RESUMO
Campylobacter is regarded as the most common bacterial cause of gastroenteritis throughout the world and most cases of human campylobacteriosis can be traced back to the consumption of poultry meat. In Brazil, few studies evaluated the genetic relatedness among Campylobacter isolates. The aim of this research was to evaluate the genetic diversity of Campylobacter spp. isolated from poultry meat products sold on the retail market in Southern Brazil. The presumptive identification of Campylobacter was performed using traditional microbiological analysis, followed by molecular confirmation by PCR. The genetic diversity of isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Campylobacter spp. was isolated from 91.7% (33/36) of the samples, totaling 48 isolates. Campylobacter jejuni was the most prevalent species isolated (90.8%). PFGE data revealed 26 pulsotypes and 18 PFGE patterns composed of only 1 isolate. Campylobacter isolates exhibited high genetic diversity; however, some clones were recurrent in the poultry meat products sold on the retail market. As the south region of Brazil is an important producer and exporter of chicken meat, our results highlight the need to control this pathogen in the food chain in this area of the world to reduce the risks of exposing consumers to campylobacteriosis.
Assuntos
Campylobacter coli/genética , Campylobacter jejuni/genética , Microbiologia de Alimentos , Variação Genética , Produtos Avícolas/microbiologia , Animais , Brasil , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Galinhas , Eletroforese em Gel de Campo Pulsado/veterinária , Reação em Cadeia da Polimerase/veterináriaRESUMO
Bovine tuberculosis (BTB) remains an important economic and zoonotic problem in Latin America. Traditionally, the fight against BTB is initiated by the implementation of routine diagnostic tests for certification of free properties. The diagnosis of BTB can be made by direct and indirect methods, in which we can mention clinical, post mortem, histopathological, immunological, bacteriological and molecular methods. The renewal of scientific interest in tuberculosis in recent year has led to develop and improve methods of diagnosis, prevention, control and eradication of BTB. The aim of this review is to present and discuss different diagnosis methods of BTB.
Assuntos
Técnicas de Laboratório Clínico/métodos , Tuberculose Bovina/diagnóstico , Animais , BovinosRESUMO
Bovine tuberculosis (BTB) remains an important economic and zoonotic problem in Latin America. Traditionally, the fight against BTB is initiated by the implementation of routine diagnostic tests for certification of free properties. The diagnosis of BTB can be made by direct and indirect methods, in which we can mention clinical, post mortem, histopathological, immunological, bacteriological and molecular methods. The renewal of scientific interest in tuberculosis in recent year has led to develop and improve methods of diagnosis, prevention, control and eradication of BTB. The aim of this review is to present and discuss different diagnosis methods of BTB.(AU)
A tuberculose bovina (BTB) continua sendo um importante problema econômico na América Latina, com potenciais consequências zoonóticas. Tradicionalmente, a luta contra a tuberculose bovina tem sido iniciada pela execução de testes de diagnóstico de rotina para a certificação de propriedades livres da doença. O diagnóstico de BTB pode ser feito através de métodos diretos e indiretos, nos quais podemos citar os métodos clínicos, post mortem, histopatológicos, imunológicos, bacteriológicos e moleculares. A renovação do interesse científico em tuberculose nos últimos anos tem levado à necessidade de desenvolver e melhorar os métodos de diagnóstico, prevenção, controle e erradicação da BTB. O objetivo deste artigo é analisar e discutir sobre os diferentes métodos de diagnóstico de BTB.(AU)
Assuntos
Animais , Bovinos , Técnicas de Laboratório Clínico/métodos , Tuberculose Bovina/diagnósticoRESUMO
Abstract Bovine tuberculosis (BTB) remains an important economic and zoonotic problem in Latin America. Traditionally, the fight against BTB is initiated by the implementation of routine diagnostic tests for certification of free properties. The diagnosis of BTB can be made by direct and indirect methods, in which we can mention clinical, post mortem, histopathological, immunological, bacteriological and molecular methods. The renewal of scientific interest in tuberculosis in recent year has led to develop and improve methods of diagnosis, prevention, control and eradication of BTB. The aim of this review is to present and discuss different diagnosis methods of BTB.
Resumo A tuberculose bovina (BTB) continua sendo um importante problema econômico na América Latina, com potenciais consequências zoonóticas. Tradicionalmente, a luta contra a tuberculose bovina tem sido iniciada pela execução de testes de diagnóstico de rotina para a certificação de propriedades livres da doença. O diagnóstico de BTB pode ser feito através de métodos diretos e indiretos, nos quais podemos citar os métodos clínicos, post mortem, histopatológicos, imunológicos, bacteriológicos e moleculares. A renovação do interesse científico em tuberculose nos últimos anos tem levado à necessidade de desenvolver e melhorar os métodos de diagnóstico, prevenção, controle e erradicação da BTB. O objetivo deste artigo é analisar e discutir sobre os diferentes métodos de diagnóstico de BTB.
Assuntos
Animais , Bovinos , Técnicas de Laboratório Clínico/métodos , Tuberculose Bovina/diagnósticoRESUMO
Heterospermic AI is commonly used in swine despite preventing precise evaluation of individual boar fertility. The present study compared the contribution of four boars (A, B, C and D) for reproductive performance and for paternity using homospermic and heterospermic (AB, AC, AD, BC, BD and CD) AI (n=204 for homospermic AI; n=307 for heterospermic AI). Blood samples from the four boars, from all sows inseminated with heterospermic doses and from the umbilical cords of their piglets, as well as tissue smears from mummified fetuses, were genotyped using single nucleotide polymorphisms (SNPs). Differences among boars were detected for the in vitro oocyte penetration rate and for the number of spermatozoa per oocyte (P<0.05), but not for sperm motility, mitochondrial functionality and integrity of the membrane, acrosome and DNA (P>0.05). Homospermic and heterospermic AI resulted in similar (P>0.05) farrowing rates (90.5% and 89.9%, respectively) and total litter size (12.4±0.4 and 12.7±0.7, respectively). Farrowing rate was lower for Boar B than for Boar C (P<0.05), but no other differences in reproductive performance among boars were observed with homospermic AI. The SNPs determined the paternity of 94.2% of the piglets sired by heterospermic AI. In the AC pool, paternity contribution per boar was similar (P>0.05), but differences between boars occurred in all other pools (P<0.05). Boar D achieved the greatest paternity contribution in all pools and parity categories (nearly 60%), whereas Boar B sired the fewest piglets (at most 40%). Reproductive performance was similar with homospermic and heterospermic AI, but differences in performance among boars undetected with homospermic AI were only evident after genotyping the piglets sired through heterospermic AI.
Assuntos
Inseminação Artificial/veterinária , Mitocôndrias/genética , Paternidade , Reprodução/genética , Espermatozoides , Animais , Feminino , Genótipo , Tamanho da Ninhada de Vivíparos/genética , Masculino , Paridade/genética , Polimorfismo de Nucleotídeo Único , Gravidez , Motilidade dos Espermatozoides , SuínosRESUMO
AIMS: The aim of the present work was to study the in vitro effect of native and recombinant Bauhinia variegata var. variegata lectins in inhibiting early adhesion of Streptococcus mutans, Streptococcus sanguis and Streptococcus sobrinus to experimentally acquired pellicle. METHODS AND RESULTS: Native lectin from B. variegata (BVL) was purified by affinity chromatography of extract of seeds. The recombinant lectin (rBVL-I) was expressed in E. coli strain BL21 (DE3) from a genomic clone encoding the mature B. variegata lectin gene using the vector pAE-bvlI. Recombinant protein deposited in inclusion bodies was solubilized and subsequently purified by affinity chromatography. The rBVL-I was compared to BVL for agglutination of erythrocytes and initial adherence of oral bacteria on a saliva-coated surface. The results revealed that rBVL-I acts similarly to BVL for agglutination of erythrocytes. Both lectins showed adhesion inhibition effect on Step. sanguis, Step. mutans and Step. sobrinus. CONCLUSION: We report, for the first time, the inhibition of early adhesion of oral bacteria by a recombinant lectin. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results support the proposed biotechnological application of lectins in a strategy to reduce development of dental caries by inhibiting the initial adhesion and biofilm formation.
Assuntos
Aderência Bacteriana/efeitos dos fármacos , Bauhinia/química , Escherichia coli/metabolismo , Lectinas/farmacologia , Streptococcus/efeitos dos fármacos , Animais , Biofilmes/efeitos dos fármacos , Cromatografia de Afinidade , Testes de Hemaglutinação , Humanos , Lectinas/isolamento & purificação , Extratos Vegetais/química , Coelhos , Proteínas Recombinantes/farmacologia , Saliva/química , Sementes/química , Streptococcus/fisiologiaRESUMO
The gene expression of Bax, Bcl-2, survivin and p53, following in vitro maturation of equine oocytes, was compared in morphologically distinct oocytes and cumulus cells. Cumulus-oocyte complexes (COC) were harvested and divided into two groups: G1 - morphologically healthy cells; and G2 - less viable cells or cells with some degree of atresia. Total RNA was isolated from both immature and in vitro matured COC and real-time reverse transcription polymerase chain reaction (qRT-PCR) was used to quantify gene expression. Our results showed there was significantly higher expression of survivin (P < 0.05) and lower expression of p53 (P < 0.01) in oocytes compared with cumulus cells in G1. No significant difference in gene expression was observed following in vitro maturation or in COC derived from G1 and G2. However, expression of the Bax gene was significantly higher in cumulus cells from G1 (P < 0.02).
Assuntos
Apoptose/genética , Técnicas de Maturação in Vitro de Oócitos , Oócitos/fisiologia , Animais , Proteínas Reguladoras de Apoptose/genética , Células do Cúmulo/citologia , Células do Cúmulo/fisiologia , Feminino , Regulação da Expressão Gênica , Genes p53 , Cavalos/genética , Oócitos/citologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Proteína X Associada a bcl-2/genéticaRESUMO
Erythropoietin (EPO) gene therapy can be used for several purposes; however, its effects on reproductive performance are unknown. The aim of this study was to evaluate the toxicological effects of non-viral (EPO) gene transfer on sperm motility, viability, morphology and concentration. Rabbit EPO cDNA was cloned into a pTarget mammalian expression vector. Rabbits were administered with: (1) pTarget/EPO vector, (2) recombinant human EPO (rHuEpo) and (3) saline (control). Both pTarget/EPO and rHuEpo significantly increased (P < 0.05) hematocrit levels 1 week after injection and they remained significantly higher than the control for up to 5 weeks (P < 0.05), showing that both EPO treatments were effective in stimulating the production of red blood cells in rabbits. The EPO gene transfer or rHuEPO administration had no significant effect (P > 0.05) on sperm motility, vigor, viability, concentration or morphology in the testis.
Assuntos
Eritropoetina/genética , Terapia Genética/veterinária , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/citologia , Espermatozoides/fisiologia , Animais , Clonagem Molecular , Terapia Genética/métodos , Células HeLa , Humanos , Masculino , Coelhos , TestículoRESUMO
Porcine enzootic pneumonia (PEP), which is caused by the fastidious bacterium Mycoplasma hyopneumoniae, is one of the most economically important diseases in the pig industry worldwide. Commercial bacterins provide only partial protection; therefore, the development of more efficient vaccines against PEP is necessary. In this study, the cellular and humoral immune responses elicited by DNA and recombinant subunit vaccines based on the P37, P42, P46 and P95 antigens of M. hyopneumoniae were evaluated after the intramuscular inoculation of BALB/c mice. The expression of the cytokines INFγ, TNFα and IL1 was evaluated by real-time RT-PCR in splenocytes from vaccinated mice. All antigens delivered as subunit vaccines, especially P42 and P95, and the pcDNA3/P46 DNA vaccine were able to elicit strong immune responses. These vaccines induced cellular immune responses and the production of antibodies able to react with native M. hyopneumoniae proteins. Because both cellular and humoral immune responses were induced, P42 and P95 are promising candidates for a recombinant subunit vaccine and P46 is a promising candidate for a DNA vaccine against PEP.
Assuntos
Antígenos de Bactérias/imunologia , Imunização/métodos , Mycoplasma hyopneumoniae/imunologia , Mycoplasma hyopneumoniae/patogenicidade , Vacinas de DNA/imunologia , Animais , Camundongos , Vacinas de DNA/uso terapêuticoRESUMO
Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), a chronic respiratory disease which causes significant economic losses to the swine industry worldwide. More efficient strategies for controlling this disease are necessary. In this study, we cloned17 genes coding for transmembrane proteins from M. hyopneumoniae, among which six were successfully expressed in Escherichia coli and had their immunogenic and antigenic properties evaluated. All proteins were immunogenic in mice and sera from naturally infected pigs reacted with the recombinant proteins, suggesting that they are expressed during infection. These antigens may contribute for the development of new recombinant vaccines and diagnostic tests against EP.