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

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

Antibodies against Leptospira spp. in Free-living and Captive Broad-snouted Caiman (Caiman latirostris) and Free-living Yacare Caiman (Caiman yacare) in Brazil.

We evaluated antibodies against Leptospira spp. in both free-living and captive Caiman latirostris from Atlantic Forest, and free-living Caiman yacare from Pantanal, Brazil, by using a microscopic agglutination test. Overall seropositivity was 17%, with rates of 36% in captive C. latirostris (n=4/11) and 18% in free-living C. yacare (n=4/22).

1H NMR spectroscopy as a tool to probe potential biomarkers of the drying-salting process: A proof-of-concept study with the Amazon fish pirarucu.

Dry-salted pirarucu (Arapaima gigas) plays an important cultural role in the Amazon region - South America. In this study, we explored the changes in the chemical composition of pirarucu meat following the drying-salting process via 1H NMR spectroscopy. Combining multivariate and univariate statistical analyses yielded a robust differentiation of metabolites involved in the process. VIP score (>1), p-value (<0.05), and AUC (>0.7) were considered to selecting compounds that had significant fluctuations in their contents along the process. Our results pointed out acetate, lactate, succinate, and creatinine as metabolites undergoing significant changes during the drying-salting process. Creatinine was not detected in fresh samples. The investigation of multiple components delves deeper into the molecular nuances of the salting-drying process's impact on fish meat, providing a more comprehensive understanding of the possible chemical transformations and how the matrix's quality control and nutritional aspects should be addressed.

Pork belly quality variation and its association with fatness level.

Belly is a very popular pork cut composed of different layers of fat and muscle tissue. This work aims to investigate the effect of belly fatness on the morphological, mechanical (firmness) and compositional characteristics of fresh pork bellies and the distribution of the fat within the belly slice. A total of 182 bellies, selected to ensure variability of fatness, sexes and genotypes, were scanned by computed tomography (CT) to determine the fat content which, together with the genotype, led to the formation of 5 classes: F1 class below 26%, F2 class from 26% to 33.9%, and F3 class above 33.9% of fat content from common commercial crossbred pigs; F4 class with an average fatness of 47.3% from pure Duroc pigs; and last, F5 class with 62.6% average fat content from Iberian×Duroc pigs. The distribution of the fat in the central belly slice obtained by CT revealed important differences by region although the fat content was proportional to the overall fatness of the belly. Both belly weight and belly firmness increased with higher fatness. In bellies from common commercial pigs, an increase of SFA and MUFA and a decrease of PUFA as fatness increased was observed. This study highlights variations in belly characteristics among different fat classes, indicating considerable differences in the quality of bellies currently available in the market. This may influence producers and consumers acceptability such that fat content could be considered as a quality criterion to pre-classify bellies and better match the raw product with its final destination.

Global proteome of the saprophytic strain Leptospira biflexa and comparative analysis with pathogenic strain Leptospira interrogans uncover new pathogenesis mechanisms.

Leptospira is a genus of bacteria that includes free-living saprophytic species found in water or soil, and pathogenic species, which are the etiologic agents of leptospirosis. Besides all the efforts, there are only a few proteins described as virulence factors in the pathogenic strain L. interrogans. This work aims to perform L. biflexa serovar Patoc1 strain Paris global proteome and to compare with the proteome database of pathogenic L. interrogans serovar Copenhageni strain Fiocruz L1-130. We identified a total of 2327 expressed proteins of L. biflexa by mass spectrometry. Using the Get Homologues software with the global proteome of L. biflexa and L. interrogans, we found orthologous proteins classified into conserved, low conserved, and specific proteins. Comparative bioinformatic analyses were performed to understand the biological functions of the proteins, subcellular localization, the presence of signal peptide, structural domains, and motifs using public softwares. These results lead to the selection of 182 low conserved within the saprophyte, and 176 specific proteins of L. interrogans. It is anticipated that these findings will indicate further studies to uncover virulence factors in the pathogenic strain. This work presents for the first time the global proteome of saprophytic strain L. biflexa serovar Patoc, strain Patoc1. SIGNIFICANCE: The comparative analysis established an array of specific proteins in pathogenic strain that will narrow down the identification of immune protective proteins that will help fight leptospirosis.

NMR and multivariate methods: Identification of chemical markers in extracts of pedra-ume-caá and their antiglycation, antioxidant, and enzymatic inhibition activities.

INTRODUCTION: In Brazil, the plant group popularly known as "pedra-ume-caá" is used in folk medicine for the treatment of diabetes, and its raw material is commonly sold. OBJECTIVE: The aim of the study was to apply a method for chemical identification of extracts of dry pedra-ume-caá leaves using HPLC-high-resolution mass spectrometry (HRMS) and NMR and develop a multivariate model with NMR data to authenticate commercial samples. In addition, to evaluate the biological activities of the extracts. MATERIALS AND METHODS: Dry extracts of Myrcia multiflora, Myrcia amazonica, Myrcia guianensis, Myrcia sylvatica, Eugenia punicifolia leaves, and 15 commercial samples (sold in Manaus and Belém, Brazil) were prepared by infusion. All the extracts were analysed using HPLC-high-resolution mass spectrometry (HRMS), NMR, principal component analysis (PCA), and hierarchical cluster analysis (HCA). The antidiabetic effect of extracts was evaluated according to enzymatic inhibition. Their content of total phenols, cell viability, and antioxidant and antiglycation activities were also determined. RESULTS: HPLC-HRMS and NMR analysis of these extracts permitted the identification of 17 compounds. 1H NMR data combined with multivariate analyses allowed us to conclude that catechin, myricitrin, quercitrin, and gallic and quinic acids are the main chemical markers of pedra-ume-caá species. These markers were identified in 15 commercial samples of pedra-ume-caá. Additionally, only the extracts of M. multiflora and E. punicifolia inhibited α-glucosidase. All the extracts inhibited the formation of advanced glycation end products (AGEs) and showed free-radical-scavenging activity. These extracts did not present cytotoxicity. CONCLUSION: This study revealed the chemical markers of matrices, and it was possible to differentiate the materials marketed as pedra-ume-caá. Moreover, this study corroborates the potential of these species for treating diabetes.

Structural Relationships between Asthma and Dental Caries in Children: A Birth Cohort Study in Southern Brazil.

INTRODUCTION: There is an inconclusive causal association between asthma symptoms and dental caries in the primary dentition. This study aimed to investigate, using SEM (structural equation modeling), a possible causal relation between asthma and dental caries in the primary dentition. METHODS: Using data from the 2004 Pelotas Birth Cohort Study, a sub-sample of 1,303 individuals was selected. Dental caries was clinically evaluated at 5 years old based on decayed, missing, and filled tooth (dmft) index criteria. Asthma-related symptoms (wheezing and shortness of breath) at 1- and 4-year-olds composed a latent variable and were the main exposures to caries occurrence. SEM was used to identify possible direct, indirect, and mediated effects of asthma in primary dentition dental caries. RESULTS: The general prevalence of caries at age 5 was 1.95 (SD: 3.56). When comparing the dmft values for children with asthma symptoms and those without, they presented similar values in both periods where asthma symptoms were evaluated (1- and 4-year-old). SEM analysis showed that asthma was neither directly nor indirectly related to dental caries. CONCLUSION: Asthma, using a latent variable constructed based on asthma symptoms, showed no causal effect on dental caries occurrence in the primary dentition.

Global proteome of the saprophytic strain Leptospira biflexa and comparative analysis with pathogenic strain Leptospira interrogans uncover new pathogenesis mechanisms

Leptospira is a genus of bacteria that includes free-living saprophytic species found in water or soil, and pathogenic species, which are the etiologic agents of leptospirosis. Besides all the efforts, there are only a few proteins described as virulence factors in the pathogenic strain L. interrogans. This work aims to perform L. biflexa serovar Patoc1 strain Paris global proteome and to compare with the proteome database of pathogenic L. interrogans serovar Copenhageni strain Fiocruz L1–130. We identified a total of 2327 expressed proteins of L. biflexa by mass spectrometry. Using the Get Homologues software with the global proteome of L. biflexa and L. interrogans, we found orthologous proteins classified into conserved, low conserved, and specific proteins. Comparative bioinformatic analyses were performed to understand the biological functions of the proteins, subcellular localization, the presence of signal peptide, structural domains, and motifs using public softwares. These results lead to the selection of 182 low conserved within the saprophyte, and 176 specific proteins of L. interrogans. It is anticipated that these findings will indicate further studies to uncover virulence factors in the pathogenic strain. This work presents for the first time the global proteome of saprophytic strain L. biflexa serovar Patoc, strain Patoc1.

Effect of glycosaminoglycans on the structure and composition of articular cartilage and bone of broilers.

This study aimed to assess the influence of glycosaminoglycan (chondroitin and glucosamine sulfates) supplementation in the diet of broilers on the expression of matrix metallopeptidase 9 (MMP-9) and metallopeptidase inhibitor 2 (TIMP-2) genes, the synthesis of proteoglycans, collagen type II and chondrocytes, bone and cartilage macroscopy, bone mineral densitometry, bone breaking strength and mineral profile. A completely randomized design was carried out in a 3 × 3 factorial scheme (3 levels of chondroitin sulfate: 0.00, 0.05, and 0.10%; and 3 levels of glucosamine sulfate: 0.00, 0.15, and 0.30%), totaling 9 treatments. At 21 and 42 d of age, broilers were slaughtered, and tibias and femurs were collected for evaluation. There was an interaction (P < 0.05) of sulfates for the expression of MMP-9 and its inhibitor TIMP-2 in femur articular cartilage, as well as for the number of chondrocytes, collagen type II and proteoglycans in tibia articular cartilage, bone and cartilage macroscopy and mineral profile (P < 0.05), with better results obtained with the inclusion of chondroitin and/or glucosamine sulfates in the feed. In conclusion, chondroitin and glucosamine sulfates can be used in broiler diets in order to favor the development of the structure of the locomotor system (bones and joints), thus preventing locomotion problems.

Emergence of livestock-associated Mammaliicoccus sciuri ST71 co-harbouring mecA and mecC genes in Brazil.

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 ß-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.

Chapter 9.4: Operative Treatment and Monitoring of Coronal Caries in Daily Practice.

After the caries lesion reaches a certain extent of tooth structure loss, a restoration is often needed to repair the defect. Operative interventions in cariology aim to aid biofilm removal and lesion arrest by cavity sealing, avoid pulpal damage, and restore form, function, and esthetics. There are no clear evidence-based parameters to determine the most appropriate treatment option for each clinical situation. Despite of this, direct composite resins have been the preferable restorative treatment. Scientific literature shows that composites and adhesive strategies play a minor role in treatment success. Patient-related risk factors (mainly those associated with lifestyle and health choices), in addition to the dentist's decision-making process, play a significant role in longevity of the restorations, which tend to fail for the same reasons that lead to the need for restoration (dental caries, tooth/restoration fracture, and esthetics). Therefore, monitoring old restorations in clinical service, even if those present clear signs of degradation, is possible and reasonable within the concept of minimal intervention in dentistry. Unnecessary reinterventions are harmful and costly to health systems, and the clinician's efforts must be directed to eliminating or reducing the etiological factors that can cause the restoration to fail. Thus, patient risk factors assessment is a crucial point in monitoring restorations. Clinician should - whenever possible - postpone operative reinterventions, monitoring the etiological factors that may compromise the restoration's longevity. Also, when operative reintervention is necessary, refurbishment, polishing, and repair should be prioritized over replacement.

Constant testing for Pestivirus in cell lines reveals different routes of contamination.

Pestivirus can contaminate cell cultures and sera and cause serious problems that evolve the integrity of studies, confidence in diagnostic results, and safety of human and animal vaccines. Contaminations by Pestivirus and other viruses may occur at any time and regular assays of monitoring in cell cultures and your supplies are necessary. This study aimed to analyze the phylogeny of Pestivirus detected from cell cultures, calf serum, and standard strains of three laboratories in Brazil that carry out frequent tests for the monitoring of cellular contaminations. These samples were submitted to phylogenetic analysis to understand the genetic relationship between contaminants occurring in these facilities. As result, the Pestivirus found in samples were Bovine viral diarrhea virus (BVDV-1 and BVDV-2), Hobi-like viruses (often named BVDV-3), and Classical swine fever virus (CSFV), and the phylogenetic analysis help us to infer at three possible routes of contamination in this work.

Bovine-associated staphylococci and mammaliicocci trigger T-lymphocyte proliferative response and cytokine production differently.

We examined whether distinct staphylococcal and mammaliicoccal species and strains trigger B- and T-lymphocyte proliferation and interleukin (IL)-17A and interferon (IFN)-γ production by peripheral blood mononuclear cells in nulliparous, primiparous, and multiparous dairy cows. Flow cytometry was used to measure lymphocyte proliferation with the Ki67 antibody, and specific monoclonal antibodies were used to identify CD3, CD4, and CD8 T lymphocyte and CD21 B lymphocyte populations. The supernatant of the peripheral blood mononuclear cell culture was used to measure IL-17A and IFN-γ production. Two distinct, inactivated strains of bovine-associated Staphylococcus aureus [one causing a persistent intramammary infection (IMI) and the other from the nose], 2 inactivated Staphylococcus chromogenes strains [one causing an IMI and the other from a teat apex), as well as an inactivated Mammaliicoccus fleurettii strain originating from sawdust from a dairy farm, and the mitogens concanavalin A and phytohemagglutinin M-form (both specifically to measure lymphocyte proliferation) were studied. In contrast to the "commensal" Staph. aureus strain originating from the nose, the Staph. aureus strain causing a persistent IMI triggered proliferation of CD4+ and CD8+ subpopulations of T lymphocytes. The M. fleurettii strain and the 2 Staph. chromogenes strains had no effect on T- or B-cell proliferation. Furthermore, both Staph. aureus and Staph. chromogenes strains causing persistent IMI significantly increased IL-17A and IFN-γ production by peripheral blood mononuclear cells. Overall, multiparous cows tended to have a higher B-lymphocyte and a lower T-lymphocyte proliferative response than primiparous and nulliparous cows. Peripheral blood mononuclear cells of multiparous cows also produced significantly more IL-17A and IFN-γ. In contrast to concanavalin A, phytohemagglutinin M-form selectively stimulated T-cell proliferation.

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

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

Hypoglycemic effect of the dry leaf extract of Myrcia multiflora in streptozotocin-induced diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Myrcia multiflora (Lam) DC. is a medicinal plant used in folk medicine for diabetes control, mainly in the Brazilian Amazon. The leaves of this species has already demonstrated antidiabetic properties; however, in mice with type 2 diabetes (DM2), the cumulative effect of the consumption of the dry extract of M. multiflora leaves (Mm) has not yet been reported. AIM OF THE STUDY: To investigate the effect of the dry extract obtained from the infusion of the dried leaves of M. multiflora on the blood glucose levels of diabetic mice. MATERIALS AND METHODS: DM2 was induced in Swiss male mice by a single intraperitoneal injection of streptozotocin [150 mg/kg body weight (bw)]. The animals were divided into two control groups (healthy and diabetic without treatment) and three sample groups that received Mm (25 and 50 mg/kg bw) and acarbose (200 mg/kg bw) by gavage once daily for 28 days (D28). Additionally, biochemical parameters, thiobarbituric acid reactive species (TBARS) levels in the liver, and histopathological analyses of the kidneys and liver were performed. RESULTS: On the seventh day of treatment, a 74.7% reduction in glucose levels were observed in the group of diabetic animals treated with Mm (50 mg/kg bw) when compared to the beginning of the treatment. At D28, the hypoglycemic effect was maintained. The results of the biochemical and histopathological parameters and the TBARS levels suggest that this dry extract exerts nephro- and hepatoprotective effects. CONCLUSIONS: The findings demonstrate the potential that this extract has to inhibit the α-glucosidase enzyme, and it acts similarly to the positive control acarbose. Furthermore, this extract is nephro- and hepatoprotective. Therefore, this dry extract has the potential to be an adjuvant for DM2, which corroborates its use in folk medicine.

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

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

Maternal risk factors for congenital anomalies of the kidney and urinary tract: A case-control study.

BACKGROUND: Congenital anomalies of the kidney and urinary tract (CAKUT) are significant causes of pediatric morbidity and mortality. The spectrum of CAKUT can be part of a syndrome, but most of these abnormalities occur as isolated and sporadic forms. The etiology of human CAKUT is unknown in the majority of cases. This case-control study aimed to investigate the association between maternal characteristics and the occurrence of CAKUT and specific CAKUT phenotypes. METHODS: In this case-control study, 29,653 newborns were evaluated consecutively in a tertiary neonatal unit using the Latin American Collaborative Study of Congenital Malformations (ECLAMC) registry. Newborns without congenital anomalies were matched to CAKUT cases by sex, date, and place of birth at a ratio of 3:1. For analysis purposes, the cases were stratified into four subgroups: upper tract abnormalities (UTA), including ureteropelvic junction obstruction, vesicoureteral reflux, primary megaureter and others (n = 239), lower urinary tract obstruction (LUTO) (n = 79), cystic diseases (n = 59) and agenesis/hypodysplasia (n = 28). Multivariable logistic regression analyses were used to calculate crude and adjusted odds ratios (ORs) with 95% confidence intervals (CIs) for associations between the maternal risk factors and the presence of CAKUT. RESULTS: The prevalence of non-syndromic CAKUT in our sample was 13 per 1000 live births. Data records allowed the analysis of 405 cases and 1208 controls. After adjustment by the binary regression logistic, three covariates remained associated as risk factors for the entire spectrum of CAKUT: consanguinity (Odds ratio [OR], 7.1, 95%CI, 2.4-20.4), family history of CAKUT (OR, 6.4, 95%CI, 1.9-21.3), and maternal chronic hypertension (OR, 14.69, 95%CI, 3.2-67.5) (Figure). These risk factors persisted consistently across the various CAKUT phenotypes with minor variations. Consanguinity was the only factor consistently associated with almost all CAKUT phenotypes. Maternal hypertension was associated with all phenotypes except for the agenesis/hypodysplasia group. The prevalence of CAKUT cases was 15 times higher in hypertensive mothers (3%) compared to normotensive mothers (0.2%). CONCLUSION: Our study suggests that an increased risk of CAKUT is associated with consanguinity, a positive family history of CAKUT, and maternal hypertension. However, the prevalence of these risk factors in our cohort was rare and most cases presented as sporadic forms.

Emergence of livestock-associated Mammaliicoccus sciuri ST71 co-harbouring mecA and mecC genes in Brazil

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.

Staphylococcus aureus-Cure-Associated Antigens Elicit Type 3 Immune Memory T Cells.

BACKGROUND: Staphylococcus aureus is one of the most frequently major mastitis pathogens that cause clinical and subclinical mastitis worldwide. Current antimicrobial treatments are usually ineffective, and the commercially available vaccines lack proven effectiveness. The immunological response elicited by the recombinant S. aureus-cure-associated proteins phosphoglycerate kinase (PGK), enolase (ENO), and elongation factor-G (EF-G) in combination with the granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccination was studied in this work. METHODS: Here, twenty-three C57BL/6 mice were divided into four groups and vaccinated with: G1: none (control); G2: GM-CSF DNA plasmid DNA vaccine; G3: the combination of EF-G+ENO+PGK; and G4: the combinations of EF-G+ENO+PGK proteins plus GM-CSF plasmid DNA vaccine. After 44 days, spleen cells were collected for immunophenotyping and lymphocyte proliferation evaluation by flow cytometry upon S. aureus stimulus. RESULTS: Immunization with the three S. aureus recombinant proteins alone resulted in a higher percentage of IL-17A+ cells among CD8+ T central memory cells, as well as the highest intensity of IL-17A production by overall lymphocytes indicating that the contribution of the combined lymphocyte populations is crucial to sustaining a type 3 cell immunity environment. CONCLUSION: The immunization with three S. aureus-cure-associated recombinant proteins triggered type 3 immunity, which is a highly interesting path to pursue an effective bovine S. aureus mastitis vaccine.

A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis.

The zoonotic disease leptospirosis is caused by pathogenic species of the genus Leptospira and was recently included in the list of Neglected Diseases by the World Health Organization. Leptospirosis burden is estimated to have over a million human cases and cause 60 thousand deaths annually, in addition to its economic impact and veterinary concern. The microscopic agglutination test (MAT), recommended by the World Health Organization, exhibits reduced sensitivity at the beginning of the disease, in addition to being technically difficult. New recombinant antigens are being pursued for rapid and specific serodiagnostic tests, especially in the initial phase of the disease, and chimeric multiepitope proteins are a strategy with a great potential to be implemented in serology. Based on previous subproteomic results, we designed a synthetic construct comprising 10 conserved leptospiral surface antigens, and the recombinant protein was purified and evaluated regarding its diagnostic potential. The protein termed rChi2 was recognized by antibodies in serum from patients both at the onset (MAT-) and in the convalescent (MAT+) phase in 75 and 82% of responders, respectively. In addition, rChi2 immunization in hamsters elicited a strong humoral response, and anti-rChi2 antibodies recognized several immobilized intact Leptospira species, validating its potential as an early, broad, and cross-reactive diagnostic test.