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Weissella is a genus containing Gram-positive, heterofermentative bacteria belonging to the lactic acid bacteria (LAB) group. These bacteria are endowed with promising technological and antimicrobial attributes. Weissella cibaria W25 was isolated from a dairy environment where raw milk cheeses are produced. Therefore, we sequenced and assembled the W25 draft genome sequence, which consists of 41 contigs totaling ~2.4 Mbp, with a G + C content of 45.04%. Then we carried out a comprehensive comparative genomic analysis with W. cibaria 110, known to produce the weissellicin 110 bacteriocin, and four other non-bacteriocin-producing W. cibaria strains.

The aim of this work was to verify the occurrence, quantification, pulse types, and antimicrobial susceptibility profiles of Salmonella sp. isolated from chicken meat produced and marketed in the state of Paraná, considered to be the state with the highest production of poultry meat in Brazil. Ninety-five of 300 (31.5%) frozen cuts of chicken were found to contain Salmonella sp., and 98 different isolates of Salmonella sp. were cultured from the positive samples. Quantification showed low Salmonella sp. loading, ranging from 0.12 to 6.4 MPN/g. The antimicrobial resistance test was performed against 16 agents from 6 different classes. All isolates were sensitive to meropenem, imipenem, chloramphenicol, and amikacin. The highest resistance rates were observed for nalidixic acid (95%), tetracycline (94%), doxycycline (94%), ampicillin (87%), amoxicillin with clavulanic acid (84%), ceftriaxone (79%), and ciprofloxacin (76%). A total of 84 (85.7%) of the isolates were identified with a multidrug resistant profile, 13 of which were found to have encoding genes extended-spectrum beta-lactamase (ESBL), especially blaCTX-M-2 e blaTEM-1. The major serovars identified were S. Typhimurium (43%) and S. Heidelberg (39%). The third most isolated serovar was S. Ndolo (6%), without previous reports of its presence in poultry meat in Brazil. Molecular characterization of S. Typhimurium and S. Heidelberg isolates by pulsed field gel electrophoresis (PFGE) showed a clonal relationship between all isolates of the same serovar (genetic similarity greater than 80%). Isolates of S. Typhimurium and S. Heidelberg with 100% similarity were found in up to five different geographic regions of the state, showing the potential for the spread of this pathogen in the Paraná poultry chain. Epidemiological surveys like this are important to understand the dynamics of dissemination and to monitor the prevalence of pathogens in the final products of poultry chains. In addition, to know the resistance profile of strains of Salmonella sp. present in food that contributes to the adoption of faster and more effective therapeutic measures, when necessary.

Antibiotic resistance has become a major concern for human and animal health. As fluoroquinolones have been extensively used in human and veterinary medicine, there has also been the rapid emergence and spread of antimicrobial resistance around the world. Here, we analysed the microbiome of goat milk using samples from healthy goats and those diagnosed with persistent mastitis and treated using the antibiotic enrofloxacin with 16S rRNA amplicon sequencing. We selected a group of 11 goats and 22 samples of milk that did not respond clinically to enrofloxacin treatment. Milk samples were evaluated before and after treatment to verify changes of the microbiota; the three first lactating goats were selected from the healthy control group. The milk samples from the healthy control animals presented a larger abundance of different species of bacteria of the Staphylococcus genus, but a smaller number of different genera, which indicated a more specific niche of resident bacteria. The Firmicutes phylum was predominantly different between the studied groups. Samples from before-treatment animals had a higher number of new species than those from the control group, and after being treated again. These microbiota received new bacteria, increasing the differences in bacteria even more in relation to the control group. Genotypes such as Trueperella and Mannheimia , between other genera, had a high abundance in the samples from animals with persistent mastitis. The dysbiosis in this study, with marked evidence of a complex microbiota in activity in cases of the failure of antimicrobial treatment for persistent chronic mastitis, demonstrates a need to improve the accuracy of pathogen identification and increases concern regarding antibiotic treatments in milk production herds.

This research was carried out to investigate the differences in adhesion and growth during biofilm formation of L. monocytogenes from different sources and clonal complexes. Biofilm by L. monocytogenes (isolates CLIST 441 and 7: both lineage I, serotype 1/2b, CC3; isolates 19 and 508: both lineage II, serotype 1/2c, CC9) was grown on stainless steel coupons under different stressing conditions (NaCl, curing salts and quaternary ammonium compounds—QAC), to determine the expression of different genes involved in biofilm formation and stress response. CLIST 441, which carries a premature stop codon (PMSC) in agrC, formed high-density biofilms in the presence of QAC (7.5% w/v) or curing salts (10% w/v). Reverse Transcriptase-qPCR results revealed that L. monocytogenes isolates presented differences in transcriptional profile of genes related to biofilm formation and adaptation to environmental conditions. Our results demonstrated how L. monocytogenes can survive, multiply and form biofilm under adverse conditions related to food processing environments. Differences in transcriptional expression were observed, highlighting the role of regulatory gene networks for particular serotypes under different stress responses.

In this study, we aimed to characterize the distribution of Yersinia enterocolitica in a pork production chain in Brazil, as well as the virulence profile and antibiotic resistance of the obtained isolates. Samples from 10 pig lots obtained from finishing farms (water, feed, and barn floors, n = 30), slaughterhouse (lairage floors, carcasses at four processing steps, tonsils, and mesenteric lymph nodes, n = 610), and processing (end cuts, processing environment, n = 160) were obtained in Paraná state, Brazil, and subjected to Y. enterocolitica detection by ISO 10,273. The obtained isolates were identified based on biochemical and molecular features (16 s rRNA, inv, bioserotyping) and subjected to PCR assays to detect virulence (ail, ystA, ystB, virF, myfA, fepA, fepD, fes, tccC, ymoA, hreP, and sat) and multidrug resistance–related genes (emrD, yfhD, and marC). Also, isolates were subjected to disk diffusion test to characterize their resistance against 17 antibiotics from 11 classes and to pulsed field gel electrophoresis (PFGE) after XbaI macro-restriction. Y. enterocolitica was detected in a single sample (tonsil), and the obtained three isolates were characterized as serotype O:3, harboring ail, ystA, virF, myfA, tccC, ymoA, hreP, emrD, yfhD, and marC, and resistant to all tested antibiotics. The three isolates presented identical macro-restriction profiles by PFGE, also identical to isolates obtained from Minas Gerais, other Brazilian state; one selected isolate was identified as biotype 4. Despite the low occurrence of Y. enterocolitica in the studied pork production, the virulence potential and the antibiotic resistance profiles of the isolates demonstrated their pathogenic potential, and the macro-restriction profiles indicate strains descending from a common subtype in the pork production chain of two Brazilian States.

Pestiviruses, including bovine viral diarrhea virus (BVDV), are significant pathogens in veterinary medicine, posing diagnostic and biosecurity challenges in livestock. This study investigated the seroprevalence of BVDV-1 and BVDV-2 in pigs from the micro-region of Ponte Nova, Minas Gerais, Brazil, a significant swine-producing area. A total of 400 serum samples were collected from sows and finished pigs, across nine farms practicing intensive farrow-to-finish systems. Virus neutralization assays were performed to detect neutralizing antibodies against BVDV-1 and BVDV-2. The results revealed a seroprevalence of 0.25% (BVDV-1) and 4.5% (BVDV-2), predominantly in finished pigs. The findings suggested variable exposure or susceptibility across the production stages and highlighted the potential antigenic cross-reactivity with the classical swine fever virus (CSFV). Despite the region’s CSFV-free status, the absence of routine surveillance, could hinder the early detection of potential incursions. This study provided a valuable baseline data for BVDV circulation in Brazilian swine populations and underscores the importance of continued pestivirus surveillance. Enhanced diagnostic protocols and biosecurity measures are recommended to mitigate interspecies transmission risks and to improve disease control strategies in mixed-species farming environments. RESUMO: Os pestivírus, incluindo o vírus da diarreia viral bovina (BVDV), são patógenos significativos na medicina veterinária, representando desafios diagnósticos e de biossegurança em animais de produção. Este estudo investiga a soroprevalência de BVDV-1 e BVDV-2 em suínos da microrregião de Ponte Nova, Minas Gerais, Brasil, uma área significativa de produção de suínos. Um total de 400 amostras de soro foram coletadas de matrizes e suínos de terminação em nove granjas que praticam sistemas intensivos de ciclo completo. Ensaios de neutralização viral foram realizados para detectar anticorpos neutralizantes contra BVDV-1 e BVDV-2. Os resultados revelaram uma baixa soroprevalência de BVDV-1 (0,25%) e uma prevalência relativamente maior de BVDV-2 (4,5%), predominantemente em suínos de terminação. Os achados sugerem exposição ou suscetibilidade variável entre os estágios de produção e destacam a potencial reatividade cruzada antigênica com o vírus da peste suína clássica (PSC). Apesar do status da região livre de PSC, a ausência de vigilância de rotina pode dificultar a detecção precoce de possíveis incursões. Este estudo fornece dados básicos valiosos sobre a circulação de BVDV em populações suínas brasileiras e reforça a importância da vigilância contínua de pestivírus. Protocolos diagnósticos aprimorados e medidas de biossegurança são recomendados para mitigar os riscos de transmissão interespecífica e melhorar as estratégias de controle de doenças em ambientes de produção com múltiplas espécies.

Ticks and fleas are essential vectors of pathogens that affect humans and animals, and among their hosts, synanthropic animals such as the black-eared opossum, Didelphis aurita, play a role in public health due to their ability to move between urban centers and forested areas in Brazil. This study aimed to assess the ectoparasite fauna of D. aurita, as well as the presence of pathogens and endosymbionts in ticks and fleas. Opossums (n = 58) captured in Tomahawk livetraps were examined for ectoparasites, and their blood sampled for further analysis. Additionally, spleen samples were collected in individuals found dead. Samples were PCR screened for Rickettsia spp., Borrelia spp., Anaplasmataceae, and Babesia spp. Two tick species were morphologically identified as Ixodes loricatus 24/58 (41.4%) and Amblyomma sculptum 1/58 (1.7%). For fleas, Ctenocephalides felis was detected in 60.3% (35/58) of the animals, and Xenopsylla cheopis in 5.2% (3/58). PCR analysis detected Anaplasmataceae DNA in 34% (16/47) of pooled samples of C. felis, and in 66.7% (2/3) pooled samples of X. cheopis. Sequence analysis revealed Wolbachia pipientis symbiont in all positive samples. Tick, blood and spleen samples were all negative for the microorganisms assessed. These findings suggest that these arthropods circulate among wildlife and urban environments, which may implicate in their participation in the cycle of zoonotic pathogens among opossums, humans and companion animals.

Toxoplasmosis is a worldwide-distributed zoonotic disease of great relevance to public health. Many wildlife species, including marsupials of the genus Didelphis, are considered hosts for Toxoplasma gondii , which makes them to have a possible role in the dispersion and maintenance of this parasite in nature. This study provides data on the molecular detection of T. gondii in D. aurita opossums from an urbanized area of Southeastern Brazil. Animals were captured and blood and/or spleen samples were collected. Real Time PCR was performed for the detection of T. gondii . From the opossums captured, 26.3% (n = 15/57) scored positive, with a frequency of 21.6% (n = 11/51) in blood, and 66.7% (n = 6/9) in spleen samples. BLAST analysis demonstrated 100% identity and 100% cover query with sequences of T. gondii available in GenBank database. Data herein reported present great public health importance, since Didelphis spp. are usually observed inhabiting close to human dwellings, which facilitates their contact with people and domestic animals, and consequently, the transmission of zoonotic agents. However, further studies are needed to elucidate whether these opossums play an important role in the zoonotic cycle of T. gondii in urban areas of Brazil.

Currently, a great proportion of the emerging infectious human diseases are zoonotic, with most of the pathogens originated from wildlife. In this sense, synanthropic animals such as marsupials play important role in the dissemination of pathogens due to their proximity to human dwellings. These hosts are affected by many gastrointestinal parasites, including species with zoonotic potential. The aim of this study was to assess the diversity of gastrointestinal parasites infecting the black-eared opossum D. aurita captured in urban areas of Southeastern, Brazil. In addition, the potential risk for the human population based on the One Health perspective has been discussed. Forty-nine marsupial specimens were captured with Tomahawk live traps and fecal samples were collected. The samples were evaluated by parasitological procedures. Eggs and oocysts were analyzed at different magnifications (400 × and 1000 ×), and their identification, together with adult nematodes, was established on morphological and morphometric data. Forty-three hosts (87.76%) scored positive for at least one gastrointestinal parasite, being 83.67% (41/49) for helminths, and 65.30% (32/49) for protozoa. For Cryptosporidium sp., only 13 samples were evaluated due to insufficient amount of feces obtained of some animals. A prevalence of 23.08% (3/13) was reported for this parasite. PCR analysis revealed Ancylostomatidae eggs to belong to the genus Ancylostoma . Our results demonstrated that multiparasitism is frequently found in these animals and a high percentage of potentially zoonotic parasites are observed, implying that D. aurita may be involved in zoonotic cycles in urban environments.

The black-eared opossum (Didelphis aurita) is a South American synanthropic marsupial. The presence of opossums in domestic spaces is relevant in the One-Health context since they are hosts of pathogens and ectoparasites that may affect the health of domestic animals and humans. In this study, we aim to determine the occurrence of hemoplasmas and selected tick-borne pathogens in free-ranging black-eared opossums, along with their molecular characterization, hematological and biochemical evaluation and factors associated with infection, in the municipality of Viçosa, State of Minas Gerais, southeastern Brazil. Thirty black-eared opossums were trapped between March 2021 and June 2022. Ectoparasites were collected. Hematological and biochemical analyses were performed. DNA from EDTA-blood samples were analyzed by PCR and qPCR assays. By molecular analyses, ‘Candidatus Mycoplasma haemoalbiventris’ was the most prevalent hemoparasite (73.3%), followed by Hepatozoon sp. (22.2%). Significant differences were observed in the number of platelets, and in the concentration of protein and globulins in the animals infected by ‘Ca. M. haemoalbiventris’ when compared with the negative group. This is the first report of ‘Ca. M. haemoalbiventris’ infection in D. aurita.