Enteropathogenic and Multidrug-Resistant blaCTX-M-Carrying E. coli Isolates from Dogs and Cats.

Enteropathogenic Escherichia coli (EPEC) are pathogens associated with gastrointestinal illnesses. Dogs and cats can harbor EPEC, and antimicrobial resistance may impair necessary treatments. This study characterized E. coli strains from dogs and cats, focusing on phylogroup classification, virulence factors, and antimicrobial resistance profiles. Ninety-seven E. coli isolates from fecal samples of 31 dogs and 3 cats were obtained from a private diagnostic laboratory in Botucatu, Brazil, from March to October 2021. The antimicrobial susceptibility was assessed using the disk diffusion method. Polymerase chain reaction (PCR) was employed to screen for blaCTX-M and genes encoding virulence factors, as well as to classify the isolates into phylogroups. Twenty isolates were positive for intimin encoding gene eae and, consequently, these isolates were classified as EPEC (20.62%). Notably, 5.1% (5/97) of the isolates exhibited extended-spectrum ß-lactamase (ESBL) production and 13.4% (13/97) were identified as multidrug-resistant bacteria. Phylogroups A and B2 were the most prevalent, comprising 29.9% (29/97) and 26.8% (26/97) of the bacterial isolates, respectively. This characterization highlights the prevalence of EPEC in domestic animals, emphasizing the potential risk they pose to public health and highlighting the urgency of responsible antimicrobial use in veterinary practices and the important role of laboratories in the surveillance of pathogenic multidrug-resistant bacteria.

Investigating antimicrobial-resistant bacteria from exotic domestic birds - a One Health concern.

Antimicrobial resistance is a natural mechanism in microorganisms, making the treatment of infections more complex in human and veterinary medicine. Global exotic and ornamental bird markets have significantly increased, and the close relationship between pets and humans makes exploring the potential role of these birds as vectors for the spread of antimicrobial-resistant bacteria imperative. This study aimed to use culture-dependent methods to investigate cloacal bacteria and the presence of antibiotic-resistant bacteria in four breeding stocks of ornamental birds. Cloacal swab samples were collected from 53 birds (canaries = 32, cockatiels = 17, and budgies = 4) and used for culturing and isolating facultative anaerobic and/or obligatory aerobic Gram-positive and Gram-negative bacteria. The antimicrobial susceptibility profile of each isolate was determined by the disk diffusion method. Thirty-four isolates were obtained, most of which belonged to the Staphylococcus genus. Bacterial richness was higher in canaries and in one of the breeding stockings, where Gram-negative bacteria were more abundant than in the others. In addition, canaries exhibited a predominance of resistant isolates, particularly multidrug-resistant strains, probably due to prophylactic antimicrobial usage. Most Gram-negative bacteria were resistant to at least one drug tested. A vancomycin-resistant Enterococcus faecalis strain was isolated. Most Staphylococcus strains were resistant to gentamycin, followed by penicillin. Eight strains were cefoxitin-resistant, including oxacillin-resistant S. epidermidis, in which the mecA gene was detected. Understanding the prevalence of resistance in avian species is crucial in the collaborative pursuit of maintaining antibiotic effectiveness and strengthening public health defense against emerging infectious risks.

A resistência antimicrobiana é um mecanismo natural dos microrganismos, complicando o tratamento de infecções na medicina humana e veterinária. O mercado global de aves exóticas e ornamentais cresceu significativamente, e a relação próxima entre esses animais e humanos destaca a necessidade de investigar o papel das aves na disseminação de bactérias resistentes. Este estudo utilizou métodos dependentes de cultura para examinar bactérias cloacais e a presença de resistência a antibióticos em quatro plantéis de aves ornamentais. Amostras de suabe cloacal foram coletadas de 53 aves (canários = 32, calopsitas = 17, periquitos = 4) e usadas para cultivar e isolar bactérias Gram-positivas e Gram-negativas, facultativas anaeróbias e aeróbias obrigatórias. A suscetibilidade antimicrobiana foi determinada pelo método de difusão em disco. Foram obtidos 34 isolados, principalmente do gênero Staphylococcus. A riqueza bacteriana foi maior nos canários e em um dos plantéis, onde houve aumento de Gram-negativos. Canários mostraram predominância de isolados resistentes, especialmente cepas multirresistentes, provavelmente devido ao uso profilático de antimicrobianos. A maioria das bactérias Gram-negativas foi resistente a pelo menos um fármaco testado. Um Enterococcus faecalis resistente à vancomicina foi isolado. A maioria dos Staphylococcus foi resistente à gentamicina e penicilina; oito cepas foram resistentes à cefoxitina, incluindo S. epidermidis resistente à oxacilina com o gene mecA detectado. Compreender a prevalência de resistência em aves é crucial para manter a eficácia dos antibióticos e fortalecer a saúde pública contra riscos infecciosos emergentes.

Genomic analysis of the first multidrug-resistant ESBL-producing high-risk clonal lineage Klebsiella pneumoniae ST147 isolated from a cat with urinary tract infection.

Urinary tract infections (UTIs) are pervasive in human and veterinary medicine, notably affecting companion animals. These infections frequently lead to the prescription of antibiotics, contributing to the rise of antimicrobial-resistant bacteria. This escalating concern is underscored by the emergence of a previously undocumented case: a high-risk clone, broad-spectrum cephalosporin-resistant K. pneumoniae ST147 strain, denoted USP-275675, isolated from a cat with UTI. Characterized by a multidrug-resistant (MDR) profile, whole genome sequencing exposed several antimicrobial-resistance genes, notably blaCTX-M-15, blaTEM-1B, blaSHV-11, and blaOXA-1. ST147, recognized as a high-risk clone, has historically disseminated globally and is frequently associated with carbapenemases and extended-spectrum ß-lactamases. Notably, the core-genome phylogeny of K. pneumoniae ST147 strains isolated from urine samples revealed a unique aspect of the USP-276575 strain. Unlike its counterparts, it did not cluster with other isolates. However, a broader examination incorporating strains from both human and animal sources unveiled a connection between USP-276575 and a Portuguese strain from chicken meat. Both were part of a larger cluster of ST147 strains spanning various geographic locations and sample types, sharing commonalities such as IncFIB or IncR plasmids. This elucidates the MDR signature inherent in widespread K. pneumoniae ST147 strains carrying these plasmids, highlighting their pivotal role in disseminating antimicrobial resistance (AMR). Finally, discovering the high-risk clone K. pneumoniae ST147 in a domestic feline with a UTI in Brazil highlights the urgent need for thorough AMR surveillance through a One Health approach.

Alterations of rumen and fecal microbiome in growing beef and dairy steers fed rumen-protected Capsicum oleoresin.

The microbiome has been linked to animal health and productivity, and thus, modulating animal microbiomes is becoming of increasing interest. Antimicrobial growth promoters (AGP) were once a common technology used to modulate the microbiome, but regulation and consumer pressure have decreased AGP use in food animals. One alternative to antimicrobial growth promoters are phytotherapeutics, compounds derived from plants. Capsaicin is a compound from the Capsicum genus, which includes chili peppers. Capsaicin has antimicrobial properties and could be used to manipulate the gastrointestinal microbiome of cattle. Both the rumen and fecal microbiomes are essential to cattle health and production, and modulation of either microbiome can affect both cattle health and productivity. We hypothesized that the addition of rumen-protected capsaicin to the diet of cattle would alter the composition of the fecal microbiome, but not the rumen microbiome. To determine the impact of rumen-protected capsaicin in cattle, four Holstein and four Angus steers were fed rumen-protected Capsicum oleoresin at 0 (Control), 5, 10, or 15 mg kg-1 diet dry matter. Cattle were fed in treatment groups in a 4 × 4 Latin Square design with a 21-d adaptation phase and a 7-d sample collection phase. Rumen samples were collected on day 22 at 0-, 2-, 6-, 12-, and 18-h post-feeding, and fecal swabs were collected on the last day of sample collection, day 28, within 1 h of feeding. Sequencing data of the 16s rRNA gene was analyzed using the dada2 pipeline and taxa were assigned using the SILVA database. No differences were observed in alpha diversity among fecal or rumen samples for either breed (P > 0.08) and no difference between groups was detected for either breed in rumen samples or for Angus steers in fecal samples (P > 0.42). There was a difference in beta diversity between treatments in fecal samples of Holstein steers (P < 0.01), however, a pairwise comparison of the treatment groups suggests no difference between treatments after adjusting for multiple comparisons. Therefore, we were unable to observe substantial overall variation in the rumen or fecal microbiomes of steers due to increasing concentrations of rumen-protected capsaicin. We do, however, see a trend toward increased concentrations of capsaicin influencing the fecal microbiome structure of Holstein steers despite this lack of significance.

The microbiome is the collection of microbes present in an animal's body and has been discovered to be directly connected to animal health and productivity. In production animals, such as feedlot cattle, the microbiome can be modulated by antimicrobials to promote growth, but increasing consumer pressure to reduce antimicrobial use has producers seeking alternatives. Capsaicin is a phytotherapeutic derived from chili peppers that can be used to modulate the microbiome due to its antimicrobial properties. Eight steers were fed rumen-protected Capsicum oleoresin to determine its effect on average daily gain. In addition, rumen and fecal samples were collected for microbiome testing. No differences were detected in the rumen microbiomes between cattle fed capsaicin (treatment) or those that received no capsaicin (control). While no overall effect was observed on the fecal microbiome of cattle fed different doses of capsaicin or control, we did observe changes in fecal beta diversity due to capsaicin treatment in Holstein steers fed greater doses. The fecal microbiome structure of Holsteins fed greater dosages of capsaicin differed from those fed control or low doses, as observed by the presence of two distinct clusters. This observation suggests an impact of greater doses of capsaicin treatment on microbiome structure.

Bats (Mammalia: Chiroptera) in urban-rural interfaces: community structure associated with pathogen screening in São Paulo-the largest metropolitan region in Brazil.

Little is known about the influence of the urban environments on bat species 'ecology. The urbanization process potentially lead to critical ecological changes in bat communities' intra and interspecific pathogenic transmissions dynamics. To date, the monitoring of pathogens in bats in Brazil has only been done with bats found dead or alive in households, from rabies surveillance systems. The present work aimed to investigate how urbanization influenced bat richness, relative abundance and pathogen occurrence. Most captured bats were Phyllostomidae, especially Sturnira lilium, Artibeus lituratus, A. fimbriatus, Glossophaga soricina, and Platyrrhinus lineatus, among others. From preserved-rural towards urban areas the lesser the bat richness, the higher the relative abundance of the captured bats. Noise level, luminosity and relative humidity correlated with bat abundance. The proportion of genders, sexually active bats and their size (weight, right forearm length, and body condition index) were stable throughout the investigation. Still, the proportion of pregnant females was higher in Spring and the number of juveniles in Summer, evidencing the seasonality of reproduction. Several Enterobacteria were isolated, evidencing a significant role of bats in the circulation of pathogens of medical and veterinary interest. These results are crucial in the pursuit of a harmonious coexistence between humans, bats and domestic animals in areas with different levels of anthropization.

Bats Are Carriers of Antimicrobial-Resistant Staphylococcaceae in Their Skin.

Bats have emerged as potential carriers of zoonotic viruses and bacteria, including antimicrobial-resistant bacteria. Staphylococcaceae has been isolated from their gut and nasopharynx, but there is little information about Staphylococcaceae on bat skin. Therefore, this study aimed to decipher the Staphylococci species in bat skin and their antimicrobial susceptibility profile. One hundred and forty-seven skin swabs were collected from bats during the spring and summer of 2021 and 2022. Bats were captured in different areas of the Metropolitan Region of São Paulo, Brazil, according to the degree of anthropization: Area 1 (Forested), Area 2 (Rural), Area 3 (Residential-A), Area 4 (Slum-- up to two floors), Area 5 (Residential-B-condo buildings), and Area 6 (Industrial). Swabs were kept in peptone water broth at 37 °C for 12 h when bacterial growth was streaked in Mannitol salt agar and incubated at 37 °C for 24 h. The disc-diffusion test evaluated antimicrobial susceptibility. Staphylococcaceae were isolated from 42.8% of bats, mostly from young, from the rural area, and during summer. M. sciuri was the most frequent species; S. aureus was also isolated. About 95% of isolates were resistant to at least one drug, and most strains were penicillin resistant. Eight isolates were methicillin resistant, and the mecA gene was detected in one isolate (S. haemolyticus). Antimicrobial resistance is a One Health issue that is not evaluated enough in bats. The results indicate that bats are carriers of clinically meaningful S. aureus and antimicrobial-resistant bacteria. Finally, the results suggest that we should intensify action plans to control the spread of resistant bacteria.

Genomic analysis of a multidrug methicillin-resistant staphylococcus epidermidis recovered from the urine of a guinea pig (Cavia porcellus) with suspected pyelonephritis.

Staphylococcus epidermidis is an emerging pathogen causing infant pyelonephritis. There is a lack of genomic data on Staphylococcus epidermidis as the etiology of pyelonephritis and its resistant determinants. In this study, we have conducted a genomic and microbiologic investigation of an S. epidermidis recovered from the urine of a guinea pig with suspected pyelonephritis in Brazil. The genome was sequenced using the Illumina MiSeq platform and de novo assembled using SPades. Resistome, virulome, and plasmidome were in silico predicted using bioinformatics tools. Data analysis revealed that S. epidermidis USP-LZB-G06 belonged to sequence type ST332. Strikingly, a broad resistome (antibiotics, hazardous heavy metals, and biocides) was predicted, including the presence of the clinically relevant mecA, blaZ, and qacA efflux pump genes. SNP-based analysis revealed that strain USP-LZB-G06 was clustered along mecA positive S. epidermidis strains of ST332 isolated between 2008 and 2016 from humans in Australia and the United States of America. Our results indicate that the detection of this microorganism should be considered as a urinary tract infection agent in exotic pets, particularly guinea pigs. In addition, there is an urgent need to update veterinarians regarding the detection and therapeutic management of these microorganisms.

The first Mycoplasma ovipneumoniae recovered from a sheep with respiratory disease in Brazil - draft genome and genomic analysis.

Mycoplasma ovipneumoniae is an important etiological agent of sheep respiratory disease worldwide. Here, we describe the first isolation and draft genome sequence of M. ovipneumoniae strain USP-BR2017 retrieved from tracheobronchial lavage of a sheep showing clinical signs of respiratory disease in the Rio de Janeiro State, Brazil. The culture of tracheobronchial lavage resulted in glucose-fermenting fried egg colonies, which were identified as M. ovipneumoniae by polymerase chain reaction. The genome was sequenced using the Illumina NextSeq 2000 and de novo assembled using SPAdes. The genome of the sequenced organism presented an approximate size of 1,122,253 bp. The annotation revealed 773 coding DNA sequences (CDSs), 806 genes, three rRNAs, and 30 tRNAs. Data analysis revealed M. ovipneumoniae strain USP-BR2017 contains a few virulence genes, including the hemolysing C gene (hlyC). In addition, strain USP-BR2017 showed high identity over the 16S rRNA gene with other sheep isolates from China and United States. This first description of M. ovipneumoniae in diseased Brazilian sheep demonstrates the importance of continuous surveillance and diagnostics of pathogens causing respiratory disease in sheep in Brazil.

Bacteria associated with infectious keratoconjunctivitis outbreak in confined lambs.

Introduction. Infectious keratoconjunctivitis is a significant ocular disease found in confined sheep. Little information about the aetiological agents and their antimicrobial susceptibility is available.Gap Statement. There is limited information on the aetiological agents involved in keratoconjunctivitis outbreaks in sheep.Aim. The present research aimed to determine the bacterial aetiological factors involved in an outbreak of infectious keratoconjunctivitis in confined lambs.Methodology. Ocular swabs were collected from 23 randomly selected lambs, which were classified into three groups according to the severity of the lesion: group I (N=6; no ocular involvement), group II (N=8; less severe injuries) and group III (N=9; more severe injuries). Isolation of aerobic bacteria and antimicrobial susceptibility tests were carried out. Molecular detection of Mollicutes was performed, and positive samples were tested to confirm the presence of the following species: Mycoplasma conjunctivae, Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri.Results. Moraxella sp. and Mollicutes were detected in all groups, but we inferred that Moraxella sp. are only significant in the early stages of the disease. M. conjunctivae was detected in all tested groups, while M. agalactiae was detected in samples of group III only. One strain of Moraxella sp. was resistant to erythromycin and showed intermedite resistance to tetracycline.Conclusion. The presence of these species confirms their importance in the aetiology of this disease, and the low resistance profile observed in the studied farm suggested an increased cure success rate.

Leptospira sp. infection in bats: A systematic review and meta-analysis.

Bats are the only flying mammalian animals and are distributed worldwide. Bats are well-known hosts of several zoonotic viruses and bacteria, including Leptospira sp. Here, we performed a systematic review and a meta-analysis of evidence of Leptospira sp. infection in bats by examining studies published between 1964 and 2021. We reported the frequencies of various species and serogroups on all continents, several species and feeding habits of bats, and different diagnostic tools. Together, 33 papers from all continents with seven to 2077 individuals from one to 31 species were included. Molecular detection was conducted in most studies, followed by MAT (Microscopic Agglutination Test) and isolation and identification. Molecular characterization of Leptospira sp. revealed L. borgpetersenii as the most frequent species. Moreover, 179 positive samples for MAT contained the most likely infecting serogroups described, particularly the Australis serogroup. The percentage of positive tests in isolation and identification ranged between 0% and 0.5%. The highest frequency of Leptospira infection among the continents was observed in Asia, whereas South America had the lowest percentage. Finally, Nycteridae and Rhinonycteridae were the most frequently infected bat families. Our study provides valuable information about the epidemiology of Leptospira sp. infection in bats.

Genomic features of a multidrug-resistant and mercury-tolerant environmental Escherichia coli recovered after a mining dam disaster in South America.

Mining dam disasters contribute to the contamination of aquatic environments, impacting associated ecosystems and wildlife. A multidrug-resistant Escherichia coli strain (B2C) was isolated from a river water sample in Brazil after the Mariana mining dam disaster. The genome was sequenced using the Illumina MiSeq platform, and de novo assembled using Unicycler. Resistome, virulome, and plasmidome were predicted using bioinformatics tools. Data analysis revealed that E. coli B2C belonged to sequence type ST219 and phylogroup E. Strikingly, a broad resistome (antibiotics, hazardous heavy metals, and biocides) was predicted, including the presence of the clinically relevant blaCTX-M-2 extended-spectrum ß-lactamase (ESBL) gene, qacE∆1 efflux pump gene, and the mer (mercury resistance) operon. SNP-based analysis revealed that environmental E. coli B2C was clustered along to ESBL-negative E. coli strains of ST219 isolated between 1980 and 2021 from livestock in the United States of America. Acquisition of clinically relevant genes by ST219 seems to be a recent genetic event related to anthropogenic activities, where polluted water environments may contribute to its dissemination at the human-animal-environment interface. In addition, the presence of genes conferring resistance to heavy metals could be related to environmental pollution from mining activities. Antimicrobial resistance genes could be essential biomarkers of environmental exposure to human and mining pollution.

Choice of Commercial DNA Extraction Method Does Not Affect 16S Sequencing Outcomes in Cloacal Swabs.

As the applications of microbiome science in agriculture expand, laboratory methods should be constantly evaluated to ensure optimization and reliability of downstream results. Most animal microbiome research uses fecal samples or rectal swabs for profiling the gut bacterial community; however, in birds, this is difficult given the unique anatomy of the cloaca where the fecal, urinary, and reproductive tracts converge into one orifice. Therefore, avian gut microbiomes are usually sampled from cloacal swabs, creating a need to evaluate sample preparation methods to optimize 16S sequencing. We compared four different DNA extraction methods from two commercially available kits on cloacal swabs from 10 adult commercial laying hens and included mock communities and negative controls, which were then subjected to 16S rRNA amplicon sequencing. Extracted DNA yield and quality, diversity analyses, and contaminants were assessed. Differences in DNA quality and quantity were observed, and all methods needed further purification for optimal sequencing, suggesting contaminants due to cloacal contents, method reagents, and/or environmental factors. However, no differences were observed in alpha or beta diversity between methods. Importantly, multiple bacterial contaminants were detected in each mock community and negative control, indicating the prevalence of laboratory and handling contamination as well as method-specific reagent contamination. We found that although the extraction methods resulted in different extraction quality and yield, overall sequencing results were not affected, and we did not identify any method that would be an inappropriate choice in extracting DNA from cloacal swabs for 16S rRNA sequencing. Overall, our results highlight the need for careful consideration of positive and negative controls in addition to DNA isolation method and lend guidance to future microbiome research in poultry.

Detecting Mollicutes by PCR in goats in southwestern Bahia, Brazil.

Brazil has a herd of over 9 million goats, and the northeast of Brazil is home to over 93% of this herd. Caprine mycoplasmosis are widely disseminated worldwide, being highly contagious with high rates of morbidity and mortality, causing considerable economic loss to goat herders. In addition, there has been a lack of research using molecular testing to monitor the health and detect Mollicutes in this herd in Brazil. Therefore, the aim of this study is to associate animal management with the presence of the caprine origin Mollicutes in goats, in the southwest region of the state of Bahia, Brazil. A cross-sectional study was conducted on twelve farms, and statistical analyses were performed to identify associations between the presence of Mollicutes and the management of goats. Molecular testing identified Mollicutes class, Mycoplasma agalactiae (Ma) and M. conjunctivae (Mc), in the samples analyzed. Statistical associations were observed between animals from intensive livestock facilities and the presence of Mollicutes in nasal samples and dairy ranch animals and the presence of Mollicutes in ocular samples and animals from extensive ranching sites and positive results of Mollicutes in genital samples. We conclude that mycoplasmas are present in goat herds in the southwestern region of Bahia, which supports the need for more focused studies of mycoplasmas throughout the country. Our research also demonstrated the presence of two important opportunistic bacteria, Mc and Ma, and, to the best of our knowledge, this is the first time that M. conjunctivae was detected in Brazilian goats by molecular testing.

A Cross-Sectional Study of Dairy Cattle Metagenomes Reveals Increased Antimicrobial Resistance in Animals Farmed in a Heavy Metal Contaminated Environment.

The use of heavy metals in economic and social development can create an accumulation of toxic waste in the environment. High concentrations of heavy metals can damage human and animal health, lead to the development of antibiotic resistance, and possibly change in bovine microbiota. It is important to investigate the influence of heavy metals in food systems to determine potential harmful effects environmental heavy metal contamination on human health. Because of a mining dam rupture, 43 million cubic meters of iron ore waste flowed into the Doce river basin surrounding Mariana City, Brazil, in 2015. Following this environmental disaster, we investigated the consequences of long-term exposure to contaminated drinking water on the microbiome and resistome of dairy cattle. We identified bacterial antimicrobial resistance (AMR) genes in the feces, rumen fluid, and nasopharynx of 16 dairy cattle 4 years after the environmental disaster. Cattle had been continuously exposed to heavy metal contaminated water until sample collection (A) and compared them to analogous samples from 16 dairy cattle in an unaffected farm, 356 km away (B). The microbiome and resistome of farm A and farm B differed in many aspects. The distribution of genes present in the cattle's nasopharynx, rumen, and feces conferring AMR was highly heterogeneous, and most genes were present in only a few samples. The relative abundance and prevalence (presence/absence) of AMR genes were higher in farm A than in farm B. Samples from farm A had a higher prevalence (presence) of genes conferring resistance to multiple drugs, metals, biocides, and multi-compound resistance. Fecal samples had a higher relative abundance of AMR genes, followed by rumen fluid samples, and the nasopharynx had the lowest relative abundance of AMR genes detected. Metagenome functional annotation suggested that selective pressures of heavy metal exposure potentially skewed pathway diversity toward fewer, more specialized functions. This is the first study that evaluates the consequences of a Brazilian environmental accident with mining ore dam failure in the microbiome of dairy cows. Our findings suggest that the long-term persistence of heavy metals in the environment may result in differences in the microbiota and enrichment of antimicrobial-resistant bacteria. Our results also suggest that AMR genes are most readily detected in fecal samples compared to rumen and nasopharyngeal samples which had relatively lower bacterial read counts. Since heavy metal contamination has an effect on the animal microbiome, environmental management is warranted to protect the food system from hazardous consequences.

Use of a commercial ELISA kit specific for glycoprotein E peptides to indirectly detect Caprine Herpesvirus 1 (CpHV-1) in the state of São Paulo, Brazil / Uso de kit ELISA comercial específico para peptídeos da glicoproteína E para detecção indireta de Herpesvírus Caprino tipo 1 (CpHv-1) no estado de São Paulo, Brasil

Caprine herpesvirus 1 (CpHV-1) infection is associated with clinical manifestations related to animal age, with high mortality in kids and infertility in adults. Given the scarcity of research about the epidemiological situation of this infection in Brazilian flocks, we aimed to conduct a cross-sectional descriptive study to detect antibodies against CpHV-1 in goats in the state of São Paulo, Brazil. Fifty-five male and female goats ­ kids and adult ­ were assessed in this study. Blood serum was analyzed by a commercial ELISA kit to detect antibodies against CpHV-1, which had not been used in Brazil before. No animals were reactive. Brazil lacks information about CpHV-1 infection in goat flocks. Continuing the study is crucial to understand the epidemiological situation of the disease and establish protocols for infection control.(AU)

A infecção pelo Herpesvírus Caprino tipo 1 (CpHv-1) está associada a manifestações clínicas relacionadas à idade dos animais, com alta mortalidade em filhotes e infertilidade em adultos. Diante da escassez de estudos sobre situação epidemiológica dessa infecção nos rebanhos brasileiros, a presente pesquisa teve como objetivo realizar um estudo transversal e descritivo para a detecção de anticorpos anti-Herpesvírus Caprino tipo 1 em caprinos do estado de São Paulo, Brasil. Foram avaliados 55 caprinos machos e fêmeas, filhotes e adultos. O soro sanguíneo foi analisado por um kit ELISA comercial para detecção de anticorpos contra CpHv-1, de utilização inédita no Brasil. Nenhum animal estudado foi sororreagente. O Brasil carece de informações acerca da infecção pelo Herpesvírus Caprino tipo 1 nos rebanhos caprinos do país. A continuidade do estudo é imprescindível para compreender a situação epidemiológica da enfermidade e estabelecer protocolos para controle da infecção.(AU)

Bacteria isolated from the lower respiratory tract of sheep and their relationship to clinical signs of sheep respiratory disease / Bactérias detectadas no trato respiratório inferior de ovinos e a relação com as manifestações clínicas da doença respiratória ovina

Respiratory diseases are among the most important diseases in sheep flocks. Herein was studied the bacterial etiology of respiratory disease and the clinical signs of 99 female and male sheep breed in the states of São Paulo (SP) and Rio de Janeiro (RJ), Brazil. After physical examination of animals, tracheobronchial flushing samples were obtained. The usual bacteria and Mycoplasma spp. were searched, as well as their association with the clinical status and clinical signs of sheep with respiratory disease. The main observed signs were: tachypnea (75%), increase of rectal temperature (09.4%), mucopurulent/purulent nasal discharge (21.9%), cough (25%), dyspnea (31.2%), changes of lung sounds at auscultation (87.5%) and chest percussion (28.1%) in pneumonic sheep. Non-fermenting gram-negative bacteria and Bacillus sp. were the most isolated bacteria. Microorganisms of the Mollicutes class were molecularly (PCR) detected in 33.3% of the animals. In addition, the specific detection of M. mycoides subsp. capri was described for the first time in sheep from the state of São Paulo, Brazil.(AU)

A doença respiratória é uma das doenças mais importantes em rebanhos ovinos. Esta pesquisa teve como objetivo determinar a etiologia bacteriana da doença respiratória e sua relação com sinais clínicos em ovinos criados nos estados de São Paulo e Rio de Janeiro, Brasil. Noventa e nove ovelhas machos e fêmeas dos Estados de São Paulo (SP) e Rio de Janeiro (RJ) foram estudadas. Após o exame físico, amostras de lavagem traqueobrônquica foram obtidas. A presença de bactérias aeróbias e Mycoplasmaspp. foram estudados, assim como a associação entre os microrganismos e estado clínico e sinais clínicos de doença respiratória em ovinos. As principais manifestações clínicas observadas foram: taquipneia (75%), alta temperatura retal (09,4%), secreção nasal mucopurulenta/purulenta (21,9%), tosse (25%), dispneia (31,2%), sons pulmonares alterados na ausculta (87,5%) e na percussão torácica (28,1%) em ovelhas pneumônicas. Bactérias gram-negativas não fermentadoras e Bacillus sp. foram as bactérias mais isoladas. Microrganismos da classe Mollicutes foram detectados molecularmente (PCR) em 33,3% dos ovinos. Além disso, descreve-se pela primeira vez no estado de São Paulo, Brasil, a detecção do M. mycoides subsp. capri na espécie ovina utilizando a reação de polimerase em cadeia.(AU)

Serological investigation of antibodies against respiratory viruses in calves from Brazilian family farming and their relation to clinical signs of bovine respiratory disease / Investigação sorológica de anticorpos contra vírus respiratórios em bezerros de propriedades familiares brasileiras e sua relação com os sinais clínicos da doença respiratória dos bovinos

Bovine respiratory disease (BRD) is responsible for economic losses in cattle production. Viruses are categorized as primary etiological agents. The aims of this study were to evaluate the presence of antibodies against bovine viral diarrhea virus (BVDV), bovine herpes virus type 1 (BoHV-1), and bovine respiratory syncytial virus (BRSV) in healthy and BRD calves from family farming in relation to clinical signs of BRD. Hundred and forty-five calves were randomly selected and physical examination was performed. Only 123 animals were classified as healthy and BRD calves. Antibodies were evaluated by virus neutralization test. Person's Chi-square test and Fisher's exact test were performed as univariate analysis. Binary Logistic Regression was applied as multivariate analysis. Variables with P<0.10 were considered statistically significant. Variables with 0.15<P<0.10 were considered as statistical tendencies. Antibodies against BoHV-1, BVDV, and BRSV were detected in 32%, 23% and 37% animals. Antibodies against both three viruses were detected in 26.8% of calves. The presence of antibodies against BRSV were associated to normal heart rates (P=0.018) and unilateral airflow (P=0.035). Tendency was observed to unilateral airflow (P=0.06) Statistical tendencies were observed to Ab-BoHV-1 and body temperature (P=0.119) and breathing pattern (P=0.123). The profile of antibodies against respiratory viruses in calves from Brazilian family farming was firstly described. The results confirmed the importance of some clinical signs to viral infection.(AU)

A doença respiratória dos bovinos (DRB) é responsável por importantes perdas econômicas para a produção bovina, com maior impacto na agricultura familiar. Os vírus são comumente caracterizados como agentes etiológicos primários devido a mudanças na mucosa respiratória, na produção de citocinas e no funcionamento do sistema imune. Os objetivos deste estudo transversal foram avaliar a presença de anticorpos contra o vírus da diarreia viral bovina (VDVB), herpes vírus bovino tipo 1 (HVbo-1) e vírus respiratório sincicial bovino (VRSB) em bezerros sadios e com DRB de assentamentos em associação com a presença sinais clínicos de DRB. Cento e quarenta e cinco animais foram randomicamente selecionados e exame físico foi realizado. Somente 123 animais foram classificados em sadios e com DRB. Anticorpos foram identificados pelo teste de virusneutralização. Teste de qui-quadrado e teste exato de Fisher foram utilizados como análise univariada. A análise multivariada foi realizada pela regressão binária logística com o método Backward conditional. Variáveis com P<0,10 foram considerados significantes. Variáveis com 0,15<P<0,10 foram consideradas tendências. Anticorpos contra HVbo-1, VDVB e VRSB foram detectados em 32%, 23% e 37% dos animais. A presença concomitante de anticorpos contra os três vírus foi detectada em 26,8% dos bezerros. A presença de anticorpos contra VRSB foi associada à frequência cardíaca normal (P=0,018) e fluxo de ar nasal unilateral (P=0,035). Tendência estatística observada para anticorpos contra VDVB temperatura corporal (P=0,119) e padrão respiratório (P=0,123). Ao nosso conhecimento, este é o primeiro estudo que objetivou a avaliação da presença de anticorpos contra VDVB, VRSB e HVbo-1 em bezerros de assentamentos do estado de São Paulo, Brasil e sua relação com as manifestações clínicas da DRB.(AU)

Bacterial pathogens of the lower respiratory tract of calves from Brazilian rural settlement herds and their association with clinical signs of bovine respiratory disease / Patógenos bacterianos do trato respiratório inferior de bezerros criados em rebanhos de assentamentos brasileiros e sua associação com os sinais clínicos da doença respiratória dos bovinos

Bovine respiratory disease (BRD) is considered the major cause of economic losses in dairy and beef cattle production. The study aimed to detect the most important bacteria related to respiratory disease in tracheobronchial fluid samples of healthy and dairy calves with clinical signs of BRD in Brazilian rural settlements. Hundred and forty-one mongrel dairy calves were randomly selected from 42 family farm dairy herds from Brazilian settlements. Physical examination was performed and calves were classified as healthy (n=100) and BRD (n=41). Tracheobronchial fluid samples were collected. Isolation and molecular detection of Mycoplasma dispar, M. bovis and M. mycoides subsp. mycoides SC besides isolation of other aerobic bacteria were performed. Abnormal lung sounds (crackle/snoring/whistle), mucopurulent/purulent nasal discharge, body temperature >39.5°C and respiratory rate >40 breaths/min were higher in BRD calves compared to healthy calves (P<0.05). Bacillus sp., Staphylococcus intermedius and non-fermentative Gram-negative were the most prevalent bacteria isolated. Non-identified species from Enterobacteriaceae family was higher in BRD calves compared to healthy calves (P<0.05). Mollicutes were isolated in 7.4% of samples and only M. dispar was detected. Mollicutes was associated with purulent/mucopurulent nasal discharge (P=0.017). Pantoea agglomerans was associated to tachypnea (P=0.020), and Streptococcus spp. was associated with hyperthermia. Statistical tendencies were observed to M. dispar and tachypnea (P=0.066), and P. agglomerans and tachycardia (P=0.066). The obtained results describe the microorganisms found in tracheobronchial fluid of calves with BRD in some herds of Brazilian family farming and their relation to clinical signs of BRD.(AU)

A doença respiratória dos bovinos (DRB) é considerada a principal causa de perdas econômicas nas produções de leite e carne. O objetivo deste estudo foi detectar as mais importantes bactérias relacionadas a doença respiratória presentes em amostras de lavado traqueobrônquico de bezerros sadios e com sinais clínicos da DRB de assentamentos brasileiros. Cento e quarenta e um bezerros leiteiros sem raça definida foram randomicamente selecionados de 42 rebanhos leiteiros de assentamentos brasileiros. Exame físico foi realizado e os animais foram classificados em sadios (n=100) e com DRB (n=41). Amostras de lavado traqueobrônquico foram coletadas. Foram realizados o isolamento e a detecção molecular de Mycoplasma dispar, M. bovis e M. mycoides subsp. mycoides SC além de isolamento de outras bactérias aeróbias. Ruídos pulmonares anormais (crepitação/ ronco/sibilo), secreção nasal mucopurulenta/purulenta, temperatura corporal >39.5°C e frequência respiratória >40 movimentos respiratórios/min foram observados com maior frequência em bezerros com DRB comparado aos animais sadios (P<0.05). Bacillus sp, Staphylococcus intermedius e bactérias Gram-negativas não fermentadoras foram as bactérias mais prevalentes. Bactérias da família Enterobacteriaceae cuja espécie não fora identificada foram mais frequentes em bezerros com DRB comparado aos bezerros sadios (P<0.05). Mollicutes foram isolados em 7,4% das amostras e somente M. dispar foi detectado. Mollicutes foi associado à secreção nasal purulenta/mucopurulenta (P=0.017). Pantoea agglomerans foi associada a taquipneia (P=0.020), e Streptococcus spp. Foi associado a hipertermia. Tendência estatística foi observada para M. dispar e taquipneia (P=0.066), e P. agglomerans e taquicardia (P=0.066). Os resultados obtidos descrevem os micro-organismos encontrados no lavado traqueobrônquico de bezerros com DRB em rebanhos de agricultura familiar brasileira e sua relação com as manifestações clínicas da DRB.(AU)

Bovine leukaemia virus genotypes 5 and 6 are circulating in cattle from the state of São Paulo, Brazil.

PURPOSE: Enzootic bovine leucosis (EBL) is a silent disease caused by a retrovirus [bovine leukaemia virus (BLV)]. BLV is classified into almost 10 genotypes that are distributed in several countries. The present research aimed to describe two BLV gp51 env sequences of strains detected in the state of São Paulo, Brazil and perform a phylogenetic analysis to compare them to other BLV gp51 env sequences of strains around the world. METHODOLOGY: Two bovines from different herds were admitted to the Bovine and Small Ruminant Hospital, School of Veterinary Medicine and Animal Science, University of São Paulo, Brazil. In both, lymphosarcoma was detected and the presence of BLV was confirmed by nested PCR. The neighbour-joining algorithm distance method was used to genotype the BLV sequences by phylogenetic reconstruction, and the maximum likelihood method was used for the phylogenetic reconstruction. The phylogeny estimates were calculated by performing 1000 bootstrap replicates. RESULTS: Analysis of the partial envelope glycoprotein (env) gene sequences from two isolates (25 and 31) revealed two different genotypes of BLV. Isolate 25 clustered with ten genotype 6 isolates from Brazil, Argentina, Thailand and Paraguay. On the other hand, isolate 31 clustered with two genotype 5 isolates (one was also from São Paulo and one was from Costa Rica). The detected genotypes corroborate the results of previous studies conducted in the state of São Paulo, Brazil. The prediction of amino acids showed substitutions, particularly between positions 136 and 150 in 11 out of 13 sequences analysed, including sequences from GenBank. CONCLUSION: BLV is still important in Brazil and this research should be continued.

Transmissibilidade de Lentivírus de Pequenos Ruminantes para cabritos e cabras adultas por meio de sêmen infectado experimentalmente / Transmissibility of Small Ruminants Lentivirus in kids by experimentally infected semen

A Artrite Encefalite Caprina se caracteriza por ser multissistêmica e infecciosa, causada por um lentivírus. O estudo teve como objetivo avaliar a transmissibilidade do Lentivírus Caprino, para fêmeas e sua prole, por meio de sêmen infectado experimentalmente. Para tanto, onze fêmeas livres de CAEV foram inseminadas artificialmente com sêmen de bode livre de CAEV ao qual foi adicionado CAEV-Cork para obter título infectante com carga viral em 105 TCID50/ml. (grupo experimental 1). Destas, seis obtiverem prenhez confirmada, e a sua prole (n=6) constituiu o grupo experimental 2. Duas cabras livres de CAEV foram inseminadas artificialmente com sêmen do mesmo bode, sem o inócuo viral, constituindo-se o grupo controle. O diagnóstico da infecção pelo Lentivírus Caprino, foi realizado por IDGA, cELISA e nested-PCR. As fêmeas foram monitoradas durante 210 dias pós inseminação artificial. Já as proles foram imediatamente separadas das mães após o nascimento, e monitoradas nos momentos hora zero, aos quinze dias de idade e mensalmente, até doze meses de idade. Em relação às cabras, 56,96%(9/158) apresentaram positividade para cELISA, 24,05% (38/158) foram positivas a IDGA e nenhuma para nested-PCR. Em relação aos cabritos, 11,28% (15/133) amostras positivas para nested-PCR, 5,26% (7/133) amostras positivas para IDGA e nenhum para cELISA. As proles do grupo controle apresentaram resultados negativos para as três técnicas. A positividade encontrada em nested-PCR pode indicar grande importância para identificação de animais infectados, porém soronegativos, em situações de soroconversão tardia. De acordo com os resultados, concluiu-se que há a transmissão do Lentivírus caprino para a prole e para as mães pelo sêmen infectado.(AU)

Caprine Arthritis Encephalitis is a multisystemic infectious disease, caused by a lentivirus. The objective of this study was to evaluate the transmissibility of caprine lentivirus to goats and their offspring, through experimentally infected semen. Therefore, eleven free-CAEV goats were artificially inseminated using semen from a free-CAEV buck experimentally infected with CAEV-Cork strain (experimental group one). Pregnancy was confirmed in only six goats and their offspring (n=6) constituted the experimental group two. Two free-CAEV females were artificially inseminated with semen from the same seronegative buck, without viral inoculum to constitute the control group. The diagnosis of caprine lentivirus infection was performed using AGID, cELISA and nested-PCR. All females were monitored for 210 days after artificial insemination. Kids were immediately separated from their mothers after birth, and monitored at zero time, 15 days old and monthly until 12 months old. Regarding goat samples, 56.96% (9/159) were positive in cELISA, 24.05% (38/158) were positive in IDGA and none was positive in nested-PCR. Regarding to the offspring samples, 11.28% (15/133) and 5.26% (7/133) were positive in nested-PCR and IDGA, respectively, while no sample was positive in cELISA. The control group showed no positives in the three techniques. The positivity observed to nested-PCR may show its importance to identify infected, but seronegative animals, in late seroconversion situations. According to results, the transmission of caprine lentivirus to offspring and their mothers through infected semen is possible.(AU)