Considering the increasing popularity of reptiles as pets and their possible role as reservoirs of pathogenic microorganisms, the aim of this study was to isolate Escherichia coli, Salmonella spp., Clostridium perfringens, and C. difficile strains from reptiles in Brazil and to characterize the isolated strains. The characterization was based on phylogenetic typing of E. coli, identification of virulence genes of E. coli, C. perfringens, and C. difficile, serotyping of Salmonella spp., ribotyping and MLST of C. difficile and antimicrobial susceptibility test of pathogenic strains. Cloacal swabs were collected from 76 reptiles, of which 15 were lizards, 16 chelonians, and 45 snakes, either living in captivity, in the wild, or as companion animals. E. coli was isolated from 52 (68.4%) reptiles, of which 46 (88.4%) were characterized as phylogroup B1. The virulence factor CNF1 of E. coli was found in seven (9.2%) sampled animals, whereas the gene of EAST1 was found in isolates from two (2.6%) reptiles. Three isolates positive for CNF1 were resistant to cephalothin, one of which was also resistant to ciprofloxacin, trimethoprim/sulfamethoxazole, and chloramphenicol, being then classified as multidrug resistant strain (MDR). Salmonella enterica was identified in 26 (34.2%) reptiles, of which 13 belonged to the subspecies enterica. Serotypes such as S. Mbandaka, S. Panama, S. Infantis, S. Heidelberg, and S. Anatum were identified. One isolate of S. enterica subsp. houtenae was resistant to cephalothin and ciprofloxacin. C. perfringens type A was isolated from six (7.8%) animals. C. difficile was isolated from three (3.9%) reptiles. Two of these isolates were toxigenic and classified into ribotypes/MLST 081/ST9 and 106/ST42, which have been previously reported to infect humans. In conclusion, reptiles in Brazil can harbor toxigenic C. difficile and potentially pathogenic E. coli and Salmonella enterica subsp. enterica, thus representing a risk to human and animal health.

Skin loss can be caused by accident, burn, trauma, chronic wounds, and diseases, which is severely aggravated by multidrug‐resistant bacterial infections. Soft hybrids based on biopolymers combined with silver nanoparticles (AgNPs) have potential applications as wound dressing supports and skin tissue repair. Thus, our study focused on the design, green synthesis, and comprehensive characterization of carboxymethyl cellulose (CMC–AgNP) nanocomposites for producing hydrogel membranes, with tunable physicochemical properties, cytocompatibility, and biocidal activity for potential application as wound dressing and skin repair. These nanocomposites were prepared using CMC with two degrees of carboxymethylation, distinct concentrations of citric acid (CA) crosslinker, and AgNPs by in situ chemical reduction, forming hybrid membranes by the solvent casting method. The results demonstrated that superabsorbent hydrogels were produced with swelling and degradation behaviors dependent on the concentration of CA crosslinker, degree of carboxymethylation of CMC, and content of AgNP in the matrices. Moreover, the Fourier transform infrared spectroscopy analysis evidenced that the CMC functional groups (e.g., COOH and OH) were directly involved in the chemical reactions for the formation of AgNPs and hydrogel crosslinking pathway. These nanocomposites were cytocompatible using in vitro 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyltetrazolium bromide cell viability assay with of human embryonic kidney cells. Conclusively, the CMC–AgNP nanohybrids demonstrated to be simultaneously non‐toxic combined with highly effective antibacterial activity against gram‐positive multi‐resistant wound/skin pathogens (Staphylococcus aureus) and moderate effect towards gram‐negative strains (Escherichia coli and Pseudomonas aeruginosa). © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 45812.

The present study aimed to explore the motivations of Brazilian dog owners and their knowledge about the risks related to raw meat-based diets (RMBD) as well as to evaluate important enteropathogens such as Salmonella spp., C. perfringens, and C. difficile, in feces of dogs fed different diets. The majority of the pet owners (69.3%) reported to have chosen this diet for their dogs, considering it to be more "natural". A large number of owners declared that RMBD do not pose health risks for their animals (87.9%) or humans (98.8%), even though almost one third of the respondents (34.8%) declared having at least one individual at high risk of infection in contact with RMBD-fed dogs. Stool samples from 46 RMBD-fed dogs and 192 dogs fed commercial dry feed were collected. The present study revealed that dogs fed raw meat diets were almost 30 times more likely to be positive for Salmonella spp. than dogs on a conventional diet. Some of the serovars detected were commonly associated with human salmonellosis, such as S. Typhimurium and S. Saintpaul, and were multidrug resistant. RMBD-fed dogs were more likely to be positive for C. perfringens type A (p = 0.008) and one C. perfringens type F was isolated from these animals. Two toxigenic strains (4.3%) of C. difficile were isolated only from raw meat-fed dogs, all of which were under antibiotic therapy. These toxigenic C. difficile isolates were classified as RT106/ST54 and RT600/ST149, previously associated with infection in dogs and humans. The present work revealed that the owners have a tendency to ignore or are unaware of the risks associated with raw meat diets for dogs. Also, the higher fecal shedding of important enteropathogens in dogs fed RMBD suggests that this diet poses a risk for the animals and the people in contact with them. OPEN ACCESS Citation: Viegas FM, Ramos CP, Xavier RGC, Lopes EO, Júnior CAO, Bagno RM, et al. (2020) Fecal shedding of Salmonella spp., Clostridium perfringens, and Clostridioides difficile in dogs fed raw meat-based diets in Brazil and their owners' motivation. PLoS ONE 15(4): e0231275. https://

The practice of feeding dogs raw meat-based diets (RMBDs) is growing in several countries, and the risks associated with the ingestion of pathogenic and antimicrobial-resistant Escherichia coli in dogs fed these diets are largely unknown. We characterized E. coli strains isolated from dogs fed either an RMBD or a conventional dry feed, according to the phylogroup, virulence genes, and antimicrobial susceptibility profiles of the bacteria. Two hundred and sixteen E. coli strains were isolated. Dogs fed RMBDs shed E. coli strains from the phylogroup E more frequently and were positive for the E. coli heat-stable enterotoxin 1-encoding gene. Isolates from RMBD-fed dogs were also frequently positive for multidrug-resistant E. coli isolates including extended-spectrum beta-lactamase (ESBL) producers. Whole-genome sequencing of seven ESBL-producing E. coli strains revealed that they predominantly harbored blaCTX-M-55, and two strains were also positive for the colistin-resistant gene mcr-1. These results suggest that feeding an RMBD can affect the dog’s microbiota, change the frequency of certain phylogroups, and increase the shedding of diarrheagenic E. coli. Also, feeding an RMBD seemed to be linked with the fecal shedding of multidrug-resistant E. coli, including the spread of strains harboring mobilizable colistin resistance and ESBL genes. This finding is of concern for both animal and human health.

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The present study aimed to describe and characterize a nosocomial outbreak caused by a multidrug resistant Salmonella Typhimurium in hospitalized calves at a veterinary medical teaching hospital from Brazil. Sixty-three (96.9%) calves showed lethargy, hyperthermia and profuse diarrhea and despite treatment, 26 (41.2%) animals died. Five animals were necropsied and stool samples of six calves were collected. The isolated strains were subjected to antimicrobial susceptibility test by disc-difusion method and were fingerprinted by ERIC-PCR. Macroscopic lesions suggestive of salmonellosis, such as fibrinonecrotic enteritis and hepatosplenomegaly were observed. Salmonellosis was confirmed by isolation of S. Typhimurium from stool samples and organs from seven affected animals. Six out of seven isolates of S. Typhimurium, exhibited 100% of similarity at ERIC-PCR, suggesting occurrence of nosocomial transmission of S. Typhimurium among the hospitalized calves. All but one S. Typhimurium isolated were resistant to marbofloxacin, enrofloxacin, florfenicol, oxytetracycline and trimethoprim/sulfamethoxazole, antimicrobial agents largely used for humans and animal treatment. This is the first study of a nosocomial outbreak of multidrug resistant S. Typhimurium in a veterinary hospital in Brazil and highlighted the need for preventive measures to reduce the risks for inpatients and humans in contact with animals.RESUMO: O objetivo do presente estudo é descrever e caracterizar um surto nosocomial provocado por S. Typhimurium multirresistente em bezerros hospitalizados em um hospital escola de medicina veteriária localizado no Brasil. Sessenta e três (96,9%) bezerros apresentaram letargia, hipertermia e diarreia profusa e, apesar do tratamento, vinte e seis animais (41,2%) morreram. Cinco animais foram necropsiados e amostras fecais de seis bezerros foram coletadas. As estirpes isoladas foram submetidas a testes de susceptibilidade a antimicrobianos pelo método de disco-difusão e foram genotipadas pelo ERIC-PCR. Lesões macroscópicas sugestivas de salmonelose, como enterite fibrinonecrótica e hepatoesplenomegalia, foram observadas. Salmonelose foi confirmada pelo isolamento de S. Typhimurium em amostras fecais e órgãos de sete animais. Dos sete isolados, seis apresentaram 100% de similaridade ao ERIC-PCR, sugerindo ocorrẽncia de transmissão nosocomial de S. Typhimurium entre os bezerros hospitalizados. Com excessão de uma estirpe, todas foram resistentes a marbofloxacina, enrofloxacina, florfenicol, oxitetraciclina e trimetoprima/sulfametoxazol, agentes antimicrobianos amplamente utilizados para o tratamento humano e animal. Esse é o primeiro estudo que demonstra um surto nosocomial de estirpes de S. Typhimurium resistentes a múltiplas drogas em um hospital veterinário no Brasil, enfatizando a necessidade de medidas preventivas que reduzam os riscos aos animais hospitalizados e a pessoas que entrarem em contato com esses animais.

The present study aimed to describe and characterize, for the first time, two outbreaks of salmonellosis caused by Salmonella Ndolo in foals and calves in Brazil and compare the isolated strains with S. Ndolo previously identified in asymptomatic reptiles. The affected calves and foals presented fever, lethargy, and profuse diarrhea. Isolated strains were subjected to antimicrobial susceptibility testing, characterized according to virulence genes, and fingerprinted by ERIC-PCR. Salmonella Ndolo was identified in fecal samples from two foals and four calves. One isolate from a calf was resistant to amoxicillin/clavulanic acid, trimethoprim/sulfamethoxazole, and florfenicol. Strains from two other calves were resistant to oxytetracycline. All virulence genes tested were present in the isolates, and two major clusters of closely related strains were identified by ERIC-PCR, each per outbreak. This is the first report of Salmonella Ndolo infection in domestic and symptomatic animals. Previously, this serovar had been identified only in human infections. The presence of relevant virulence genes in all Salmonella Ndolo isolates and the detection of antimicrobial multi-resistant strains highlighted the importance of monitoring serovars associated with salmonellosis in domestic animals. RESUMO:O objetivo do presente estudo foi descrever e caracterizar, pela primeira vez, dois surtos de salmonelose causados por Salmonella Ndolo em potros e bezerros do Brasil e comparar esses isolados com Salmonella Ndolo previamente identificada em répteis assintomáticos. Os animais infectados apresentaram febre, letargia e diarreia profusa. Os isolados foram submetidos a testes de susceptibilidade a antimicrobianos e foram caracterizados conforme a presença de genes de virulência e diversidade genética, utilizando-se o ERIC-PCR. Salmonella Ndolo foi identificado em amostras fecais de dois potros e quatro bezerros. Um isolado de bezerro foi resistente a amoxicilina/ácido clavulanico, trimethoprima/sulfametoxazol e florfenicol. Estirpes de dois outros bezerros foram resistentes a oxitetraciclina. Todos os genes de virulência testados foram identificados nos isolados e dois grandes grupos de estirpes geneticamente relacionadas foram identificados pelo ERIC-PCR, um para cada surto. Esse é o primeiro relato de Salmonella Ndolo em animais domésticos e sintomáticos. Previamente, esse sorovar foi identificado apenas em infecções humanas. A presença de fatores de virulência relevantes em todos os isolados e a detecção de estirpes multirresistentes a antimicrobianos destaca a importância do monitoramento de sorovares associados a salmonelose em animais domésticos.

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