Antimicrobial resistance and public and animal health risks associated with pathogenic Escherichia coli isolated from calves.

We aimed to determine the antimicrobial susceptibility profile of pathogenic Escherichia coli strains isolated from fecal samples of calves and buffalo calves (2008-2013), in Minas Gerais, Brazil, as well as the frequency of O157 gene and strains carrying extended-spectrum beta-lactamases (ESBL) and mobile colistin resistance (mcr) genes. E. coli strains (n=518) were tested for susceptibility against ten antimicrobials. Tetracycline was the antimicrobial with the highest resistance rate (382/518), followed by ampicillin (321/518), sulfamethoxazole/trimethoprim (312/518), chloramphenicol (192/518), gentamicin (126/518), ciprofloxacin (148/518), cefazolin (89/518), colistin (54/518) and cefoxitin (34/518). Multidrug resistance (MDR) was observed in 381/518 isolates. No strain harbored mcr or O157 genes, whereas 19/99 were ESBL positive. The most prevalent pathotype and phylogroup were STEC and B1, respectively. Age, EHEC pathotype and resistance to aminoglycoside and cephem were significantly associated with MDR in the multivariate model. Overall, E. coli strains showed high rates of resistance to penicillin, tetracyclines and folate inhibitors, in addition to an alarming rate of MDR and ESBL-producing strains.

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.

Immunomodulatory effect of long-term oclacitinib maleate therapy in dogs with atopic dermatitis.

BACKGROUND: Canine atopic dermatitis (cAD) is a chronic disease characterised by hypersensitivity to environmental allergens. Oclacitinib maleate selectively inhibits pro-inflammatory mediators associated with cAD. However, the impact of chronic oclacitinib use on immunocompetence requires further investigation. OBJECTIVES: Herein, we examined the potential immunomodulatory effects of prolonged oclacitinib treatment in dogs. ANIMALS: Thirteen privately owned dogs with cAD, treated with 0.4-0.6 mg/kg oclacitinib for 12 months. METHODS AND MATERIALS: Pruritus level was evaluated using a pruritus Visual Analog Scale (pVAS) and the canine atopic dermatitis extent and severity index, 4th iteration (CADESI IV). Peripheral blood samples were collected for routine laboratory assays and lymphocyte subtypes were analysed using flow cytometry. Antigen-specific intracellular cytokine production from CD4+ and CD8+ T lymphocytes was analysed following in vitro stimulation by Dermatophagoides farinae antigens. RESULTS: Oclacitinib treatment significantly reduced pVAS and CADESI-04 scores, by 51% and 86.7%, respectively. Flow cytometric analysis revealed increased CD4+ and CD14+ lymphocyte populations. The cytokine profile at 360 days after treatment initiation was similar to that before treatment and was not associated with clinical relapse. CONCLUSION: Oclacitinib, when administered at the currently labelled dose for one year, is associated with a significant increase in circulating CD4+ T cells, but does not alter cytokine production from antigen-stimulated T cells. The results reported do not support evidence for immunosuppression mediated by the mechanisms evaluated in this study.

FML/QuilA-Vaccinated Dogs Naturally Infected with Leishmania infantum: Serum Cytokines, Clinicopathological Profile, and Parasitological Parameters.

Dogs are the main reservoir of Leishmania infantum in endemic regions. Canine leishmaniasis, caused by L. infantum, can progress to a chronic disease resulting in death. Vaccines have been developed with a certain degree of success. The pathogenesis of this disease is not completely understood, especially in previously vaccinated dogs. We herein described clinical data, parasite load, serum levels of cytokines, and the reservoir potential in vdogs vaccinated with the fucose-mannose ligand (FML)/QuilA saponin vaccine (Leishmune™) naturally infected (Vi) and compared to vaccinated not infected dogs (Vn). Thirty-four dogs from private owners were divided into two groups: vaccinated/infected and vaccinated/uninfected. Clinical evaluation, hematological and biochemical parameters, and serum levels of cytokines were measured by conventional methods. The parasite burden in the bone marrow was measured by quantitative real-time PCR, and the transmissibility of parasites to sand flies was assessed by xenodiagnosis. Clinical, biochemical, and hematological parameters of vaccinated infected dogs were mostly normal. Vi dogs developed mild disease with low clinical scores. Serum levels of IL-10 were higher in Vi dogs, and a strong correlation was observed in IL-4 levels and the A/G ratio in Vi dogs. These results suggest a role of TH2 response in Vi dogs, although more data is needed to better understand the disease in vaccinated dogs.

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.

Serological study of feline leishmaniasis and molecular detection of Leishmania infantum and Leishmania braziliensis in cats (Felis catus).

Blood samples and swabs from ocular conjunctiva and mouth were obtained from 64 cats. Of 64 serum samples, 19 were positive for Leishmania antibodies by ELISA (29.80%). Eight cats were positive by PCR (12.5%) in swab samples from mouth and/or ocular mucosa. Poor kappa agreement between serological and molecular results (k = 0.16) was obtained. From five positive PCR samples one was L. braziliensis and four were L. infantum. Phylogenetic analysis performed with the five isolates of Leishmania, showed that samples of L. infantum isolated from the cats were phylogenetically close to those isolated from domestic dogs in Brazil, while the L. braziliensis is very similar to the one described in humans in Venezuela. The study demonstrated that, despite high seropositivity for Leishmania in cats living in the study region, poor agreement between serological and molecular results indicate that positive serology is not indicative of Leishmania infection in cats. Parasite DNA can be detected in ocular conjunctiva and oral swabs from cats, indicating that such samples could be used for diagnosis. Results of phylogenetic analyzes show that L. infantum circulating in Brazil is capable of infecting different hosts, demonstrating the parasite's ability to overcome the interspecies barrier.

Seroprevalence of Brucella ovis-epididymitis, smooth-Brucella, leptospirosis, toxoplasmosis, and Maedi-Visna in sheep slaughtered in Minas Gerais State, Brazil / Soroprevalência de Brucella ovis, Brucella lisa, leptospirose, toxoplasmose e Maedi-Visna em ovinos abatidos em Minas Gerais, Brasil

The present study aimed to estimate the prevalence of antibodies against Brucella ovis-epididymitis, smooth-Brucella, leptospirosis, toxoplasmosis and Maedi-visna in sheep slaughtered in Minas Gerais, Brazil and to study their simultaneous occurrence, including caseous lymphadenitis, at sheep and flock levels. The study was conducted at a sheep slaughterhouse with Federal Inspection Service. Sera from 594 animals from 21 flocks were collected, in 2007. The agar gel immunodiffusion (AGID) was employed to detect anti-B. ovis and anti-Maedi Visna antibodies, whereas Rose Bengal (RB) and the 2-mercaptoethanol test (2ME) were used to test anti-smooth Brucella antibodies. For the detection of anti-Leptospiraantibodies, sera were examined by microscopic agglutination test (MAT), while for the detection of IgG antibodies to Toxoplasma gondii ELISA was used. Prevalence of antibodies against smooth Brucella, B. ovis-epididimitis, Leptospiraspp., toxoplasmosis and Maedi-Visna found in sheep from Minas Gerais was 0.00%, 24.04%, 25.96%, 10.46% and 3.08%, respectively; whereas the seroprevalence in flocks was 0.00%, 80.95%, 90.48%, 71.43% and 23.81%, respectively. Moreover, when data on antibodies anti-Corynebacterium pseudotuberculosis, previously obtained, were included, about 60% of the flocks showed animals that were exposed to four or more of the studied agents. However, only 25.47% of the sheep exhibited simultaneously antibodies against more than one pathogen. Thus, data from the present study on sheep slaughtered in Minas Gerais, Brazil, showed no antibodies to smooth-Brucella and a low frequency of antibodies anti-Maedi Visna lentivirus, and a high and widespread seroprevalence of B. ovis, Leptospira spp., and T. gondii among animals and flocks.(AU)

O presente estudo teve como objetivo estimar a prevalência de anticorpos contra Brucella ovis (epididimite ovina), Brucellalisa, leptospirose, toxoplasmose e Maedi-visna em ovinos abatidos em Minas Gerais, Brasil, e estudar sua ocorrência simultânea, incluindo linfadenite caseosa, nos ovinos e nos rebanhos. O estudo foi realizado em um abatedouro de ovinos com Serviço de Inspeção Federal. Soros de 594 animais de 21 rebanhos foram coletados, em 2007. A imunodifusão em gel de ágar (IDGA) foi empregada para detectar anticorpos anti-B. ovis e anticorpos anti-Maedi Visna, enquanto o teste do antígeno acidificado tamponado (AAT) e o teste de 2-mercaptoetanol (2ME) foram utilizados para testar anticorpos anti-Brucella lisa. Para a detecção de anticorpos anti-Leptospira, os soros foram examinados pelo teste de aglutinação microscópica (MAT), enquanto que para a detecção de anticorpos IgG para Toxoplasma gondii, foi usado o ELISA. A prevalência de anticorpos anti-Brucella lisa, B. ovis, Leptospira spp., toxoplasmose e Maedi-Visna encontrados em ovinos de Minas Gerais foi de 0,00%, 24,04%, 25,96%, 10,46% e 3,08%, respectivamente; enquanto a soroprevalência em rebanhos foi de 0,00%, 80,95%, 90,48%, 71,43% e 23,81%, respectivamente. Além disso, quando dados de anticorpos anti-Corynebacterium pseudotuberculosis, previamente obtidos, foram incluídos, cerca de 60% dos rebanhos apresentaram animais expostos a quatro ou mais dos agentes estudados. No entanto, apenas 25,47% dos ovinos exibiram simultaneamente anticorpos contra mais de um patógeno. Assim, os dados do presente estudo sobre ovinos abatidos em Minas Gerais, Brasil, mostram que ausência de anticorpos anti-Brucella lisa e baixa frequência de anticorpos anti-Maedi Visna, e uma soroprevalência alta e generalizada de B. ovis, Leptospira spp. e T. gondii entre animais e rebanhos.(AU)

Serological study of feline leishmaniasis and molecular detection of Leishmania infantum and Leishmania braziliensis in cats (Felis catus) / Pesquisa sorológica da leishmaniose felina e detecção molecular de Leishmania infantum e Leishmania braziliensis em gatos (Felis catus)

Abstract Blood samples and swabs from ocular conjunctiva and mouth were obtained from 64 cats. Of 64 serum samples, 19 were positive for Leishmania antibodies by ELISA (29.80%). Eight cats were positive by PCR (12.5%) in swab samples from mouth and/or ocular mucosa. Poor kappa agreement between serological and molecular results (k = 0.16) was obtained. From five positive PCR samples one was L. braziliensis and four were L. infantum. Phylogenetic analysis performed with the five isolates of Leishmania, showed that samples of L. infantum isolated from the cats were phylogenetically close to those isolated from domestic dogs in Brazil, while the L. braziliensis is very similar to the one described in humans in Venezuela. The study demonstrated that, despite high seropositivity for Leishmania in cats living in the study region, poor agreement between serological and molecular results indicate that positive serology is not indicative of Leishmania infection in cats. Parasite DNA can be detected in ocular conjunctiva and oral swabs from cats, indicating that such samples could be used for diagnosis. Results of phylogenetic analyzes show that L. infantum circulating in Brazil is capable of infecting different hosts, demonstrating the parasite's ability to overcome the interspecies barrier.

Resumo Amostras de sangue e swabs da conjuntiva ocular e oral foram obtidas de 64 gatos. Das 64 amostras de soro, 19 foram positivas para anticorpos contra Leishmania por ELISA (29,80%). Oito gatos foram positivos por PCR (12,5%) em amostras de swab da boca e / ou mucosa ocular. Demonstrou-se baixa concordância kappa entre os resultados sorológicos e moleculares (k = 0,16). Das cinco amostras positivas para PCR, uma era L. braziliensis e quatro eram L infantum. A análise filogenética realizada com os cinco isolados de Leishmania, mostrou que amostras de L. infantum, isoladas dos gatos, eram filogeneticamente próximas às isoladas de cães domésticos do Brasil enquanto L. braziliensis era muito semelhante ao descrito em humanos na Venezuela. O estudo demonstrou que, apesar da alta soropositividade para Leishmania, em gatos que vivem na região do estudo, pouca concordância entre os resultados sorológicos e moleculares indica que a sorologia positiva não é indicativa de infecção por Leishmania em gatos. O DNA do parasita pode ser detectado na conjuntiva ocular e nas zaragatoas orais de gatos, indicando que essas amostras podem ser usadas para o diagnóstico. . Resultados de análises filogenéticas mostram que L. infantum, circulando no Brasil, é capaz de infectar diferentes hospedeiros, demonstrando a capacidade do parasita de superar a barreira interespécies.

Avaliação microbiológica e sensorial de alfajor vegano e sem glúten elaborado a partir da farinha de okara de amendoim / Microbiological analysis and sensory evaluation of a vegan gluten free alfajor produced with okara peanut flour

O veganismo cresceu nos últimos anos e é cada vez maior o número de pessoas que praticam uma dieta com restrições alimentares. O objetivo do trabalho foi desenvolver um alfajor vegano sem glúten, utilizando farinha de okara de amendoim e avaliar suas características microbiológicas e aceitação sensorial. As amostras foram submetidas a testes sensoriais de aceitação e microbiológicos para coliformes, bolores e leveduras. Os testes para coliformes foram realizados utilizando-se Caldo Lauril Sulfato Triptose e para a análise de bolores e leveduras foi realizada a contagem total por plaqueamento em superfície nos meios Ágar Dicloran Rosa de Bengala Cloranfenicol e Ágar Batata Dextrose. A formulação obteve boa aceitação e os resultados das análises microbiológicas mostraram que o produto possui qualidade microbiológica satisfatória de pelo menos seis dias. Logo, a utilização da farinha de okara de amendoim se mostrou interessante para o desenvolvimento de um alfajor venago e sem glúten.

Resultados das conformidades e não conformidades dos parâmetros microbiológicos de queijo Minas Artesanal Canastra / Conformity and non-conformity results of the microbiological parameters of Minas Artesanal Canastra cheese

O objetivo do trabalho foi analisar a qualidade microbiológica do Queijo Minas Artesanal produzido por queijarias certificadas da microrregião Canastra durante os anos de 2016 e 2017, utilizando os resultados das análises microbiológicas do queijo nestes anos. Foram encontradas não conformidades nos queijos estudados em 7,84% das amostras para Coliformes totais, 3,92% para coliformes termotolerantes e 9,8% para Staphylococcus coagulase positivo, porém nenhuma análise apresentou presença de Listeria spp. ou Salmonella spp. Apesar das principais bactérias patogênicas não terem sido encontradas, as não conformidades indicam a necessidade da orientação do produtor sobre as Boas Práticas de Fabricação e de Ordenha. Além disto, é imprescindível a orientação sobre as exigências do órgão fiscalizador e a realização da análise do queijo periodicamente, tornando possível o acompanhamento da qualidade do queijo produzido na região e a correção das não conformidades encontradas.

Causas de condenações em bovinos em um abatedouro com inspeção municipal de Minas Gerais / Causes of bovine condemnation in an slaughter under municipal inspection services in the Minas Gerais staction

A identificação de condenações em frigoríficos é importante para fornecer melhorias no processo produtivo e dados de possíveis fontes de contaminação na linha de abate. O objetivo foi identificar as causas de condenação de carcaças e vísceras bovinas, em um frigorífico de inspeção municipal em Minas Gerais, e discutir fatores que favorecem a ocorrência das mesmas. Foram analisados os registros de condenações no período de julho de 2017 a setembro de 2018. Do total de 3.723 animais abatidos, 101 (2,71%) apresentaram algum tipo de condenação. Foram condenados principalmente fígado, coração, fragmentos de caraças e carcaças inteiras. Abscessos hepáticos foram a causa mais frequente de condenação. Observamos que a frequência de descarte de carcaças e vísceras variaram entre os Serviços de Inspeção e estão relacionadas com a região estudada.

Virulence factors and phylotyping of Escherichia coli isolated from non-diarrheic and diarrheic water buffalo calves / Caracterização dos fatores de virulência e filogrupos de Escherichia coli isoladas de bezerros bubalinos sem diarreia e diarréicos

ABSTRACT: This study aimed to determine the virulence factors, phylogenetic groups, and the relationships between pathovars and phylogenetic groups of E. coli strains isolated from feces of buffalo calves. A total of 217 E. coli strains were obtained from feces after culture and were screened by PCR for detection of virulence factors EAST-1, enterohemolysin, Saa, CNF2, F41, F5, STa, intimin, Stx1 and Stx2. One hundred and thirty-four isolates were positive for one or more virulence factors: eighty-four from diarrheic animals, and fifty from non-diarrheic calves. The pathovars of E. coli identified in diarrheic feces were ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), and EAEC (EAST-1+) (33/84). Pathovars identified in non-diarrheic animals were NTEC (21/50), STEC (17/50), EHEC (1/50) and EAEC (7/50). E. coli strains positive for EAST-1 (P=0.008) and phylogroup C (P = 0.05) were associated with the presence of diarrhea. Phylogenetic analysis showed that 58.95% of the isolates belonged to phylogroup B1, followed by E (9.70%), B2 (5.90%), C (5.90%), D (5.22%), A (2.24%), and F (1.50%). Phylogroup B1 predominated in pathogenic E. coli isolated from water buffalo, and phylogroup C constituted an enteropathogenic E. coli for water buffalo calves.

RESUMO: O objetivo foi determinar os fatores de virulência, os grupos filogenéticos e as possíveis relações entre os patovares e os grupos filogenéticos identificados de cepas de Escherichia coli isoladas de fezes de bezerros bubalinos. Um total de 217 amostras de E. coli foram identificadas a partir de cultura das fezes e submetidas a reação em cadeia da polimerase (PCR) para detecção dos fatores de virulência EAST-1, enterohemolisina, Saa, CNF2, F41, F5, STa, intimina, Stx1 e Stx2. Foram identificadas 134 cepas positivas para um ou mais fatores de virulência: 84isoladas de bezerros bubalinos diarreicos e 50 de bezerros bubalinos saudáveis. Os patovares de E. coli obtidos de fezes diarreicas foram ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), e EAEC (EAST-1+) (33/84). Os patovares isolados de fezes não diarreicas foram NTEC (21/50), STEC (17/50), EHEC (1/50) e EAEC (7/50). Cepas de E. coli positivas para EAST-1 (P = 0,008) e filogrupo C (P = 0,05) foram associadas com a presença de diarreia. A análise de filogrupos revelou que 58,95% dos isolados pertencem ao filogrupo B1, seguido por E (9,70%), B2 (5,90%), C (5,90%), D (5,22%), A (2,24%) e F (1,50%). O filogrupo B1 predomina em cepas de E. coli patogênicas isoladas de bezerros búfalos e o filogrupo C constitui um filogrupo de E. coli patogênica entérica para bezerros.

Serological, molecular, and microscopic detection of Leishmania in cats (Felis catus) in Belo Horizonte, Minas Gerais State, Brazil / Detecção sorológica, molecular e microscópica de Leishmania em gatos (Felis catus) em Belo Horizonte, Minas Gerais, Brasil

Abstract The role of cats in the epidemiological cycle of leishmaniasis remains unclear. To better understand the occurrence of leishmaniasis in cats, we studied the frequency of Leishmania in serum samples of 100 cats living in an endemic region for canine and human leishmaniasis by serological, parasitological, and molecular methods. Of the 100 cats, 54 were seropositive for Leishmania antibodies by immunofluorescence antibody test. None of the bone marrow aspirates collected from these cats tested positive for the parasite in culture or upon polymerase chain reaction (PCR) analysis. Biopsy samples of the ears also tested negative for Leishmania upon PCR analysis. These findings may indicate that the region is endemic for canine leishmaniasis and cats are infected by Leishmania; or that cross-reaction with antibodies against other parasites increases the frequency of seropositivity; or that cats respond to Leishmania infection by producing antibodies when few or no parasites are present in bone marrow and tissue samples. Overall, our results suggest that cats can be infected by Leishmania ; however, we failed to demonstrate feline parasitosis. These findings highlight the need to study leishmaniasis in cats, since sandflies feed on cats, these animals may act as a reservoir for the parasite.

Resumo O papel dos gatos no ciclo epidemiológico da leishmaniose ainda não está claro. Para entender melhor a ocorrência de leishmaniose em gatos, estudou-se a frequência de Leishmania em amostras de soro de 100 gatos, os quais vivem em uma região endêmica para leishmaniose canina e humana, por métodos sorológicos, parasitológicos e moleculares. Dos 100 gatos, 54 foram soropositivos para anticorpos de Leishmania por teste de anticorpos de imunofluorescência. Nenhum dos aspirados de medula óssea coletados desses gatos mostrou-se positivo para o parasita em cultura, ou após a realização da reação em cadeia da polimerase (PCR). Amostras de biópsia das orelhas também foram negativas para Leishmania submetidas a PCR. Esses achados indicam que na região estudada endêmica para leishmaniose canina, os gatos podem se infectar por Leishmania; ou que a reação cruzada com anticorpos contra outros parasitas aumenta a frequência de soropositividade; ou que os gatos respondem à infecção por Leishmania produzindo anticorpos quando poucos ou nenhum parasita estão presentes na medula óssea e em amostras de tecido. Em geral, os resultados sugerem que os gatos podem ser estar infectados por Leishmania spp. No entanto, não foi possível demonstrar parasitismo felino. Essas descobertas evidenciam a necessidade de estudar a leishmaniose em gatos, uma vez que, como os flebotomíneos se alimentam em gatos, e esses animais podem atuar como um reservatório para o parasita.

Serological, molecular, and microscopic detection of Leishmania in cats (Felis catus) in Belo Horizonte, Minas Gerais State, Brazil.

The role of cats in the epidemiological cycle of leishmaniasis remains unclear. To better understand the occurrence of leishmaniasis in cats, we studied the frequency of Leishmania in serum samples of 100 cats living in an endemic region for canine and human leishmaniasis by serological, parasitological, and molecular methods. Of the 100 cats, 54 were seropositive for Leishmania antibodies by immunofluorescence antibody test. None of the bone marrow aspirates collected from these cats tested positive for the parasite in culture or upon polymerase chain reaction (PCR) analysis. Biopsy samples of the ears also tested negative for Leishmania upon PCR analysis. These findings may indicate that the region is endemic for canine leishmaniasis and cats are infected by Leishmania; or that cross-reaction with antibodies against other parasites increases the frequency of seropositivity; or that cats respond to Leishmania infection by producing antibodies when few or no parasites are present in bone marrow and tissue samples. Overall, our results suggest that cats can be infected by Leishmania ; however, we failed to demonstrate feline parasitosis. These findings highlight the need to study leishmaniasis in cats, since sandflies feed on cats, these animals may act as a reservoir for the parasite.

The incidence of Clostridioides difficile and Clostridium perfringens netF-positive strains in diarrheic dogs.

The aim of this study was to examine the incidence of Clostridioides (previously Clostridium) difficile and Clostridium perfringens in the feces of diarrheic and non-diarrheic dogs. Also, the presence of other common canine enteropathogens was examined. Toxigenic C. difficile and C. perfringens positive for the NetF-encoding gene (netF) were detected in 11 (11.9%) and seven (7.6%) diarrheic dogs, respectively. Three dogs were diagnosed simultaneously with toxigenic C. difficile and netF-positive C. perfringens. Among other enteropathogens, Giardia sp. was the most common agent detected in dogs positive for toxigenic C. difficile or netF-positive C. perfringens. The results suggest that C. difficile and C. perfringens occur more frequently as a primary cause of diarrhea.

Detection of virulence genes and the phylogenetic groups of Escherichia coli isolated from dogs in Brazil / Detecção de genes de virulência e grupos filogenéticos de amostras Escherichia coli isoladas de cães no Brasil

ABSTRACT: This study identified the virulence genes, pathovars, and phylogenetic groups of Escherichia coli strains obtained from the feces of dogs with and without diarrhea. Virulence genes and phylogenetic group identification were studied using polymerase chain reaction. Thirty-seven E. coli isolates were positive for at least one virulence factor gene. Twenty-one (57.8%) of the positive isolates were isolated from diarrheal feces and sixteen (43.2%) were from the feces of non-diarrheic dogs. Enteropathogenic E. coli (EPEC) were the most frequently (62.2%) detected pathovar in dog feces and were mainly from phylogroup B1 and E. Necrotoxigenic E. coli were detected in 16.2% of the virulence-positive isolates and these contained the cytotoxic necrotizing factor 1 (cnf1) gene and were classified into phylogroups B2 and D. All E. coli strains were negative for the presence of enterotoxigenic E. coli (ETEC) enterotoxin genes, but four strains were positive for ETEC-related fimbriae 987P and F18. Two isolates were Shiga toxin-producing E. coli strains and contained the toxin genesStx2 or Stx2e, both from phylogroup B1. Our data showed that EPEC was the most frequent pathovar and B1 and E were the most common phylogroups detected in E. coli isolated from the feces of diarrheic and non-diarrheic dogs.

RESUMO: Este estudo pesquisou genes de virulência, patovares e grupos filogenéticos de amostras de E. coli isoladas de fezes de cães com e sem diarreia. Os genes de virulência e a identificação de grupos filogenéticos foram estudados pela técnica de reação em cadeia da polimerase (PCR). 37 isolados de E. coli foram positivos para pelo menos um fator de virulência na análise de PCR. Destes, 21 (57,8%) foram isolados de fezes de cães com diarreia e 16 (43,2%) de fezes de cães não diarreicos. E. coli enteropatogênica (EPEC) (23/37, 62,2%) foi o patovar mais frequente detectado em fezes de cães e foram classificados principalmente como filogrupos B1 e E. E. coli necrotoxigênica (NTEC) positivos para CNF1 foram detectados (6/37, 16,2%) e classificados como B2 e D. Todas as amostras de E. coli foram negativas quanto à presença de genes de enterotoxinas de E. coli enterotoxigênica (ETEC), mas quatro amostras foram positivas para fimbrias relacionadas ao ETEC, 987P (2) e F18 (2). As amostras de E. coli (STEC) produtora de toxina Shiga foram positivas para a toxina Stx2 (1/37) e Stx2e (1/37), ambas do filogrupo B1. Nossos resultados indicaram que EPEC foi o patovar mais frequente e B1 e E foram os filogrupos mais comuns detectados em amostras E. coli isoladas de fezes de cães diarreicos e não diarreicos.

Frequency and antimicrobial susceptibility of Staphylococcus pseudintermedius in dogs with otitis externa / Frequência e sensibilidade antimicrobiana de Staphylococcus pseudintermedius isolados de cães com otite externa

ABSTRACT: Infections by Staphyloccocus pseudintermedius in the skin of dogs have been object of studies, since such microorganisms often present multiple resistance to antibiotics. This study aimed to identify and evaluate the antimicrobial susceptibility of Staphylococcus pseudintermedius (SP) strains isolated from dogs with otitis. Swabs from both ears of 52 dogs with otitis (n=104) were included. Bacteria were cultured using Muller-Hinton agar (supplemented with 5% equine blood and incubated at 37°C for 24 to 48 hours. All colonies underwent biochemical evaluation for identification of staphylococci. The identity of colonies as SP was confirmed by polymerase chain reaction. The antimicrobial susceptibility of SP strains was evaluated by disk diffusion. The presence of the gene mecA was evaluated in all SP isolates by PCR. Forty-four SP strains were isolated from swabs of 31 dogs (31/52, 59.6%). Seventy-five percent of the strains were susceptible to cephalexin and 93.2% to amoxicillin plus clavulanic acid. Less than 23% of the strains were susceptible to penicillin. For non-beta-lactam antimicrobials, 63.6% of the strains showed resistance to sulfamethoxazole-trimethoprim, 61.4% to tetracycline, and 38, 64% to enrofloxacin. Aminoglycoside resistance rate was 27.3% for gentamicin. Resistance to oxacillin in vitro was detected in 13 of the 44 strains (29, 55%). A total of 12 strains (27.3%) were positive for mecA gene and five of these 12 strains were susceptible to in vitro oxacillin. Twenty-six (59, 1%) strains were resistant to three or more classes of antimicrobials, and classified as multi resistant. Our results showed high frequency of SP and multi resistant isolates to antimicrobials commonly used in veterinary.

RESUMO: Infecções por Staphyloccocus pseudintermedius (SP) na pele de cães tem sido objeto de estudos, uma vez que esses microrganismos geralmente apresentam resistência múltipla à antibióticos. Este estudo teve como objetivo identificar e avaliar a susceptibilidade antimicrobiana das cepas de SP isoladas de cães com otite. Amostras de ambas orelhas de 52 cães com otite (n=104) foram incluídas. As bactérias foram cultivadas em ágar Muller-Hinton suplementado com 5% de sangue equino e incubadas a 37ºC por 24 a 48h. Todas as colônias foram submetidas à avaliação bioquímica para identificação de estafilococos. A identificação das colônias como SP foi confirmada pela reação em cadeia da polimerase. A susceptibilidade antimicrobiana das cepas SP foi avaliada pela técnica de difusão em disco. A presença do gene mecA foi avaliada em todos os isolados de SP por PCR. 44 estirpes de SP foram isoladas de 31 cães (31/52, 59,6%). 75% das cepas foram suscetíveis a cefalexina e 93,2% à amoxicilina mais ácido clavulânico. Menos de 23% das estirpes eram suscetíveis à penicilina. Para antimicrobianos não beta-lactâmicos, 63,6% apresentaram resistência ao sulfametoxazol-trimetoprim, 61,4% à tetraciclina e 38,64% à enrofloxacina. A frequência de resistência a aminoglicosídeos foi de 27,3% para gentamicina. A resistência a oxacilina in vitro foi detectada em 13 das 44 estirpes (29,55%). Um total de 12 amostras (27,3%) foram positivas para mecA, cinco das quais foram suscetíveis à oxacilina na difusão em disco. 26 amostras (59,1%) foram resistentes a três ou mais classes de antimicrobianos e classificadas como multirresistentes. Nossos resultados mostram elevada frequência de isolados SP e multiresistentes para os antimicrobianos comumente usados em veterinária.

Characterization of virulence factors and phylogenetic group determination of Escherichia coli isolated from diarrheic and non-diarrheic calves from Brazil.

This study aimed to detect virulence factors, pathovars, and phylogenetic groups of Escherichia coli strains obtained from feces of calves with and without diarrhea up to 70 days old and to determine the association between occurrence of diarrhea, phylogenetic groups, and pathovars. Phylo-typing analysis of the 336 E. coli strains isolated from calves with Clermont method showed that 21 (6.25 %) belong to phylogroup A, 228 (67.85 %) to phylogroup B1, 2 (0.6 %) to phylogroup B2, 5 (1.49 %) to phylogroup C, 57 (16.96 %) to phylogroup E, and 3 (0.9 %) to phylogroup F. Phylogroup D was not identified and 20 strains (5.95 %) were assigned as "unknown." The distribution of phylogenetic groups among pathovars showed that NTEC belong to phylogroups B1 (17) and C (4); EPEC to phylogroups B1 (6) and E (8); STEC to phylogroups A (5), B1 (56), B2 (2), C (1), and E (15); EHEC to phylogroups B1 (95) and E (5); and ETEC to phylogroups A (3), B1 (7), and E (10). The EAST-1 strains were phylogroups A (13), B1 (47), E (19), and F (3); E. coli strains of "unknown" phylogroups belonged to pathovars EPEC (1), EHEC (2), STEC (7), and EAST-1 strains (6). ETEC was associated with diarrhea (P = 0.002). Our study did not find association between the phylogenetic background and occurrence of diarrhea (P = 0.164) but did find some relationship in phylogenetic group and pathovar. The study showed that EHEC and STEC are classified as phylogroup B1, EAST-1 phylogroup A, ETEC, and EPEC phylogroup E.

Phylogenetic Group Determination of Escherichia coli Isolated from Animals Samples.

This study analyzes the occurrence and distribution of phylogenetic groups of 391 strains of Escherichia coli isolated from poultry, cattle, and water buffalo. The frequency of the phylogroups was A = 19%, B1 = 57%, B2 = 2.3%, C = 4.6%, D = 2.8%, E = 11%, and F = 3.3%. Phylogroups A (P < 0.001) and F (P = 0.018) were associated with E. coli strains isolated from poultry, phylogroups B1 (P < 0.001) and E (P = 0.002) were associated with E. coli isolated from cattle, and phylogroups B2 (P = 0.003) and D (P = 0.017) were associated with E. coli isolated from water buffalo. This report demonstrated that some phylogroups are associated with the host analyzed and the results provide knowledge of the phylogenetic composition of E. coli from domestic animals.

Longitudinal study of Salmonella spp., diarrheagenic Escherichia coli, Rotavirus, and Coronavirus isolated from healthy and diarrheic calves in a Brazilian dairy herd.

This prospective longitudinal study investigated the epidemiology of enteric disease associated with infections in calves aging up to 70 days. A total of 850 fecal samples were collected from 67 calves. Seventeen isolates of Salmonella spp. were recovered from feces of 11 calves (16.4%), and statistical analysis revealed no association between the presence of Salmonella spp. and clinical signs of diarrhea or age. Virulence factors of Escherichia coli were identified in 103 strains: eae (7), K99/STa (7), Stx1 (7), Stx1/eae (36), Stx1/Stx2/eae (2), Stx2 (43), and Stx2/eae (1). There was statistical association between diarrheic animals carrying E. coli Stx1/eae (+) in their feces at 2 and 4 weeks of age (P = 0.003) and E. coli Stx2 (+) at 5 weeks of age (P = 0.03). Rotavirus was detected in 49 (5.76%) fecal samples collected from 33 calves (49.2%). The presence of rotavirus was correlated with diarrheic feces (P < 0.0001) rather than feces with normal consistency. There was a significant relationship between age group and diarrhea (P = 0.001). Bovine coronavirus (BCoV) was detected in 93 fecal samples collected from 46 calves (68.6%). There was an association (P < 0.0001) between diarrheic animals positive for BCoV and age groups. The results demonstrate the importance of the pathogens studied in the etiology of diarrhea in calves.