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Mycoplasma hyopneumoniae ( M. hyopneumoniae ) is considered the primary causative agent of porcine enzootic pneumonia (EP), a chronic contagious respiratory disease that causes economic losses. Obtaining new pathogenic isolates and studying the genome and virulence factors are necessary. This study performed a complete sequencing analysis of two Brazilian strains, UFV01 and UFV02, aiming to characterize the isolates in terms of the virulence factors and sequence type. The complete genome analysis revealed the main virulence genes ( mhp385 , mhp271 , MHP_RS03455 , p102 , p97 , p216 , MHP_RS00555 , mhp107 ) and ST-123, the presence of three toxin-related genes ( tlyC , PLDc_2 and hcnC ), and some genetic groups specific to these two isolates. Subsequently, the pathogenicity of the isolates was evaluated via an experimental infection conducted in a swine model. The study was divided into three groups, namely a negative control group ( n  = 4) and two test groups ( n  = 8), totaling 20 animals. They were challenged at 35 days of age with 10 7 CCU (Color Changing Units) M. hyopneumoniae via the intratracheal route. The UFV01 group showed earlier and higher seroconversion (IgG) (100%), while only 50% of the UFV02 group seroconverted. The same trend was observed when analyzing the presence of IgA in the bronchoalveolar lavage fluid (BALF) at 35 days post-infection (dpi). The UFV01 group had a mean macroscopic lesion score of 11.75% at 35 dpi, while UFV02 had 3.125%. Microscopic lesions were more severe in the UFV01 group. Based on laryngeal swab samples evaluated by qPCR, and the detection began at 14 days. The UFV01 group showed 75% positivity at 14 dpi. The UFV02 group also started excreting at 14 dpi, with a positivity rate of 37.5%. The results indicate that the UFV01 isolate exhibits higher virulence than UFV02. These findings may aid in developing new vaccines and diagnostic kits and establishing experimental models for testing., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Toledo, de Souza, Pereira, Polveiro, Bressan, Yamatogi, Jeong, Marks, Diamantino, de Carvalho, Malcher, Petri, de Oliveira, Moreira and Silva-Júnior.)

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 .

Mastitis, mainly caused by bacterial intramammary infections, is the main problem in the breeding of dairy animals. The inflammations of the mammary gland is separated by types of mastitis, being subclinical, clinical, and the most severe, gangrenous mastitis. Here, we used 16S rRNA amplicon sequencing to characterize the bacterial microbiota of goat milk in the different types of goat mastitis caused by bacteria. We used 72 goat milk samples from a region of the state of Minas Gerais in Brazil, of which 12 were from clinically healthy animals, 42 from animals diagnosed with subclinical mastitis, 16 from animals with clinical mastitis, and 2 from animals with gangrenous mastitis. The group related to gangrenous mastitis was the most divergent in terms of alpha and beta diversity. The most abundant genus among samples of the groups was Staphylococcus spp., and we found a high abundance of Mycoplasma sp. in the milk of animals diagnosed with clinical mastitis. The most statistically relevant microorganisms among the groups were Prevotella sp., Ruminococcaceae , Prevotella ruminicola sp., and Providencia sp. We highlight a new association of bacterial agents in gangrenous mastitis among Escherichia sp./ Shigella sp. and Enterococcus sp. and provide the second report of the genus Alkalibacterium sp., in milk samples. Only the taxa Staphylococcus sp., Bacteroides sp., Enterococcus , and Brevidabacterium sp., were present in all groups. The superpathway of L-tryptophan biosynthesis metabolites and the sucrose degradation III (sucrose invertase) pathway were the most prominent ones among the groups. In this study, we demonstrate how a rich microbiota of goat milk from healthy animals can be altered during the aggravation of different types of mastitis, in addition to demonstrating new bacterial genera in milk not previously detected in other studies as well as new associations between agents., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Polveiro, Vidigal, de Oliveira Mendes, Yamatogi, da Silva, Fujikura, Da Costa and Moreira.)

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.

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., (© 2021. Sociedade Brasileira de Microbiologia.)

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., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Indian Society for Parasitology 2020.)

The objective of this study was to evaluate the effects of autolyzed yeast on performance, cecal microbiota, and leukogram of weaned piglets. A total of 96 piglets of commercial line weaned at 21-day-old were used. The experimental design was a randomized block design with four treatments (diets containing 0.0%, 0.3%, 0.6%, and 0.9% autolyzed yeast), eight replicates, and three animals per pen in order to evaluate daily weight gain, daily feed intake, and feed conversion in periods of 0 to 15, 0 to 26, and 0 to 36 days. Quadratic effects of autolyzed yeast inclusion were observed on the feed conversion from 0 to 15 days, on daily weight gain from 0 to 15 days, 0 to 26 days and, 0 to 36 days, indicating an autolyzed yeast optimal inclusion level between 0.4% and 0.5%. No effect from autolyzed yeast addition was observed on piglet daily feed intake, cecal microbiota, and leukogram; however, i.m. application of E. coli lipopolysaccharide reduced the values ​​of total leukocytes and their fractions (neutrophils, eosinophils, lymphocytes, monocytes, and rods). Therefore, autolyzed yeast when provided at levels between 0.4% and 0.5% improved weaned piglets' performance.

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., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Indian Society for Parasitology 2020.)

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.

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 bla CTX-M-2 e bla TEM-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.

Abstract: Listeria monocytogenes contamination was assessed in different steps of a pork production chain. Ten lots of pigs were sampled at termination barns, at slaughter (after bleeding, after buckling, after evisceration, and after final washing), at processing (knives, deboning tables, and employees' hands), and of end products (ribs, shoulder, ham, and sausage). All samples (n = 670) were subjected to L. monocytogenes detection, and the obtained isolates (n = 18, identified as Listeria spp.) were characterized by their biochemical characteristics, serogroups, virulence genes, pulsed-field gel electrophoresis profiles, antibiotic resistances (ampicillin, penicillin, gentamicin, and sulfamethoxazole-trimethoprim), and adhesion abilities. The results revealed the low occurrence of Listeria spp. in the evaluated pork production chain. However, four tested sausage samples (40%) were positive for Listeria spp., with L. monocytogenes identified in two (20%) of these samples. Ten isolates were identified as L. monocytogenes (eight from serogroup 1/2a or 3a and two from serogroup 4b, 4d, or 4e): all isolates were also positive for the virulence-related genes hlyA, iap, plcA, actA, inlA, inlB, inlC, and inlJ and susceptible to the tested antibiotics. One sausage sample was contaminated by both serogroups 1/2a or 3a and 4b, 4d, or 4e. Isolates from serogroup 1/2a or 3a obtained during visits 5 and 6 presented distinct genetic profiles by pulsed-field gel electrophoresis, indicating that contamination may come from different sources. The adhesion potential exhibited by Listeria spp. isolates (n = 18) ranged from weak (serogroup 4b, 4d, or 4e) to moderate (L. innocua and L. monocytogenes serogroup 1/2a or 3a). Despite the low occurrence of L. monocytogenes, pathogenic serogroups were detected in sausages, demanding control measures by the industry., (Copyright ©, International Association for Food Protection.)

Considering the growing consumption of artisanal foods worldwide, we aimed to evaluate the microbial safety of Serro artisanal cheese (SAC), produced in Minas Gerais State, Brazil. This cheese is produced with raw milk using 1 of 2 natural starter cultures: "pingo" and "rala." A total of 53 SAC samples (pingo = 8; rala = 45) were obtained from different farmers and subjected to conventional and molecular assays to detect and enumerate Listeria monocytogenes, Salmonella spp., coagulase-positive staphylococci (CPS), diarrheagenic Escherichia coli, Mycobacterium tuberculosis, and Brucella abortus. The SAC samples were also subjected to an ELISA to detect classical staphylococcal enterotoxins (CSE: SEA, SEB, SEC, SED, SEE) and to PCR assays to detect staphylococcal enterotoxin-related genes (sea, seb, sec, sed, see). Coagulase-positive staphylococci isolates were obtained and tested by the same assays to detect their potential in CSE production and presence of CSE-related genes. None of the SAC samples showed any of the screened food-borne pathogens and zoonotic agents, and none showed the presence of CSE by phenotypic and genotypic approaches. Despite the absence of microbial hazards, mean counts of CPS in SAC samples were 5.2 log cfu/g (pingo starter) and 4.6 log cfu/g (rala starter), indicating poor hygiene practices during production. None of the tested CPS isolates (n = 116) produced CSE or presented CSE-related genes. Despite the relative microbial safety, hygienic conditions during SAC production must be improved to meet official guidelines established in Brazil., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Atypical enteropathogenic (serotypes O4:H16, O8:H25, O68:H2, O105:H7, and OR:H25) and Shigatoxigenic (ONT:H46) Escherichia coli were isolated from samples of ground beef and poultry breast purchased in Botucatu, Brazil. Phenotypic and molecular characterization indicated the potential of these isolates to adhere to host epithelial cells and cause damage.

Introduction:: Brazilian native species are reemerging as increasingly free-ranging populations., Methods:: Sera from 31 capybaras (Hydrochoerus hydrochaeris) and 28 peccaries (Pecari tajacu and Tayassu pecari) were tested for anti-Leptospira and anti-Toxoplasma gondii antibodies using microscopic seroagglutination test., Results:: Nineteen percent of free-ranging and 10.0% of captive capybaras, along with 31.8% of collared peccaries, were seropositive for T. gondii. None was seropositive for Leptospira sp., Conclusions:: The present findings indicated low risk of disease, particularly among capybaras and white-lipped peccaries; however, active surveillance programs are important for monitoring wildlife health and public health once they are in public parks around cities.

Background: Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake ( Python molurus ) and one native green snake ( Philodryas olfersii ) from the Sorocaba Zoo, São Paulo state, Brazil., Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry., Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense ., Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.

Poultry products are important in the transmission of zoonotic pathogens, mainly Salmonella. This genus causes millions of foodborne diseases worldwide every year. Cross-contamination by food sources in human cases of salmonellosis and the increase in resistant strains have become important issues. A qualitative and quantitative Salmonella detection method was utilized in a poultry slaughterhouse in São Paulo State, Brazil. We collected 33 samples from different batches of carcasses. Each sample was analyzed at three process points: postbleeding, postdefeathering, and postchilling. A fourth point, retail simulation, was added to simulate retail market storage at 5°C for 72 h. The qualitative methods revealed 100% (33 samples) contamination at postbleeding, 39% (13 samples) contamination at postdefeathering, 58% (19 samples) contamination at postchilling, and 30% (10 samples) contamination at the retail simulation. The quantitative results, determined by the most-probable-number (MPN) technique, ranged from <0.03 to >2,400 MPN/g. We identified 23 Salmonella serovars; the most prevalent were Mbandaka, Senftenberg, and Enteritidis. Resistance to nalidixic acid was significantly more common (P < 0.05) than resistance to other antimicrobial agents. Five multidrug-resistant strains were identified. This study contributes important epidemiological data and demonstrates the need to improve sanitary conditions in slaughterhouses.

Introduction: Salmonella is a major cause of foodborne disease, and poultry products are important contributors to the transmission of this zoonotic pathogen. Although considered to be rare in most countries, Salmonella Corvallis has been reported in specific geographic areas isolated from both human and non-human sources. The aim of this study was to report the occurrence, the antimicrobial resistance profiles including the extended-spectrum beta-lactamase (ESBL) production, and the clonal relatedness of S. Corvallis strains., Methodology: A total of 132 fragments of poultry carcasses from a slaughterhouse in São Paulo State, Brazil, were collected at different stages of the manufacturing process (post-bleeding, post-plucking, and post-chilling) and analyzed for the presence of Salmonella. Antimicrobial resistance was determined by disc diffusion method and Etest. Clonal relatedness was determined by pulsed-field gel electrophoresis (PFGE)., Results: Among the 272 Salmonella strains recovered, fourteen were S. Corvallis. Ten (71.4%) showed ESBL production and resistance to at least three antimicrobial agents. Nalidixic acid resistance and reduced ciprofloxacin susceptibility was verified in four (28.6%) strains. PFGE analyses showed that all the S. Corvallis strains belonged to the same pulsotype., Conclusion: This study identified genetically related S. Corvallis strains exhibiting ESBL production and reduced susceptibility to quinolone. The results suggest the need to improve the sanitary conditions in the slaughterhouse. Moreover, from a public health perspective, continuous surveillance on Salmonella is needed to control the dissemination of this important zoonotic pathogen and its resistance determinants.