Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region / Diversidade genética e levantamento molecular de Trypanosoma (Megatrypanum) theileri em bovinos na região da Amazônia Ocidental do Brasil

Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region.

Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Identification of Pasteurella multocida transcribed genes in porcine lungs through RNAseq.

Pasteurella multocida is one of the most important pathogen that causes pneumonia in swine. Although several virulence factors are known, the pathogenesis of this bacterium is not well-studied. Therefore, to study the pathogenesis of P. multocida infection in porcine lung, next-generation RNA sequencing was used to compare the transcriptomes of P. multocida grown in vivo and in vitro, respectively. After P. multocida infection a total of 704 genes were expressed in vitro, 1422 genes were expressed in vivo, and 237 genes were differentially expressed based on statistical analyses, padj of ≤0.1. Genes encoding ribosomal proteins or other products that function in the regulation of transcription and translation were downregulated, whereas genes whose products affected cellular processes (protein transport and RNA degradation) and metabolic pathways, such as those of amino acid metabolism and nucleotide metabolism, were upregulated in vitro compared with in vivo. This study shows that differentially expressed genes in P. multocida regulate pathways that operate during stress, iron capture, heat shock, and nitrogen regulation. However, extensive investigation of the pathogenic mechanism of P. multocida is still required.

Molecular detection of tick-borne protozoan parasites in a population of domestic cats in midwestern Brazil.

Some tick-borne pathogens that infect domestic cats have been considered emergent in veterinary medicine. Occurrences of Hepatozoon spp., Babesia spp. and Cytauxzoon spp. have been described in several regions of Brazil. This paper offers a comprehensive analysis of the 18S rRNA gene of a Hepatozoon sp. strain detected in domestic cats in the metropolitan area of Cuiabá, in Midwestern Brazil. Based on a molecular analysis, we detected the presence of Hepatozoon species circulating among cats in this region. The aforementioned strain is closely related to other isolates of H. felis detected in wild felids. Moreover, a phylogenetic analysis indicates that this genotype is grouped into a clade of 18S rRNA sequences previously described for the genus Hepatozoon in wild felids around the world. Hepatozoon felis strains detected in cats from Spain and Israel showed, respectively, 98% and 97% identity to our sequence and are clustered on a separate branch of the phylogenetic tree. This finding suggests a high diversity of Hepatozoon genotypes occurring in cats in Europe and South America. None of the analyzed cats were positive for Babesia spp. or Cytauxzoon spp. by PCR analysis.

Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA / Caracterização molecular de isolados de Pasteurella multocida de pulmões de suínos através da técnica Randonly Amplified Polymorphic DNA

ABSTRACT: Swine respiratory diseases such as atrophic rhinitis and bronchopneumonia caused by Pasteurella (P.) multocida cause important economic losses to the modern swine industry. The purpose of this study was to characterize P. multocida strains isolated from swine lungs by RAPD (Randomly Amplified Polymorphic DNA) to demonstrate their genetic diversity. Ninety-four samples of fragments from lungs with pneumonia and sixty one samples without pneumonia were collected in slaughterhouses in Mato Grosso during the period from December 2009 to March 2010. Clinical cases in 2012 and 2013 were also included in this study. Among the lung fragments with macroscopic lesions, without macroscopic lesions and clinical samples, 40.42%, 4.49% and 100% were positive for P. multocida, respectively. Bacterial identification culturing was confirmed by PCR (polymerase chain reaction) by means of the amplification of the gene kmt1. RAPD technique was performed for 46 isolates, and in every isolate, a total of 7 to 11 amplification bands were detected, composed of 8 clusters based on genetic similarity. Thus, treatment, control and preventive measures should consider the genetic diversity of P. multocida populations in swine herds in order to improve the development of new protocols to produce antimicrobials and vaccines.

RESUMO: As doenças respiratórias suínas como a rinite atrófica e broncopneumonia, associada a Pasteurella (P.) multocida causam importantes perdas econômicas na suinocultura moderna. O objetivo deste trabalho foi caracterizar isolados de P. multocida de pulmão suíno através do Randomly Amplified Polymorphic DNA (RAPD) para demonstrar a diversidade genética. Noventa e quatro amostras de fragmentos de pulmões com lesões de pneumonia e sessenta e uma amostras sem lesão foram coletadas em frigoríficos no Estado do Mato Grosso, durante o período de dezembro de 2009 a março de 2010. Amostra de casos clínicos ocorridos em 2012 e 2013 também foram inlcuídos. Amostras de pulmões com lesões macroscópicas, sem lesões macroscópicas e amostras clínicas apresentaram presença de 40,42%, 4,49% e 100% de isolamento para P. multocida, respectivamente. Os isolados foram todos confirmados através da PCR (Polymerase Chain Reaction) pela amplificação do gene kmt1. A técnica de RAPD foi realizada em 46 amostras e em cada isolado foi detectado 7 a 11 bandas, que foram agrupadas em 8 grupos baseados em suas similaridades genéticas. Dessa forma, tratamento, controle e medidas preventivas deveriam considerar a diversidade genética da população de P. multocida em rebanhos suínos para melhorar o desenvolvimento de novos protocolos para produção de antimicrobianos e vacinas.

Prevalência de Streptococcus suis tipo 2 por meio da técnica de reação em cadeia da polimerase em suínos abatidos no Estado do Mato Grosso / Prevalence of Streptococcus suis type 2 using the polimerase chain reaction in slaughtered pigs in the State of Mato Grosso, Brazil

Streptococcus suis é um patógeno que a afeta a produção industrial de suínos em todo o mundo. É de extrema importância, pois está associado a doenças em suínos e humanos. O objetivo deste estudo foi determinar a prevalência do Streptococcus suis tipo 2 em 201 amostras de tonsilas de animais clinicamente sadios a partir da técnica de PCR. As amostras positivas foram submetidas à pesquisa do gene codificador do fator extracelular (ef). Os resultados demonstraram que a prevalência (23,38 por cento) foi maior que em outro estudo recentemente realizado no mesmo Estado, indicando que a PCR é um método mais sensível em relação ao isolamento bacteriano. Houve baixa ocorrência do gene ef* (1,49 por cento), o que mostra uma grande importância para população analisada, pois cepas negativas são potencialmente menos virulentas que cepas positivas.

Streptococcus suis is a pathogen that affects the industrial production of swine worldwide. It is extremely important, because it is associated with pigs and humans diseases. The aim of this study was to determine the prevalence of Streptococcus suis type 2 in 201 samples of tonsils from clinically healthy animals by the PCR technique. The samples positive for S. suis type 2 were tested for the gene encoding extracellular factors (ef). The results showed that the prevalence (23.38 percent) was higher than other recent survey in the State, demonstrating that the PCR is a more sensitive method in relation to the bacterial isolation. There was a low occurrence of ef* gene in samples (1.49 percent) showing great importance to local swine population, because negative strains are potentially less virulent that positive strains.