RESUMO: O objetivo do presente estudo foi caracterizar (fenotipicamente e genotipicamente) duas cepas de Brucella abortus identificadas como pertencentes ao biovar 4 isolada de bovinos no Brasil. As cepas foram isoladas de bursite cervical de bovinos dos estados do Pará e Rio Grande do Sul, respectivamente. Na identificação fenotípica, os isolados foram positivos na exigência de CO2, produziram H2S, foram resistentes à fucsina básica (20 µg / mL) e sensíveis à tionina (20 µg / mL e 40 µg / mL) e apresentaram antígeno de superfície M, mas o antígeno de superfície A foi ausente. Os isolados foram positivos na PCR para o gene bcsp31 (gênero específico) e na PCR - AMOS, ambos os isolados apresentaram perfil de banda consistente com B. abortus biovar 1, 2 ou 4. Além disso, ambos os isolados também apresentaram padrões de restrição idêntica à cepa de referência quando testada pelo omp2b PCR-RFLP. Na genotipagem usando Multiple Locus Variable Number of Tandem Repeat (VNTR) - MLVA (MLVA16), os isolados apresentaram diferenças em vários loci (Bruce42, Bruce19, Bruce04, Bruce16 e Bruce30); no Multiple Locus Sequence Typing (MLST), os isolados também exibiram diferenças na sequência tipo (ST), amostra 16/02 ST1 (2-1-1-2-1-3-1-1-1) e amostra 128/11 ST (22-1-1-8-9-3-1-1-1). A extensa tipagem de cepas de B. abortus isoladas de bovinos no Brasil por diferentes abordagens confirmou a rara ocorrência de B. abortus biovar 4 no país.

ABSTRACT: The aim of the present study was to characterize (phenotypically and genotypically) two strains of Brucella abortus identified as belonging to biovar 4 isolated from cattle in Brazil. The strains were isolated from cervical bursitis from cattle in the states of Pará and Rio Grande do Sul, respectively. In the phenotypic identification, the isolates were positive in CO2 requirement, produced H2S, were resistant to basic fuchsin (20 µg / mL) and sensitive to thionin (20 µg / mL and 40 µg / mL) and presented M surface antigen, but A surface antigen is absent. The isolates were positive in the PCR for the bcsp31 gene (genus-specific) and in the AMOS-enhanced PCR, both isolates showed a band profile consistent with B. abortus biovar 1, 2 or 4. Moreover, both isolates also showed restriction patterns identical to the reference strain when tested by the omp2b PCR-RFLP. In genotyping using Multiple Locus Variable Number of Tandem Repeat (VNTR) Analysis - MLVA (MLVA16), the isolates showed differences in several loci (Bruce42, Bruce19, Bruce04, Bruce16 and Bruce30); by Multiple Locus Sequence Typing (MLST), they also exhibited differences in sequence type (ST), strain 16/02 ST1 (2-1-1-2-1-3-1-1-1) and strain 128/11 ST (22-1-1 -8-9-3-1-1-1). The extensive typing of B. abortus strains isolated from cattle in Brazil using different approaches confirmed the occurrence of rare B. abortus biovar 4 in the country.

RESUMO: Os objetivos do presente estudo foram (i) genotipar amostras de Corynebacterium pseudotuberculosis, C. silvaticum e C. auriscanis usando Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR), bem como (ii) analisar as relações epidemiológicas entre os isolados de acordo com biovar (Equi e Ovis), espécie, hospedeiro e origem geográfica das amostras de C. pseudotuberculosis. Sessenta e oito isolados de C. pseudotuberculosis, nove C. silvaticum, um C. auriscanis, C. pseudotuberculosis ATCC® 19410 ™ e a amostra vacinal atenuada C. pseudotuberculosis 1002 foram tipificadas por ERIC 1 + 2-PCR. As amostras de campo foram isoladas de diferentes hospedeiros (bovinos, búfalos, ovinos, caprinos, equinos, cães e suínos) em seis países (México, Portugal, Brasil, Guiné Equatorial, Egito e Israel). Uma alta diversidade genética foi observada entre os isolados de Corynebacterium spp., agrupados em vinte e quatro genótipos com um índice de diversidade Hunter & Gaston (HGDI) de 0,937. A análise da minimal spanning tree (MST) de Corynebacterium spp. revelou três complexos clonais, cada um associado a uma espécie bacteriana. Vinte e dois genótipos foram observados entre isolados de C. pseudotuberculosis, com um HGDI de 0,934. Na análise da MST, três grandes complexos clonais foram formados, agrupando-se em torno da origem geográfica dos isolados de C. pseudotuberculosis. Esses resultados reforçam a alta tipabilidade, concordância epidemiológica e poder discriminatório do ERIC-PCR como método consistente de genotipagem para C. pseudotuberculosis, podendo ser útil como ferramenta epidemiológica no controle da linfadenite caseosa. Além disso, os resultados também indicam o grande potencial de ERIC 1 + 2-PCR para genotipagem de espécies do gênero Corynebacterium além de C. pseudotuberculosis.

ABSTRACT: The aims of the present study were (i) to genotype Corynebacterium pseudotuberculosis, C. silvaticum, and C. auriscanis strains using enterobacterial repetitive intergenic consensus (ERIC-PCR), and (ii) to analyze the epidemiological relationships among isolates according to biovar (Equi and Ovis), species, host, and geographical origin of the C. pseudotuberculosis strains. Sixty-eight C. pseudotuberculosis, nine C. silvaticum, and one C. auriscanis, C. pseudotuberculosis ATCC® 19410™ strain and the attenuated C. pseudotuberculosis 1002 vaccinal strain were fingerprinted by ERIC 1+2-PCR. Field strains were isolated from various hosts (cattle, buffaloes, sheep, goats, horses, dogs, and pigs) in six countries (Mexico, Portugal, Brazil, Equatorial Guinea, Egypt, and Israel). High genetic diversity was found among the studied Corynebacterium spp. isolates, clustering in 24 genotypes with a Hunter & Gaston diversity index (HGDI) of 0.937. The minimal spanning tree of Corynebacterium spp. revealed three clonal complexes, each associated with one bacterial species. Twenty-two genotypes were observed among C. pseudotuberculosis isolates, with an HGDI of 0.934. Three major clonal complexes were formed at the minimal spanning tree, grouped around the geographic origin of C. pseudotuberculosis isolates. These results reinforce the high typeability, epidemiological concordance, and discriminatory power of ERIC-PCR as a consistent genotyping method for C. pseudotuberculosis, which could be useful as an epidemiological tool to control caseous lymphadenitis. Moreover, our results also indicate the potential of ERIC 1+2-PCR for the genotyping of other species of Corynebacterium other than C. pseudotuberculosis.

Abstract The chemotaxis of C. fetus subsp. venerealis and C. fetus subsp. fetus was determined in the presence of bovine cervical mucus and bovine placental extract. Some reported substances and ion in those materials, such amino acids, ferrous iron, hormones, sugars and organic acids were also investigated. Bovine cervical mucus, bovine placenta extracts and some substances and ion of these materials namely L–fucose, L– aspartate, L–glutamate, L–serine, ferrous iron, fumarate, pyruvate and succinate were chemoattractants. The chemottraction was significantly larger in higher concentrations of the tested substances and ion and significant differences among tested strains were also observed. Meso-erythritol and hormones bovine placental lactogen, 17β-estradiol, and progesterone did not elicit chemotactical response. In conclusion, this chemotactic behavior may guide the C. fetus navigation in the bovine host's genital tract and be an important cofactor of tissue tropism for this bacterium.

RESUMO: Campilobacteriose Genital Bovina (CGB) é uma doença venérea e subclínica que causa problemas reprodutivos em rebanhos, causada por Campylobacter fetus subsp. venerealis. Este trabalho teve como objetivo selecionar peptídeos miméticos ao agente da CGB de uma biblioteca de fagos. Phage display é uma técnica que aplica bibliotecas de bacteriófagos que expõem peptídeos fusionados ao capsídeo viral em seleções biológicas contra proteínas alvo. Biopannings foram realizados para seleção biológica na biblioteca de fagos por meio de soro hiperimune de coelho e extrato proteico de C. fetus subsp. venerealis. Cinco heptapeptídeos selecionados foram considerados miméticos para Cfv-NCTC 10354 a partir de análises de bioinformática e ensaios com soro hiperimune e muco cérvico-vaginal de novilhas. ALASLPL e LSYLFPP foram os peptídeos mais reativos e considerados promissores como possíveis imunógenos miméticos para C. fetus subsp. venerealis.

ABSTRACT: Bovine genital campylobacteriosis (BGC) is a venereal and subclinical disease that affects the fertility of cattle herds, and it is caused by Campylobacter fetus subsp. venerealis . This study selected peptides mimetic to the BGC-causing agent from a phage library. Phage display is a technique that applies bacteriophage libraries that reveal peptides fused to the viral capsid in biological selections against target proteins. Biopannings were performed for biological selection in the phage library using rabbit hyperimmune serum and C. fetus subsp. venerealis protein extract. Five selected heptapeptides were considered mimetic to Cfv-NCTC 10354 based on the results of bioinformatics analysis and assays with hyperimmune serum and cervicovaginal mucus obtained from heifers. ALASLPL and LSYLFPP were the most reactive peptides and considered promising as possible mimetic immunogens for C. fetus subsp. venerealis.

RESUMO: O objetivo foi determinar os fatores de virulência, os grupos filogenéticos e as possíveis relações entre os patovares e os grupos filogenéticos identificados de cepas de Escherichia coli isoladas de fezes de bezerros bubalinos. Um total de 217 amostras de E. coli foram identificadas a partir de cultura das fezes e submetidas a reação em cadeia da polimerase (PCR) para detecção dos fatores de virulência EAST-1, enterohemolisina, Saa, CNF2, F41, F5, STa, intimina, Stx1 e Stx2. Foram identificadas 134 cepas positivas para um ou mais fatores de virulência: 84isoladas de bezerros bubalinos diarreicos e 50 de bezerros bubalinos saudáveis. Os patovares de E. coli obtidos de fezes diarreicas foram ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), e EAEC (EAST-1+) (33/84). Os patovares isolados de fezes não diarreicas foram NTEC (21/50), STEC (17/50), EHEC (1/50) e EAEC (7/50). Cepas de E. coli positivas para EAST-1 (P = 0,008) e filogrupo C (P = 0,05) foram associadas com a presença de diarreia. A análise de filogrupos revelou que 58,95% dos isolados pertencem ao filogrupo B1, seguido por E (9,70%), B2 (5,90%), C (5,90%), D (5,22%), A (2,24%) e F (1,50%). O filogrupo B1 predomina em cepas de E. coli patogênicas isoladas de bezerros búfalos e o filogrupo C constitui um filogrupo de E. coli patogênica entérica para bezerros.

ABSTRACT: This study aimed to determine the virulence factors, phylogenetic groups, and the relationships between pathovars and phylogenetic groups of E. coli strains isolated from feces of buffalo calves. A total of 217 E. coli strains were obtained from feces after culture and were screened by PCR for detection of virulence factors EAST-1, enterohemolysin, Saa, CNF2, F41, F5, STa, intimin, Stx1 and Stx2. One hundred and thirty-four isolates were positive for one or more virulence factors: eighty-four from diarrheic animals, and fifty from non-diarrheic calves. The pathovars of E. coli identified in diarrheic feces were ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), and EAEC (EAST-1+) (33/84). Pathovars identified in non-diarrheic animals were NTEC (21/50), STEC (17/50), EHEC (1/50) and EAEC (7/50). E. coli strains positive for EAST-1 (P=0.008) and phylogroup C (P = 0.05) were associated with the presence of diarrhea. Phylogenetic analysis showed that 58.95% of the isolates belonged to phylogroup B1, followed by E (9.70%), B2 (5.90%), C (5.90%), D (5.22%), A (2.24%), and F (1.50%). Phylogroup B1 predominated in pathogenic E. coli isolated from water buffalo, and phylogroup C constituted an enteropathogenic E. coli for water buffalo calves.

RESUMO: Este estudo pesquisou genes de virulência, patovares e grupos filogenéticos de amostras de E. coli isoladas de fezes de cães com e sem diarreia. Os genes de virulência e a identificação de grupos filogenéticos foram estudados pela técnica de reação em cadeia da polimerase (PCR). 37 isolados de E. coli foram positivos para pelo menos um fator de virulência na análise de PCR. Destes, 21 (57,8%) foram isolados de fezes de cães com diarreia e 16 (43,2%) de fezes de cães não diarreicos. E. coli enteropatogênica (EPEC) (23/37, 62,2%) foi o patovar mais frequente detectado em fezes de cães e foram classificados principalmente como filogrupos B1 e E. E. coli necrotoxigênica (NTEC) positivos para CNF1 foram detectados (6/37, 16,2%) e classificados como B2 e D. Todas as amostras de E. coli foram negativas quanto à presença de genes de enterotoxinas de E. coli enterotoxigênica (ETEC), mas quatro amostras foram positivas para fimbrias relacionadas ao ETEC, 987P (2) e F18 (2). As amostras de E. coli (STEC) produtora de toxina Shiga foram positivas para a toxina Stx2 (1/37) e Stx2e (1/37), ambas do filogrupo B1. Nossos resultados indicaram que EPEC foi o patovar mais frequente e B1 e E foram os filogrupos mais comuns detectados em amostras E. coli isoladas de fezes de cães diarreicos e não diarreicos.

ABSTRACT: This study identified the virulence genes, pathovars, and phylogenetic groups of Escherichia coli strains obtained from the feces of dogs with and without diarrhea. Virulence genes and phylogenetic group identification were studied using polymerase chain reaction. Thirty-seven E. coli isolates were positive for at least one virulence factor gene. Twenty-one (57.8%) of the positive isolates were isolated from diarrheal feces and sixteen (43.2%) were from the feces of non-diarrheic dogs. Enteropathogenic E. coli (EPEC) were the most frequently (62.2%) detected pathovar in dog feces and were mainly from phylogroup B1 and E. Necrotoxigenic E. coli were detected in 16.2% of the virulence-positive isolates and these contained the cytotoxic necrotizing factor 1 (cnf1) gene and were classified into phylogroups B2 and D. All E. coli strains were negative for the presence of enterotoxigenic E. coli (ETEC) enterotoxin genes, but four strains were positive for ETEC-related fimbriae 987P and F18. Two isolates were Shiga toxin-producing E. coli strains and contained the toxin genesStx2 or Stx2e, both from phylogroup B1. Our data showed that EPEC was the most frequent pathovar and B1 and E were the most common phylogroups detected in E. coli isolated from the feces of diarrheic and non-diarrheic dogs.

RESUMO: Com o objetivo de detectar e identificar Campylobacter spp. em carcaças de frango de corte utilizando três metodologias distintas - ensaio imunoenzimático VIDAS®30, Reação em Cadeia da Polimerase (PCR) e PCR em tempo real - foram coletadas e analisadas 43 carcaças de frango resfriadas e 43 congeladas. Quatro carcaças refrigeradas (4,6%) foram consideradas positivas para Campylobacter spp. pelo VIDAS®30 e nenhuma amostra positiva foi identificada quando utilizada a técnica de PCR. Porém, ao analisar as carcaças pela metodologia da PCR em tempo real, foi observada uma maior incidência de Campylobacter spp., com 45 amostras (52,3%) positivas, sendo que Campylobacter jejuni foi a espécie mais frequentemente encontrada nas amostras (88,8%). Não foi observada diferença na frequência do micro-organismo entre carcaças de frangos resfriadas e congeladas. Concluiu-se que a técnica de PCR em tempo real apresentou maior sensibilidade na detecção de Campylobacter spp. em carcaças de frangos de corte e que foi encontrada elevada presença de carcaças contaminadas, especialmente por C. jejuni.

ABSTRACT: In order to detect and identify Campylobacter spp. in broiler chicken carcasses, and to compare detection methods, 43 chilled and 43 frozen carcasses were collected and analyzed. Three methodologies were evaluated: an automated Enzyme Linked Fluorescent Assay (ELFA) VIDAS®30, Polymerase Chain Reaction (PCR) and real-time PCR. Only four chilled carcasses (4.6%) were considered positive for Campylobacter spp. by VIDAS®30 and no sample was positive when the conventional PCR technique was used. However, real-time PCR showed a higher incidence of contamination by Campylobacter spp. in broiler carcasses, with 45 (52.3%) positive samples. C. jejuni was the species most frequently reported in the samples (88.8%). No differences in the frequencies of Campylobacter spp. were observed between the chilled and frozen broiler carcasses. In conclusion, real-time PCR was the most sensitive method for the detection of Campylobacter spp. in chilled or frozen broiler carcasses, which were mainly contaminated by C. jejuni.

Um inquérito sorológico em quatis (Procyonidae: Nasua nasua) e lobinhos (Canidae: Cerdocyon thous) de vida livre foi conduzido na sub-região da Nhecolândia do Pantanal Sulmatogrossense para avaliar a presença de anticorpos anti-Brucella (fenótipo liso). A pesquisa de anticorpos anti-Brucella foi realizada pelo teste do antígeno acidificado tamponado, utilizado como teste de triagem e pelo teste de polarização fluorescente, utilizado como teste confirmatório. A frequência de animais soropositivos para anticorpos anti-Brucella foi de 13,2% (5/38, IC 95%: 4,4% - 28,1%) para lobinhos e de 8,8% (3/34, IC 95%: 1,9% - 23,7%) para coatis. Não foi encontrada associação entre a soropositividade para brucelose e gênero ou idade. Os resultados deste estudo sugerem exposição a ou infecção de lobinhos e quatis do Pantanal Brasileiro por Brucella spp

A serological survey in free-ranging crab-eating foxes (Canidae: Cerdocyon thous) and brown-nosed coatis (Procyonidae: Nasua nasua) was performed in the Nhecolândia sub-region of the Brazilian Pantanal to evaluate the presence of anti-smooth Brucella antibodies on those wild populations. The detection of anti-smooth Brucella antibodies was performed by the Rose Bengal Test (RBT) as screening test and the Fluorescence Polarization Assay (FPA) as a confirmatory test. The frequency of smooth Brucella seropositive animals were 13.2% (5/38, 95% CI: 4.4% - 28.1%) for crab-eating foxes and 8.8% (3/34, 95% CI: 1.9% -23.7%) for brown-nosed coatis. No association was found between seropositivity for brucellosis and gender or age. The results of this study suggest exposure to or infection of crab-eating fox and brown-nosed coati from the Brazilian Pantanal by Brucella spp

Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.

We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

O conhecimento sobre a fisiologia e regulação dos eletrólitos e do equilíbrio ácido-base é necessário, pois a saúde do animal depende diretamente da composição normal dos fluídos nos diversos compartimentos corporais. Esta regulação está intimamente entrelaçada em um rigoroso e preciso controle, de modo que uma anormalidade em um destes compartimentos reflete na alteração dos demais. A avaliação do equilíbrio eletrolítico e ácido-base pode fornecer informações fundamentais para o diagnóstico, prognóstico e decisão terapêutica de diversas enfermidades nos bovinos, tais como diarreias neonatais, hemoparasitoses (anaplasmose e a babesiose), deslocamento de abomaso e processos de sépticos. As correções destas alterações com medidas terapêuticas adequadas são pontos chave para minimizar as perdas por mortalidade causadas por estas enfermidades.

The knowledge of the physiology and regulation of electrolytes and blood gas parameters is necessary because the health of the animal depends directly on the normal composition of fluid in body compartments. The regulation of these is closely intertwined with a rigorous and precise control so that an abnormality in one of these compartments reflects the change in the other. The evaluation of electrolyte and acid-base balance can provide key information for diagnosis, prognosis and therapeutic decision for several diseases in animals such as neonatal diarrhea, anaplasmosis and babesiosis, displacement of abomasum and septic processes. The corrections of these changes with appropriate treatment are key to minimizing losses due to mortality in these diseases.

Aborto infeccioso é uma causa significativa de falhas reprodutivas e perdas econômicas na bovinocultura. O objetivo deste estudo foi detectar ácidos nucleicos de vários agentes infecciosos reconhecidos como causadores de aborto, incluindo-se Arcanobacterium pyogenes, Herpesvirus bovino tipo 1, Brucella abortus, Campylobacter fetus subsp. venerealis, Chlamydophila abortus, Leptospira sp., Listeria monocytogenes, Salmonella sp., Mycoplasma bovis, Mycoplasma bovigenitalium, Neospora caninum e Tritrichomonas foetus. Homogenados de tecidos de 42 fetos e tecidos incluídos em parafina de 28 fetos e 14 placentas/endométrio foram incluídos neste estudo. Brucella abortus foi detectada em 14,2% (12/84) das amostras. DNA de Salmonella sp. foi amplificado de dois fetos e houve um feto positivo para Neospora caninum e outro para Listeria monocytogenes. Essa metodologia baseada em PCR resultou na identificação da etiologia em 19% das amostras ou 20% se considerados somente os tecidos fetais.

Infectious abortion is a significant cause of reproductive failure and economic losses in cattle. The goal of this study was to detect nucleic acids of several infectious agents known to cause abortion including Arcanobacterium pyogenes, Bovine Herpesvirus 1, Brucella abortus, Campylobacter fetus subsp. venerealis, Chlamydophila abortus, Leptospira sp., Listeria monocytogenes, Salmonella sp., Mycoplasma bovis, Mycoplasma bovigenitalium, Neospora caninum, and Tritrichomonas foetus. Tissue homogenates from 42 fetuses and paraffin-embedded tissues from 28 fetuses and 14 placentas/endometrium were included in this study. Brucella abortus was detected in 14.2% (12/84) of the samples. Salmonella sp. DNA was amplified from 2 fetuses, and there was one positive for Neospora caninum, and another for Listeria monocytogenes. This PCR-based approach resulted in identification of the etiology in 19% of samples, or 20% if considered fetal tissues only.

A maioria dos estudos epidemiológicos envolvendo Haemophilus parasuis isolados de rebanhos suínos brasileiros se baseia em sorotipagem. Entretanto, uma alta porcentagem de amostras não é sorotipável. Técnicas moleculares têm sido utilizadas com sucesso para caracterizar a diversidade dos isolados de H. parasuis e a epidemiologia das infecções por esse agente dentro e entre rebanhos. Esta revisão enfoca aspectos gerais da infecção por H. parasuis, principalmente em relação às técnicas de epidemiologia molecular.

The majority of epidemiological studies involving Haemophilus parasuis from Brazilian herds are based on serotyping. However, a high percentage of isolates are non-typable. Recently, molecular-based techniques were successfully used to characterize the diversity of H. parasuis isolates and the epidemiology of H. parasuis infections within and among herds. This review tackles general aspects of H. parasuis infection, mainly regarding to techniques of molecular epidemiology.

Foi estudado o perfil de susceptibilidade aos antimicrobianos de 25 amostras de Campylobacter sp isoladas de bezerros com e sem diarréia (7 C. coli, 16 C. fetus e 2 C. jejuni). Foram testados pelo método de difusão 11 agentes antimicrobianos: amicacina, ampicilina, canamicina, cloranfenicol, eritromicina, gentamicina, neomicina, nitrofurantoína, penicilina G, tetraciclina e sulfametoxazole-trimetoprim. Todas as amostras de Campylobacter sp foram susceptíveis a amicacina, ampicilina, cloranfenicol, eritromicina, gentamicina, neomicina e nitrofurantoína. Três amostras foram moderadamente sensíveis à canamicina (2 C. coli e 1 C. fetus). Todas as amostras foram resistentes à penicilina G. Duas amostras de C. fetus foram moderadamente sensíveis a sulfametoxazole-trimetoprim e 1 C. coli, 9 C. fetus e 2 C. jejuni foram resistentes. Duas amostras de C. fetus foram moderadamente sensíveis à tetraciclina e 3 de C. coli, 2 de C. fetus e 1 de C. jejuni foram resistentes. Onze amostras apresentaram multirresistência (2 C. coli, 8 C. fetus e 1 C. jejuni). Não houve correlação entre resistência de amostras de Campylobacter sp aos antimicrobianos e a ocorrência de diarréia nos bezerros. A frequência de resistência e, principalmente, a multirresistência encontradas nas amostras Campylobacter sp isoladas de bezerros em Minas Gerais, Brasil, foram altas. O perfil de resistência de amostras de Campylobacter sp observado está relacionado aos agentes antimicrobianos mais utilizados em bovinos no Brasil.

The antimicrobial susceptibility of 25 Campylobacter sp strains isolated from calves with and without diarrhea - 7 C. coli, 16 C. fetus and 2 C. jejuni was studied by the disk diffusion method. Eleven antimicrobial agents were tested amikacin, ampicillin, kanamycin, chloramphenicol, erythromycin, gentamicin, neomycin, nitrofurantoin, penicillin G, tetracycline and sulfamethoxazole-trimethoprim. All Campylobacter sp strains were susceptible to amikacin, ampicillin, chloramphenicol, erythromycin, gentamicin, neomycin and nitrofurantoin. Three strains were moderately susceptible to kanamycin (2 C. coli and 1 C. fetus). All the strains were resistant to penicillin G. Two C. fetus strains were moderately susceptible to sulfamethoxazole-trimethoprim and 1 C. coli, 9 C. fetus and 2 C. jejuni strains were resistant. Two C. fetus strains were moderately susceptible to tetracycline and 3 C. coli, 2 C. fetus and 1 C. jejuni strains were resistant. Eleven strains showed multidrug resistance (2 C. coli, 8 C. fetus and 1 C. jejuni). There was no correlation between resistance of Campylobacter sp strains to antimicrobials and the occurrence of diarrhea in calves. The frequency of resistance and, most importantly, multi drug resistance found among Campylobacter sp strains isolated from calves in Minas Gerais, Brazil, were high and the patterns of resistance observed are related to the antimicrobials agents most largely used in cattle in Brazil.

Polyvinyl alcohol (PVA), PVA crosslinked with glutaraldehyde hydrogels (PVA/GA), PVA with tetraethylorthosilicate (PVA/TEOS) and PVA/GA/TEOS hybrids with recombinant MPB70 protein (rMPB70) incorporated were chemically characterized by Fourier transform infrared spectroscopy (FTIR). FTIR spectra of PVA hydrogel samples showed the absorption regions of the specific chemical groups associated with poly(vinyl alcohol) (-OH, -CO, -CH2) and PVA/GA confirming the formation of crosslinked hydrogel (duplet -CH). It was observed C-H broad alkyl stretching band (n = 2850-3000 cm-1) and typical strong hydroxyl bands for free alcohol (nonbonded -OH stretching band at n = 3600-3650 cm-1), and hydrogen bonded band (n = 3200-3570 cm-1). The most important vibration bands related to silane alcoxides have been verified on FTIR spectra of PVA/TEOS and PVA/GA/TEOS hybrids (Si-O-Si, n = 1080 and n = 450 cm-1; Si-OH, n = 950 cm-1). FTIR spectra of f PVA hydrogel with rMPB70 incorporated have indicated the specific groups usually found in protein structures, such as amides I, II and III, at 1680-1620 cm-1, 1580-1480 cm-1 and 1246 cm-1, respectively. These results have given strong evidence that recombinant protein rMPB70 was successfully adsorbed in the hydrogels and hybrids networks. These PVA based hydrogels and hybrids were further used in immunological assays (Enzyme-Linked Immunosorbent Assay - ELISA). Tests were performed to detect antibodies against rMPB70 protein in serum samples from bovines that were positive in the tuberculin test. Corresponding tests were carried out without PVA samples in microtiter plates as control. Similar results were found for commercially available microplates and PVA based hydrogels and hybrids developed in the present work regarding to immunoassay sensitivity and specificity response.