RESUMO

Babesia bovis and Theileria annulata are tick-borne hemoprotozoans that impact bovine health and are responsible for considerable fatalities in tropical and subtropical regions around the world. Both pathogens infect the same vertebrate host, are closely related, and contain similar-sized genomes; however, they differ in invertebrate host specificity, absence vs. presence of a schizont stage, erythrocyte invasion mechanism, and transovarial vs. transstadial transmission. Phylogenetic analysis and bidirectional best hit (BBH) identified a similar number of aspartic, metallo, and threonine proteinases and nonproteinase homologs. In contrast, a considerably increased number of S54 serine rhomboid proteinases and S9 nonproteinase homologs were identified in B. bovis, whereas C1A cysteine proteinases and A1 aspartic nonproteinase homologs were found to be expanded in T. annulata. Furthermore, a single proteinase of families S8 (subtilisin-like protein) and C12 (ubiquitin carboxyl-terminal hydrolase), as well as four nonproteinase homologs, one with dual domains M23-M23 and three with S9-S9, were exclusively present in B. bovis. Finally, a pronounced difference in species-specific ancillary domains was observed between both species. We hypothesize that the observed degradome differences represent functional correlates of the dissimilar life history features of B. bovis and T. annulata. The presented improved classification of piroplasmid proteinases will facilitate an informed choice for future in-depth functional studies.

RESUMO

The low-land tapir (Tapirus terrestris) is the largest wild terrestrial mammal found in Brazil. Although T. terrestris has been already reported as a host of hemoparasites, the occurrence and genetic identity of Piroplasmida agents in this species is still cloudy. Although it is reported that Theileria equi, an endemic equid-infective agent in Brazil, is occurring in lowland tapirs, these reports are probably misconceived diagnoses since they are solely based on small fragments of 18S rRNA that may not achieve accurate topologies on phylogenetic analyses. The present study aimed to detect and investigate the identity of Theileria spp. in tapirs from Pantanal and Cerrado biomes. Blood-DNA samples from tapirs were screened for a partial (~800 bp) 18S rRNA gene fragment from Piroplasmida and 64 (64/122; 52.46% CI: 43.66-61.11%) presented bands of expected size. Samples were submitted to different protocols for molecular characterization, including near-full length 18S rRNA gene (~1500 bp), and the ema-1 gene from T. equi. Eight sequences were obtained for extended fragments (1182-1473 bp) from the 18S rRNA gene. Moreover, three sequences from partial cox-1 and five from partial hsp70 gene were obtained. None of the samples presented amplifications for the ema-1 gene. Phylogenetic and distance analyses from the 18S rRNA sequences obtained demonstrated a clear separation from tapirs' Theileria spp. and T. equi. Phylogenetic analyses of cox-1 and hsp70 sequences obtained herein also showed a unique clade formed by tapir's Theileria spp. Theileria terrestris sp. nov. is positioned apart from all other Theileria species in 18S rRNA, cox-1, and hps70 phylogenetic analyses. This novel proposed species represents a new Piroplasmida clade, yet to be characterized regarding biological features, vectors involved in the transmission cycles, additional vertebrate hosts, and pathogenicity.

RESUMO

Experimental studies have demonstrated that Rhipicephalus (Boophilus) microplus transmits Theileria equi to horses. However, the degree and dynamics of this protozoan infection in the vector's organism have not been fully elucidated. Therefore, this study aimed to evaluate the infection rate and parasitic load of T. equi in R. (B.) microplus, the infection dynamics in this arthropod during experimental infestation in a horse chronically infected with T. equi, and to evaluate the trans-stadial and intrastadial transmission competence of T. equi by R. (B.) microplus. The experimental infestation period of R. (B.) microplus on the horse was 33 days, but males were found on the animal up to 60 days post-infestation. After the fifth day post-infestation, ticks and equine blood were collected every two days. Whole ticks from the same developmental stage collected in the same day were pooled. Adult ticks were dissected to extract salivary glands and gut. DNA extraction was performed for all the samples, and they were then submitted to qPCRs for T. equi diagnosis. Freshly molted nymphs collected as larvae in the horse and freshly molted males and females collected as nymphs in the horse showed equal to or greater than 75% positivity for T. equi, indicating a strong possibility of trans-stadial transmission. The longest permanence of the male ticks on the horse associated with the high positivity rate of this type of sample for T. equi indicate that the male may play a role in the intrastadial transmission of T. equi to infection-free horses. The salivary glands displayed 77.78% positivity for T. equi and presented a higher infection rate at the end of the experimental period (100% from 29 to 33 days post-infection). This study shows that R. (B.) microplus has high T. equi infection rates and that the infection rate and parasitic load increased over the experimental period. These findings confirm the importance of chronically infected horses with T. equi as a source of infection for R. (B.) microplus.

RESUMO

Equine piroplasmosis is the most important tick-borne disease to affect horses in Brazil. Theileria equi is one of the causative agents of equine piroplasmosis. Chronic cases are expected, in which the animals show no apparent signs of infection and remain asymptomatic but constitute a source of the infectious agent that ticks can spread. This study was conducted across 81 ranches located in the municipality of Sinop, State of Mato Grosso, Brazil. A sample calculation was performed to estimate the apparent prevalence of T. equi among horses. A total of 1,853 animals were included in the sampling analysis based on the information available from the Institute of Agricultural and Livestock Defense of Mato Grosso State. The serological analysis of 367 serum samples using an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-T. equi antibodies revealed that 337 animals were positive, representing a frequency of 90.70%. The molecular analysis to amplify the EMA-1 gene showed positivity in 20 of 89 tested samples. The fragments of four samples were sequenced and analyzed to determine their similarities to sequences from other species, based on sequences deposited at GenBank. All showed 100% similarity with T. equi. Our study represents the first report of T. equi antibodies among the equids in north-central region of Mato Grosso, revealing the widespread distribution of seropositive animals.(AU)

A piroplasmose equina é a doença transmitida por carrapatos mais importante em cavalos no Brasil. Theileria equi é um dos agentes causadores da piroplasmose equina. São esperados casos crônicos, nos quais os animais não apresentam sinais aparentes de infecção e permanecem assintomáticos, mas constituem uma fonte de infecção e disseminação por carrapatos. Este estudo foi realizado em 81 fazendas localizadas no município de Sinop, Estado de Mato Grosso, Brasil. Um cálculo amostral foi realizado para estimar a prevalência aparente de T. equi entre cavalos. No total, 1.853 animais foram incluídos na análise amostral com base nas informações disponíveis no Instituto de Defesa Agropecuária do Estado de Mato Grosso. A análise sorológica de 367 amostras de soro por meio de ensaio imunoenzimático indireto (ELISA) para detecção de anticorpos anti-T. equi revelou que 337 animais eram positivos, representando uma frequência de 90,70%. A análise molecular para o gene EMA-1 mostrou positividade em 20 das 89 amostras testadas. Os fragmentos de quatro amostras foram sequenciados e analisados para determinar suas semelhanças com sequências de outras espécies, a partir das sequências depositadas no GenBank. Todos mostraram 100% de similaridade com T. equi. Nosso estudo representa o primeiro relato de anticorpos contra T. equi entre os equídeos na região centro norte de Mato Grosso, revelando a ampla distribuição de animais soropositivos.(AU)

Assuntos

RESUMO

Equine theileriosis, caused by the Theileria equi protozoan, is a disease of worldwide importance. T. equi expresses surface proteins, of which the EMA-2 protein is a promising antigen for vaccine use. The aim of this study was to evaluate the immune response of adult horses, pregnant mares, and foals to an experimental EMA-2 protein of recombinant T. equi vaccine. A total of 46 horses were used in this study for vaccine trials and challenges. Twelve geldings, 14 pregnant mares, and 14 foals were divided into vaccinated and control groups. Total serum specific anti-rEMA-2 IgG, IgG subclasses, and transcription of cytokines related to the immune response were evaluated. For the vaccine challenge, six six-month-old foals were divided into vaccinated and control groups. For the challenge, blood from a horse with theileriosis was transfused to the foals. Geldings and pregnant mares maintained anti-rEMA-2 IgG levels at 130 and 140 days after vaccination, respectively. The most-detected IgG subclasses in vaccinated were IgG3/5, IgG4/7, and IgG1. IL2, IL10, IL12, IL17, IFN-γ, and TNF-α were the most-transcribed cytokines in PBMCs of vaccinated horses stimulated with rEMA-2. Challenge with T. equi demonstrated that vaccinated foals had an increase of 33% in total IgG four days after blood transfusion, while control foals had no significant response, suggesting that vaccine antibodies may have recognized EMA-2 protein of the native T. equi antigen. T. equi recombinant EMA-2 was shown to be a promising vaccine antigen by inducing humoral and cellular immunity similar to that observed in natural parasite infections.

Assuntos

RESUMO

This study aimed to serologically and molecularly survey Babesia caballi and Theileria equi in thoroughbred horses from racecourses in Chile. Additionally, the genetic diversity of the positive samples was assessed. A total of 286 thoroughbred horses from the Santiago and Valparaíso racecourses had their serum samples submitted to an ELISA for B. caballi and T. equi, and 457 samples (from the Santiago, Valparaíso, and Concepción racecourses) were tested with nested PCRs for the B. caballi 48 KDa rhoptry protein (RAP-1) and T. equi 18S rRNA genes. Selected RAP-1 and 18S positive products were sequenced to perform phylogenetic and haplotype analyses. An overall seroprevalence of 35.6% was observed for these Chilean racecourses: 23.7% for T. equi, 8.4% for B. caballi, and 3.5% for both agents. Overall, a 53.6% occurrence by nPCR was detected for the three Chilean racecourses: 44.2% for T. equi, 5.4% for B. caballi, and 3.9% for both agents. Phylogenetic analysis of T. equi and B. caballi showed genetic proximity with sequences previously detected in other countries. Haplotype analysis revealed a low diversity among the Chilean sequences, which may have originated from those reported in Brazil, Israel, or Cuba. Babesia caballi and T. equi were detected for the first time in Chilean thoroughbred horses.

RESUMO

Equine piroplasmosis is the most important tick-borne disease to affect horses in Brazil. Theileria equi is one of the causative agents of equine piroplasmosis. Chronic cases are expected, in which the animals show no apparent signs of infection and remain asymptomatic but constitute a source of the infectious agent that ticks can spread. This study was conducted across 81 ranches located in the municipality of Sinop, State of Mato Grosso, Brazil. A sample calculation was performed to estimate the apparent prevalence of T. equi among horses. A total of 1,853 animals were included in the sampling analysis based on the information available from the Institute of Agricultural and Livestock Defense of Mato Grosso State. The serological analysis of 367 serum samples using an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-T. equi antibodies revealed that 337 animals were positive, representing a frequency of 90.70%. The molecular analysis to amplify the EMA-1 gene showed positivity in 20 of 89 tested samples. The fragments of four samples were sequenced and analyzed to determine their similarities to sequences from other species, based on sequences deposited at GenBank. All showed 100% similarity with T. equi. Our study represents the first report of T. equi antibodies among the equids in north-central region of Mato Grosso, revealing the widespread distribution of seropositive animals.

A piroplasmose equina é a doença transmitida por carrapatos mais importante em cavalos no Brasil. Theileria equi é um dos agentes causadores da piroplasmose equina. São esperados casos crônicos, nos quais os animais não apresentam sinais aparentes de infecção e permanecem assintomáticos, mas constituem uma fonte de infecção e disseminação por carrapatos. Este estudo foi realizado em 81 fazendas localizadas no município de Sinop, Estado de Mato Grosso, Brasil. Um cálculo amostral foi realizado para estimar a prevalência aparente de T. equi entre cavalos. No total, 1.853 animais foram incluídos na análise amostral com base nas informações disponíveis no Instituto de Defesa Agropecuária do Estado de Mato Grosso. A análise sorológica de 367 amostras de soro por meio de ensaio imunoenzimático indireto (ELISA) para detecção de anticorpos anti-T. equi revelou que 337 animais eram positivos, representando uma frequência de 90,70%. A análise molecular para o gene EMA-1 mostrou positividade em 20 das 89 amostras testadas. Os fragmentos de quatro amostras foram sequenciados e analisados para determinar suas semelhanças com sequências de outras espécies, a partir das sequências depositadas no GenBank. Todos mostraram 100% de similaridade com T. equi. Nosso estudo representa o primeiro relato de anticorpos contra T. equi entre os equídeos na região centro norte de Mato Grosso, revelando a ampla distribuição de animais soropositivos.

Assuntos

RESUMO

Equine piroplasmosis is the most important tick-borne disease to affect horses in Brazil. Theileria equi is one of the causative agents of equine piroplasmosis. Chronic cases are expected, in which the animals show no apparent signs of infection and remain asymptomatic but constitute a source of the infectious agent that ticks can spread. This study was conducted across 81 ranches located in the municipality of Sinop, State of Mato Grosso, Brazil. A sample calculation was performed to estimate the apparent prevalence of T. equi among horses. A total of 1,853 animals were included in the sampling analysis based on the information available from the Institute of Agricultural and Livestock Defense of Mato Grosso State. The serological analysis of 367 serum samples using an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-T. equi antibodies revealed that 337 animals were positive, representing a frequency of 90.70%. The molecular analysis to amplify the EMA-1 gene showed positivity in 20 of 89 tested samples. The fragments of four samples were sequenced and analyzed to determine their similarities to sequences from other species, based on sequences deposited at GenBank. All showed 100% similarity with T. equi. Our study represents the first report of T. equi antibodies among the equids in north-central region of Mato Grosso, revealing the widespread distribution of seropositive animals.

A piroplasmose equina é a doença transmitida por carrapatos mais importante em cavalos no Brasil. Theileria equi é um dos agentes causadores da piroplasmose equina. São esperados casos crônicos, nos quais os animais não apresentam sinais aparentes de infecção e permanecem assintomáticos, mas constituem uma fonte de infecção e disseminação por carrapatos. Este estudo foi realizado em 81 fazendas localizadas no município de Sinop, Estado de Mato Grosso, Brasil. Um cálculo amostral foi realizado para estimar a prevalência aparente de T. equi entre cavalos. No total, 1.853 animais foram incluídos na análise amostral com base nas informações disponíveis no Instituto de Defesa Agropecuária do Estado de Mato Grosso. A análise sorológica de 367 amostras de soro por meio de ensaio imunoenzimático indireto (ELISA) para detecção de anticorpos anti-T. equi revelou que 337 animais eram positivos, representando uma frequência de 90,70%. A análise molecular para o gene EMA-1 mostrou positividade em 20 das 89 amostras testadas. Os fragmentos de quatro amostras foram sequenciados e analisados para determinar suas semelhanças com sequências de outras espécies, a partir das sequências depositadas no GenBank. Todos mostraram 100% de similaridade com T. equi. Nosso estudo representa o primeiro relato de anticorpos contra T. equi entre os equídeos na região centro norte de Mato Grosso, revelando a ampla distribuição de animais soropositivos.

Assuntos

RESUMO

Theileriosis is an important disease of economic significance which badly affects the equine husbandry of developing countries. The present study was planned to investigate the molecular prevalence of theileriosis, associated risk factors, and alterations in hematological parameters of donkeys and mules from district Mianwali, Punjab, Pakistan. Blood samples from 150 equids (n = 75 donkeys; n = 75 mules) were examined microscopically, and the genomic DNA from each sample was processed for the amplification of the 18S rRNA gene of Theileria. The polymerase chain reaction confirmed isolates were purified followed by sequencing. The data regarding the analysis of risk factors were collected in a predesigned questionnaire and statistically analyzed by logistic regression analysis. An overall prevalence of 17.33% was noted in this study. Donkeys showed more prevalence followed by mules being 20.0% and 14.7%, respectively. The study isolates showed high resemblance (99%) with isolates from the United States of America, Spain, Brazil, Israel, Cuba, France, South Africa, Korea, Turkey, Tunisia, India, E. Caribbean, and Nigeria. The potential risk factors found to be significantly associated (P < .05) with disease dynamics were tick infestation on study animals, previous tick history, and house hygiene. A significant (P < .05) decrease in the number of platelets, erythrocytes, hemoglobin level, and packed cell volume was observed in donkeys and mules suffering from theileriosis compared with the healthy ones. The study is the first report regarding the molecular characterization of theileriosis in donkeys and mules in Pakistan. The findings will be effectual in designing effective control strategies for this disease in Punjab, Pakistan.

Assuntos

RESUMO

Ticks and the vast array of pathogens they transmit, including bacteria, viruses, protozoa, and helminths, constitute a growing burden for human and animal health worldwide. In Cuba, the major tropical island in the Caribbean, ticks are an important cause of vector-borne diseases affecting livestock production, pet animal health and, to a lesser extent, human health. The higher number of tick species in the country belong to the Argasidae family and, probably less known, is the presence of an autochthonous tick species in the island, Ixodes capromydis. Herein, we provide a comprehensive review of the ticks and tick-borne pathogens (TBPs) affecting animal and human health in Cuba. The review covers research results including ecophysiology of ticks, the epidemiology of TBPs, and the diagnostic tools used currently in the country for the surveillance of TBPs. We also introduce the programs implemented in the country for tick control and the biotechnology research applied to the development of anti-tick vaccines.

RESUMO

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.(AU)

Este estudo foi desenvolvido para detectar piroplasmose equina usando a técnica molecular na província de Al-Najaf durante o período do ano com maior ocorrência de carrapatos. Foram coletadas amostras de sangue de 110 cavalos que apresentaram mais de dois sinais de piroplasmose. Após a extração do DNA, o produto foi examinado por reação em cadeia da polimerase para amplificar o 18SrRNA. Os resultados mostraram que a porcentagem geral de teileriose equina foi de 38, 18%. De acordo com o sexo, o percentual de infecção foi 43,48% e 29,27% no sexo feminino e masculino, respectivamente. Apareceram variações significativas entre os cavalos infectados de acordo com a idade, e a porcentagem de infecção foi 50% e 35,22% em menos de 2 anos e mais de 2 anos, respectivamente. Além disso, as porcentagens de infecção foram 62, 5% e 19, 35% em animais com e sem acariasis, respectivamente. Também foram observadas variações significativas na teileriose dos equídeos, de acordo com as áreas geográficas, e o maior percentual foi relatado no distrito de Hera (60, 87%), enquanto o menor percentual foi no centro de Al-Najaf (21,43%). Essa diferença pode ser devido à distribuição diferente do vetor da doença (carrapato), que pode ser a disponibilidade do clima adequado que ajuda na multiplicação dos vetores intermediários. Em conclusão, este estudo provou as variações nas ocorrências de piroplasmose eqüina de acordo com sexo, idade e áreas geográficas.(AU)

Assuntos

RESUMO

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the var

Este estudo foi desenvolvido para detectar piroplasmose equina usando a técnica molecular na província de Al-Najaf durante o período do ano com maior ocorrência de carrapatos. Foram coletadas amostras de sangue de 110 cavalos que apresentaram mais de dois sinais de piroplasmose. Após a extração do DNA, o produto foi examinado por reação em cadeia da polimerase para amplificar o 18SrRNA. Os resultados mostraram que a porcentagem geral de teileriose equina foi de 38, 18%. De acordo com o sexo, o percentual de infecção foi 43,48% e 29,27% no sexo feminino e masculino, respectivamente. Apareceram variações significativas entre os cavalos infectados de acordo com a idade, e a porcentagem de infecção foi 50% e 35,22% em menos de 2 anos e mais de 2 anos, respectivamente. Além disso, as porcentagens de infecção foram 62, 5% e 19, 35% em animais com e sem acariasis, respectivamente. Também foram observadas variações significativas na teileriose dos equídeos, de acordo com as áreas geográficas, e o maior percentual foi relatado no distrito de Hera (60, 87%), enquanto o menor percentual foi no centro de Al-Najaf (21,43%). Essa diferença pode ser devido à distribuição diferente do vetor da doença (carrapato), que pode ser a disponibilidade do clima adequado que ajuda na multiplicação dos vetores inter

RESUMO

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the var

Este estudo foi desenvolvido para detectar piroplasmose equina usando a técnica molecular na província de Al-Najaf durante o período do ano com maior ocorrência de carrapatos. Foram coletadas amostras de sangue de 110 cavalos que apresentaram mais de dois sinais de piroplasmose. Após a extração do DNA, o produto foi examinado por reação em cadeia da polimerase para amplificar o 18SrRNA. Os resultados mostraram que a porcentagem geral de teileriose equina foi de 38, 18%. De acordo com o sexo, o percentual de infecção foi 43,48% e 29,27% no sexo feminino e masculino, respectivamente. Apareceram variações significativas entre os cavalos infectados de acordo com a idade, e a porcentagem de infecção foi 50% e 35,22% em menos de 2 anos e mais de 2 anos, respectivamente. Além disso, as porcentagens de infecção foram 62, 5% e 19, 35% em animais com e sem acariasis, respectivamente. Também foram observadas variações significativas na teileriose dos equídeos, de acordo com as áreas geográficas, e o maior percentual foi relatado no distrito de Hera (60, 87%), enquanto o menor percentual foi no centro de Al-Najaf (21,43%). Essa diferença pode ser devido à distribuição diferente do vetor da doença (carrapato), que pode ser a disponibilidade do clima adequado que ajuda na multiplicação dos vetores inter

RESUMO

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.(AU)

Este estudo foi desenvolvido para detectar piroplasmose equina usando a técnica molecular na província de Al-Najaf durante o período do ano com maior ocorrência de carrapatos. Foram coletadas amostras de sangue de 110 cavalos que apresentaram mais de dois sinais de piroplasmose. Após a extração do DNA, o produto foi examinado por reação em cadeia da polimerase para amplificar o 18SrRNA. Os resultados mostraram que a porcentagem geral de teileriose equina foi de 38, 18%. De acordo com o sexo, o percentual de infecção foi 43,48% e 29,27% no sexo feminino e masculino, respectivamente. Apareceram variações significativas entre os cavalos infectados de acordo com a idade, e a porcentagem de infecção foi 50% e 35,22% em menos de 2 anos e mais de 2 anos, respectivamente. Além disso, as porcentagens de infecção foram 62, 5% e 19, 35% em animais com e sem acariasis, respectivamente. Também foram observadas variações significativas na teileriose dos equídeos, de acordo com as áreas geográficas, e o maior percentual foi relatado no distrito de Hera (60, 87%), enquanto o menor percentual foi no centro de Al-Najaf (21,43%). Essa diferença pode ser devido à distribuição diferente do vetor da doença (carrapato), que pode ser a disponibilidade do clima adequado que ajuda na multiplicação dos vetores intermediários. Em conclusão, este estudo provou as variações nas ocorrências de piroplasmose eqüina de acordo com sexo, idade e áreas geográficas.(AU)

Assuntos

RESUMO

Equine piroplasmosis is a tick-borne disease caused by the protozoan parasites Babesia caballi and Theileria equi. We report a case of abortion in association with transplacental Theileria equi infection in a mare from the State of Espírito Santo, southeast Brazil. An apparently healthy mare aborted at the tenth month of gestation. At necropsy, the subcutaneous tissue, skeletal muscles, and visible mucosae of the aborted fetus were pale, and there was moderate hydrothorax and marked splenomegaly. Microscopic findings included splenic lymphoid hyperplasia and nephrosis. Merozoite-infected erythrocytes were found within blood vessels of all organs examined and were most numerous in the brain. DNA extracted from the spleen, liver, kidney, and thymus was used as a template for PCR. Generic primers were employed for the detection of piroplasm 18S ribosomal gene. All samples were positive for piroplasm DNA by PCR. Amplicons were purified and then sequenced. Sequencing analysis of these amplicons revealed 98% identity to T. equi sequences. Based on our findings, we suggest that abortion in this case resulted from transplacental Theileria infection.(AU)

A piroplasmose equina é transmitida por carrapatos, sendo causada pelos protozoários Babesia caballi e Theileria equi. O objetivo deste trabalho foi relatar um caso de aborto equino associado à piroplasmose fetal. Uma égua gestante, aparentemente saudável, abortou no 10º mês de gestação. Na necropsia do feto, foi observada palidez de subcutâneo, da musculatura esquelética e das mucosas visíveis, moderado hidrotórax e acentuada esplenomegalia. Histologicamente, havia hiperplasia linfoide esplênica e nefrose. Muitos merozoítos foram observados em eritrócitos nos vasos sanguíneos de todos os órgãos examinados, com maior intensidade no encéfalo. Amostras de DNA do baço, fígado, rim e timo foram utilizadas para PCR com primers genéricos para detecção de sequências do gene 18S ribossomal de piroplasmas, com resultado positivo para todas as amostras. Em seguida, os amplicons das amostras de baço foram purificados, sequenciados e, após análise, mostraram 98% de identidade com a sequência de T. equi. Portanto, as alterações microscópicas, os resultados de PCR e sequenciamento confirmam aborto associado à T. equi por transmissão transplacentária.(AU)

Assuntos

RESUMO

Equine piroplasmosis is a tick-borne disease caused by the protozoan parasites Babesia caballi and Theileria equi. We report a case of abortion in association with transplacental Theileria equi infection in a mare from the State of Espírito Santo, southeast Brazil. An apparently healthy mare aborted at the tenth month of gestation. At necropsy, the subcutaneous tissue, skeletal muscles, and visible mucosae of the aborted fetus were pale, and there was moderate hydrothorax and marked splenomegaly. Microscopic findings included splenic lymphoid hyperplasia and nephrosis. Merozoite-infected erythrocytes were found within blood vessels of all organs examined and were most numerous in the brain. DNA extracted from the spleen, liver, kidney, and thymus was used as a template for PCR. Generic primers were employed for the detection of piroplasm 18S ribosomal gene. All samples were positive for piroplasm DNA by PCR. Amplicons were purified and then sequenced. Sequencing analysis of these amplicons revealed 98% identity to T. equi sequences. Based on our findings, we suggest that abortion in this case resulted from transplacental Theileria infection.(AU)

A piroplasmose equina é transmitida por carrapatos, sendo causada pelos protozoários Babesia caballi e Theileria equi. O objetivo deste trabalho foi relatar um caso de aborto equino associado à piroplasmose fetal. Uma égua gestante, aparentemente saudável, abortou no 10º mês de gestação. Na necropsia do feto, foi observada palidez de subcutâneo, da musculatura esquelética e das mucosas visíveis, moderado hidrotórax e acentuada esplenomegalia. Histologicamente, havia hiperplasia linfoide esplênica e nefrose. Muitos merozoítos foram observados em eritrócitos nos vasos sanguíneos de todos os órgãos examinados, com maior intensidade no encéfalo. Amostras de DNA do baço, fígado, rim e timo foram utilizadas para PCR com primers genéricos para detecção de sequências do gene 18S ribossomal de piroplasmas, com resultado positivo para todas as amostras. Em seguida, os amplicons das amostras de baço foram purificados, sequenciados e, após análise, mostraram 98% de identidade com a sequência de T. equi. Portanto, as alterações microscópicas, os resultados de PCR e sequenciamento confirmam aborto associado à T. equi por transmissão transplacentária.(AU)

Assuntos

RESUMO

The present study aims to determine the frequencies of Theileria equi and Anaplasma phagocytophilum antibodies among horses from the state of Rio de Janeiro, Brazil, and to detect the presence of DNA of these pathogens through molecular methods. A total of 98 serum samples of horses from the municipality of Seropedica were tested by indirect immunofluorescence antibody (IFA) to detect anti-A. phagocytophilum and anti-T. equi IgG antibodies. In addition, quantitative real-time PCR (qPCR) was used to detect these pathogens in the DNA extracted from the whole blood and buffy coat of horses. Bivariate analysis and odds ratio were performed to verify the possible association between positivity and characteristics related to the horses. As evaluated by IFA and qPCR, the frequency of animals that tested positive for T. equi was 89.8% (n = 88/98) and 91.8% (n = 90/98), whereas A. phagocytophilum was 17.4% (n = 17/98) and 1.0% (n = 1/98), respectively. Serological evidence of exposure to A. phagocytophilum and T. equi was observed in 16.3% (n = 16/98) of the horses; however, exposure was confirmed by qPCR in only 1.0% (n = 1/98). No statistical association was found in the bivariate and odds ratio analysis. This is the first study reporting the molecular detection of A. phagocytophilum DNA in horses from the state of Rio de Janeiro, and also the coinfection of A. phagocytophilum and T. equi in a horse from Brazil confirmed by molecular methods. Equine granulocytic anaplasmosis is circulating in Brazilian horses, together with T. equi, and should be included in the differential diagnosis of tick-borne diseases.

RESUMO

Theileria equi is one of the etiologic agents of the equine piroplasmosis. This infectious disease is transmitted by ticks and is a worldwide problem in the international horse movement. The 18S rRNA gene of T. equi is often used for genotyping and phylogenetic purpose. This study aimed to analyze the degree of the heterogeneity of the 18S rRNA gene of T. equi in horses from the state of Rio de Janeiro, Brazil. The complete T. equi 18S rRNA sequences were obtained from twenty naturally infected horses. The PCR amplicons were cloned and sequenced. The phylogenetic analyses were performed using a set of T. equi 18S rRNA sequences and other related organisms available in ARB-Silva database. There were twelve distinct T. equi 18S rRNA gene sequences circulating in horses in the state of Rio de Janeiro, Brazil. Monophyletic clades with 2% evolutionary divergence between clades and high bootstrap value were the support to divide T. equi sequences in three distinct clades. The sequences from this study grouped into clades I (70%, n=14/20) and II (30%, n=6/20). All of the T. equi sequences grouped within a node other than the theileriids. This study reported a clear division of two distinct genotypes of T. equi 18S rRNA sequences in state of Rio de Janeiro, Brazil, and it demonstrates that distinct isolates of T. equi can coexist in the same geographic region.

Assuntos

RESUMO

Theileria equi is the causative agent of worldwide piroplasmosis, an important tick-borne disease of equids associated to a lifetime carrier state of infected horses. Since rapid, accessible and reliable tests have been posted as a challenge for clinicians, the aim of the present study was to verify the agreement between an in-house immunofluorescence antibody assay (IFA) and a commercial competitive inhibition enzyme-linked immunosorbent assay (cELISA) in a population of 198 carthorses from Southern Brazil. The horse seroreactivity for T. equi revealed 152/198 (76.8%) positive samples by IFA and 155/198 (78.3%) by cELISA. Using cELISA as gold standard, IFA has shown a sensitivity of 91.6% (95% CI = 86.18-95.03%) and specificity of 76.7% (95% CI = 62.26-86.85%), with a substantial degree of agreement (k = 0.8445). In conclusion, the in-house IFA may be used as a screening test for diagnosis of equine piroplasmosis.(AU)

Theileria equi é o agente causador da piroplasmose no mundo, é uma importante doença transmitida por carrapatos nos equídeos em todo o mundo, e está associada ao estado de portador nos equinos infectados. Uma vez que testes rápidos, acessíveis e confiáveis são considerados um desafio para os clínicos, o objetivo do presente estudo foi avaliar a concordância entre os testes de imunofluorescência indireta (RIFI) e um ensaio imunoenzimático por competição comercial (cELISA) em uma população de 198 equinos de tração na região Sul do Brasil. A sororreatividade dos cavalos para T. equi mostrou 152/198 (76.8%) amostras positivas para RIFI e 155/198 (78.3%) amostras positivas para cELISA. Utilizando o cELISA como padrão ouro, a RIFI demonstrou sensibilidade de 91.6% (95% CI = 86.18-95.03%) e especificidade de 76.7% (95% CI = 62.26-86.85%), com uma concordância de grau substancial (k = 0.8445). Em conclusão, a RIFI pode ser utilizada como um teste de triagem para o diagnóstico de piroplasmose equina.(AU)

Assuntos

RESUMO

Theileria equi is the causative agent of worldwide piroplasmosis, an important tick-borne disease of equids associated to a lifetime carrier state of infected horses. Since rapid, accessible and reliable tests have been posted as a challenge for clinicians, the aim of the present study was to verify the agreement between an in-house immunofluorescence antibody assay (IFA) and a commercial competitive inhibition enzyme-linked immunosorbent assay (cELISA) in a population of 198 carthorses from Southern Brazil. The horse seroreactivity for T. equi revealed 152/198 (76.8%) positive samples by IFA and 155/198 (78.3%) by cELISA. Using cELISA as gold standard, IFA has shown a sensitivity of 91.6% (95% CI = 86.18-95.03%) and specificity of 76.7% (95% CI = 62.26-86.85%), with a substantial degree of agreement (k = 0.8445). In conclusion, the in-house IFA may be used as a screening test for diagnosis of equine piroplasmosis.

Theileria equi é o agente causador da piroplasmose no mundo, é uma importante doença transmitida por carrapatos nos equídeos em todo o mundo, e está associada ao estado de portador nos equinos infectados. Uma vez que testes rápidos, acessíveis e confiáveis são considerados um desafio para os clínicos, o objetivo do presente estudo foi avaliar a concordância entre os testes de imunofluorescência indireta (RIFI) e um ensaio imunoenzimático por competição comercial (cELISA) em uma população de 198 equinos de tração na região Sul do Brasil. A sororreatividade dos cavalos para T. equi mostrou 152/198 (76.8%) amostras positivas para RIFI e 155/198 (78.3%) amostras positivas para cELISA. Utilizando o cELISA como padrão ouro, a RIFI demonstrou sensibilidade de 91.6% (95% CI = 86.18-95.03%) e especificidade de 76.7% (95% CI = 62.26-86.85%), com uma concordância de grau substancial (k = 0.8445). Em conclusão, a RIFI pode ser utilizada como um teste de triagem para o diagnóstico de piroplasmose equina.

Assuntos