PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Cattle trypanosomiasis imposes significant economic burdens on the global livestock industry. The causative agents of this disease belong to the protozoan Trypanosoma genus. This study aims to perform detection (parasitological and molecular) and genetic characterization to analyze Trypanosoma spp. in cattle from 15 municipalities in the state of Rio de Janeiro, focusing on the 18S rDNA and Cathepsin-L (CatL) gene of Trypanosoma vivax and Trypanosoma theileri. A total of 389 blood samples from 15 dairy cattle farms in the state of Rio de Janeiro were collected, and DNA was extracted for subsequent PCR amplification of Trypanosoma spp. 18S rDNA and CatL genes. The resulting amplicons underwent sequencing and alignment for phylogenetic analysis, with comparisons made to GenBank isolates. Concerning parasitological analysis, blood smears presented 4.4% of positive cattle (n=17/389) for T. vivax and did not show any trypomastigote forms of T. theileri. The absolute frequency of Trypanosoma spp. through molecular detection targeting 18S rDNA was 11.6% (45/389). However, when performing species-specific PCRs, the T. vivax frequency, determined through CatL gene PCR, was 12.8%, and the T. theileri frequency was 3.6%. Phylogenetic analysis based on 18S rDNA revealed low diversity among T. vivax sequences, suggesting potential host segregation. This study emphasizes the high frequency of positive samples by PCR when compared to direct parasitological exams. Additionally, T. vivax phylogeny targeting 18S rDNA hints at sequence clustering related to host species. Importantly, this investigation unveils, for the first time in Rio de Janeiro’s cattle, the circulation of T. theileri lineage ThI, encompassing genotypes IIB and IF. This discovery expands our understanding of this parasite’s geographical distribution and genetic diversity.

• Cattle trypanosomiasis Trypanosoma spp. genetic diversity phylogenetic analysis protozoan pathogens

Abstract in Portuguese:

A tripanossomíase bovina impõe significativos ônus econômicos à indústria pecuária global. Os agentes causadores dessa doença pertencem a protozoários do gênero Trypanosoma. Objetivou-se, com este estudo, realizar detecção (parasitológica e molecular) e caracterização genética de Trypanosoma spp. em bovinos de 15 municipalidades do estado do Rio de Janeiro, com foco na sequência 18S rDNA e no gene Cathepsin-L (CatL) de Trypanosoma vivax e Trypanosoma theileri. Um total de 389 amostras de sangue de 15 fazendas leiteiras no estado do Rio de Janeiro foram coletadas, e o DNA foi extraído para subsequente amplificação por PCR dos genes 18S rDNA e CatL de Trypanosoma spp. Os amplicons resultantes foram submetidos a sequenciamento e alinhamento para análise filogenética, com comparações realizadas com isolados do GenBank. No que se refere à análise parasitológica, os esfregaços de sangue apresentaram 4,4% de bovinos positivos (n=17/389) para T. vivax e não mostraram nenhuma forma tripomastigota de T. theileri. A frequência absoluta de Trypanosoma spp. através da detecção molecular visando 18S rDNA foi de 11,6% (45/389). No entanto, ao realizar PCRs específicos de espécies, a frequência de T. vivax, determinada por PCR do gene CatL foi de 12,8%, e a frequência de T. theileri foi de 3,6%. A análise filogenética com base no 18S rDNA revelou baixa diversidade entre as sequências de T. vivax, sugerindo uma possível segregação de hospedeiros. Este estudo enfatiza a alta frequência de amostra positiva pela PCR quando comparada com a parasitológica direta. Além disso, a filogenia de T. vivax direcionada ao 18S rDNA sugere agrupamento de sequências relacionado à espécie hospedeira. Importante destacar que esta investigação revela, pela primeira vez no gado do Rio de Janeiro, a circulação da linhagem ThI de T. theileri, abrangendo os genótipos IIB e IF. Esta descoberta amplia nosso entendimento sobre a distribuição geográfica e diversidade genética desse parasito.

• Tripanossomíase bovinos Trypanosoma spp. diversidade genética análise filogenética protozoários patogênicos

Abstract in English:

This study aimed to identify the presence of Trypanosoma vivax DNA in the colostrum of infected goats and to explore the possibility of transmission for neonates fed using colostrum collected from infected goats. We used twelve goats in the final third of gestation with an age of approximately 24 months. Six goats were inoculated intravenously with 0.5mL of blood containing approximately 1.25x105 trypomastigotes of T. vivax, and six remained uninfected. The presence of T. vivax in colostrum was evaluated by Polymerase Chain Reaction (PCR). The possibility of T. vivax transmission by colostrum was assessed by feeding six neonates born of serologically negative goats using colostrum from infected goats. Peripheral blood from neonates was collected daily for thirty days to assess the T. vivax presence through the examination of Giemsa-stained smears of leukocyte layers with the buffy coat technique (BCT) and by PCR. The results of a direct examination of colostrum were negative, but PCR confirmed the presence of T. vivax DNA in all infected goats. Additionally, lactogenic transmission by colostrum was not demonstrated once both BCT and PCR of neonate peripheral blood were negative.

• Experimental infection Trypanosoma vivax goats DNA lactogenic transmission PCR colostrum small ruminants trypanosomosis Brazil

Abstract in Portuguese:

Este estudo teve como objetivo identificar a presença de DNA de Trypanosoma vivax no colostro de cabras infectadas experimentalmente e verificar a possibilidade de transmissão para neonatos alimentados com colostro coletado de cabras infectadas. Foram utilizadas doze cabras no terço final de gestação com idade aproximada de 24 meses. Seis cabras foram inoculadas intravenosamente com 0,5mL de sangue contendo aproximadamente 1,25x105 tripomastigotas de T. vivax, e seis permaneceram não infectadas. A presença de T. vivax no colostro foi avaliada por Reação em Cadeia da Polimerase (PCR). A possibilidade de transmissão de T. vivax pelo colostro foi avaliada através da alimentação de seis neonatos nascidos de cabras sorologicamente negativas com colostro de cabras infectadas. Foi coletado diariamente o sangue periférico dos neonatos, por trinta dias para avaliar a presença de T. vivax através do exame de esfregaços de camadas leucocitárias coradas por giemsa, pela técnica BCT e por PCR. Os resultados do exame direto do colostro foram negativos, mas a PCR confirmou a presença de DNA de T. vivax no colostro em todas as cabras infectadas. Além disso, a transmissão lactogênica pelo colostro não foi demonstrada, uma vez que tanto a BCT quanto a PCR do sangue periférico do neonato foram negativas

• Infecção experimental Trypanosoma vivax caprinos DNA transmissão lactogênica PCR colostro pequenos ruminantes tripanossomíase Brasil

Abstract in English:

ABSTRACT.- Guerra N.R., Monteiro M.F.M., Sandes H.M.M., Cruz N.L.N., Ramos C.A.N., Santana V.L.A., Souza M.M.A. & Alves L.C. 2013. [Detection of IgG antibodies against Trypanosoma vivax in cattle by indirect immunofluorescence test.] Detecção de anticorpos IgG anti-Trypanosoma vivax em bovinos através do teste de Imunofluorescência indireta. Pesquisa Veterinária Brasileira 33(12):1423-1426. Laboratório de Doenças Parasitárias dos Animais Domésticos, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manuel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: leucioalves@gmail.com Trypanosoma vivax infects a wide range of wild and domestic ungulates, causing important losses for the livestock industry. The aim of the present study was to assess the detection of IgG antibodies against T. vivax in cattle from the state of Pernambuco, Brazil. Therefore, we analyzed 2.053 blood serum samples from cattle herds of municipalities in Pernambuco, what was made by Immunofluorescence Assay. The overall seroprevalence of IgG antibodies against T. vivax in cattle was 13.93% (286/2053). The frequencies, by region, varied from 11.90% to 15.99%. Thus, the data obtained allowed to characterize the state of Pernambuco as an area of enzootic instability for T. vivax. The frequency herein reported (i.e., 13.93%) indicates that Pernambuco is an endemic area for T. vivax, this parasite being spread throughout the state.

• Trypanosoma vivax trypanosomiasis tripanossomíase

Abstract in Portuguese:

RESUMO.- Guerra N.R., Monteiro M.F.M., Sandes H.M.M., Cruz N.L.N., Ramos C.A.N., Santana V.L.A., Souza M.M.A. & Alves L.C. 2013. [Detection of IgG antibodies against Trypanosoma vivax in cattle by indirect immunofluorescence test.] Detecção de anticorpos IgG anti-Trypanosoma vivax em bovinos através do teste de Imunofluorescência indireta. Pesquisa Veterinária Brasileira 33(12):1423-1426. Laboratório de Doenças Parasitárias dos Animais Domésticos, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manuel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: leucioalves@gmail.com Trypanosoma vivax infecta uma grande variedade de animais ungulados selvagens e domésticos, podendo causar grande impacto na produção de ruminantes. Este trabalho teve como objetivo avaliar a detecção de anticorpos IgG anti-Trypanosoma vivax em bovinos provenientes do estado de Pernambuco, Brasil. Para tanto, foram analisadas 2,053 amostras de soro sanguíneo de bovinos provenientes de rebanhos de municípios do estado de Pernambuco, os quais foram analisados através da Reação de Imunofluorescência Indireta. Das amostras testadas 13,93% (286/2.053) foram reagentes para anticorpos IgG anti-Trypanosoma vivax. As freqüências, por mesorregião, variaram de 11,90% a 15,99%. Assim, os dados obtidos permitiram a caracterização do estado de Pernambuco como uma área de instabilidade enzoótica e sugere que o estado Pernambuco é área endêmica para Trypanosoma vivax e este parasito está distribuído por todo o estado.

Abstract in English:

Rodrigues A., Fighera R.A., Souza T.M., Schild A.L., Soares M.P., Milano J. & Barros C.S.L. 2005. [Outbreaks of trypanosomiasis in horses by Trypanosoma evansi in the state of Rio Grande do Sul, Brazil: epidemiological, clinical, hematological, and pathological aspects.] Surtos de tripanossomíase por Trypanosoma evansi em eqüinos no Rio Grande do Sul: aspectos epidemiológicos, clínicos, hematológicos e patológicos. Pesquisa Veterinária Brasileira 25(4):239-249. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Cases of trypanosomiasis by Trypanosoma evansi were diagnosed in horses in the state of Rio Grande do Sul, Brazil, between 2003 and 2004. In one stud farm (Farm A) with 125 horses, 52 died. Additionally, around 80 mares were sent to Farm A to be bred. Of those, 66 became ill and 56 died after being returned to their farms of origin. Twenty one horses clinically affected by the disease were observed. Clinical signs included loss of weight (despite voracious appetite), lethargy, incoordination and instability of hindlimbs, atrophy of the large muscles of the hindlimbs, muscle weakness and paleness of mucosae. Specimens of T. evansi were detected in the blood drawn from four affected horses. Normocytic normochromic anemia with PCVs ranging from 15 to 31%, leucocytosis due to lymphocytosis associated to large atypical lymphocytes was observed in several affected horses. High levels of antibodies against T. evansi were detected in the serum of six horses from Farm A. Eight horses presented encephalic neurological signs such as circling, ataxia, blindness, excitation, falls, listlessness, proprioception deficits and head tilt. One horse assumed a “dog-seating position”. Necropsy findings included muscle atrophy, enlargement and lymphoid hyperplasia of the spleen and lymphnodes, edema and softening of the white and grey matter of the brain. Histologically, an overwhelming necrotizing panencephalitis was observed in the seven horses with encephalic signs. This panencephalitis was characterized by marked edema, demyelination and necrosis and perivascular infiltrates of 6-10 layers of lymphocytes and plasm cells affecting both the white and gray matter. Several plasm cells in the inflammatory infiltrate contained numerous eosinophilic globules in their cytoplasm (Mott cells). Similar histological lesions were observed in the spinal cord of the horse with the “dog-seating position”. The brains of five horses with the encephalic signs were submitted to immunohistochemistry stain by the streptavidin-biotin technique. In all of those five brains moderate to abundant specimens of T. evansi in the perivascular spaces and neuropile were marked by the specific antibody. Epidemiological, clinical, hematological, and pathological aspects of equine trypanosomiasis caused by T. evansi are discussed.

• Trypanosomiasis Trypanosoma evansi infectious diseases encephalitis hematology pathology diseases of horses.

Abstract in Portuguese:

Rodrigues A., Fighera R.A., Souza T.M., Schild A.L., Soares M.P., Milano J. & Barros C.S.L. 2005. [Outbreaks of trypanosomiasis in horses by Trypanosoma evansi in the state of Rio Grande do Sul, Brazil: epidemiological, clinical, hematological, and pathological aspects.] Surtos de tripanossomíase por Trypanosoma evansi em eqüinos no Rio Grande do Sul: aspectos epidemiológicos, clínicos, hematológicos e patológicos. Pesquisa Veterinária Brasileira 25(4):239-249. Depto Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Cases of trypanosomiasis by Trypanosoma evansi were diagnosed in horses in the state of Rio Grande do Sul, Brazil, between 2003 and 2004. In one stud farm (Farm A) with 125 horses, 52 died. Additionally, around 80 mares were sent to Farm A to be bred. Of those, 66 became ill and 56 died after being returned to their farms of origin. Twenty one horses clinically affected by the disease were observed. Clinical signs included loss of weight (despite voracious appetite), lethargy, incoordination and instability of hindlimbs, atrophy of the large muscles of the hindlimbs, muscle weakness and paleness of mucosae. Specimens of T. evansi were detected in the blood drawn from four affected horses. Normocytic normochromic anemia with PCVs ranging from 15 to 31%, leucocytosis due to lymphocytosis associated to large atypical lymphocytes was observed in several affected horses. High levels of antibodies against T. evansi were detected in the serum of six horses from Farm A. Eight horses presented encephalic neurological signs such as circling, ataxia, blindness, excitation, falls, listlessness, proprioception deficits and head tilt. One horse assumed a “dog-seating position”. Necropsy findings included muscle atrophy, enlargement and lymphoid hyperplasia of the spleen and lymphnodes, edema and softening of the white and grey matter of the brain. Histologically, an overwhelming necrotizing panencephalitis was observed in the seven horses with encephalic signs. This panencephalitis was characterized by marked edema, demyelination and necrosis and perivascular infiltrates of 6-10 layers of lymphocytes and plasm cells affecting both the white and gray matter. Several plasm cells in the inflammatory infiltrate contained numerous eosinophilic globules in their cytoplasm (Mott cells). Similar histological lesions were observed in the spinal cord of the horse with the “dog-seating position”. The brains of five horses with the encephalic signs were submitted to immunohistochemistry stain by the streptavidin-biotin technique. In all of those five brains moderate to abundant specimens of T. evansi in the perivascular spaces and neuropile were marked by the specific antibody. Epidemiological, clinical, hematological, and pathological aspects of equine trypanosomiasis caused by T. evansi are discussed.