DISTRIBUTION AND ANTIMICROBIAL RESISTANCE OF STAPHYLOCOCCI ISOLATED FROM FREE-LIVING SOUTH AMERICAN COATI (NASUA NASUA) IN AN URBAN PARK IN MINAS GERAIS, BRAZIL.

The scientific information regarding staphylococci in procyonids is scarce. Hence, the aim of this study was to evaluate the frequency, distribution, and pattern of antimicrobial resistance of staphylococcal species isolated from free-roaming coatis (Nasua nasua) in an urban park in Belo Horizonte, Minas Gerais, Brazil. Rectal swabs from 55 free-living coatis were plated onto mannitol salt agar for isolating staphylococci, and species were identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry, polymerase chain reaction (PCR) of nuc, and sequencing of 16S rRNA and rpoB when needed. Antimicrobial susceptibility was investigated using the disk diffusion method, and the presence of the mecA gene was investigated by PCR. A total of 72.7% of the animals tested positive for staphylococci. Nine different species were identified, and Staphylococcus intermedius (60.4%) and S. delphini (20.9%) were the most frequently isolated species. Most of the isolates were susceptible to most of the antimicrobials evaluated, with a resistance pattern seen for penicillin (13.9%). One isolate was multidrug-resistant (MDR). The present study suggests that coatis are natural hosts of S. intermedius and S. delphini and, despite living in a heavily anthropized environment, the Staphylococcus spp. isolates showed a low incidence of drug resistance.

Occurrence and characterization of methicillin-resistant Staphylococcus spp. in diseased dogs in Brazil.

Staphylococcus pseudintermedius is a major commensal bacterium of the skin and mucosae of dogs and an opportunistic agent responsible for several clinical infections, such as pyoderma, otitis, and surgical wound infections. The emergence of methicillin-resistant S. pseudintermedius (MRSP) has become a problem of great concern in veterinary and human medicine because it is multidrug resistant (MDR) and can also infect humans. This study aimed to identify the occurrence of Staphylococcus spp. in infected patients and investigate the antimicrobial resistance profiles and molecular structure of MRSP isolates. Samples were obtained from two different veterinary clinics; suggestive colonies were submitted to matrix-assisted laser desorption ionization-time of flight (MALDI-ToF) mass spectrometry and confirmed at the species level by polymerase chain reaction (PCR). Sequencing of the 16S rRNA and rpoB genes were used in selected samples that were not identified by MALDI-ToF and by the species-specific PCR. Antimicrobial susceptibility and PCR detection of mecA were performed. MRSP isolates were subjected to multilocus sequence typing. Of all the clinical staphylococci (n = 131), 98 (74.8%) were identified as S. pseudintermedius. Multidrug resistance (resistance to ≥3 classes of antimicrobials) was observed in 63.2% of S. pseudintermedius isolates, and 24.5% of S. pseudintermedius isolates were methicillin-resistant. Half of the MRSP isolates were isolated from surgical site infections. Among the ten sequence types (ST) identified, nine were novel. ST71 was the most prevalent and associated with resistance to fluoroquinolones. Prior antimicrobial therapy, hospitalization, and surgical site infections seemed to be risk factors for MRSP acquisition. The present study showed a high rate of MDR staphylococci in infected dogs. MRSP was isolated from different clinical conditions, mainly surgical site infections. Additionally, this is the first study to extensively investigate the population structure of MRSP in Brazil, which revealed the dispersion of CC71 and nine novel ST. These findings raise concerns for both animal and human health due to the zoonotic potential of this species and limited therapeutic options available for MRSP infections.

Fecal Shedding of Multidrug Resistant Escherichia coli Isolates in Dogs Fed with Raw Meat-Based Diets in Brazil.

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.

Fecal shedding of Salmonella spp., Clostridium perfringens, and Clostridioides difficile in dogs fed raw meat-based diets in Brazil and their owners' motivation.

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.

Antimicrobial susceptibility and molecular characterization of Salmonella serovar Ndolo isolated from outbreaks in cattle and horses / Susceptibilidade antimicrobiana e caracterização molecula de isolados de Salmonella sorovar Ndolo de bovinos e equinos

ABSTRACT: 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.