Potentially Pathogenic Multidrug-Resistant Escherichia coli in Lamb Meat.

Extended-spectrum cephalosporin (ESC) resistance remains a threat since ESC are important antimicrobials used to treat infections in humans and animals. Escherichia coli is an important source of ESC-resistance genes, such as those encoding extended-spectrum ß-lactamases (ESBLs). E. coli is a common commensal of lambs. Reports that contaminated food can be a source of ESC-resistant bacteria in humans and that ESBL-producing E. coli are found in sheep in Brazil led us to survey their presence in retail lamb meat. Twenty-five samples intended for human consumption were screened for ESC-resistant E. coli, and the isolates were characterized. IncI1-blaCTX-M-8 and IncHI2-blaCTX-M-2 were the main plasmids responsible for ESC resistance. The plasmids harbored common ESBL genes in Enterobacteriaceae from food-producing animals in Brazil. IncI1-blaCTX-M-14 and IncF-blaCTX-M-55 plasmids, associated with human infections, were also detected. Few CTX-M-producing E. coli have been clustered by typing methods, and some may be genetically pathogenic. The findings indicate the presence of diverse strains of E. coli, harboring important ESBL genes, in lamb meat in Brazil. Surveillance of ESC-resistant bacteria could reduce the spread of antimicrobial resistance through the food chain.

Dissemination of Multidrug-Resistant Commensal Escherichia coli in Feedlot Lambs in Southeastern Brazil.

Antimicrobial resistance (AR) is a public health issue since it limits the choices to treat infections by Escherichia coli in humans and animals. In Brazil, the ovine meat market has grown in recent years, but studies about AR in sheep are still scarce. Thus, this study aims to investigate the presence of AR in E. coli isolated from lambs during feedlot. To this end, feces from 112 lambs with 2 months of age, after weaning, were collected on the first day of the animals in the feedlot (day 0), and on the last day before slaughtering (day 42). Isolates were selected in MacConkey agar supplemented with 4 mg/L of ceftiofur and identified by biochemical methods. Isolates were submitted to an antimicrobial susceptibility test by disc-diffusion and PCR to investigate genes for phylogenetic group, virulence determinants and resistance to the several antimicrobial classes tested. The genetic localization of the bla genes detected was elucidated by S1-PFGE followed by Southern blot-hybridizations. The isolates were typed by XbaI-PFGE and MLST methods. Seventy-eight E. coli were isolated from 8/112 (7.1%) animals on day 0, and from 55/112 (49.1%) animals on day 42. Since only fimH was present in almost all E. coli (97.4%) as a virulence gene, and also 88.5% belonged to phylogroups B1 or A, we consider that isolates represent intestinal commensal bacteria. The dendrogram separated the 78 non-virulent isolates in seven clusters, two of which comprised 50 E. coli belonging to ST/CC 1727/446 or ST 3994 recovered on day 42 commonly harboring the genotype bla CMY -2-aac(3)-IIa -tetA-sul1-sul2-floR-cmlA. Special attention should be given to the presence of bla CTX-M-15, a worldwide gene spread, and bla CTX-M-14, a hitherto undetected gene in Enterobacteriaceae from food-producing animals in Brazil. Importantly, E. coli lineages and plasmids carrying bla genes detected here have already been reported as sources of infection in humans either from animals, food, or the environment, which raises public health concerns. Hence, two types of commensal E. coli carrying important AR genes clearly prevailed during feedlot, but lambs are also reservoirs of bacteria carrying important AR genes such as bla CTX-M-14 and bla CTX-M-15, mostly related to antimicrobial treatment failure.

Characterization of the microtranscriptome of macrophages infected with virulent, attenuated and saprophyte strains of Leptospira spp.

Leptospirosis is a bacterial zoonosis, caused by Leptospira spp., that leads to significant morbidity and mortality worldwide. Despite considerable advances, much is yet to be discovered about disease pathogenicity. The influence of epigenetic mechanisms, particularly RNA-mediated post-transcriptional regulation of host immune response has been described following a variety of bacterial infections. The current study examined the microtranscriptome of macrophages J774A.1 following an 8h infection with virulent, attenuated and saprophyte strains of Leptospira. Microarray analysis revealed that 29 miRNAs were misregulated following leptospiral infection compared to control macrophages in a strain and virulence-specific manner. Pathway analysis for targets of these differentially expressed miRNAs suggests that several processes involved in immune response could be regulated by miRNAs. Our data provides the first evidence that host miRNAs are regulated by Leptospira infection in macrophages. A number of the identified miRNA targets participate in key immune response processes. We suggest that post-transcriptional regulation by miRNAs may play a role in host response to infection in leptospirosis.

Transcriptome datasets of macrophages infected with different strains of Leptospira spp.

The datasets reported herein provide information about microarray experiment of macrophage cell line J774A.1 infected with three different strains of Leptospira spp. Transcriptomic profiles were generated using Affymetrix® Mouse Gene 2.1 ST Array Strip. Data was normalized and statically process, p-value < 0.01, FDR < 0.05 and log2 fold change (± 2). The microarray raw data are available in Gene Expression Omnibus (GEO) under accession number GSE105141.

IL-6 and TNF-alpha production during active canine visceral leishmaniasis.

We investigated the production of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) during canine visceral leishmaniasis (VL) to gain a better understanding of the role of such multi-functional cytokines in parasite resistance. IL-6 and TNF-alpha levels were measured by capture ELISA in sera from 8 healthy dogs from a non-endemic area (control group) and in sera from 16 dogs from Araçatuba, SP, Brazil, an area endemic for leishmaniosis. The dogs from the endemic area were selected by positive ELISA serology against total Leishmania chagasi antigen, positive spleen imprints for Leishmania, and the presence of at least three clinical signs associated with active visceral leishmaniasis (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachexia, locomotory difficulty, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy). Enhanced systemic IL-6 production was found in sera from dogs with the active disease compared to healthy dogs (t-test, P<0.05). In contrast, TNF-alpha did not differ between the two groups studied. There was no correlation between IL-6 production and anti-leishmanial antibody titers in the sera. Our findings suggest that IL-6 is a good marker of active disease during leishmaniasis, and that other cytokines may be involved in the hypergammaglobulinemia characteristic of canine visceral leishmaniasis.

Proteinograma sérico de bezerros recém-nascidos alimentados com colostro de vacas com mastite / Serum protein concentration in newborn calves fed with colostrum from cows wirh mastitis

O objetivo do presente estudo foi o de avaliar o proteinograma sérico de bezerros alimentados com colostro oriundo de vacas sadias (n = 10 ), com mastite assintomática (n = 10 ) e mastite clínica (n =10 ). As vacas foram alocadas em seus respectivos grupos de acordo com o exame macroscópico da secreção colostral, contagem de células somáticas, CMT e isolamento microbiano. As amostras de sangue dos conceptos foram colhidas logo após o nascimento, 24 e 48 horas após a ingestão do colostro dos quartos infectados e dos sadios. Foi avaliada a concentração de proteína total pelo método do biureto e as concentrações de imunoglobulina A (IgA), imunoglobulina G (IgG), transferrina, albumina e haptoglobina por meio da eletrofoerese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Não foram observadas diferenças entre os grupos nas concentrações de albumina, proteína total e IgA. Os bezerros alimen- tados com colostro de vacas com mastite assintomática e clínica apresentaram teores de haptoglobina superiores aos animais sadios. As concentrações de IgG e transferrina foram significativamente inferiores nos bezerros tratados com colostro de vacas com mastite clínica. Concluiu-se que a ingestão de colostro de quartos sadios e infectados de vacas que pariram com mastite (GII e GIII) não resulta em falha de transferência da imunidade passiva.

The aim of this study was to evaluate the serum protein concentration in newborns fed with colostrum derived from healthy cows (n = 10), cows with subclinical mastitis (n = 10) and cows with clinical mastitis (n = 10). 30 Holstein cows were assigned to their respective groups according to macroscopic examination of colostral secretion, somatic cell count, CMT and presence of bacteria in colostrum samples. Blood samples of the calves were collected immediately after birth, at 24 and 48 hours after ingestion of colostrum. The total protein was measured by the biuret method and the concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG), transferrin, albumin and haptoglobin was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). No differences were observed amongst groups in the concentrations of albumin, total protein and IgA. In animals from cows with subclinical and clinical mastitis haptoglobin concentrations were higher than those of healthy animals. The concentrations of IgG and transferrin were significantly lower in calves from cows with mastitis. We concluded that the ingestion of colostrum from infected and uninfected glands from cows with mastitis (GII e GIII) is unlikely to be an important contributor to the high rate of failure of passive transfer of immunoglobulins in calves.

Glicemia, proteinograma e perfil de alguns componentes bioquímicos séricos de cabras da raça Bôer no pós-parto / Glycemic, protein profile and serum biochemical profile in Bôer goat in the postpartum period

Objetivou-se testar a hipótese de que ocorre variação de alguns componentes proteicos, bioquímicos e glicêmicos do sangue de cabras no período pós-parto. Utilizaram-se 11 cabras para avaliar as seguintes variáveis séricas: proteína total (PT), albumina, α-globulina, β-globulina, γ-globulina, imunoglobulina G (IgG), aspartatoaminotransferase (AST), fosfatase alcalina (FA), gamaglutamiltransferase (GGT), creatinina,ureia e glicemia. Esses componentes foram determinados nos momentos zero (imediatamente após o parto), dois, sete, 15, 30 e 75 dias pós-parto. A β-globulina, IgG, creatinina e uréia das cabras não apresentaram variações significativas; a glicemia no momento zero foi maior que os valores basais fisiológicos devido ao estímulo da glicogênese determinado pelo aumento de cortisol no parto. Os constituintes sanguíneos das cabras evidenciaram variações no período até 75 dias pós-parto em consequência de causas fisiológicas ligadas ao parto e puerpério.

The objective was to test the hypothesis that there is variation of some protein components, biochemical and glucose from the blood of goats in the postpartum period. ElevenBôer goats were used to evaluate the serum following variables: total protein (TP), albumin, α-globulin, β-globulin, γ-globulin, immunoglobulin G (IgG), aspartate (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine, urea and glucose. These components were determined in moments zero (immediately after delivery), two, seven, 15, 30 and 75 days postpartum. The β-globulin, IgG, creatinine and urea from the goats did not show significant variations, the blood glucose at time zero was greater than the baseline values due to physiological stimulation of glycogenesis determined by the increase of cortisol in delivery. The blood constituents of goats showed variations in the period to 75 days postpartum as a result physiological causes related to delivery and postpartum.

Glicemia, proteinograma e perfil de alguns componentes bioquímicos séricos de cabritos da raça Bôer / Glycemic, protein profile and serum biochemical profile in Bôer goat kids

Os cabritos recém-nascidos sofrem diversas adaptações à vida extrauterina que afetam diferentes funções orgânicas, pois precisam produzir calor, iniciar a atividade muscular e buscar alimento. Todos estes eventos levam a modificações em diversos constituintes sanguíneos, como proteínas e parâmetros bioquímicos séricos. Outros estudos demonstraram estas variações, mas sem se estender além do período neonatal até a fase de animais jovens. Objetivou-se testar a hipótese de que ocorre variação doproteinograma, dos componentes bioquímicos e da glicemia de cabritos desde o nascimento até 75 dias de vida em função da adaptação à vida extrauterina. Para atingir estes objetivos, foram colhidas amostras de sangue de 25 cabritos nascidos de partos normais, independentemente do sexo. As variáveis séricas proteína total (PT), albumina, α-globulina, β-globulina, γ-globulina, onde se incluem a imunoglobulina G (IgG), aspartatoaminotransferase (AST), fosfatase alcalina (FA), gamaglutamiltransferase (GGT), creatinina, ureia, e glicemia foram determinadas nos momentos zero (logo após o nascimento), dois, sete, 15, 30 e 75 dias pós-parto. Observaram-se diferenças significativas em todas as variáveis entre os momentos, mas somente a concentração de creatinina foi maior que aquelas dos demais momentos, nos tempos zero e de dois dias pós-nascimento, devido, provavelmente, à imaturidade da função renal em animais neonatos. Os constituintes sanguíneos dos cabritos tiveram variações no período avaliado, relacionados a causas fisiológicas e nutricionais.

The newborn goat kids suffer several adjustments to life outside the uterus that affect different body functions that they need to produce heat, muscle activity and start searching for food. All these events lead to changes in several blood constituents such as proteins and serum biochemical parameters. Other studies have shown these variations, but not extend beyond the neonatal period to phase in young animals. The aim was to test the hypothesis that there is variation of the protein profile, and biochemical components of blood glucose in goat kids from birth to 75 days of life in terms of adaptation to extra uterine life. To achieve these objectives have been collected blood samples from 25 goats born by normal delivery, regardless of their sex. The variables serum total protein (TP), albumin, α-globulin, β-globulin, γ-globulin, which includes the immunoglobulin G (IgG), aspartate (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine and urea, and glucose were determined at times zero (immediately after birth), two, seven, 15, 30 and 75 days old. We observed significant differences in all variables between times, but only the creatinine concentration was higher than those of other moments in time zero and two days old, probably due to immaturity of renal function in newborn animals. The blood constituents of the kids had variations in the study period, related to physiological and nutritional causes.

Estudo soroepidemiológico de leishmaniose em equinos na região de Araçatuba-SP, Brasil, área endêmica para leishmaniose visceral / A seroepidemiological study of leishmaniasis in horses in the region from Araçatuba-SP, Brazil, an endemic area for visceral leishmaniasis

O presente estudo teve como objetivo pesquisar a presença de anticorpos anti-Leishmania sp. em equinos provenientes do município de Araçatuba-São Paulo, área endêmica para leishmaniose visceral canina e humana. Dos 466 equinos testados para a presença de IgG anti-Leishmania chagasi pelo método de ELISA, 68 (14,59%) apresentaram sororeatividade, com títulos variando entre 0,324 e 0,813. As amostras de soro dos equinos positivos por ELISA foram testadas também por imunocromatografia, e 19/466 (4,08%) apresentaram resultado positivo. Os resultados obtidos no presente estudo indicaram que esta espécie animal está em contato e deve atrair flebotomíneos transmissores da doença, e salientam a necessidade de investigação mais acurada sobre o real papel dos equinos residentes em áreas endêmicas, de forma a auxiliar nas medidas de controle da doença.

The aim of the present study was to perform a leishmaniasis survey in horses from Araçatuba, São Paulo, an endemic area of Brazil. Of the 466 horses tested for the presence of anti-Leishmania chagasi titers by ELISA, 68 (14.59%) were seropositive, with titers varying between 0.324 and 0.813. ELISA positive samples were also tested by immunocromatography and 19/466 (4.08%) were positive. The results of the present study indicated that equines are in contact and can attract phlebotomines, and highlight the necessity of a more accurate investigation on the role played by the horses living in endemic areas, in order to help to control the spread of the illness.

Erythrogram and erythrocytes measurement of ostriches (Struthio camelus) in São José do Rio Preto-SP, Brazil / Eritrograma e medição dos eritrócitos de avestruzes (Struthio camelus) em São José do Rio Preto-SP, Brasil

Ostriches’ erythrocytic parameters help on the diagnosis of specific pathologies and serve as basic knowledge for studies incomparative avian pathology. To obtain reference values of erythrocyte indices for ostriches (Struthio camelus) raised in acommercial system in Brazil and verify if there are differences between gender and age groups, 240 healthy from both sexesanimals were bled. Heparinized blood samples were analyzed using standard techniques to determine the red blood cell(RBC) count, hemoglobin concentration, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscularhemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) values, besides the red cell morphometryanalysis using a computer software that calculates the greater and the smaller diameters of the erythrocytes. Prior to dataanalysis, ostriches were divided into three different age groups: from four to 13 months, from 13 to 23 months and from23 to 30 months the. Younger ostriches presented lower erythrocyte indices than the older ones. The age group effect wasonly significant in females for the erythrocyte, hemoglobin, MCV, MCH, and MCHC variables. The female ostrichespresented PCV, MCV, MCH, and MCHC values significantly higher than males in some age groups. It was observed thatthe erythrocytes of the female ostriches are more elongated and larger than males. It was concluded that erythrocyticparameters of ostriches in São José do Rio Preto-SP, Brazil are influenced by gender and age, highlighting the importanceof consider besides these factors also the geoclimatic conditions to an adequate interpretation of the erythrogram.

Os parâmetros eritrocitários de avestruzes auxiliam no diagnóstico de afecções específicas, além de servir comoconhecimento básico no estudo de patologia comparativa das aves. Para obtenção de valores de referência dos índiceseritrocitários de avestruzes (Struthio camelus) criados em um sistema comercial no Brasil e verificar se existemdiferenças relativas ao sexo e faixas etárias, foram colhidas amostras sanguíneas de 240 animais saudáveis e de ambos ossexos. Amostras sanguíneas heparinizadas foram analisadas utilizando técnicas-padrão para determinar a contagem deeritrócitos, concentração de hemoglobina, hematócrito, volume corpuscular médio (VCM), hemoglobina corpuscularmédia (HCM), concentração de hemoglobina corpuscular média (CHCM) além da análise morfométrica dos eritrócitosutilizando um programa de computador que calcula os diâmetros maior e menor dos eritrócitos. Para a análise dos dadosos avestruzes foram divididos em três diferentes faixas etárias: de quatro a 13 meses; de 13 a 23 meses e de 23 a 30 meses.De modo geral, avestruzes jovens apresentaram índices eritrocitários inferiores aos dos animais mais velhos. Diferençasrelativas à idade só foram significativamente relevantes em fêmeas para os valores de eritrócitos, hemoglobina, VCM,HCM e CHCM. As avestruzes fêmeas apresentaram valores de hematócrito, VCM, HCM e CHCM significantementemaiores que os machos em algumas faixas etárias. Os eritrócitos de avestruzes fêmeas são mais compridos e largosdo que os de machos. Pode-se concluir que os parâmetros eritrocitários de avestruzes em São José do Rio Preto-SP, Brasil estão sob influência do sexo e da idade, ressaltando a importância de considerar além desses fatores, também ascondições geoclimáticas para uma interpretação adequada do eritrograma.

Araçatuba virus: a vaccinialike virus associated with infection in humans and cattle.

We describe a vaccinialike virus, Araçatuba virus, associated with a cowpoxlike outbreak in a dairy herd and a related case of human infection. Diagnosis was based on virus growth characteristics, electron microscopy, and molecular biology techniques. Molecular characterization of the virus was done by using polymerase chain reaction amplification, cloning, and DNA sequencing of conserved orthopoxvirus genes such as the vaccinia growth factor (VGF), thymidine kinase (TK), and hemagglutinin. We used VGF-homologous and TK gene nucleotide sequences to construct a phylogenetic tree for comparison with other poxviruses. Gene sequences showed 99% homology with vaccinia virus genes and were clustered together with the isolated virus in the phylogenetic tree. Araçatuba virus is very similar to Cantagalo virus, showing the same signature deletion in the gene. Araçatuba virus could be a novel vaccinialike virus or could represent the spread of Cantagalo virus.

Avaliação da transferência de imunidade passiva em cordeiros com até 30 dias de idade / Passive transfer immunity valuation in lambs with until 30 days of life

O objetivo deste estudo foi determinar as concentrações séricas de imunoglobulina G, PT e de sua fração eletroforética gama globulina, assim como a atividade enzimática de GGT e ALP. Estabeleceu-se a possibilidade da atividade destas enzimas ser utilizada indiretamente como indicadoras de um adequado nível de proteção humoral e/ou como falha de transferência de imunidade passiva em cordeiros. Foi pesquisada a correlação entre as variáveis em quatro grupos de idade divididos em até 30 dias de vida. Para tanto, foram coletadas 191 amostras de sangue de cordeiros em 21 propriedades da região de Araçatuba - São Paulo. Realizaram-se os testes de imunodifusão radial, espectrofotometria e eletroforese para obtenção dos valores de IgG, PT e γ globulina, respectivamente. Para os valores da atividade de GGT e ALP, foram utilizados kits comerciais. Notou-se correlação significativa entre ALP e GGT, fato observado também com PT, IgG e GGT. A gama-globulina mostrou-se correlacionada com GGT, IgG e PT. A atividade de FA não deve ser utilizada como indicadora de transferência de imunidade passiva em cordeiros.

The aim of this study was to determine the serum concentrations of immunoglobulin G, TP and the gamma globulin electrophoretic fraction, as well as the enzymatic activities of GGT and ALP. This was done in order to explore the possibility of using changes in activities of these enzymes as indirect indicators of adequate humoral immunity and/ or failure of immune passive transfer in lambs. Pearson correlation was performed between variable of four groups of age during the first 30 days of life. Blood samples (191) from lambs from 21 different farms in Araçatuba region- São Paulo. The following tests were perfomed: radial immunodiffusion, spectrophotometry and electrophoresis to stain the amount of IgG, TP and gamma globulin, respectively. GGT and ALP values were determinate using commercial kits. There was a statistically significant correlation between ALP and GGT. The same correlation was observed from TP, IgG and GGT. A positive gamma globulin correlation was found between GGT, IgG and TP. ALP activity cannot be used as an indicator of immune passive transfer.

Concentração de imunoglobulinas G e M no soro sanguíneo de bezerros da raça Holandesa até os 90 dias de idade / Serum immunoglobulins G and M concentration in the serum of holstein calves until 90 days old

Avaliou-se os níveis de imunoglobulinas G e M no soro sangüíneo de 32 bezerros da raça Holandesa desde o nascimento até os 90 dias de vida, levando-se em consideração a forma de ingestão (mamada natural na mãe ou em mamadeira) e a quantidade de colostro administrado. Os animais foram distribuídos em quatro grupos. O grupo 1 recebeu quatro litros de colostro fornecido na mamadeira; o grupo 2 recebeu dois litros de colostro também fornecido através da mamadeira; o grupo 3 ingeriu colostro diretamente nas mães e o grupo 4 foi tratado exclusivamente com leite administrado através de mamadeira. Os resultados obtidos para as concentrações de IgG sérica não apresentaram diferenças estatísticas significativas quanto a forma de fornecimento do colostro (mamada na mãe ou em mamadeira), porém os níveis dessa imunoglobulina sempre foram mais elevados nos animais que mamaram nas mães, seguidos pelos que receberam quatro litros de colostro. Nos animais que não mamaram colostro, a produção de imunoglobulinas G e M é mais precoce, respectivamente, a partir de 30 e 15 dias de vida.

This study evaluated the G and M concentrations in serum of 32 calves until 90 days old, considering the way (direct suckling from mother or by bottle) and the volume of ingested colostrum. The calves were divided into 4 groups. Group 1: four liters of colostrum fed by bottle; Group 2: two liters of colostrum fed by bottle; Group 3: direct suckling from mother and and Group 4: two liters of milk by bottle. Serum IgG concentrations obtained in this study had no significant difference in the way of colostrum feeding (direct suckling from mother or by bottle), but their levels were higher in the former way of administration of colostrum, followed by those which received 4 liters of colostrum. Those animals which were not fed with colostrum presented IgG and IgM levels earlier, from 30 and 15 days old, respectively.