Staphylococcus spp. as part of the microbiota and as opportunistic pathogen in free-ranging black-tuffed marmosets (Callithrix penicillata) from urban areas: Epidemiology, antimicrobial resistance, and pathology.

BACKGROUND: Marmosets (Callithrix sp.), including black-tuffed marmosets (C. penicillata), are neotropical primates that can be highly adapted to urban environments, especially parks and forested areas near cities. Staphylococcus spp. are part of the microbiota of many different hosts and lead to opportunistic severe infection. Isolates from wild animals can be resistant to antimicrobial drugs. However, there are a few studies that evaluated Staphylococcus spp. in neotropical primates. The goal of this study was to evaluate Staphylococcus spp. isolated from free-ranging black-tuffed marmosets. METHODS: Marmosets were captured in six urban parks. After sedation, skin and rectal swabs and feces were sampled. Staphylococcus spp. isolates were identified by MALDI-ToF and their antimicrobial susceptibility was determined. RESULTS: Over 30% of captured individuals were positive for Staphylococcus spp., and S. aureus was the most isolated species followed by Mammaliicoccus (Staphylococcus) sciuri. With the exception of the marmoset subjected to necropsy, none of the other had lesions, which supports that notion that Staphylococcus spp. are members of the microbiota, but also opportunistic pathogens. Most isolates were susceptible to all antimicrobials tested; however, one isolate of S. epidermidis was resistant to multiple antimicrobials (penicillin, cefoxitin, ciprofloxacin, clindamycin, and erythromycin). We considered S. aureus as the main staphylococci to colonize black-tuffed marmosets. CONCLUSIONS: Black-tuffed marmosets can be colonized by several Staphylococcus species, most frequently by S. aureus, and the majority of isolates were sensible to the antimicrobials tested. One S. epidermidis isolate was considered multidrug resistant.

Fecal shedding of Clostridioides difficile in calves in Sao Paulo state, Brazil.

OBJECTIVE: This study aimed to evaluate the fecal shedding of C. difficile in calves on farms in Sao Paulo State, Brazil. MATERIALS AND METHODS: Fecal samples (n = 300) were collected from diarrheic (n = 78) and nondiarrheic (n = 222) calves less than 60 days of age from 20 farms. Fecal samples were inoculated into enrichment broth supplemented with taurocholate and cultured under anaerobic conditions. Colonies suspected to be C. difficile were harvested for DNA extraction and then multiplex PCR for the detection of genes encoding toxins A and B and binary toxins. All toxigenic isolates were ribotyped and tested for antimicrobial susceptibility, and five selected strains were subjected to whole-genome sequencing to determine their sequence type. RESULTS AND DISCUSSION: C. difficile was isolated from 29.3 % (88/300) of the samples. All toxigenic isolates (17/88, 19.3 %) were classified as ribotypes RT046 (13/17-79.47 %, A+B+ CDT-) and RT126 (4/17 = 20.53 %, A+B+ CDT+). The sequenced strains from RT046 were classified as ST35 (Clade 1), while those from RT126 were classified as ST11 (Clade 5). No associations between the epidemiological factors in any of the groups and C. difficile isolation were observed. Most of the toxigenic isolates (16/17 = 94.41 %) were classified as multidrug-resistant. Calves can be an important source of toxigenic C. difficile strains, including multidrug-resistant isolates from ribotypes commonly observed in humans.

Occurrence of Clostridium perfringens in Wild Mammals in the Amazon Biome.

The objective of this study was to evaluate the occurrence of Clostridium perfringens in stool samples and swabs collected from wild mammals in the Amazon biome. Sixty-five faecal and swab samples were collected in situ and ex situ from 16 species and three genera of wild mammals, some of which were in good health and some of which had diarrhoea. After pre-enrichment, the samples were plated on selective agar for C. perfringens. Characteristic colonies were subjected to multiplex PCR for the detection of genes encoding the main C. perfringens toxins (alpha, beta, epsilon, and iota toxin and enterotoxin). Among the 65 samples, 40 (61.5%) were positive for the gene encoding the alpha toxin and were classified as type A, 36 of which were asymptomatic animals and four were diarrheal. No other toxinotypes were found. The findings of this study suggest that C. perfringens type A is commonly found in mammal species of the Amazon biome. This seems to be the first study to identify C. perfringens type A in species such as B. variegatus (common ground sloth), C. didactylus (two-toed sloth), P. flavus (Jupará), T. tetradactyla (anteater), S. collinsi (squirrel monkey), S. niger (black marmoset), and S. apella (Guyana capuchin) and in the genus Didelphis sp. (opossum).

Production and purification of Clostridium perfringens type C beta-toxin and IgG and IgY antitoxins.

OBJECTIVES: This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY). METHODS: Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY. RESULTS: All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %). CONCLUSIONS: C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.

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.

Bacterial microbiome changes after fecal transplantation for recurrent Clostridioides difficile infection in the Brazilian center.

Clostridioides difficile infection (CDI) poses a significant global health threat owing to its substantial morbidity and associated healthcare costs. A key challenge in controlling CDI is the risk of multiple recurrences, which can affect up to 30% of patients. In such instances, fecal microbiota transplantation (FMT) is increasingly recognized as the optimal treatment. However, few related studies have been conducted in developing countries, and the microbiota composition of Brazilian patients and its dynamic modification post-FMT remain largely unexplored. This study aimed to evaluate the changes in the bacterial gut microbiome in Brazilian patients with recurrent CDI post-FMT. Ten patients underwent FMT, and the primary and overall CDI resolution rates were 80% and 90% after the first and second FMT, respectively. FMT was associated with an early increase in Shannon's diversity, evident as soon as 1 week post-FMT and persisting for at least 25 days post-treatment. Post-treatment, the abundance of Firmicutes increased and that of Proteobacteria decreased. Specifically, the abundance of the genera Ruminococcus, Faecalibacterium, Lachnospira, and Roseburia of the Firmicutes phylum was significantly higher 1 week post-transplantation, with Ruminococcus and Faecalibacterium remaining enriched 25 days post-transplantation. This study is the first of its kind in Brazil to evaluate the microbiota of a donor and patients undergoing FMT. Our findings suggest that FMT can induce remarkable changes in the gut microbiota, characterized by an early and sustained increase in diversity lasting at least 25 days. FMT also promotes enrichment of genera such as Ruminococcus spp., Faecalibacterium spp., and Roseburia spp., essential for therapeutic success.

Non-vaccination of dogs is linked to uncompleted COVID-19 vaccination protocols of owners.

Despite the known importance of vaccines as one of the greatest achievements in medical science, vaccine hesitancy has been increasing in the last decade and has become a major threat to global health. The growth of vaccine hesitancy worldwide became more evident with the onset of COVID-19 and raised the fear that this hesitancy would also impact companion animal vaccination. This study aimed to characterize vaccine-hesitant groups regarding companion animals and human vaccinations, as well as to explore the possible association between vaccine hesitancy in pets and their owners. An anonymous online survey containing 27 closed questions was conducted, including questions about dog health, such as vaccination, neutering, and homeopathy use, as well as questions about the COVID-19 vaccination status of the owner and motivations for vaccinating or not vaccinating their pets. Most participants (81.5 %) reported receiving three or more doses of the COVID-19 vaccine. Not vaccinating against COVID-19 or having an incomplete vaccination protocol was associated with an increase in the age range of participants (odds ratio [OR] = 1.43), not having higher education (OR = 7.70), and being in extreme income ranges (less than two minimum wages [OR = 7.57] and more than 10 [OR = 5.20]). The use of homeopathy in humans (OR = 3.24) and dogs (OR = 3.74) was associated with non-vaccination against COVID-19. Owners who were not fully vaccinated against COVID-19 were almost six times more likely to not vaccinate their dogs (OR = 5.94). Non-vaccination of dogs was also associated with non-neutering (OR = 3.56), keeping the dog in contact with other dogs (OR = 2.09), and an increase in the number of dogs in the house increased the chance of not vaccinating the animals (OR = 1.30). The present study revealed a strong association between non-vaccination against COVID-19 and non-vaccination in companion dogs, raising the hypothesis that vaccination hesitancy is a growing challenge in veterinary medicine. In addition, the characteristics of Brazilians who are reluctant to be vaccinated against COVID-19 or immunize their companion animals are described here for the first time.

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.

CLINICAL OUTCOME AND SEVERITY OF CLOSTRIDIOIDES (CLOSTRIDIUM) DIFFICILE INFECTION AT A TERTIARY REFERRAL HOSPITAL IN BRAZIL.

•The outcomes of CDI were evaluated in 65 patients with CDI in a Brazilian tertiary hospital. •Lack of clinical improvement after treatment and the severity score (ATLAS) increased the risk of death. •The use of multiple antimicrobial agents was associated with longer hospital stays. •Patients with high Charlson comorbidity index (>7) were more likely to recur. Background - Clostridioides difficile infection (CDI) is a potentially severe disease that can present with refractoriness, recurrence, and evolution to death. In Brazil, the epidemiology of CDI seems to differ from that of the United States and most European countries, with only one ribotype (RT) 027-related case and a high prevalence of RT106. Objective - The aim of this study was to evaluate the outcomes of CDI and its possible association with ribotypes at a university hospital in Brazil. Methods - A total of 65 patients with CDI were included and stool samples were submitted to A/B toxin detection and toxigenic culture, and toxigenic isolates (n=44) were also PCR ribotyped. Results - Patients' median age was 59 (20-87) years and there were 16 (24.6%) deaths. The median Charlson comorbidity index (CCI) was 4 (0-15) and 16.9% of the patients had CCI ≥8. The ATLAS score and non-improvement of diarrhea were related to higher mortality. A longer length of hospitalization was related to the enteral nutrition and use of multiple antibiotics. The period between CDI diagnosis and hospital discharge was longer in those who received new antibiotics after diagnosis, multiple antibiotics, and required intensive care treatment. Recurrence was associated with CCI >7. Twenty ribotypes were identified and RT106 was the most frequently detected strain (43.2%). No relationship was observed between the ribotypes and outcomes. CDI was present in patients with more comorbidities. Conclusion - Risk factors for higher mortality, longer hospital stay and recurrence were identified. A diversity of ribotypes was observed and C. difficile strains were not related to the outcomes.

Desfechos clínicos e gravidade da infecção pelo Clostridioides (Clostridium) difficile em um hospital terciário de referência no Brasil / Clinical outcome and severity of clostridioides (clostridium) difficile infection at a tertiary referral hospital in brazil

ABSTRACT Background: Clostridioides difficile infection (CDI) is a potentially severe disease that can present with refractoriness, recurrence, and evolution to death. In Brazil, the epidemiology of CDI seems to differ from that of the United States and most European countries, with only one ribotype (RT) 027-related case and a high prevalence of RT106. Objective: The aim of this study was to evaluate the outcomes of CDI and its possible association with ribotypes at a university hospital in Brazil. Methods: A total of 65 patients with CDI were included and stool samples were submitted to A/B toxin detection and toxigenic culture, and toxigenic isolates (n=44) were also PCR ribotyped. Results: Patients' median age was 59 (20-87) years and there were 16 (24.6%) deaths. The median Charlson comorbidity index (CCI) was 4 (0-15) and 16.9% of the patients had CCI ≥8. The ATLAS score and non-improvement of diarrhea were related to higher mortality. A longer length of hospitalization was related to the enteral nutrition and use of multiple antibiotics. The period between CDI diagnosis and hospital discharge was longer in those who received new antibiotics after diagnosis, multiple antibiotics, and required intensive care treatment. Recurrence was associated with CCI >7. Twenty ribotypes were identified and RT106 was the most frequently detected strain (43.2%). No relationship was observed between the ribotypes and outcomes. CDI was present in patients with more comorbidities. Conclusion: Risk factors for higher mortality, longer hospital stay and recurrence were identified. A diversity of ribotypes was observed and C. difficile strains were not related to the outcomes.

RESUMO Contexto: A infecção pelo Clostridioides difficile (ICD) é uma doença potencialmente grave que pode se apresentar com refratariedade, recidiva e evoluir para óbito. No Brasil, a epidemiologia da ICD parece diferir da dos Estados Unidos e da maioria dos países europeus, com apenas um caso relacionado ao ribotipo (RT) 027 e alta prevalência do RT106. Objetivo: Avaliar os desfechos da ICD e sua possível associação com ribotipos em um hospital universitário do Brasil. Métodos: Um total de 65 pacientes com ICD foram incluídos e amostras de fezes foram submetidas à detecção de toxina A/B e cultura toxigênica e as cepas toxigênicas isoladas (n=44) também foram ribotipadas por PCR. Resultados: A idade mediana dos pacientes foi de 59 (20-87) anos e houve 16 (24,6%) óbitos. A mediana do índice de comorbidade de Charlson (ICC) foi de 4 (0-15) e 16,9% dos pacientes apresentaram ICC ≥8. O escore ATLAS e a não melhora da diarreia foram relacionados a maior mortalidade. Maior tempo de internação esteve relacionado à nutrição enteral e ao uso de múltiplos antibióticos. O período entre o diagnóstico de ICD e a alta hospitalar foi maior naqueles que receberam novos antibióticos após o diagnóstico, múltiplos antibióticos e necessitaram de tratamento intensivo. A recorrência foi associada com ICC >7. Vinte ribotipos foram identificados e o RT106 foi a cepa mais frequentemente detectada (43,2%). Não foi observada relação entre os ribotipos e os desfechos. ICD esteve presente em pacientes com mais comorbidades. Conclusão: Foram identificados fatores de risco para maior mortalidade, maior tempo de internação e recorrência. Uma diversidade de ribotipos foi observada e cepas de C. difficile não foram relacionadas aos desfechos.

Chylothorax associated with pulmonary compressive atelectasis in an emperor tamarin (Saguinus imperator).

Chylothorax is the accumulation of lymph in the thoracic cavity, and it has never been reported in neotropical primates. An emperor tamarin died and at necropsy chylothorax associated with pulmonary compressive atelectasis was diagnosed. Idiopathic chylothorax can be a cause of respiratory insufficiency and death in tamarins.

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.

Identification and characterization of pathogenic and multidrug-resistant bacteria in feral pigeons surrounding a veterinary hospital in Minas Gerais, Brazil / Identificação de bactérias patogênicas e multirresistentes a antimicrobianos em pombos urbanos no entorno de um Hospital Veterinário em Belo Horizonte, Minas Gerais, Brasil

Pigeons are known for their capacity to harbor and spread several zoonotic agents. Studies have suggested that pigeons are also relevant disseminators of multidrug-resistant strains. In this study, pigeons surrounding a veterinary hospital were sampled and tested for the presence of pathogenic Escherichia coli, Salmonella spp., Staphylococcus spp., and Clostridioides (Clostridium) difficile. E. coli isolates from 19 (40.4%) pigeons tested positive for the E. coli heat-stable enterotoxin 1 (EAST1)-encoding gene. The intimin-encoding gene (eae) of enteropathogenicE. coli (EPEC) was found in one isolate (2.1%). Salmonella spp. were found in nine (19.1%) pigeons, all from the first capture event (P < 000.1). S. Typhimurium and S. Heidelberg were isolated from six and three pigeons, respectively. Enterobacterial repetitive intergenic consensus (ERIC-PCR) of the Salmonella spp. isolates suggested that eight of the nine strains had a high genetic similarity, supporting the hypothesis of an outbreak of salmonellosis in these pigeons. Twenty (42.5%) staphylococcal isolates were recovered from 18 (38.3%) pigeons. Eight different species were detected, with S. xylosus being the most frequent. Two (4.3%) C. difficile strains were isolated. Three isolates, one each of S. Typhimurium, S. aureus, and C. difficile, were classified as multidrug-resistant strains. The present research suggested that pigeons residing in urban areas can act as reservoirs and disseminators of pathogenic bacteria, including nosocomial pathogens, such as diarrheagenicE. coli and multidrug-resistant Staphylococcus spp., C. difficile, and Salmonella spp.

Pombos urbanos são conhecidos pela sua capacidade de carrear e disseminar diversos agentes zoonóticos. Estudos tem sugerido que pombos são também relevantes na disseminação de estirpes resistentes a múltiplas drogas. No presente estudo, pombos no ambiente de um hospital veterinário foram amostrados em três diferentes períodos e testados para a presença de Escherichia coli patogênica, Salmonella spp., Staphylococcus spp. e Clostridioides (Clostridium) difficile. Isolados de E. coli de 19 pombos (40.4%) foram positivos para o gene codificador da toxina EAST1. O gene codificador de intimina (eae) do patotipo E. coli enteropatogênica foi encontrada em um isolado (2.1%). Salmonella spp. foi encontrada em nove pombos (19.1%), sendo todos isolados do primeiro período de captura (P < 000.1). S. Typhimurium foi isolado de seis animais e S. Heidelberg de três. A tipagem molecular de isolados de Salmonella spp. por ERIC-PCR demonstrou que oito estirpes possuíam alta similaridade genética entre si, sugerindo a ocorrência de um surto de salmonelose nos animais carreadores. Vinte Staphylococcus (42.5%) foram isolados de 18 animais (38.3%). Oito diferentes espécies foram detectadas, sendo S. xylosus a mais frequente. Duas estirpes de C. difficile não-toxigênica (4.3%) foram isoladas. Uma estirpe de S. Typhimurium, uma de S. aureus e um isolado de C. difficile foram classificados como resistentes a múltiplas drogas antimicrobianas. O presente estudo sugere que pombos capturados no ambiente do hospital veterinário podem atuar como reservatórios e disseminadores de bactérias patogênicas e envolvidas em infecção hospitalar, incluindo E. coli diarreiogênica e Staphylococcus sp., C. difficile e Salmonella spp multirresistente.

Urethral obstruction secondary to Staphylococcus felis chronic urethritis and hepatoid gland adenoma in a captive ocelot (Leopardus pardalis)

A male adult ocelot (Leopardus pardalis) kept under human care developed anuria, which progressed to death. Grossly, the urinary bladder was markedly dilated and filled with red discolored urine containing blood clots. In addition, the animal had a hepatoid cell adenoma adjacent to the urethra, which likely caused partial urethral occlusion. Microscopically, there was a predominantly neutrophilic, fibrinous and hemorrhagic urethritis, cystitis, and pyelonephritis with intralesional gram-positive cocci. Microbiologic culture followed by MALDI-TOF MS analysis resulted in the identification of isolates from the urine and urethra as Staphylococcus felis.(AU)

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.

First description of bacillary hemoglobinuria in a cattle in Minas Gerais, Brazil

Bacillary hemoglobinuria (BH) is a histotoxic infection caused by Clostridium haemolyticum that affects mostly cattle parasitized with trematodes. The aim of present study was to describe an atypical case of BH in a cow without access to flooded areas and without parasitism by Fasciola hepatica. After eight days of sternal decubitus and apathy, a Nellore cow was euthanized and necropsied. During postmortem examination, mild jaundice, black urine and multifocal lesions in the liver were observed. Histopathology revealed multifocal coagulation foci of necrosis in liver and basophilic bacillary structures, which were confirmed as C. haemolyticum by bacterial isolation, PCR and sequencing techniques. This is the first description of BH in cattle in Minas Gerais state, Brazil and highlight the need for of inclusion of BH as differential diagnosis even in animals not parasitized by trematodes.(AU)

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.

Characterization of the virulence of three novel clade 2 Clostridioides (Clostridium) difficile strains and a two-year screening in animals and humans in Brazil.

Clostridioides (Clostridium) difficile infection (CDI) is an evolving global healthcare problem, and owing to the diverse and dynamic molecular epidemiology of C. difficile, new strains continue to emerge. In Brazil, only two cases of CDI due to the so called hypervirulent PCR ribotype (RT) 027 belonging to clade 2 have ever been reported, whereas incidence of CDI due to another "hypervirulent" RT078 (clade 5) has not yet been reported. In contrast, novel clade 2 strains have been identified in different hospitals. To better understand the epidemiology of CDIs in Brazil, this study aimed to genotypically and phenotypically characterize three novel Brazilian clade 2 strains (RT883, 884, and 885) isolated from patients with confirmed CDI. In addition, to better understand the circulating RTs, a two-year sampling was conducted in patients from the same hospital and in several domestic and wild animal species. The three strains examined showed lower production of A/B toxins than the control RT027, although two of these strains harbored a truncated tcdC gene. All strains showed swimming motility similar to that of RT027, while RT883 showed higher spore production than the reference strain. In the in vivo hamster model, the lethality of all strains was found to be similar to that of RT027. Both cgMLST and cgMLSA analyses revealed a high genetic similarity among the three-novel clade 2 isolates. In the two-year survey in animals and humans, RT883, 884, and 885 were not detected; however, three new RTs (RT988, RT989, and RT990) were isolated, two of which were genetically related to the three previously reported clade 2 strains. RT106 and RT126 were most frequently detected in humans (47.9%) and animals (57.9%), respectively. Furthermore, RT027 and RT078 were not detected in humans. The results of this study suggest that these novel clade 2 strains have virulence potential and that new strains from clade 2 continue to emerge in our setting, indicating the need for long-term local surveillance.