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Objective: Heat stroke is a life-threatening illness characterized by an increased core body temperature as a result of exposure to high ambient temperature. Despite advances in supportive care, heat stroke is often fatal, and no specific and effective therapies exist. The pathophysiological responses to heat stroke involve a systemic inflammatory response and a disseminated intravascular coagulation in the host, which lead to a multiorgan dysfunction syndrome. Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear DNA-binding protein that has been shown to play a relevant role in cell necrosis and organ failure in various diseases associated with inflammation. Therefore, we set out to investigate whether inhibition of PARP activity might affect the heat stroke-induced injury. Design: Controlled animal study. Setting: Research laboratory of an academic institution. Subjects: PARP-1-deficient mice (Parp-1(-/-)) and wild-type mice (C57BL/6J). Interventions: Wild-type mice untreated or treated with either PJ34 or 3-AB, two generic PARP inhibitors, and Parp-1(-/-) mice were subjected to heat exposure as a model to study heat stroke. Measurements and Main Results: We measured rectal temperature, serum interleukin-1beta and interleukin-6, liver histology, and heat shock proteins expression. We found that the heat stroke-induced injury was attenuated in mice lacking PARP-1 and was markedly reduced in wild-type mice treated with PARP inhibitors. Interestingly, heat-induced expression of heat shock proteins 27 and 70 was boosted after PARP inhibition. Indeed, PARP inhibition increased expression of heat shock proteins 27 and 70 even in the absence of heat exposure. Accordingly, PARP inhibition increased thermal tolerance that may contribute to attenuate the clinical effects of heat stroke, resulting in increased survival. Conclusions: Our results find a new protective function of PARP inhibitors and support their potential therapeutic application in the treatment of heat stroke. [ABSTRACT FROM AUTHOR]

Contents The aim of this study was to estimate the prevalence of genomic DNA of Toxoplasma gondii in semen samples from commercial rams in artificial insemination centres in Brazil, as well as in fresh semen from rams in the northeast of Brazil. In total, 108 semen samples were obtained from artificial insemination centres, and genomic DNA of T. gondii was detected in 24 of 108 (22.2%). The prevalence of antibodies anti- Toxoplasma gondii among sheep on rural properties was 9.2% (10/109), and 100% of the semen samples of these animals were positive in the PCR for T. gondii DNA. The molecular identity was confirmed through sequencing, which indicated 99.9% similarity with the T. gondii DNA sequences stored in the Gen Bank. This study reports the first occurrence of T. gondii DNA in the semen of rams, which came from artificial insemination centres in Brazil, as well as the occurrence of T. gondii DNA in the fresh semen of naturally infected rams in the northeast of Brazil. [ABSTRACT FROM AUTHOR]

This study aimed to assess the impact of S. aureus as well as Non-aureus Staphylococci and Mammaliicocci (NASM) subclinical mastitis-causing in the economic return in dairy herds. Data were gathered from the databases of five dairy herds located in the Agreste region of Pernambuco state, Northeastern Brazil, over a period of three consecutive months. A total of 155 mammary quarters from 155 healthy cows were categorized into the healthy group. Meanwhile, 257 mammary quarters from 187 cows that tested positive for either S. aureus or NASM were categorized into the infected group. The effect of mammary quarter infection in economic return was estimated using milk payment criteria on milk samples from healthy vs. infected cows based on a linear mixed model. Milk yield and milk price influenced the economic return, and both varied according to factors like herd, parity, stage of lactation, period of analysis, and the type of pathogen causing subclinical mastitis. There was a reduction in the average economic return caused by NASM infection (by 0.41 to 0.65 US$/cow/day) and S. aureus infection (by 0.25 to 0.36 US$/cow/day), when considering the effect of the pathogen over time for 1 and ≥ 2 infected quarters. Although no significant differences were observed in economic return between healthy cows and those infected with NASM or S. aureus, it is important to collect data on these pathogens for a more precise assessment of the economic impact of subclinical mastitis and for developing enhanced approaches for prevention and control., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The present study aimed to evaluate the antimicrobial and modulating activity of the ethanol extract obtained from the leaves, stems, and roots of Cnidoscolus urens in multiresistant bacteria. The Minimum Inhibitory Concentration (MIC) values obtained for the extracts of leaves, stems, and roots were greater than 1024 µg/mL for all isolates. In the antimicrobial resistance modulation test, the extract of the leaves of C. urens showed a better modulating effect than that of the stems and roots for gentamicin, highlighting the reduction of MIC for Escherichia coli, Lactococcus garvieae and Staphylococcus sciuri. For erythromycin, a reduction of MIC was observed in L. garvieae, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus agalactiae. The extract from the leaves of C. urens has an important modulating effect on resistance in multiresistant bacteria, especially with gentamicin and erythromycin., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

This study aimed to evaluate the presence and viability of Toxoplasma gondii in chickens intended for human consumption in the Pernambuco State, Brazil. Blood and tissue samples were collected from 25 chickens sold in markets in Recife, Pernambuco. Samples were evaluated by indirect immunofluorescence assay (IFA) to detect antibodies to T. gondii . Pools of brain and heart of seropositive chickens were subjected to bioassay in two Swiss Webster mice, which were evaluated for 45 days then tested by IFA to detect seroconversion. The mice were euthanized, and their brains were evaluated for cysts. Peritoneal lavage was also conducted in mice that exhibited clinical signs. Brains containing cysts or peritoneal lavage with tachyzoites were inoculated into MA-104 cells. Brains of mice inoculated with the same tissue were pooled and analysed by ITS1-PCR. We obtained a frequency of antibodies to T. gondii of 68.00% (17/25) in chickens, and a seroconversion rate of 70.58% (24/34) in mice. Detection of Toxoplasma ITS1 DNA confirmed an isolation rate of 41.1% (7/17). Three isolates were characterized by mnPCR-RFLP as genotypes ToxoDB#36 and ToxoDB#114. We highlight the occurrence of ToxoDB#36 in chickens in Pernambuco State and the parasites' viability in chickens intended for human consumption., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Given their role as Co-Editor, Frank Katzer had no involvement in the peer-review of this article and has no access to information regarding its peer-review. Full responsibility for the editorial process for this article was delegated to Professor Aneta Kostadinova (Editor-in-Chief)., (© 2024 The Authors.)

This study evaluated the reproductive, productive and financial consequences of chronic Trypanosoma vivax infection in a dairy cattle herd located in a region without the cyclic vector during two years. Animals were categorized as either positive (chronically infected) or negative for T. vivax antibodies using a commercial rapid test. Additionally, serum samples from cows were analyzed for the presence of anti-Neospora caninum antibodies. Pregnancy diagnoses were performed through rectal palpation and ultrasonography after 30, 60 and every 21 days until the 144th day of pregnancy. If an abortion occurred in the final trimester, serology and cPCR were performed on calves for T. vivax and N. caninum. The breeding period, calving interval and pregnancy losses were recorded. The milk production of each animal during the 305 days of lactation was measured, and the annual financial impact of milk production was calculated using a revenue minus feed cost (RMFC) indicator. Out of 177 cows, 71.75 % were chronically infected, and 13.50 % were T. vivax-negative. No correlation (p = 0.8854) of co-infection between T. vivax and N. caninum was observed. Negative cows required fewer (p≤0.05) artificial inseminations than chronically infected ones. T. vivax was not significantly associated (p = 0.7893) with pregnancy loss up to 81 days of pregnancy. Cows chronically infected by T. vivax had 4-fold greater chance (p = 0.0280) of experiencing pregnancy loss between 82 and 144 days of gestation. Eighteen cows aborted, two were positive for T. vivax antibodies, and one for N. caninum antibodies. The calves were negative for T. vivax and N. caninum. Chronically infected cows and negative cows for T. vivax that experienced pregnancy loss (82-144 days of pregnancy) had a longer (p≤0.05) breeding period to become pregnant, and consequently a longer calving interval compared to cows that maintained pregnancy. The difference (p≤0.05) in milk production was evident when pregnancy loss occurred between 82 and 144 days of gestation in cows chronically infected by T. vivax. The RMFC indicated a negative impact of 38.2 % on the farm's annual milk revenue due to the presence of chronically infected cows., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cat-transmitted sporotrichosis is caused by the emerging fungal pathogen Sporothrix brasiliensis and constitutes a significant public health issue that affects people living in resource-poor urban centers in Brazil. The lack of knowledge about transmission dynamics makes it difficult to propose public health policies to contain the advance of sporotrichosis. We describe the recent emergence of 1,176 cases of sporotrichosis in cats (2016 to 2021) in the metropolitan region of Recife, Brazil, leading to significant zoonotic transmission and an overwhelming occurrence of S. brasiliensis as the etiological agent. Most cases were from cats in the cities of Olinda (408/1,176; 34.70%), Jaboatão dos Guararapes (332/1,176; 28.23%), and Recife (237/1,176; 20.15%). Molecular typing using amplified fragment length polymorphism (EcoRI-GA/MseI-AG) revealed low polymorphic information content (PIC = 0.2499) and heterozygosity (H = 0.2928), typical of an outbreak scenario. Dendrogram and multivariate cluster analysis revealed that isolates from Pernambuco are closely related to Rio de Janeiro isolates. We report a substantial occurrence of MAT1-2 idiomorphs in the metropolitan region of Recife (0:60 ratio; χ 2  = 60.000, P < 0.0001). The limited population differentiation and genetic diversity of the isolates from Pernambuco suggest a recent introduction, possibly via a founder effect, from the parental population in Rio de Janeiro. Our findings emphasize the critical importance of molecular surveillance of S. brasiliensis for outbreak response. A comprehensive one-health strategy is mandatory to control the spread of cat-transmitted sporotrichosis driven by S. brasiliensis, encompassing sanitary barriers, quick diagnosis, and treatment., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to diagnose Mycobacterium avium subsp. paratuberculosis (MAP) infections in sheep in the state of Pernambuco, Brazil. A total of 276 blood samples were analyzed using the enzyme-linked immunosorbent assay IDEXX Paratuberculosis Screening kit, and 261 fecal samples were submitted for bacterial culture and polymerase chain reaction tests. An animal-level sero-frequency of 0.72% (n = 2/276) and a farm-level sero-frequency of 20% (n = 2/10) were found. All fecal sample cultures were negative, and molecular analyses were also negative. To the best of our knowledge, this is the first study of MAP infection in sheep in the state of Pernambuco and one of the pioneers in the country. It is an asymptomatic disease that is difficult to diagnose in this species because the susceptibility of sheep to the organism is lower than that of other ruminant species. However, the sero-frequency found reveals that there is MAP exposure in sheep flocks in the region. In addition, serological monitoring can contribute to the observation of the organism's behavior in herds. Our results support the potential risk of MAP infection in sheep in the state of Pernambuco, Brazil., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to research Toxoplasma gondii DNA in 102 samples of raw bovine milk from expansion tanks, in small properties located in different cities of the Midwest region of São Paulo, Brazil. For this, polymerase chain reaction (PCR) was performed with the primers TOX4/TOX5 for cPCR (conventional PCR), TgNP1/TgNP2 gene for nested PCR and the Tg18s58F/Tg18s348R for nested PCR. It was possible to detect T. gondii DNA in 18 (17.65 %) milk samples from the 102 tanks, corresponding to 4.90 % for TOX4/TOX5 primers, 12.74 % for TgNP1/TgNP2 gene and 0.98 % for Tg18s58F/Tg18s348R gene. The results showed that the TgNP1 and TgNP2 genes were more efficient to detect T. gondii DNA, and also indicated the importance of raw bovine milk as a source of human infections caused by this protozoan, being a public health problem. It is important to continue studies involving T. gondii from bovine milk considering the need for proper pasteurization, and for better comprehension regarding the epidemiology of this protozoan., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The objective of this study was to characterise a Toxoplasma gondii -induced abortion outbreak on a goat farm in the State of Paraíba, Northeast Region of Brazil. From a herd of 10 does, seven experienced abortions and one gave birth to twins (one stillborn and the other weak and underdeveloped). Serum samples from all of the does were analysed by indirect fluorescent antibody test (IFAT). Samples of colostrum and placenta from two does, along with lung, heart, brain and umbilical cord samples from four of the foetuses, were screened by nested ITS1 PCR specific for T. gondii . The positive samples were then analysed by multiplex nested PCR-RFLP. All ten does tested positive by IFAT for anti- T. gondii IgG (titrations ranging from 1:4096 to 1:65,536). The ITS1 PCR screening revealed T. gondii DNA in the placenta (2/2), colostrum (2/2), umbilical cord (2/4), lung (1/4), heart (1/4), and brain (1/4). Four samples produced complete RFLP genotyping results, identifying a single genotype, ToxoDB #13. In conclusion, we demonstrated a high rate of abortion caused by T. gondii in a goat herd, highlighting the pathogenicity of genotype #13, one of the most prevalent genotypes of T. gondii in Brazil., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Given their role as Co-Editor, Frank Katzer had no involvement in the peer review of this article and has no access to information regarding its peer review. Full responsibility for the editorial process for this article was delegated to Editor-in-Chief Aneta Kostadinova., (© 2023 The Author(s).)

This study aimed to genotype isolates of Toxoplasma gondii obtained from samples of brain, diaphragm and heart of goats and sheep intended for human consumption in the State of Paraíba, Brazil. Tissue samples from 14 animals, goats ( n  = 5) and lambs ( n  = 9), were sourced from public slaughterhouses in seven cities and bio-assayed in mice. The brains of the mice were utilized for DNA extraction. Genotyping was carried out by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) using 10 markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico). A total of 10 isolates were fully genotyped (i.e. at all loci), three from goats and seven from sheep, revealing five distinct genotypes: #13 ( n  = 4); #48 ( n  = 3); #57 ( n  = 1); #273 ( n  = 1); and one new genotype that had not been previously described. Genotype #13 is frequently found in the Northeast of Brazil and represents a clonal lineage circulating in this region and was the most prevalent genotype identified ( n  = 4). Moreover, in the present study genotypes #13, #48, #57, and #273 were documented for the first time in sheep from Brazil, and the novel genotype was isolated from a goat. Our findings align with previous studies on T. gondii from Brazil, where new genotypes are continuously being identified, highlighting a high level of genetic diversity of T. gondii isolates in the country., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Given their role as Co-Editor, Frank Katzer had no involvement in the peer review of this article and has no access to information regarding its peer review. Full responsibility for the editorial process for this article was delegated to Professor Aneta Kostadinova (Editor-in-Chief)., (© 2023 The Authors.)

The objective was to describe the seroprevalence of anti-Neospora caninum antibodies in goats and sheep slaughtered in the state of Paraíba and to identify possible associated factors with the infections. Two hundred twenty-nine samples from goats and two hundred five from sheep were analyzed by Indirect Immunofluorescence Reaction (IFAT) using a cutoff point of 1:50. The presence of anti-N. caninum antibodies was identified in 28.4% (65/ 229; 95% Confidence Interval: 22.6-34.2) of the goat samples and in 12.7% (26/ 205; 95% CI: 8.2 - 17.2) of the sheep samples. Contact between goats and dogs (Odds ratio 4.81; CI 1.13 - 2.67; p = 0.041) and cattle (OR. 1.87; CI 1.13 - 2.67; p = 0.002) was identified as a risk factor for goats and contact between sheep and dogs (OR 2.32; CI 1.58 - 3.14; p = 0.026) and history of abortion (OR 1.94; CI 1.28 - 2.90; p = 0.001) was considered a risk factor for sheep. The study revealed a high seroprevalence of anti-N. caninum antibodies in slaughtered goats and sheep in Paraíba. Risk factors such as contact with dogs/cattle and abortion history underscore the need for preventive measures to control infection and enhance animal health management.

The aim of the study was to track the spread of antimicrobial resistance among the different sectors of One Health through the detection of Multidrug-Efflux-System in multidrug-resistant Staphylococcus aureus isolates. Multidrug-resistant (MDR) and methicillin-resistant (MRSA) S. aureus isolates were selected: 25 of human, one of animal and eight of food origin. The efflux system genes norA, norB, norC, LmrS, tet38 and msrA were screened by PCR. The activity of the efflux systems was determined by the minimum inhibitory concentration (MIC) of tetracycline and ciprofloxacin in the presence and absence of CCCP and in the quantification of ethidium bromide efflux. Furthermore, biofilm formation was determined in the presence and absence of the CCCP. The molecular epidemiology of the isolates was traced with the aid of PFGE. The gene norC was the most prevalent, detected in all isolates and msrA was the least prevalent, detected in only two isolates from humans. There was no difference in the MICs of tetracycline and ciprofloxacin in the presence of CCCP, but 55.9% of isolates showed ethidium bromide efflux. The presence of CCCP decreased the biofilm formation. Regarding the molecular epidemiology, in three clusters was a mixture of the isolates from different origins. Therefore, S. aureus MDR with active multidrug efflux systems are circulating between One Health domains and it is necessary to consider strategies to decrease this circulation in order to prevent the dissemination of resistance mediated by MES., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Aims: This study aimed to evaluate matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial subtyping for the rapid detection of biomarkers in Staphylococcus aureus from subclinical bovine mastitis., Methods and Results: A total of 229 S. aureus isolates were obtained from milk samples collected from cows with subclinical mastitis using microbiological culture. Staphylococcus aureus isolates were also submitted to PCR analysis targeting the mecA and mecC genes, which are indicative of methicillin resistance. Confirmation of the species was achieved through MALDI-TOF MS analysis. To analyze antimicrobial resistance patterns, the MALDI BioTyper Compass Explorer and ClinProTools Bruker software were employed, and dendrograms were generated using Bionumerics software., Conclusions: MALDI-TOF MS successfully identified S. aureus at the species level, but no methicillin resistance was observed. Moreover, spectral typing displayed limited similarity when compared to pulsed-field gel electrophoresis (PFGE)., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

An adult male free-living yellow armadillo (Euphractus sexcinctus) was found by hunters and referred for clinical evaluation because of a tumour-like lesion on the carapace. The animal was lethargic and weak with severe dehydration, enophthalmos and cachexia, and was euthanized because of its very poor clinical condition. Necropsy revealed a whitish, exophytic, irregular and moist mass (7 × 6.5 × 1.5 cm) in the caudal third of the carapace. On cut section, the mass was hard, compact, irregular and whitish. Histopathology revealed a densely cellular, expansive, poorly delimited neoplasm composed of malignant mesenchymal cells arranged in islands and cords, interspersed by numerous areas of irregularly mineralized osteoid matrix. Neoplastic cells were intensely immunolabelled for vimentin. The diagnosis of a high-grade osteosarcoma of osteoblastic subtype was based on the clinical, gross, histopathological and immunohistochemical findings. This first report of an osteosarcoma in an armadillo expands the list of armadillo diseases and will assist the management of these animals by veterinarians., Competing Interests: Declaration of competing interest The authors declared no conflicts of interest in relation to the research, authorship or publication of this manuscript., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Antimicrobial resistance is a threat to public health. The emergence of antibiotic-resistant Staphylococcus aureus represents a priority for the implementation of preventive measures. The objective was to isolate S. aureus in humans, animals, and animal health care environment, and to characterize the genotypic and phenotypic profile of antimicrobial resistance in these isolates. We isolated S. aureus from staff, animals, and environment of a veterinary hospital, and identified their antimicrobial resistance profiles. Samples were collected from 20 humans, 13 animals, 14 surfaces, 8 mobile phones, and 7 veterinarians' stethoscopes by using sterile swabs. S. aureus was isolated by culturing on mannitol salt agar and preliminary identification was done by Gram staining and catalase test. Subsequently, a polymerase chain reaction was performed for species confirmation and investigating their antimicrobial-resistant genotypic profiles. Phenotypic profiles of resistant isolates were determined using the disk-diffusion technique. Ten S. aureus isolates were recovered from 5/20 humans (25%), it was also recovered from 2/13 animals (15.38%), including 1 dog and 1 cat, and from 1/14 of surfaces (7.14%). The oxacillin-susceptible mecA-positive Staphylococcus aureus phenotype was identified in a feline. Most of the isolates carried at least two resistance genes of different antimicrobial classes, with 90% (9/10) presenting the gene blaZ, with 10% (1/10) presenting the gene mecA, 20% (2/10) presenting tet38, 10% (1/10) presenting tetM, 90% (9/10) presenting norA, 50% (5/10) presenting norC, 10% (1/10) presenting ermA, and 60% (6/10) presenting ermB. In antibiograms, resistance to penicillin was identified in all the isolates, resistance to erythromycin was identified in 80% (8/10), and all the isolate's resistance to erythromycin presented erythromycin-induced resistance to clindamycin. Antimicrobial resistance in the veterinary hospital requires attention due to the risk of interspecies transmission, gene transfer between bacteria that colonize companion animals and humans and, can make antimicrobial therapy difficult., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The core objective of this study was to genetically and phenotypically characterize subclinical mastitis-causing multidrug-resistant Staphylococcus aureus (MDRSA). In addition, risk factors associated with subclinical mastitis caused by MDRSA were investigated. Bacterial cultures were performed on 2120 mammary quarters, 40 swabs of milk utensils, 5 bulk tank milk samples, and 11 nostril and 11 hand swabs from milkers from five dairy farms. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was conducted for S. aureus identification. Antimicrobial resistance was screened phenotypically using the disk diffusion test in all S. aureus isolates. A biofilm formation assay; detection of genes associated with beta-lactam resistance, efflux pump, and biofilm formation; and pulsed-field gel electrophoresis (PFGE) were performed in all MDRSA isolates. Multi-locus sequence typing (MLST) was carried out in cefoxitin-resistant MDRSA isolates. A total of 188 S. aureus isolates from milk as well as two from milking utensils and one from bulk tank milk were identified. Most of the isolates (92.7%; 177 of 191) showed beta-lactam resistance, and 7% (14 of 191) were MDRSA. Interestingly, 36% (5 of 14) of MDRSA isolates were cefoxitin-resistant, but none carried mecA or mecC genes. Based on PFGE results, it was observed that S. aureus strains were more likely to be unique to a specific herd. Two clonal complexes were identified, CC97 (ST126; commonly livestock-associated) and CC1 (ST7440; usually community-associated). To the best of our knowledge, this is the first report of ST7440 isolated from bovine mastitis in Brazil. The risk factor results underscored the importance of considering parity, stage of lactation, SCC, milk production, and herd size when studying the risk of subclinical mastitis and antimicrobial resistance in S. aureus . Thus, to implement effective strategies to prevent subclinical mastitis in dairy herds and to minimize MDRSA spread, it is important to understand MDRSA strains' distribution and their antimicrobial resistance profile.

The discovery and tracking of antimicrobial resistance genes are essential for understanding the evolution of bacterial resistance and restraining its dispersion. Mammaliicoccus sciuri (formerly Staphylococcus sciuri) is the most probable evolutionary repository of the mecA gene, that later disseminated to S. aureus. In this study, we describe the first double mecA/mecC homologue-positive non-aureus staphylococci and mammaliicocci (NASM) from the American continent, also representing the first report of mecC-positive NASM in Brazil. Two clonally related methicillin-resistant M. sciuri strains co-carrying mecA and mecC genes were isolated from the teat skin swab and milk sample collected from an ewe's left udder half. Both M. sciuri strains belonged to the sequence type (ST) 71. Besides mecA and mecC genes, the M. sciuri strains carried broad resistomes for clinically important antimicrobial agents, including β-lactams, tetracyclines, lincosamide, streptogramin, streptomycin, and aminoglycosides. Virulome analysis showed the presence of the clumping factor B (clfB), ATP-dependent protease ClpP (ClpP) and serine-aspartate repeat proteins (sdrC and sdrE) virulence-associated genes. Phylogenomic analysis revealed that these M. sciuri strains are part of a globally disseminated branch, associated with farm and companion animals and even with food. Our findings suggest that M. sciuri is likely to emerge as a pathogen of global interest, carrying a broad repertoire of antimicrobial resistance genes with a remarkable co-presence of mecA and mecC genes. Finally, we strongly encourage to monitor M. sciuri under the One Health umbrella since this bacterial species is spreading at the human-animal-environment interface., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Extended-spectrum β-lactamase (ESBL)-producing Gram negative bacteria are becoming increasingly important in veterinary and human medicine because they can hydrolyze the third generation β-lactams, penicillins, and monobactams. The aim of this study was to identify ESBL-producing Enterobacteriaceae in raw cow milk samples from northeast Brazil. Twenty-six bacterial isolates belonging to the Enterobacteriaceae family were obtained from milk samples from 257 cows with subclinical mastitis. Using microbiological tests, 53.85% (14/26) were identified as Escherichia coli, 15.38% (4/26) as Proteus mirabilis, 26.92% (7/26) as Klebsiella spp., and 3.85% (1/26) as Citrobacter spp. Of all the isolates, 61.54% (16/26) were positive in the ESBL screening test, of which 12.5% (2/16) were positive in the double-disc synergy test using three types of cephalosporins and amoxicillin/clavulanic acid. The two isolates were identified as Klebsiella spp. Among all the isolates, 53.85% (14/26) were positive for one or both ESBL-encoding genes, blaSHV and blaTEM; among these, 71.43% (10/14) were identified as E. coli. This study demonstrates that ESBL-producing bacteria can be found in raw cow milk from northeast Brazil. Cows with subclinical mastitis should be recognized as reservoirs of these strains, which can propagate to humans., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Glanders is a contagious disease of equids caused by the Gram-negative bacterium Burkholderia mallei. In Brazil, the disease is considered to be reemerging and has been expanding, with records of equids with positive serology in most of the federative units. However, there are few reports describing the genotypic detection of the agent. This study demonstrated the detection of B. mallei by species-specific PCR directly from tissues or from bacterial cultures, followed by amplicon sequencing in equids (equines, mules, and asinines) with positive serology for glanders in all five geographic regions of Brazil. The molecular evidence of B. mallei infection in serologically positive equids in this study expands the possibility of strain isolation and the conduction of epidemiological characterizations based on molecular information. The microbiological detection of B. mallei in cultures from nasal and palate swabs, even in equids without clinical manifestations, raises the possibility of environmental elimination of the agent., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Although Toxoplasma gondii exposure has been reported in indigenous populations worldwide, a One Health approach has not been applied to date. This study concurrently assessed T. gondii exposure in indigenous populations, and their dogs, environment, and indigenous or non-indigenous healthcare professionals (HPs). Human and dog serum samples from 9 indigenous communities in Brazil were assessed by indirect immunofluorescence antibody test for anti- T. gondii antibodies. Soil samples (30 per community) were processed with PCR to amplify T. gondii DNA. Associated risk factors and seroprevalence were analyzed using logistic regression models. Human seropositivity and type of water source were assessed by generalized linear mixed model (GLMM) with binomial error distribution, and game meat consumption with chi-squared test. Overall, 225/463 (49%) indigenous persons were seropositive for anti- T. gondii antibodies. Of all the HPs, 67/168 (40%) were positive, and included 54/147 (37%) positive non-indigenous HPs. Indigenous persons more likely to be seropositive compared with non-indigenous HPs (OR: 1.63; 95% CI: 1.11-2.39). A total of 97/253 (38%) dogs were seropositive and highly associated with seropositive owners ( p  < 0.001). Based on univariate analysis for indigenous individuals, state location of community (p < 0.001), ethnicity (p < 0.001), consumption of game meat (p < 0.001), type of water source ( p  < 0.001), and educational level ( p  = 0.026) were associated with seropositivity. Logistic regression showed that indigenous seropositivity was associated with eating game meat ( p  = 0.002), drinking water from rivers ( p  < 0.001), and inversely proportional to the educational level. According to univariate analysis for non-indigenous HP, age ( p  = 0.005), frequency of visits to the indigenous populations ( p  < 0.001), consumption of water at the indigenous communities (p < 0.001), and ingestion of raw meat ( p  = 0.023) were associated with T. gondii seropositivity. Logistic regression revealed living outdoors ( p  = 0.042), habit of hunting ( p  = 0.008), and drinking river water ( p  = 0.007) as risk factors associated to seropositivity in dogs. In addition, indigenous communities lacking water treatment had higher seroprevalence for all groups including indigenous persons (GLMM; z = -7.153; p  < 0.001), their dogs (GLMM; z = -2.405; p  = 0.0162), and all HPs (GLMM; z = -2.420; p  = 0.0155). Human seropositivity was associated with that of their dogs (p < 0.001). A single soil sample, out of 270 (0.37%), was positive for T. gondii by PCR. Our results indicate water source is a risk for human and dog toxoplasmosis in indigenous communities; both share similar exposure. Moreover, quality water access was shown to be crucial to prevent toxoplasmosis in both total and non-indigenous HPs who work in these indigenous communities., Competing Interests: None declared., (© 2023 The Authors.)

Bovine neosporosis is a globally important disease, causing abortions and significant economic losses. In Brazil, studies on neosporosis in cattle are few and based on limited samples and/or from limited areas. We aimed to determine the herd and animal levels seroprevalence and associated factors for Neospora caninum infections in cattle from the state of Paraíba, Northeastern Brazil, using a planned sampling. Herds (n = 434) and cows aged ≥24 months (n = 1891) were randomly selected, and serum samples were tested with the immunofluorescence antibody test (IFAT) using as cut-off point the antibody titer 200. Herd-level and animal-level seroprevalences were 17.8% (95% CI = 14.3%-21.8%) and 18.1% (95% CI = 14.7%-22.1%), respectively. The factors associated with N. caninum infection were farm located in the Sertão mesoregion (Prevalence ratio [PR] = 2.37), mixed production (PR = 1.64), herd size of 34-111 animals (PR = 3.50) and herd size >111 animals (PR = 6.14). The results indicate high N. caninum circulation in the bovine population of the state of Paraíba, semiarid of Brazil, mainly in the Sertão mesoregion, where the highest apparent herd and animal-level prevalences of positive herds were identified. Control strategies should be adopted to mitigate the impact of disease on cattle production, as well as it's suggested the encouragement of conducting surveys in wildlife from Caatinga biome, mainly canids, to provide information on the importance of these animals on the epidemiology of bovine neosporosis., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Chronic kidney disease (CKD) has a significant impact on sickle cell disease (SCD) morbidity and mortality. Early identification of individuals at highest risk of developing CKD may allow therapeutic intervention to prevent worse outcomes. This study aimed to evaluate the prevalence and risk factors for reduced estimated glomerular filtration rate (eGFR) among adults with SCD in Brazil. Participants in the REDS-III multicenter SCD cohort with more severe genotypes aged ≥ 18 years with at least two serum creatinine values were analyzed. The eGFR was calculated using the Jamaica Sickle Cell Cohort Study GFR equation. The eGFR categories were defined according to the K/DOQI. Participants with eGFR ≥ 90 were compared to those with those with eGFR < 90. Among the 870 participants, 647 (74.4%) had eGFR ≥ 90, 211 (24.3%) had eGFR 60 to 89, six (0.7%) had eGFR 30 to 59, and six (0.7%) had ESRD. Male sex (OR: 37.3; 95%CI: 22.4-65.1), higher age (OR: 1.04; 95%CI: 1.02-1.06), higher diastolic blood pressure (OR: 1.03; 95%CI: 1.009-1.06), lower Hb (OR: 0.80; 95%CI: 0.68-0.93), and lower reticulocytes (OR: 0.94; 95%CI: 0.89-0.99) levels were independently associated with eGFR < 90. There was a trend towards higher odds of death in participants with eGFR < 90 (OR: 1.8; 95%CI: 0.95-3.32; p = 0.065). In turn, participants with eGFR < 60 had a 12.2 (95%CI: 2.1-96.9) times higher odds for death when compared to those with eGFR ≥ 60. In this study, eGFR < 90 was observed in one-quarter of adults. Older age, male sex, higher diastolic blood pressure, lower hemoglobin, and lower reticulocyte levels were associated with occurrence of eGFR < 90. Estimated GFR < 60 increased the risk of mortality., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.

African swine fever (ASF) is a transboundary infectious disease that can infect wild and domestic swine and requires enhanced surveillance between countries. In Mozambique, ASF has been reported across the country, spreading between provinces, mainly through the movement of pigs and their by-products. Subsequently, pigs from bordering countries were at risk of exposure. This study evaluated the spatiotemporal distribution and temporal trends of ASF in swine in Mozambique between 2000 and 2020. During this period, 28,624 cases of ASF were reported across three regions of the country. In total, the northern, central, and southern regions presented 64.9, 17.8, and 17.3% of the total cases, respectively. When analyzing the incidence risk (IR) of ASF per 100,000 pigs, the Cabo Delgado province had the highest IR (17,301.1), followed by the Maputo province (8868.6). In the space-time analysis, three clusters were formed in each region: (i) Cluster A involved the provinces of Cabo Delgado and Nampula (north), (ii) Cluster B involved the province of Maputo and the city of Maputo (south), and (iii) Cluster C consisted of the provinces of Manica and Sofala (central) in 2006. However, when analyzing the temporal trend in the provinces, most were found to be decreasing, except for Sofala, Inhambane, and Maputo, which had a stationary trend. To the best of our knowledge, this is the first study to evaluate the spatial distribution of ASF in Mozambique. These findings will contribute to increasing official ASF control programs by identifying high-risk areas and raising awareness of the importance of controlling the borders between provinces and countries to prevent their spread to other regions of the world.

Protozoa of the Apicomplexa phylum are worldwide distributed with capacity to infect endothermic animals. The study of these protozoa in wild birds in Brazil is scarce. This study aimed to evaluate the occurrence of apicomplexan protozoa in wild birds in the Northeast of Brazil. From October to December 2019, brain tissue samples were collected from 71 captive birds from the Wild Animal Screening Center of the Pernambuco State (CETRAS-Tangara) and 25 free-living birds from the Caatinga biome in Rio Grande do Norte, totaling 96 animals (41 species). Brain fragments were subjected to molecular diagnosis by nested PCR for the 18s rDNA gene of Apicomplexa parasites, followed by DNA sequencing. This gene was detected in 25% (24/96) of the samples, and it was possible to perform DNA sequencing of 14 samples, confirming three genera: Isospora, Sarcocystis and Toxoplasma from eight bird species (Amazona aestiva, Coereba flaveola, Egretta thula, Paroaria dominicana, Sporophila nigricollis, Cariama cristata, Columbina talpacoti, Crypturellus parvirostris). The occurrence these coccidia in wild birds provides important epidemiological information for the adoption of preventive measures for its conservation. Future studies are needed to better understand the consequence of Apicomplexa infection in birds in Caatinga and Atlantic Forest biomes.

An adult male greater bulldog bat (Noctilio leporinus) was found dead in a suburban area in the municipality of Patos, Paraiba, northeastern Brazil. At post-mortem examination, the bat was emaciated and had multifocal to coalescent grey, crusted, dry, scaly cutaneous lesions, irregularly distributed over the dorsal thoracoabdominal region, muzzle, labial commissures, ears and dorsoventral surfaces of the patagia. Histopathology revealed numerous longitudinal and transverse sections of fungal organisms, with weakly basophilic walls, associated with multifocal areas of ulceration of the epidermis, necrosis, rupture and discontinuity of collagen fibres in the dermis without any inflammatory response. Molecular identification matched the organism to Cladosporium spp, Curvularia spp, Exserohilum spp, Bipolaris spp (100%) and Alternaria spp (97%), all of which have been associated with phaeohyphomycosis. Phaeohyphomycosis should be included as a differential diagnosis of cutaneous lesions in chiropterans., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This research aimed to identify the diversity of bacterial species of the genus Staphylococcus spp. in subclinical mastitis in dairy herds in the state of Piauí, Northeastern Brazil, and to evaluate the phenotypic and genotypic resistance profile. Samples were obtained from a total of 17 dairy farms, amounting to 321 positive samples in the California Mastitis Test. Staphylococcus spp. were identified by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy. Subsequently, an antibiogram was performed, and a polymerase chain reaction was carried out to screen for resistance genes in the isolates. Among all the isolates, 59.45% (110/185) belonged to the Staphylococcus genus. Moreover, the following Staphylococcus spp. were identified Staphylococcus aureus, 68.1% (75/110); Staphylococcus chromogenes, 12.7% (14/110); Staphylococcus epidermidis, 5.4% (6/110); Staphylococcus sciuri, 4.5% (5/110); Staphylococcus warneri, 2.7% (3/110); Staphylococcus haemolyticus, 1.8% (2/110); Staphylococcus hominis, 1.8% (2/110); Staphylococcus arlettae, 0.9% (1/110); Staphylococcus capitis, 0.9% (1/110); and Staphylococcus gallinarum, 0.9% (1/110). The antibiogram showed a high frequency of resistance to penicillin and ampicillin, 70.0% (77/110) and 61.8% (68/110), respectively, and a low frequency of resistance to gentamicin and vancomycin, 10.9% (12/110) and 11.8% (13/110), respectively. In the genotypic tests for the different species of Staphylococcus spp., the occurrence of the blaZ gene was observed in 60.9% (67/110) of the isolates, followed by tetL and tetM, both with 20.0% (22/110) each, and the mecA and vanB genes were detected in 0.9% (1/110) of the samples. The identification of all Staphylococcus species isolated from subclinical mastitis cases and the phenotypic and genotypic resistance characterization in these isolates is of great importance for dairy farming in the state of Piauí, as well as for public health., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

This study aimed to determine the presence of antibodies against Mycobacterium avium subspecies paratuberculosis (MAP) in high-producing dairy cows, the presence of the pathogen in the feces, and the risk factors associated with the disease. Blood and fecal samples were collected from 708 dairy cows over 2 years from 54 herds located in five municipalities of Paraná, Brazil. The serum samples were evaluated for the presence of antibodies against MAP using enzyme-linked immunosorbent assay (ELISA). Fecal samples from 100 cows (69 seropositive and 31 seronegative) were assessed using real-time PCR (qPCR) for IS900 of MAP. The herd prevalence of antibodies against MAP was 61.1% (33/54; 95% CI 46.88-74.08), ranging from 12.5 to 80% across the municipalities, and the prevalence in the animals was 9.8% (69/708; 95% CI 7.77-12.15); it ranged from 0 to 87.5% per herd. Only one of the 69 (1.45%) fecal samples from the seropositive cows was positive for the qPCR. The factors associated with the occurrence of paratuberculosis in herds were the use of compost barn system and the type of bed, whereas only the type of bed was associated with the infection of cows. The only risk factor (OR = 2.45; 95% CI 1.03-5.85) associated with the occurrence of paratuberculosis was the introduction of animals purchased from other dairy farms. The prevalence of active infection was low; however, our results demonstrate the presence of MAP in high-producing dairy herds in Paraná state, Brazil., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Glanders is an infectious disease that causes serious damage to the equine production chain in countries where it occurs endemically and poses a risk to public health. This study aimed to conduct an integrative review of the advances in the knowledge of glanders in Brazil over the last three decades since its re-emergence. Documentary research was conducted for the period between the years 2000 and 2022. SCOPUS and PUBMED databases were used to search for scientific articles, dissertations, and thesis, in addition to the Brazilian Digital Library of Thesis and Dissertations (BDTD). A total of 41 documents were retrieved, including 12 dissertations, five theses, and 24 scientific articles. The Federal Rural University of Pernambuco (UFRPE) group provided a noteworthy amount of material on this topic (25 documents, four theses, three dissertations, and 18 scientific articles). During this period, ten dissertations, one thesis, and six scientific articles published by other groups in other states of the Federation were also identified. It was concluded that there was a significant number of scientific publications with relevant data on the clinical, epidemiological, pathological, microbiological, serological, and molecular characteristics of glanders disease in Brazil. Additionally, training of human resources regarding this disease led to an increase in the nucleation of research groups, especially in the northeast region of Brazil. Despite significant advances, new research groups and specific funding are still needed for the development of more accurate diagnostic methods, immunizing production, training of veterinarians to recognize the disease, and more robust programs to control and eradicate the disease in Brazil., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

This study aimed to evaluate Toxoplasma gondii infection in pregnant goats. The goats were divided into two groups: group one (G1) comprised of 31 pregnant goats naturally infected with T. gondii, and group two (G2) comprised of seven uninfected pregnant goats from a flock with a history of abortion due to toxoplasmosis. Serological investigation, ultrasonography, and clinical testing were performed on all goats during gestation. Serum samples from goats and their offspring (precolostral) were collected to evaluate the vertical transmission of T. gondii. Samples from placentas and aborted fetuses were also collected for molecular and histopathological analysis. Results showed that in G1, estrus recurrence occurred in 22.6% (7/31) of the goats, embryonic death in 3.3% (1/31), and abortion in 19.4% (6/31). An increase in anti-T. gondii antibodies was observed in G1 goats at day 150 of pregnancy. T. gondii DNA was detected in 42.8% (3/7) of aborted fetuses and was associated with histopathological lesions caused by this parasite. Moreover, toxoplasmosis in field conditions caused by genotype ToxoDB #1 in pregnant goats resulted in severe reproductive loss in the flock., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Toxoplasmosis is an emerging and re-emerging infectious disease that can be transmitted through a contaminated environment. Environmental contamination is an emergency health issue, and determining its occurrence is fundamental to a One Health approach. In this study, we addressed the extent of environmental contamination and viability of Toxoplasma gondii oocysts in soil in different environments on Fernando de Noronha Island, Brazil. In addition, we performed species distribution modelling to predict the environmental suitability for coccidia persistence in the studied area. Soil samples were collected in 14 neighbourhoods of the Island and in the four main squares, creating a total of 95 soil samples (five samples per site). The samples were analyzed by the polymerase chain reaction (PCR) technique for the presence of the 18S ribosomal DNA gene of Apicomplexan protozoa, followed by genetic sequencing. We obtained 4.2% (4/95) positive soil samples with 100% similarity for T. gondii sequences. Two out of four positive sites on PCR showed viability of T. gondii oocysts through the mouse bioassay technique. As a result of the application of the species distribution modelling, environmental adequacy for the coccidia was observed throughout the Island. The results confirm the contamination of the soil in this insular environment by T. gondii oocysts and the environmental suitability by modelling application. These findings are an alert for the possibility of infection in animals and humans by contaminated soil, and for contamination of the maritime environment in addition to water resources for consumption by the local population., (© 2022 Wiley-VCH GmbH.)

The aim was to detect correlations of IgG antibodies against N. caninum in serum and colostrum samples from ewes, through the IFAT, and to evaluate the presence of this immunoglobulin in the serum of newborn lambs after colostrum ingestion. Blood samples from 162 ewes that did not show any disease in the general physical examination and from their newborn lambs, not more than five days postpartum, along with 162 colostrum samples and 182 blood samples from the neonates, were analyzed. In total, 27.8% (45/162) of the mothers were positive for anti- N. caninum IgG, among which antibodies were detected in the colostrum in 46.7% (21/45). All the ewes with positive colostrum had reactive offspring. The kappa agreement for the correlation between the serological tests on the ewes and the colostrum results was 0.558. This correlation increased as the antibody titers of the mothers increased, and reached 1.000 from the titer of 1:400 from the mothers. Comparison of the antibody detection results between the offspring's serum and colostrum showed a kappa agreement of 1.000. In conclusion, there was a good agreement regarding the detection of anti- N. caninum IgG between the colostrum samples and the lambs' serum; the use of colostrum forms a noninvasive alternative for diagnosing N. caninum in sheep herds.

Manifestations of sickle cell disease (SCD) begin early in childhood and cause morbidity and decreased life expectancy. Hematopoietic stem cell transplantation (HSCT) is curative but associated with risk of mortality attributable to the transplant. This risk should be counterbalanced with SCD morbidity and mortality. A severity score using a Bayesian network model was previously validated to predict the risk of death in adult individuals with SCD. The objective of this study is to calculate the severity scores of participants in a multicenter cohort of Brazilians with SCD, using a previously published Bayesian network-derived score, associated with risk of death and then compare the severity scores between participants with and without an indication for HSCT as defined by the Brazilian Ministry of Health (MoH) criteria. This is an observational, retrospective study. We analyzed 2063 individuals with sickle cell anemia from the Recipient Epidemiology and Donor Evaluation Study-III Brazil SCD cohort and applied a Bayesian network-derived score to compare candidates and non-candidates for HSCT according to the Brazilian MoH transplant criteria. Classical statistical methods were used to analyze data and make comparisons. We compared severity scores between cohort members with (n = 431) and without (n = 1632) HSCT indications according to Brazilian MoH. Scores were not different in adult participants with ≥1 HSCT indication when compared to those with no indication (mean 0.342 versus 0.292; median 0.194 versus 0.183, P = .354) and receiver operating characteristic curves did not demonstrate an obvious threshold to differentiate participants with or without HSCT indications. Severity score may predict risk of death but does not differentiate HSCT candidates. Current indications should be evaluated to ensure that patients with more severe disease who might benefit from HSCT are appropriately identified., Competing Interests: Conflict of interest statement There are no conflicts of interest to report., (Copyright © 2022 The American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Background: The osmotin from the medicinal plant Calotropis procera (CpOsm) has characteristics similar to adiponectin, a human protein with immunoregulatory actions., Purpose: This study aimed to investigate whether recombinant osmotin inclusion bodies from C. procera (IB/rCpOsm) produced in E. coli BL21(DE3) can prevent infection-induced inflammation. A virulent strain of Listeria monocytogenes was used as an infection model., Methods: Cells of E. coli BL21(DE3) carrying the plasmid pET303-CpOsm were used to express the recombinant osmotin, which accumulated at reasonable levels as inclusion bodies (IB/rCpOsm). IB/rCpOsm were purified from induced cells and SDS-polyacrylamide gel electrophoresis followed by mass spectrometry analyses confirmed the identity of the major protein band (23 kDa apparent molecular mass) as CpOsm. Peritoneal macrophages (pMØ) from Swiss mice were cultured with IB/rCpOsm (1 or 10 µg/ml) in 96-well plates and then infected with L. monocytogenes. IB/rCpOsm (0.1, 1 or 10 mg/kg) was also administered intravenously to Swiss mice, which were then infected intraperitoneally with L. monocytogenes., Results: Pretreatment of the pMØ with IB/rCpOsm significantly increased cell viability after infection and reduced the intracellular bacterial load. The infiltration of neutrophils into the peritoneal cavity of mice pretreated with IB/rCpOsm at 10 mg/kg (but not 0.1 and 1 mg/kg) was reduced after infection. In these mice, the bacterial load was high in the peritoneal fluid and the liver, but histological damage was discrete. The treatments with IB/rCpOsm at 10 mg/kg significantly increased the expression of the anti-inflammatory cytokine IL-10., Conclusion: This study shows that recombinant osmotin inclusion bodies from C. procera were bioactive and prompted anti-inflammatory actions at therapeutic dosages in the L. monocytogenes infection model., (Copyright © 2022. Published by Elsevier GmbH.)

The aim of this study was to identify emergent pathogens associated with bovine mastitis in northeastern Brazil and to characterize them for phenotypic and genotypic resistance to antimicrobials. A total of 321 milk samples from cows with subclinical mastitis were collected, and the isolates obtained in culture were identified using matrix-associated laser desorption-ionization - time of flight mass spectrometry. Phenotypic and genotypic antimicrobial resistance tests were performed. We identified 72 bacteria considered emergent in the study region: Enterococcus faecalis (26.3%; 19/72), Streptococcus agalactiae (22.2%; 16/72), Enterococcus faecium (20.0%; 15/72), Escherichia coli (6.9%; 5/72), 6.9% (5/72) Lactococcus garvieae (6.9%; 5/72), Acinetobacter baumannii (5.5%; 4/72), Bacillus subtilis (1.3%; 1/72), Kocuria marina (1.3%; 1/72), Macrococcus caseolyticus (1.3%; 1/72), Microbacterium resistens (1.3%; 1/72), Micrococcus luteus (1.3%; 1/72), Streptococcus dysgalactiae (1.3%; 1/72), Streptococcus hyovaginalis (1.3%; 1/72) and Streptococcus pluranimalium (1.3%; 1/72). The antibiogram revealed the following resistance profiles: ampicillin (77.7%; 56/72), cefoxitin (69.4%; 50/72), erythromycin (61.1%; 44/72), oxacillin (63.8%; 46/72), penicillin (79.1%; 57/72), tetracycline (63.8%; 46/72), gentamicin (25.0%; 18/72), and vancomycin (20.8%; 15/72). Of the isolates, 83.4% (60/72) showed multiple resistance to antimicrobials. The tetM gene was identified in 43.0% (31/72) of the isolates, followed by tetL (31.9%; 23/72), and blaZ (26.3%; 19/72). 83.4% (60/72) of the isolates presented a multiple antimicrobial resistance index higher than 0,2. Emergent bacteria with zoonotic and multiresistant potential occur in cows with mastitis in northeastern Brazil. It is necessary to monitor the occurrence of these and other bacteria in livestock environments and develop control strategies to prevent their spread., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

This study aimed to describe the transmission of T. gondii in naturally infected goats in the semiarid region of northeastern Brazil, through evaluating the serological status of these animals from one to 12 months of age. Seven goat farms were selected. During the first visit, an epidemiological questionnaire was applied. Blood samples were collected from pregnant goats and from their female offspring every month, to monitor them for anti-T. gondii antibodies, using the indirect fluorescence antibody test (IFAT). Out of 66 pregnant goats evaluated, 47% (31/66) (95% CI: 34.6% - 59.7%) were positive and out of 93 female offspring, 61.3% (57/93) (95% CI: 52.4% - 69.2%) were positive, in at least one month during the evaluation period. Among the positive female offspring, 43.3% (27/57) (95% CI: 39.7% - 56.9%) presented environmental infection and 56.7% (30/57) (95% CI: 43.4% - 66.3 %) probably had only colostral antibodies or vertical transmission. It was observed that 92,6% (25/27) from environmental infections occurred until reproductive age (six months) (p<0.0001). It was also noted that there is a correlation between the titration of mothers and offspring, until four months of age (p<0.0001). Large fluctuations in the presence of antibodies were observed among the animals over the months monitored. It can be concluded that the prevalence of anti-T. gondii antibodies within the herds was greater than what has been expressed in specific prevalence studies and that most goat become infected up to the six months of life in the studied area., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Sanitary-hygienic failures in cheese making can pose health risks to consumers. This study aimed to identify multiresistant pathogens in different production stages of artisanal goat coalho cheese in Brazil and characterize their phenotypic and genotypic resistance. Eleven properties in the state of Pernambuco, Brazil, participated in the study. Samples were obtained from different stages of production and the humans involved. The samples obtained were submitted to microbiological culture, then all the isolated microorganisms were submitted to the Matrix Associated Laser Desorption-Ionization - Time of Flight technique for the microbiological identification of the species. Subsequently, Staphylococcus spp., Enterococcus spp. and Macrococcus caseolyticus were subjected to polymerase chain reaction to search for resistance genes and disc diffusion technique to evaluate the resistance profile. A total of 111 isolates were obtained and 31 species were identified, with the frequency of Staphylococcus spp. (62.20%; 69/111), Enterococcus spp. (11.60%; 13/111), Macrococcus caseolyticus (10%; 11/111), Bacillus spp. (3.60%; 4/111), Enterobacter spp. (3.60%; 4/111), Aureobasidium pullulans (1.80%; 2/111), Corynebacterium camporealensis (1.80%; 2/111), Issatchenkia occidentalis (1.80%; 2/111), Kocuria kristinae (1.80%; 2/111), Aerococcus viridans (0.90%; 1/111) and Filifactor villosus (0.90%; 1/111). Phenotypic and genotypic resistance was also detected with the occurrence of 15.90% (7/44) of the mecA gene, 4% (1/25) vanA, and 4% (1/25) vanB in Staphylococcus spp. and 20% (2/10) vanB in and Enterococcus spp. Emerging multiresistant pathogens are present in the production chain of artisanal goat cheese and humans, who exert an important role in disseminating these bacteria with imminent risks to human health., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Glanders is an infectious zoonosis caused by Burkholderia (B.) mallei that mainly affects equids. The objective of this work was to provide additional knowledge on the diversity of the strains circulating in Brazil. Six Burkholderia mallei isolates obtained during necropsies of glanderous horses between 2014 and 2017 in two different states (Pernambuco and Alagoas) were analyzed by polymerase chain reaction-high-resolution melting (PCR-HRM). While four strains (9902 RSC, BM&#95;campo 1, BM&#95;campo 3 and UFAL2) clustered in the L3B2 branch, which already includes the Brazilian 16-2438&#95;BM#8 strain, two strains (BM&#95;campo 2.1 and BM&#95;campo 2.2) clustered within the L3B3sB3 branch, which mostly includes older isolates, from Europe and the Middle East. Whole genome sequencing of two of these strains (UFAL2 and BM&#95;campo 2.1), belonging to different branches, confirmed the HRM typing results and refined the links between the strains, including the description of the L3B3Sb3Gp1SbGp1 genotype, never reported so far for contemporary strains. These results suggest different glanders introduction events in Brazil, including a potential link with strains of European origin, related to colonization or trade., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study, we determined the occurrence of Toxoplasma gondii oocysts in soil samples from public places. A total of 120 samples were collected from 24 sites, including squares, parks, university, hospitals in the city of Recife. The recovered oocysts were subjected to a nested-PCR test, and nine sites (9/24) were found to be positive for gene of apicomplexan parasites. The PCR product was sequenced, and 8.33% (10/120) of the samples showed 100% similarity to T. gondii DNA. T. gondii oocysts were detected in 75% (3/4) of the evaluated hospital soil samples and in 23.81% (5/21) soils samples from the public squares and parks. The results of this study demonstrate the potential of the soil in the areas analyzed as a source of T. gondii infection and therefore highlight the importance of devising educational strategies on the use of these sites, in addition to future cleaning protocols in public areas., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Dairy goats play a significant role in socio-economic, cultural, and nutritional development in many countries. This study aimed to identify multiresistant zoonotic pathogens causing mastitis in goats, in addition to characterizing them for the presence of resistance genes and phenotypic resistance. A total of 714 milk samples from 357 lactating goats in 12 farms in the Northeast region of Brazil were analyzed. The isolates were submitted to Matrix Associated Laser Desorption-Ionization - Time of Flight to identify bacterial species, Polymerase Chain Reaction (PCR) to search for resistance genes, and an antibiogram to evaluate the phenotypic profile of antimicrobial resistance. A total of 214 pathogens were identified and bacterial prevalence was 83.29 % (178/214) Staphylococcus spp.; 6.50 % (14/214) Micrococcus luteus; 3.73 % (8/214) Corynebacterium spp.; 2.80 % (6/214) Bacillus spp.; 1.38 % (3/214) Escherichia coli; 0.92 % (2/214) Enterobacter cloacae; 0.46 % (1/214) Aerococcus viridans; 0.46 % (1/214) Morganella morganii; and 0.46 % (1/214) Turicella otitidis. As for gene frequency, 64.60 % (115/178) of the isolates carried the blaZ gene; 37.07 % (66/178) norA; 22.47 % (40/178) tet(L); 16.85 % (30/178) tet(M); 14.04 % (25/178) norB; 8.42 % (15/178) vanA; 7.30 % (13/178) msrA; 6.41 % (5/178) tet-38; 4.49 % (8/178) norC; 2.25 % (4/178) mecA; and 0.56 % (1/178) vanB. Emerging multiresistant zoonotic pathogens are present in the goat milk production chain, especially the coagulase-negative Staphylococcus species that pose a risk to human and animal health., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Objective: 18F-sodium fluoride (18F-NaF) positron emission tomography (PET) imaging is thought to visualize active atherosclerotic plaque calcification. This is supported by the binding of 18F-NaF to plaque calcification ex vivo, but no prior studies have examined binding of 18F-NaF to human-like plaque in vivo. Our aim was to validate the specificity of 18F-NaF PET for plaque calcifications in atherosclerotic minipigs. Approach and Results: Gain-of-function PCSK9D374Y (proprotein convertase/subtilisin kexin type 9) transgenic Yucatan minipigs (n=4) were fed high-fat diet for 2.5 years to develop atherosclerosis and then subjected to 18F-NaF PET/computed tomography imaging. The heart, aorta, and iliac arteries were immediately re-scanned ex vivo after surgical extraction. Lesions from the abdominal aorta, iliac arteries, and coronary arteries were cryo-sectioned for autoradiography. Histological plaque characteristics, PET/computed tomography signal, and autoradiography were linked through regression and co-localization analysis. Arterial 18F-NaF PET signal had intensities comparable to clinical scans and colocalized moderately with calcification detected by computed tomography. Histological analysis showed calcification spanning from microcalcifications near lipid pools and necrotic core to more homogenous macrocalcifications. Comparison with arteries from autopsy cases confirmed the resemblance in localization and appearance with early human plaque calcification. Regression analysis in the abdominal aorta showed correlations with calcified plaque but could not rule out contributions from noncalcified plaque. This was resolved by autoradiography, which showed specific accumulation in plaque calcifications in all examined arteries. In the context of porcine abdominal aorta, 18F-NaF PET imaging was, however, less accurate than computed tomography for detecting small calcifications. Conclusions: 18F-NaF accumulates specifically in calcifications of atherosclerotic plaques in vivo.