.org/10.1016/j.scitotenv.2018.04.310S44945563

Do humans spread zoonotic enteric bacteria in Antarctica?

Reports of enteric bacteria in Antarctic wildlife have suggested its spread from people to seabirds and seals, but evidence is scarce and fragmentary. We investigated the occurrence of zoonotic enteric bacteria in seabirds across the Antarctic and subantarctic region; for comparison purposes, in addition to seabirds, poultry in a subantarctic island was also sampled. Three findings suggest reverse zoonosis from humans to seabirds: the detection of a zoonotic Salmonella serovar (ser. Enteritidis) and Campylobacter species (e.g. C. jejuni), typical of human infections; the resistance of C. lari isolates to ciprofloxacin and enrofloxacin, antibiotics commonly used in human and veterinary medicine; and most importantly, the presence of C. jejuni genotypes mostly found in humans and domestic animals but rarely or never found in wild birds so far. We also show further spread of zoonotic agents among Antarctic wildlife is facilitated by substantial connectivity among populations of opportunistic seabirds, notably skuas (Stercorarius). Our results highlight the need for even stricter biosecurity measures to limit human impacts in Antarctica.info:eu-repo/semantics/acceptedVersio

Chicken liver is a potential reservoir of bacteriophages and phage-derived particles containing antibiotic resistance genes

Poultry meat production is one of the most important agri-food industries in the world. The selective pressure exerted by widespread prophylactic or therapeutic use of antibiotics in intensive chicken farming favours the development of drug resistance in bacterial populations. Chicken liver, closely connected with the intestinal tract, has been directly involved in food-borne infections and found to be contaminated with pathogenic bacteria, including Campylobacter and Salmonella. In this study, 74 chicken livers, divided into sterile and non-sterile groups, were analysed, not only for microbial indicators but also for the presence of phages and phage particles containing antibiotic resistance genes (ARGs). Both bacteria and phages were detected in liver tissues, including those dissected under sterile conditions. The phages were able to infect Escherichia coli and showed a Siphovirus morphology. The chicken livers contained from 10 3 to 10 6 phage particles per g, which carried a range of ARGs (bla , bla , sul1, qnrA, armA and tetW) detected by qPCR. The presence of phages in chicken liver, mostly infecting E. coli, was confirmed by metagenomic analysis, although this technique was not sufficiently sensitive to identify ARGs. In addition, ARG-carrying phages were detected in chicken faeces by qPCR in a previous study of the group. Comparison of the viromes of faeces and liver showed a strong coincidence of species, which suggests that the phages found in the liver originate in faeces. These findings suggests that phages, like bacteria, can translocate from the gut to the liver, which may therefore constitute a potential reservoir of antibiotic resistance genes. Phage particles carrying antibiotic resistant genes have been found in chicken livers. Metagenomic analysis of chicken liver viromes (PL) and chicken faeces viromes (HP) suggests that these phage particles could translocate from chicken gut to the liver

Characterization of Campylobacter jejuni and Campylobacter coli Broiler Isolates by Whole-Genome Sequencing

Campylobacter has been the most commonly reported cause of bacterial diarrheal disease in humans in the European Union since 2005. Most broiler batches at slaughter are colonized with Campylobacter, and the major source of infection is contaminated poultry meat. The aim of this study was to characterize a selection of Campylobacter jejuni and Campylobacter coli isolates from broilers through whole-genome sequencing (WGS). A total of 16 isolates (C. jejuni = 12 and C. coli = 4) from five broiler farms from Catalonia (northeastern Spain) were analyzed. A phylogenetic analysis based on 8420 single-nucleotide polymorphisms showed two main cluster grouping strains by species. Phenotypic resistances to quinolones (100%), tetracycline (81%), streptomycin (75%), erythromycin (56%), and gentamicin (13%) were found. All the isolates carried the C257T point mutation in the subunit A of the DNA gyrase gene (Thr86Ile) conferring resistance to quinolones, while all the isolates showing resistance to tetracycline carried the tet(O) gene. The genes aph(3′)-III and aadE conferring resistance to aminoglycosides were identified in the two isolates (one C. jejuni and one C. coli) resistant to streptomycin and gentamicin. The point mutation A2075G on the 23S rDNA conferring high resistance to macrolides was detected in three C. coli isolates. The CmeABC multidrug efflux pump was also detected, both in C. jejuni and C. coli isolates. All C. jejuni and C. coli isolates were positive for most of the 34 virulence-associated genes studied related to motility, chemotaxis, adhesion, and invasion. Interestingly, the wlaN gene involved in the Guillain–Barré syndrome was found in two isolates. The results underline the power of WGS for investigation of virulence, clonality, and antimicrobial resistance in Campylobacter.info:eu-repo/semantics/acceptedVersio