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.

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.

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.

Determination of bacterial aetiologic factor on tracheobronchial lavage in relation to clinical signs of bovine respiratory disease.

This study aimed to determine the occurrence of Mannheimiahaemolytica, Pasteurella multocida and Mycoplasma spp., in relation to clinical signs of respiratory disease. Tracheobronchial lavage samples were collected from 96 (healthy and unhealthy) cattle in the State of São Paulo, Brazil. Mycoplasma spp. (12.5 %) and Pasteurellamultocida (15.50 %) were the most prevalent species. Bacillus sp., Staphylococcus sp., Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Klebsiella pneumoniae were also isolated. Mollicutes (70.83 %), Mycoplasmabovis (2.94 %) and Mycoplasma dispar (38.23 %) were identified using conventional PCR. Submassive sound on acoustic percussion of the thorax was associated with the absence of Mollicutes (P=0.025). Whistling (P=0.076) and coarse crackle (P=0.046) were associated with the absence of Mycoplasma dispar. Clear sound on acoustic percussion of the thorax was associated with the absence of Mycoplasmabovis (P=0.007). Coughing was associated with the presence of Pasteurellamultocida [P=0.035; confidence interval (CI), 1.12-26.89], but its absence was associated with mucopurulent (P=0.0215; CI, 1.55-34.5) and mucoid nasal discharge (P=0.068; CI, 19-28.5), submassive sound (P=0.031; CI, 1.23-75.5), fine crackle (P=0.058; CI, 1.23-20.1) and coarse crackle (P=0.046; CI, 2.38-70.8). The high prevalence of Pasteurella multocida and Mycoplasma spp. in unhealthy calves increases the importance of these micro-organisms in the pathogenesis of respiratory diseases. This study increases the information about the role of Mycoplasma dispar in respiratory diseases. Differences in some species in relation to clinical signs can be applied as a presumptive diagnosis.