Characterization of ESBL/AmpC-producing extraintestinal Escherichia coli (ExPEC) in dogs treated at a veterinary hospital in Brazil.

The clinical aspects and lineages involved in Extraintestinal pathogenic Escherichia coli (ExPEC) infections in dogs remain largely unknown. In this study, we investigated the antimicrobial resistance and molecular structures of ExPECs isolated from infected dogs in Brazil. Samples were obtained from dogs (n = 42) with suspected extraintestinal bacterial infections. Phylogroup B2 was predominant (65.1%). No association was observed between the site of infection, phylogroups, or virulence factors. Almost half of the isolates (44.2%) were MDR, and 20.9% were extended-spectrum ß-lactamase (ESBL)-positive. E. coli isolates that were resistant to fluoroquinolones (27.9%) were more likely to be MDR. The CTX-M-15 enzyme was predominant among the ESBL-producing strains, and seven sequence types were identified, including the high-risk clones ST44 and ST131. Single SNPs analysis confirmed the presence of two clonal transmissions. The present study showed a high frequency of ExPECs from phylogroup B2 infecting various sites and a high frequency of ESBL-producing strains that included STs frequently associated with human infection. This study also confirmed the nosocomial transmission of ESBL-producing E. coli, highlighting the need for further studies on the prevention and diagnosis of nosocomial infections in veterinary settings.

Canine Pyometra: A Short Review of Current Advances.

Pyometra, characterized by the accumulation of purulent exudate in the uterus, is the most prevalent reproductive disease in canines. While the disease often begins with mild local symptoms, it can escalate into peritonitis, sepsis, and multi-organ dysfunction, thereby posing a significant threat to life. Despite the high incidence and recognized significance of canine pyometra, gaps persist in our understanding of its epidemiology, etiology, and pathogenesis. Recent studies have, however, broadened our comprehension of this disease, shedding light on potential new infection sources, etiologies, and the application of clinical predictive biomarkers and new therapeutic protocols. This study aimed to review the current understanding of canine pyometra, with particular emphasis on the latest research concerning its etiology and epidemiology. Furthermore, it addressed key research questions and proposed directions for future investigations into various facets of canine pyometra.

Risk Factors, Genetic Diversity, and Antimicrobial Resistance of Staphylococcus spp. Isolates in Dogs Admitted to an Intensive Care Unit of a Veterinary Hospital.

Intensive Care Units (ICU) usually provide an excellent environment for the selection of pathogens associated with hospital-acquired infections (HAI), leading to increased mortality and hospitalization costs. Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is a major cause of HAI in dogs worldwide, but the risk factors and dynamics of colonization by MRSP are largely unknown. This study aimed to evaluate the risk factors associated with the acquisition of MRSP in dogs admitted to an ICU, and to report the antimicrobial resistance profiles and genetic relatedness of MRSP isolates. Sterile swabs from the nostril, axilla, and rectum were collected daily during the hospitalization of 54 dogs. Samples were subjected to Mannitol Salt Agar, and colonies were identified by MALDI-ToF, polymerase chain reaction (PCR), and sequencing of the rpoB gene. Antimicrobial susceptibility testing and PCR detection of mecA were performed. Staphylococcus spp. was isolated from 94% of the dogs, and the most frequently isolated species was S. pseudintermedius (88.2%). Carriage of multidrug resistant (MDR) staphylococci was observed in 64.4% of the dogs, and approximately 39% had methicillin-resistant Staphylococcus sp. (MRS), of which 21.6% had MRSP and 1.9% had methicillin-resistant S. aureus (MRSA). The acquisition of MRSP during ICU hospitalization was associated with sex (female), age (>7 years), and dogs that had previously been treated with antimicrobials. Animals colonized by MRSP resistant to ≥9 antimicrobial classes had longer hospital stays than those colonized by other MRS strains. Among the 13 MRSP isolates that were subjected to whole-genome sequencing, ten were classified as ST71. A single nucleotide polymorphism (SNP) analysis revealed three clones, including one that was detected in infected dogs outside the ICU. This study indicates novel risk factors associated with colonization by MRSP. The detection of the same MRSP clone causing HAI outside the ICU reinforces the need for improved infection prevention and control practices at veterinary hospitals in general and at the ICU in particular.

Evaluation of disk diffusion method for testing the rifampicin, erythromycin, and tetracycline susceptibility of Clostridioides (prev. Clostridium) difficile.

INTRODUCTION: Antimicrobial resistance (AMR) is one of the greatest threats to animal and public health. Clostridioides (prev. Clostridium) difficile is a major burden to healthcare and a relevant AMR gene reservoir. Despite the known importance of AMR in C. difficile epidemiology and treatment, antimicrobial susceptibility testing for this pathogen is still based on the determination of the minimal inhibitory concentration (MIC) by the agar dilution method, which is technically demanding and labor-intensive. In this study, the disk diffusion method was used to evaluate the susceptibility of C. difficile to erythromycin, rifampicin, and tetracycline. MATERIAL AND METHODS: A total of 155 isolates isolated between 2011 and 2022 from humans and animals in Brazil were simultaneously tested using the disk diffusion method and the epsilometer test (Etest) for these three antimicrobials on Brucella blood agar supplemented with vitamin K and hemin. RESULTS: The results suggest that disk diffusion can be an interesting routine tool to identify erythromycin- and rifampicin-resistant C. difficile isolates (≥20 mm cut-off) and wild type (WT) strains (≥28 mm). However, the disk diffusion protocol tested in this study does not seem suitable for tetracycline because of the common misclassification of resistant strains.

Polyarteritis nodosa in a captive common wooly monkey (Lagothrix lagotricha) associated with intestinal necrosis, peritonitis, and sepsis.

Polyarteritis nodosa is an idiopathic necrotizing vasculitis that affects small to medium-sized arteries. We describe a case of polyarteritis nodosa in a captive common wooly monkey (Lagothrix lagotricha) associated with transmural intestinal necrosis and secondary peritonitis. This condition must be considered for differential diagnosis of segmental arteritis in neotropical primates.

Transmission of Escherichia coli Causing Pyometra between Two Female Dogs.

Despite its clinical relevance, the pathogenesis of canine pyometra remains poorly understood. To date, it is recognized as a non-transmissible infectious disease. In this study, the simultaneous occurrence of pyometra and Escherichia coli in two cohabitant female dogs underwent in-depth investigation due to the hypothesis of transmission between these animals. Two 5-year-old Chow Chow dogs (namely, dogs 23 and 24-D23 and D24) were referred to a veterinary hospital with suspected pyometra. Both animals showed prostration, anorexia, and purulent vulvar discharge over a 1-week period. After ovariohysterectomy, uterine tissue, uterine contents, and rectal swabs were collected for histopathological and microbiological analysis. Uterine histology demonstrated purulent material and multifocal necrosis with endometrial ulceration, and a morphological diagnosis of pyometra was confirmed. Furthermore, E. coli from the same phylogroup (B2) and positive for the same virulence factors with the same antimicrobial susceptibility profile was isolated from the uterine contents of both dogs and the rectum of D23. Conversely, the E. coli strains recovered from D24 differed in phylogroup (one isolate), virulence factors (all three isolates), and antimicrobial susceptibility (all three isolates). Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) suggested that all isolates from the uterine content of both dogs and the rectal swab of D23 were 100% the same, but different from all isolates in the rectal swab of D24. One isolate from the uterine content of each animal as well as rectal swabs were subjected to whole-genome sequencing (WGS). Both whole-genome multilocus sequence typing(wgMLST) and single-nucleotide polymorphism (SNP) analysis supported the hypothesis that the isolates from the uterine content of both animals and the rectal swab of D23 were clonal. Taken together, these clinical features, pathology, microbiology, and molecular findings suggest, to the best of our knowledge, the first transmission of E. coli associated with pyometra between two animals. These results could impact the management of sites where several females cohabit in the same local area such as kennels.

Lethal acute diarrhea associated with Clostridioides difficile toxin A and B in a buffy-tufted-ear marmoset (Callithrix aurita).

This is a case of lethal acute diarrhea associated with a mild neutrophilic enteritis in a buffy-tufted-ear marmoset (Callithrix aurita) with detection of A/B toxins and isolation of a toxigenic clade 3 Clostridioides difficile strain (A+ B+ CDT+ , ST5), which should be considered as a potential cause of enteritis in this species.

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.

Characterization of Escherichia coli in Dogs with Pyometra and the Influence of Diet on the Intestinal Colonization of Extraintestinal Pathogenic E. coli (ExPEC).

Despite its high frequency and clinical relevance, the pathogenesis of canine pyometra remains poorly understood. The most accepted hypothesis is that bacteria involved ascend from the intestinal tract, causing the uterine infection. Extraintestinal pathogenic Escherichia coli (ExPEC) is the most frequent pathogen in canine pyometra, accounting for 57-100% of cases. The aim of the present study was to determine the frequency of phylogenetic groups and virulence factors in E. coli strains isolated from the uterine and rectal swabs of bitches with pyometra (n = 72) and from rectal swabs from healthy bitches fed commercial dry feed (n = 53) or a raw meat-based diet (RMBD; n = 38). A total of 512 strains of E. coli were isolated and divided into five categories according to the origin of the sample: 120 isolates from the uterine content of dogs with E. coli pyometra, 102 from the feces of bitches with E. coli pyometra, 75 from the feces of bitches without E. coli pyometra, 130 feces samples from healthy dogs fed commercial feed, and 85 feces samples from healthy dogs fed a raw meat-based diet. E. coli strains belonging to the B2 phylogroup and positive for virulence factor genes associated with adhesion (fimbriae type P [papC]) and production of toxins (α-hemolysin [hlyA] and uropathogenic specific protein [usp]) predominated in the uterine content and rectal swabs of bitches with E. coli pyometra. Interestingly, a lower growth rate of E. coli from the B2 phylogroup was observed in dogs fed a RMBD than in those fed commercial dry feed. The present study suggests that intestinal colonization by certain types of E. coli could be a risk factor for the occurrence of E. coli pyometra in bitches and that diet can influence intestinal colonization by such strains.

Mammaliicoccus (Staphylococcus) sciuri-induced suppurative meningoencephalitis and bacteremia in an infant western lowland gorilla (Gorilla gorilla gorilla).

Mammaliicoccus (Staphylococcus) sciuri has been rarely associated with infections and sepsis in humans. A 3-month-old male western lowland gorilla (Gorilla gorilla gorilla), born under human care, died after a traumatic event. Histologic, microbiologic, and molecular findings in postmortem demonstrated a suppurative meningoencephalitis and bacteremia associated with M. sciuri infection.

Virulence Genes Profile and Antimicrobial Susceptibility of Community-Acquired Bacterial Urinary Tract Infections in a Brazilian Hospital.

Urinary tract infections (UTI) are one of the most common diseases worldwide and Escherichia coli is the most common causative bacteria. Empirical treatment is challenging due to antimicrobial or multidrug-resistance. The aims of this study were to determine the uropathogens and their antimicrobial susceptibility profile, as well as to identify the phylogroups and virulence genes of E. coli strains, associated with community-acquired UTI in outpatients admitted at a Brazilian Hospital in southeast Brazil. In total, 47 bacterial strains were isolated from 47 patients, 44 women and 2 men (no gender record from one patient). The age of the patients whose urine culture were positive varied from 0 (less than one month) to 104 years. Most of the isolates were E. coli (41/47), followed by Klebsiella pneumoniae (2/47), Klebsiella variicola/Klebsiella aerogenes (1/47), Pseudomonas aeruginosa (1/47), Proteus mirabilis (1/47), and Citrobacter koseri (1/47). Most E. coli strains were classified as phylogroup B2 (15/41 = 36.59%) and B1 (12/41 = 29.27%) and the most common virulence genes among E. coli strains were fimH (31/41 = 75.61%), iutA (21/41 = 51.22%), and tratT (16/41 = 39.02%). Among the E. coli strains, 59% were multidrug-resistance and strains that were ampicillin, sulfamethoxazole/trimethoprim, or tetracycline-resistant exhibited more chance to be multidrug-resistance, with an odds ratio of 100.00 [95% confidence interval (CI) 9.44-1059.26], 22.50 (95% CI 3.95-128.30), and 12.83 (95% CI 2.68-61.45), respectively. Our results showed that E. coli was the main etiological agent identified and demonstrated high frequency of multidrug-resistance and virulence factors in bacterial strains isolated from UTIs.

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.

Identification and Characterization of Escherichia coli, Salmonella Spp., Clostridium perfringens, and C. difficile Isolates from Reptiles in Brazil.

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.

Outbreak of multidrug resistant Salmonella Typhimurium in calves at a veterinary hospital in Brazil / Surto de Salmonella Typhimurium multirresistante em bezerros em um hospital veterinário do Brasil

ABSTRACT: 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.

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.

Isolation and genotyping of Clostridium perfringens and Clostridium difficile in Capuchin Monkeys (Sapajus spp.) / Isolamento e genotipagem de Clostridium perfringens and Clostridium difficile em macacos-prego (Sapajus spp.)

ABSTRACT: The importance of Clostridium perfringens and C. difficile for most wild animal species remains unclear. This study aimed to isolate and genotype C. perfringens and C. difficile in stool samples from free-living and captive capuchin monkeys (Sapajus flavius and Sapajus libidinosus) in Brazil. Ten free-living S. flavius and 14 captive S. libidinosus were sampled for this study. To isolate C. difficile, stool samples were inoculated on plates containing cycloserine-cefoxitin fructose agar supplemented with horse blood and sodium taurocholate. Two different protocols for C. perfringens isolation were tested: direct plating onto selective agar and enrichment in brain heart infusion (BHI) broth followed by plating onto selective agar. C. difficile was not detected in the present study. The results were identical for both protocols tested for isolation of C. perfringens. Four samples (16.7%) were positive for C. perfringens type A, including one sample from a free-living animal (4.2%) and three from captive animals (12.5%), meaning there was no significant difference between these two groups. C. perfringens isolates were negative for all additional virulence factors evaluated, including enterotoxin encoding-gene (cpe) and beta-2 encoding-gene (cpb2). These results suggested that C. perfringens type A is found in the microbiota of capuchin monkeys, although it is less frequent than previously reported in domestic animals.

RESUMO: A importância de Clostridium perfringens e C. difficile para a maioria das espécies silvestres ainda não está clara. O objetivo do presente estudo foi isolar e genotipar C. perfringens e C. difficile em amostras de fezes de macacos-prego (Sapajus flavius e Sapajus libidinosus) de vida livre e criados em cativeiros no Brasil. Dez S. flavius de vida livre e 14 S. libidinosus de cativeiro foram incluídos no presente estudo. Para isolamento de C. difficile, as amostras de fezes foram inoculadas em agar cicloserina-cefoxitina frutose, suplementado com sangue e taurocolato de sódico. Para isolamento de C. perfringens, foram testados dois protocolos: plaqueamento direto em ágar seletivo e enriquecimento em caldo seguido de plaqueamento em ágar seletivo. C difficile não foi detectado no presente estudo. Os resultados foram idênticos para ambos os protocolos testados para isolamento de C. perfringens, resultando em quatro animais (16,7%) positivos para C. perfringens tipo A. Destes, uma amostra era de um animal de vida livre (4,2%) e três de animais de cativeiro (12,5%), não havendo diferença entre esses dois grupos. Os isolados de C. perfringens foram negativos para todos os fatores de virulência adicionais avaliados, incluindo o gene codificador de enterotoxina (cpe) e o gene codificador beta-2 (cpb2). O presente estudo sugere C. perfringens tipo A como parte da microbiota de macacos-prego, embora esse agente seja menos frequente como comensal, do que relatado anteriormente, em animais domésticos.