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.

Case Study of Two Post Vaccination SARS-CoV-2 Infections with P1 Variants in CoronaVac Vaccinees in Brazil.

The rapid development of efficacious and safe vaccines against coronavirus disease 2019 (COVID-19) has been instrumental in mitigating the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Moreover, the emergence of SARS-CoV-2 variants raised concerns on the efficacy of these vaccines. Herein, we report two cases of breakthrough infections with the P1 variant in patients vaccinated with CoronaVac, which is one of the two vaccines authorized for emergency use in the Brazilian immunization program. Our observations suggest that the vaccine reduced the severity of the disease and highlight the potential risk of illness following vaccination and subsequent infection with the P1 variant as well as for continued efforts to prevent and diagnose infection in vaccinated persons.

Near-Complete Genome Sequence of Feline Immunodeficiency Virus from Colombia.

Feline immunodeficiency virus (FIV) is an important pathogen of domestic and wild felids. Although serological tests suggest the presence of FIV in cats from Colombia, no molecular characterization has been reported. Here, we describe the near-complete genome of FIV subtype A from a Colombian domestic cat.

Seroepidemiological study of feline coronavirus (FCoV) infection in domiciled cats from Botucatu, São Paulo, Brazil / Estudo soroepidemiológico da infecção pelo coronavírus felino (FCOV) em gatos domiciliados de Botucatu, São Paulo, Brasil

Feline coronavirus (FCoV) is responsible for causing one of the most important infectious diseases of domestic and wild felids, the feline infectious peritonitis (FIP), which is an immune-mediated, systemic, progressive and fatal disease. FCoV is highly contagious, and infection is common in domestic feline populations worldwide. The present study aimed to determine the seropositivity of FCoV infection and its associated epidemiological variables (risk factors) in domiciled cats in Botucatu, São Paulo, Brazil. Whole blood samples (0.5-1mL) were collected from 151 cats, and sera were extracted by centrifugation. These sera were tested by an commercial enzyme-linked immunosorbent assay (ELISA) for the detection of IgG anti-FCoV antibodies. The assessed risk factors were age range, breed, gender, reproductive status, outdoor access and rearing mode (living alone or in a group). The seropositivity was 64.2% (97/151). There was no statistical significance for risk factors related to breed, gender or rearing mode. There were significant differences in seropositivity (p-values ≤0.05) for age range (p=0.0157), reproductive status (p=0.0074) and outdoor access (p=0.0001). This study verified a wide dissemination of FCoV in the studied population, with a higher than expected seropositivity for indoor cats. Among the risk factors, age range, reproductive status and outdoor access presented statistically significant differences, thus helping to establish an epidemiological profile of this population.(AU)

O coronavírus felino (FCoV) é responsável por causar uma das mais importantes doenças infecciosas que acometem os felinos domésticos e selvagens, a peritonite infecciosa felina (PIF), que é uma enfermidade imunomediada, sistêmica, progressiva e fatal. O FCoV é altamente contagioso e a infecção é comum nas populações de felinos domésticos por todo o mundo. O presente estudo objetivou determinar a soropositividade da infecção pelo FCoV e correlacionar variáveis epidemiológicas (fatores de risco) de gatos domiciliados de Botucatu, São Paulo, Brasil. Amostras de sangue total (0,5 a 1mL) foram colhidas de 151 gatos e os soros foram obtidos após centrifugação. Estes soros foram testados por um teste commercial de ELISA para detecção de anticorpos IgG anti-FCoV. Os fatores de risco avaliados foram faixa etária, raça, gênero, condição reprodutiva, acesso à rua e modo de criação (viver solitário ou em grupo). Observou-se uma soropositividade de 64,2% (97/151). Não houve significância estatística para os fatores de risco relacionados à raça, gênero e modo de criação. Houve significância estatística quanto a soropositividade (p-values ≤0,05) para os fatores de risco faixa etária (p=0,0157), condição reprodutiva (p=0,0074) e acesso à rua (p=0,0001). Através do presente estudo verificou-se que o FCoV está amplamente disseminado na população estudada, onde a soropositividade encontrada foi maior do que a esperada para gatos domiciliados. Dentre os fatores de risco, faixa etária, condição reprodutiva e acesso à rua apresentaram diferenças estatisticamente significativas, contribuindo assim, para se estabelecer um perfil epidemiológico desta população.(AU)

Proteinograma e concentração sérica de IgG em potros, do nascimento aos trinta dias de vida, tratados com plasma / Proteinogram and serum IgG concentration in newborn foals up to thirty days of life treated with plasma

Este trabalho teve por objetivo avaliar o proteinograma e concentrações séricas de IgG (após a padronização de teste ELISA) em potros do nascimento aos trinta dias de idade, antes e depois de mamarem colostro e serem tratados com plasma por via intravenosa. Foram utilizados 20 potros e suas respectivas mães, além de quatro animais doadores de plasma. Foram colhidas amostras de sangue dos potros em cinco momentos, logo após o nascimento e antes de mamar colostro (M1), dez horas após nascimento (M2), 24 horas após nascimento e previamente administração do plasma sanguíneo (M3), 48 horas de vida e 24 horas após administração do plasma sanguíneo (M4), e 30 dias após nascimento (M5). Foram colhidos sangue e colostro das éguas progenitoras no momento do parto. A concentração de proteína total (PT) e albumina foram determinadas em analisador bioquímico, a concentração de PT também foi avaliada em refratômetro manual. O fracionamento proteico foi realizado utilizando eletroforese em gel de agarose. A densidade do colostro foi avaliada com colostrômetros de refração BRIX e de densidade específica. A concentração de IgG total de todas as amostras foi determinada por teste ELISA. Com o sistema de ELISA aqui proposto foi possível determinar concentrações de IgG em amostras de soro, plasma e colostro equino com adequada repetibilidade. A média ± desvio padrão da concentração sérica de IgG dos potros ao nascer, foi de 15±8mg/dL, com dez horas de vida foi de 2.408±608mg/dL, se manteve em níveis semelhantes até 48 horas (2.364±784mg/dL) e diminuíram significativamente aos 30 dias de vida (1.414±586mg/dL). A concentração sérica e colostral de IgG nas éguas foi de 1.746±505mg/dL e 7.714±2.619mg/dL, respectivamente. A concentração plasmática de IgG dos doadores de plasma foi de 2.026±148mg/dL. Houve correlação positiva entre as concentrações séricas de IgG e PT (r=0,69 para refratômetro e r=0,76 para bioquímico), GT (r=0,81) e gamaglobulina (r=0,85). Dez horas após o nascimento foi possível verificar a transferência de imunidade passiva, possibilitando adotar medidas profiláticas e/ou terapêuticas em haras de criação de cavalos. Considerando que a proteína total, globulinas totais e fração γ-globulina apresentam correlação com IgG, estas determinações são úteis para monitorar os potros após mamarem o colostro. Um litro de plasma administrado às 24 horas de vida não foi suficiente para aumentar as concentrações séricas de IgG, 24 horas após transfusão, em potros com adequada transferência de imunidade passiva.(AU)

The aim of this study was to evaluate serum protein and serum IgG concentrations (after a direct enzyme immunoassay test ELISA optimization) in newborns foals from birth to thirty days of life before and after colostrum consumption and intravenous treatment with plasma. Twenty foals and their respective progenitors as well as four plasma donor's horses were used. Blood samples were obtained from newborn foals at five time points, immediately after birth and before colostrum intake (M1), ten hours after birth (M2), 24 hours after birth and prior administration of blood plasma (M3), 48 hours after birth and 24 hours after plasma administration (M4), and 30 days after birth (M5). Blood and colostrum samples were collected from the progenitor mares immediately postpartum. Concentration of total protein (TP) and albumin were determined using a biochemical analyzer. The TP concentration was also measured by refractometer. Fractions of total serum protein were separated using agarose gel electrophoresis. Colostrum density was evaluated using BRIX refractometer and specific density colostrometer. Total IgG concentration was determined by an enzyme-linked immunosorbent assay. With the ELISA system proposed here it was possible to determine IgG concentrations in serum, plasma, and equine colostrum samples with adequate repeatability. Serum IgG concentration in foals at birth was 15±8mg/dL (mean ± standard deviation) raising at ten hours (2,408±608mg/dL) and remaining at similar levels up to 48 hours of life (2,364±784mg/dL), and decreasing significantly at 30 days of age (1,414±586mg/dL). Serum and colostrum IgG concentrations of mares were 1,746±505mg/dL and 7,714±2,619mg/dL, respectively. The plasma IgG concentrations from donor mares were 2,026±148mg/dL. Total protein, total globulins, and γ-globulin fraction showed correlation with IgG. Ten hours post birth was an adequate time to verify the transfer of passive immunity, allowing to adoption prophylactic and/or therapeutic measures in a horse farms. One liter of plasma administered at 24 hours of life was not sufficient to raise serum IgG concentrations in foals without passive immunity transfer failure.(AU)

Histopathological, immunohistochemical, and molecular study of BHV-5 infection in the central nervous system of experimentally infected calves / Estudo histopatológico, imuno-histoquímico e molecular da infecção por BHV-5 no sistema nervoso central de bovinos experimentalmente infectados

Bovine meningoencephalitis caused by BHV-5, a double-stranded DNA enveloped virus that belongs to the family Herpesviridae and subfamily Alphaherpesvirinae, is an important differential diagnosis of central nervous diseases. The aim of this study was to describe the histological changes in the central nervous system of calves experimentally infected with BHV-5 and compare these changes with the PCR and IHC results. Formalin-fixed paraffin-embedded central nervous system samples from calves previously inoculated with BHV-5 were microscopically evaluated and tested using IHC and PCR. All the animals presented with nonsuppurative meningoencephalitis. From 18 evaluated areas of each calf, 32.41% and 35.19% were positive by IHC and PCR, respectively. The telencephalon presented more accentuated lesions and positive areas in the PCR than other encephalic areas and was the best sampling area for diagnostic purposes. Positive areas in the IHC and PCR were more injured than IHC and PCR negative areas. The animal with neurological signs showed more PCR- and IHC-positive areas than the other animals.(AU)

A meningoencefalite bovina causada pelo BHV-5, um vírus DNA fita dupla envelopado que pertence à família Herpesviridae e subfamília Alphaherpesvirinae, é um importante diagnóstico diferencial das doenças do sistema nervoso central. O objetivo deste estudo foi descrever as alterações histológicas no sistema nervoso central de bovinos experimentalmente infectados com BHV-5 e comparar estas alterações com os resultados de imunoistoquímica (IHQ) e PCR. Amostras do sistema nervoso central de bezerros previamente inoculados com BHV-5 foram microscopicamente avaliadas e submetidas à IHQ e PCR. Todos os animais apresentaram meningoencefalite não-supurativa. Das 18 áreas avaliadas de cada bezerro, 32,41% e 35,13% foram positivas na IHQ e PCR, respectivamente. O telencéfalo apresentou lesões mais acentuadas e foi mais positivo na PCR do que as demais áreas encefálicas e se apresentou como a melhor área para coleta de material para o diagnóstico. As áreas positivas na IHQ e na PCR apresentaram lesões mais acentuadas do que as áreas negativas para as mesmas técnicas. O animal com sinais neurológicos apresentou mais áreas positivas para PCR e IHQ do que os demais animais.(AU)

Characterization of methicillin-resistant coagulase-negative staphylococci in milk from cows with mastitis in Brazil.

Staphylococci are one of the most prevalent microorganisms in bovine mastitis. Staphylococcus spp. are widespread in the environment, and can infect animals and humans as opportunistic pathogens. The objective of this study was to determine the frequency of methicillin-resistance (MR) among coagulase-negative staphylococci (CoNS) previously obtained from milk of mastitic cows in Brazil and to characterize the antimicrobial resistance phenotype/genotype and the SCCmec type of MRCoNS isolates. Identification of MRCoNS was based on both biochemical and molecular methods. Susceptibility testing for eleven antimicrobials was performed by disk-diffusion agar. Antimicrobial resistance genes and SCCmec were investigated by specific PCRs. Twenty-six MRCoNS were detected (20 % of total CoNS), obtained from 24 animals, and were identified as follows: S. epidermidis (7 isolates), S. chromogenes (7), S. warneri (6), S. hyicus (5) and S. simulans (1). All MRCoNS isolates carried mecA while the mecC gene was not detected in any CoNS. The SCCmec IVa was demonstrated in nine MRCoNS, while the remaining 17 isolates harbored non-typeable SCCmec cassettes. In addition to oxacillin and cefoxitin resistance, MRCoNS showed resistance to tetracycline (n = 7), streptomycin (n = 6), tobramycin (n = 6), and gentamicin (n = 4), and harbored the genes tet(K) (n = 7), str (n = 3), ant(4') (n = 6) and aac(6')-aph(2″) (n = 4), respectively. In addition, seven strains showed intermediate resistance to clindamycin and two to streptomycin, of which two harboured the lnu(B) and lsa(E) genes and two the aad(E) gene, respectively. One isolate presented intermediate erythromycin and clindamycin resistance and harbored an erm(C) gene with an uncommon 89-bp deletion rendering a premature stop codon. MRCoNS can be implicated in mastitis of cows and they constitute a reservoir of resistance genes that can be transferred to other pathogenic bacteria.

Caracterização filogenética de amostras do vírus da imunodeficiência felina (FIV) do Estado de São Paulo / Phylogenetic characterization of feline immunodeficiency virus (FIV) isolates from the state of São Paulo

O vírus da imunodeficiência felina (FIV) é um lentivírus que causa distúrbios imunológicos em gatos domésticos. Devido à alta variabilidade genética do FIV, já foram identificados cinco subtipos (A a E) e a diversidade dentro de cada subtipo é freqüente e o seu estudo pode auxiliar no conhecimento da patogenia e epidemiologia da doença. Assim, o presente trabalho objetivou analisar filogeneticamente cepas do FIV de gatos domésticos oriundos do estado de São Paulo. Para tanto, foi realizado o seqüenciamento de 658 pares de bases do gene gag de amostras coletadas de 23 animais, cujos resultados foram analisados pelo método de substituição nucleotídica Tamura-Nei. A análise filogenética demonstrou que todas as amostras pertenciam ao subtipo B e, claramente, três subgrupos foram formados dentro deste subtipo. Adicionalmente, o resultado obtido sugeriu um ancestral comum entre as cepas do FIV oriundas do Japão e uma amostra brasileira obtida neste estudo. Em conclusão, este trabalho traz as primeiras informações sobre a diversidade genética do FIV no Estado de São Paulo. Estudos adicionais são necessários para melhor entender o cenário real e a distribuição dos tipos e subtipos do FIV na população de gatos domésticos do país.

Feline immunodeficiency virus (FIV) is a lentivirus associated with immunologic disorders in domestic cats. Due to the high genetic variability of FIV, five subtypes (A to E) have been identified and diversity within each subtype is also frequent. The study of the genetic diversity can aid the understanding the pathogenesis and epidemiology of the disease. Therefore, the present work aimed to analyze phylogenetically FIV isolates of domestic cats from the state of São Paulo, Brazil. The sequencing of 658 bp of the gag gene from 23 samples was performed and the results were analyzed using the Tamura-Nei nucleotidic substitution method. The phylogenetic analysis showed that all viruses belong to subtype B, and clearly three subgroups were present within this subtype. Additionally, these results suggest a common ancestor between the FIV strains derived from Japan and one Brazilian virus. In conclusion, this work presents the first information about the genetic diversity of FIV in the state of São Paulo. Additional studies are necessary to characterize the real scenario of the distribution of FIV subtypes in the population of Brazilian cats.

Genetic variability in geographical populations of Aedes aegypti (Diptera, Culicidae) in Brazil elucidated by molecular markers

The tropical mosquito, Aedes aegypti is the most important domestic vector of urban yellow fever and dengue. Genetic population studies on this vector are important because they may lead to new tools for surveillance. An analysis of genetic structure was conducted among populations of A. aegypti from 11 localities in four demographic regions within six Brazilian federal states. Markers included 21 random amplified polymorphic DNA (RAPD) loci. RAPD markers were detected among populations and cluster analysis revealed two main groups. We found high genetic polymorphism (H S = 0.224) and high levels of genetic differentiation between populations from different states (G ST = 0.430), as well as in populations from cities in the same state (G ST = 0.410). These results indicate significant differentiation in A. aegypti populations in Brazil. Regression analyses of geographic distances and pairwise F ST values estimated from RAPD markers showed that there is a correlation between genetic structure and geographic localization.

Tuberculosis survey of free-ranging marsh deer (Blastocerus dichotomus) in Brazil.

Esophageal-pharyngeal fluids from 53 free-ranging marsh deer (Blastocerus dichotomus) captured for a research program in the state of Mato Grosso do Sul, Brazil, were assayed for tuberculosis. Total DNA was extracted. amplified by polymerase chain reaction using specific primers for Mycobacterium tuberculosis complex (M. tuberculosis, M. bovis, M. microti, and M. africanum), and observed by agarose gel electrophoresis stained with ethidium bromide. All samples were negative. This, along with necropsy and histopathology data, suggests that these animals are not shedding and probably do not have active disease.