RESUMEN
Equine Piroplasmosis (EP) and Equine Granulocytic Anaplasmosis (EGA) are diseases that affect horses, transmitted by ixodid ticks, causing a nonspecific febrile syndrome. Equine Piroplasmosis is endemic in Brazil, and most horses are in enzootic stability. Serological and molecular studies carried out on horses in Brazil have shown the presence of Anaplasma phagocytophilum, however, the clinical relevance of this infection has not yet been established. The present study aims to evaluate the importance of Babesia caballi, Theileria equi, and A. phagocytophilum as etiological agents in horses with clinical manifestations suggestive of these diseases in the metropolitan mesoregion of Rio de Janeiro. A total of 45 animals with clinical signs were submitted to DNA extraction followed by qPCR test. Anaplasma phagocytophilum, Neorickettsia risticii and Theileria haneyi were not found in any of the horses with clinical signs, however 62.2% were infected with at least one agent of EP. Theileria equi was the most frequent etiologic agent (35.5%), followed by coinfection (15.5%) and B. caballi (11.2%). These results suggest that A. phagocytophilum has minor clinical importance in the region, while EP is frequently found in symptomatic horses, representing an important differential diagnosis in suspected cases.
Asunto(s)
Anaplasma phagocytophilum , Babesia , Babesiosis , Ehrlichiosis , Enfermedades de los Caballos , Theileria , Theileriosis , Caballos , Animales , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/epidemiología , Brasil/epidemiología , Theileria/aislamiento & purificación , Babesia/aislamiento & purificación , Anaplasma phagocytophilum/aislamiento & purificación , Theileriosis/epidemiología , Theileriosis/parasitología , Babesiosis/epidemiología , Babesiosis/parasitología , Ehrlichiosis/veterinaria , Ehrlichiosis/epidemiología , Ehrlichiosis/microbiología , Masculino , FemeninoRESUMEN
The present study was focused on the incidence of ticks and tick-borne diseases (TTBD) in cross-bred cattle (Friesian x Sahiwal) of two farms (n = 2548) in district Lahore, Pakistan. We collected total of 572 ticks (adults and nymphs) and blood samples (10 ml) for microscopic i.e., blood smear test - Giemsa Stain (BST) and molecular analysis; Reverse Line Blot-General Primer-PCR (RLB-PCR) and Specie Specific Primer PCR (SP-PCR) from infested cattle (n = 100) from months of April to September. Results: The tick specie identified was Rhipicephalus microplus at both farms, with significant difference in infestations rate amongst both farms (p< 0.0001). The cross-bred cattle having higher ratio of Friesian blood and lower ratio of Sahiwal blood were mostly infested by ticks (p < 0.0458) and haemoparasites (p <0.474) and vice versa. The SP-PCR showed higher number of haemoparasites infection than BST, which revealed 16% T. annulata (p < 0.0001 and k value 0.485, 0.0001), 51% B. bigemina (p < 0.0001 and k value 0.485, 0.0001) and 15% A. marginale (p < 0.001 and k value 0.207, 0.001), respectively. The single infection with B. bigemina was 34% (n = 34/100) and A. marginale 6% (n = 6/100). The double infection with T. annulata/B. bigemina was 8% (n = 8/100) and B. bigemina/A. marginale 1% (n = 1/100). Whereas the triple infection with T. annulata/B. bigemina/A .marginale was 8% (n = 8/100). The phylogenetic study of isolated sequence of T. annulata revealed close homology to isolates from Iran (87%), B. bigemina to isolates from Cuba (94 to 100%) and A. marginale with isolates from Pakistan (99 to 98%).
Asunto(s)
Babesia , Enfermedades de los Bovinos , Microbiota , Theileriosis , Enfermedades por Picaduras de Garrapatas , Garrapatas , Bovinos , Animales , Garrapatas/parasitología , Theileriosis/epidemiología , Theileriosis/parasitología , Filogenia , Pakistán/epidemiología , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Enfermedades por Picaduras de Garrapatas/diagnóstico , Enfermedades por Picaduras de Garrapatas/epidemiología , Enfermedades por Picaduras de Garrapatas/veterinariaRESUMEN
Equine piroplasmosis is a disease of horses, mules and donkeys, caused by the hemoprotozoans Babesia caballi and Theileria equi and transmitted by ticks of tropical and subtropical regions. Because the clinical signs are not specific, the diagnosis of equine piroplasmosis is difficult. In Mexico, where the environmental factors are conducive to the persistence of these pathogens, there is a lack of molecular studies to evaluate the occurrence of both parasites in horses. In the present study, matching serum and whole blood samples were obtained from 269 horses residing in 24 locations with tropical or subtropical climate and the presence of ticks. Testing of serum samples by ELISA demonstrated 55.7% seroprevalence of B. caballi and 68.4% prevalence of antibodies to T. equi. Blood samples analyzed with nPCR test were 7.8% positive to B. caballi and 78.8% positive to T. equi, while a duplex qPCR showed 15.24% positive samples to B. caballi and 59.11% to T. equi. From these results, 27 samples were sequenced for T. equi and 13 for B. caballi, confirming the presence of both horse parasites that cause equine piroplasmosis and suggesting that they are widespread in Mexico. This is the first study confirming the presence of B. caballi and T. equi in Mexico using both serological and molecular diagnostic methods. This study shows a high incidence of exposure to the etiological agents of equine piroplasmosis in horses in the studied areas.
Asunto(s)
Babesia , Babesiosis , Enfermedades de los Caballos , Theileria , Theileriosis , Garrapatas , Animales , Babesia/genética , Babesiosis/diagnóstico , Babesiosis/epidemiología , Bovinos , Equidae/parasitología , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/parasitología , Caballos , México/epidemiología , Estudios Prospectivos , Estudios Seroepidemiológicos , Theileria/genética , Theileriosis/diagnóstico , Theileriosis/epidemiología , Theileriosis/parasitología , Garrapatas/parasitologíaRESUMEN
Equine piroplasmosis (EP) is a tick-borne infectious disease highly prevalent in tropical and subtropical regions, such as Venezuela. EP affects wild and domestic equids leading to several clinical presentations, from asymptomatic to severely affected animals. In this study, thirty-three (33) sport horses under regular training activities and from endemic regions of north-central Venezuela were submitted to an observational survey, case-control, to describe the presence of clinical signs and natural EP infections. A conventional PCR assay targeting the SSU rRNA gene revealed EP etiologic agents in 13 out of 33 sampled horses (~ 39.4% infections). Nine (9) of these EP-positive samples were confirmed as infected with Babesia caballi (6/9 = 66.7%) or Theileria equi (3/9 = 33.3%) by DNA sequencing and BLASTN analyses. A phylogeny of SSU rRNA gene sequences revealed that these new B. caballi and T. equi sequences clustered within the worldwide distributed phylogenetic genotype A, respectively. No acute EP cases were observed in this study; however, six (6) PCR-positive animals displayed mild clinical signs compatible with EP, including a mild leukocytosis (P < 0.05). The heart rate variability frequency domain analysis in four (4) of these EP-positive infected animals revealed a significant (P < 0.05) higher low-frequency/high-frequency ratio suggesting a sympathovagal imbalance in these chronically infected animals. Other clinical and cardiovascular parameters were similar between the different groups. Sport horses are routinely submitted to intense training programs and sport-related activities that could lead to loss of the host-parasite equilibrium that characterizes enzootic regions, increasing the likelihood of infection reactivation and the risk of transmission. Heart rate variability analysis contributes to evaluate the sympathovagal balance and detecting homeostasis disturbances in sport horses. Molecular diagnostic tests for EP based on the detection of parasite DNA in equine blood samples should be included in the health programs of sport horses in endemic areas.
Asunto(s)
Babesiosis , Enfermedades de los Bovinos , Enfermedades de los Caballos , Theileria , Theileriosis , Animales , Babesiosis/diagnóstico , Babesiosis/epidemiología , Babesiosis/parasitología , Bovinos , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/epidemiología , Caballos , Filogenia , Theileria/genética , Theileriosis/diagnóstico , Theileriosis/epidemiología , Theileriosis/parasitologíaRESUMEN
To investigate the presence of Theileria equi in an endemic area of equine piroplasmosis 42 horses (Equus caballus) from Corrientes City, Argentina were sampled. Eighty-one percent (34 blood samples) of the analyzed horses were tested positive to the presence of piroplasmid 18S rDNA. All these samples could be identified as T. equi by amplifying the specific EMA-1 (merozoite antigen 1) gene of this species. Phylogenetic analysis of an obtained 18S rDNA complete sequence from one strain resulted in the identification of this sample as T. equi sensu stricto (genotype A). This study presents the first molecular detection and characterization of T. equi by the complete 18S rDNA sequence in Argentina. Based on these results further studies should be carried out to investigate the distribution and heterogeneity of presented genotypes of T. equi in Argentina, which is essential for the diagnosis, prevention and treatment of equine piroplasmosis.
Asunto(s)
Enfermedades de los Caballos/parasitología , Theileria/aislamiento & purificación , Theileriosis/parasitología , Animales , Argentina , Caballos , Filogenia , ARN Protozoario/análisis , ARN Ribosómico 18S/análisis , Theileria/clasificaciónRESUMEN
MHC peptide binding and presentation is the most selective event defining the landscape of T cell epitopes. Consequently, understanding the diversity of MHC alleles in a given population and the parameters that define the set of ligands that can be bound and presented by each of these alleles (the immunopeptidome) has an enormous impact on our capacity to predict and manipulate the potential of protein Ags to elicit functional T cell responses. Liquid chromatography-mass spectrometry analysis of MHC-eluted ligand data has proven to be a powerful technique for identifying such peptidomes, and methods integrating such data for prediction of Ag presentation have reached a high level of accuracy for both MHC class I and class II. In this study, we demonstrate how these techniques and prediction methods can be readily extended to the bovine leukocyte Ag class II DR locus (BoLA-DR). BoLA-DR binding motifs were characterized by eluted ligand data derived from bovine cell lines expressing a range of DRB3 alleles prevalent in Holstein-Friesian populations. The model generated (NetBoLAIIpan, available as a Web server at www.cbs.dtu.dk/services/NetBoLAIIpan) was shown to have unprecedented predictive power to identify known BoLA-DR-restricted CD4 epitopes. In summary, the results demonstrate the power of an integrated approach combining advanced mass spectrometry peptidomics with immunoinformatics for characterization of the BoLA-DR Ag presentation system and provide a prediction tool that can be used to assist in rational evaluation and selection of bovine CD4 T cell epitopes.
Asunto(s)
Presentación de Antígeno , Linfocitos T CD4-Positivos/inmunología , Biología Computacional/métodos , Epítopos de Linfocito T/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Péptidos/inmunología , Alelos , Animales , Secuencia de Bases , Linfocitos T CD4-Positivos/parasitología , Bovinos , Células Cultivadas , Simulación por Computador , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Antígenos de Histocompatibilidad Clase II/genética , Ligandos , Espectrometría de Masas/métodos , Unión Proteica , Theileria annulata , Theileria parva , Theileriosis/inmunología , Theileriosis/parasitologíaRESUMEN
Equine piroplasmosis is a tropical parasitic disease caused by several intraerythrocytic members of the genera Babesia and Theileria. These pathogens are transmitted by multiple tick species and are considered as important exotic microorganisms in several countries. Equine piroplasmosis causes significant economic losses due to abortions, decreased activity and even death of equines, making surveillance of these infectious disease essential. In the northern and eastern parts of Mexico, few molecular and serological reports have been made on the presence and exposure of horse to these agents. For this reason, the aim of the current work was to perform a molecular detection study of Babesia and Theileria in equines from the state of Veracruz, Mexico. A total of 100 whole blood samples were tested. Chelex-100 resin was used for DNA extraction and a fragment of 459 bp of the 18S rRNA gene of members of the genera Babesia/Theileria were identified. Of the 100 samples analysed, 18 tested positive for Babesia/Theileria, resulting in a prevalence of 18 %. Identity analyses and phylogenetic reconstruction revealed that all samples were infected with Theileria equi. This work represents the first molecular record of Babesia/Theileria in equines from the state of Veracruz, Mexico, and demonstrates the endemicity of T. equi in this region of the country.
Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/epidemiología , Enfermedades de los Caballos/epidemiología , Theileria/aislamiento & purificación , Theileriosis/epidemiología , Animales , Babesiosis/parasitología , Femenino , Enfermedades de los Caballos/parasitología , Caballos , Masculino , México/epidemiología , Filogenia , Prevalencia , ARN Protozoario/análisis , ARN Ribosómico 18S/análisis , Theileriosis/parasitologíaRESUMEN
The objective of this study was to detect natural Theileria equi infection in captive tapirs (Tapirus terrestris) in the Brazilian Amazon. Samples from 19 captive tapirs were collected from zoological and botanical gardens and conservation parks in the Pará (n = 18) and Amazonas (n = 1) states. Whole-blood samples were collected for subsequent screening of T. equi DNA by PCR using the BEC-UF2 and EQUI-R primer set. Microscopic analyses of blood smears revealed T. equi trophozoites in 37% (7/19) of the animals examined, and T. equi DNA was detected in 58% (11/19) of the blood samples analyzed. Sequencing of amplified PCR products revealed an identity with T. equi isolates obtained from horses and waterbuck available in GenBank. In conclusion, T. equi infection occurs in captive tapirs in the Brazilian Amazon, and these mammals could potentially act as reservoirs.
Asunto(s)
Perisodáctilos , Theileria/aislamiento & purificación , Theileriosis/epidemiología , Animales , Animales de Zoológico , Brasil/epidemiología , Femenino , Masculino , Prevalencia , Theileriosis/parasitologíaRESUMEN
The State of Pará has one of the largest herds of equids (horse, donkey and mule) in Brazil, most of these animals are found on cattle farms. Equine theileriosis is a tick-borne disease caused by the parasite Theileria equi and is characterized by fever, anaemia, icterus, intravascular haemolysis, haemoglobinuria, spleen and hepatomegaly, and even death. The present study aimed to determine the prevalence of antibodies against T. equi in equids in the western region of the State of Pará, Brazil, and to identify potential risk factors associated with parasite infection. A cross-sectional study was conducted with cluster sampling of farm horses from 18 municipalities. In the cities visited, samples from sport and carthorses were also included. Serum was obtained to detect T. equi-specific antibodies using an indirect enzyme-linked immunosorbent assay (iELISA) based on a crude parasite antigen. In order to identify possible risk factors of the infection which are associated with the prevalence of antibodies, a chi-squared test was carried out. Of 1,117 equids, 373 tested positive for T. equi antibodies with an overall prevalence of 33.4% (31.3%-37.0% for the 95% confidence interval). Sex, animal species and breed were found not to be associated with the presence of T. equi antibodies, whereas age, the presence of dogs or ticks were associated with seropositivity (p < 0.05). Horses with ticks were 2.4 more likely seropositive than horses without ticks. The presence of dogs in the equid habitat and the presence of ticks resulted in a higher T. equi seropositive rate probably because dogs are hosts for vector ticks of T. equi. Our study represents the first report of T. equi antibodies in equids of western Pará revealing a widespread distribution of seropositive animals.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Equidae/parasitología , Enfermedades de los Caballos/epidemiología , Theileria/inmunología , Theileriosis/epidemiología , Enfermedades por Picaduras de Garrapatas/veterinaria , Garrapatas/parasitología , Animales , Brasil/epidemiología , Estudios Transversales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Enfermedades de los Caballos/parasitología , Caballos , Masculino , Factores de Riesgo , Estudios Seroepidemiológicos , Theileriosis/parasitología , Enfermedades por Picaduras de Garrapatas/epidemiología , Enfermedades por Picaduras de Garrapatas/parasitologíaRESUMEN
Anaplasmosis and theileriosis are considered the most important tick-borne diseases for livestock production worldwide, causing significant economic losses in tropical and subtropical regions. The present study was aimed to develop a multiplex TaqMan® qPCR assay to simultaneously detect Anaplasma marginale and Theileria annulata and to applied it to investigate naturally infected cattle in Cuba. The assay was highly specific, sensible, and efficient; it was more sensitive than a well-established nested PCR and detected 1 DNA copy of each target. Consistent repeatability and reproducibility within and between multiplex qPCR runs was shown. A total of 223 blood samples collected in western Cuba were analyzed for haemoparasites infection in cattle. The multiplex qPCR assay detected A. marginale in 213 samples (95.5%; CI: 95%; 91.9%-97.5%), but all samples were negative for T. annulata. Additionally, the genetic diversity of A. marginale was assessed using 16S rRNA, MSP1a and MSP4 nucleotide and protein sequences. The MSP1a tandem repeats ranged from three to five, and twelve different MSP1a tandem repeats of A. marginale were found, which presented genotypes C, E, and G in the 5'UTR microsatellite region. Phylogenetic analysis using the msp4 gene showed that Cuban strains were closely related to others previously reported in Mexico, Brazil and Asian countries. The multiplex qPCR described here proved to be a rapid, specific and cost-effective mean for the simultaneous detection of A. marginale and T. annulata. Further epidemiological studies using this assay will improve the surveillance of the associated diseases in regions where they are endemic.
Asunto(s)
Anaplasma marginale/aislamiento & purificación , Anaplasmosis/epidemiología , Enfermedades de los Bovinos/epidemiología , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Theileria annulata/aislamiento & purificación , Theileriosis/epidemiología , Anaplasmosis/microbiología , Animales , Bovinos , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/parasitología , Cuba/epidemiología , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reproducibilidad de los Resultados , Theileriosis/parasitologíaRESUMEN
Equine piroplasmosis stands out among the diseases that affect Equidae in Brazil and the world. It is caused by the protozoa Theileria equi and Babesia caballi. The objective of the present study was to carry out the molecular characterization of T. equi using equine blood samples collected in the 5 geographic regions of Brazil. Samples from all over the country were tested for the presence of T. equi by real-time PCR. The 18S rRNA sequences (â¼1,600 bp) obtained from 23 samples taken from naturally infected horses were characterized by sequencing and analyzed to identify the genotypes and the possible sites of genetic variability. Thirteen different T. equi 18S rRNA sequences were identified, and 2 different genotypes were demonstrated to be in circulation in Brazil. Alignment entropy analysis demonstrated the existence of three hypervariable regions (V2, V4, and V8) within the 18S rRNA sequence of T. equi. The V2 region is located between nucleotides 63 and 75, V4 is located between nucleotides 524 and 586, and V8 is located between nucleotides 1,208 and 1,226. The hypervariable region V4 demonstrated the greatest variation within the 18S rRNA sequence of T. equi. Phylogenetic analysis based on the 18S rRNA sequences revealed the formation of 3 distinct clades (A, B, and C). The Brazilian samples belonged to 2 clades (A and C). The present study describes the characterization and heterogeneity of the circulating T. equi 18S rRNA sequences in Brazil. The results confirm that the country is an endemic area for the disease, and they indicate that at least 2 distinct T. equi genotypes are naturally infecting equines in Brazil.
Asunto(s)
Variación Genética , Enfermedades de los Caballos/parasitología , ARN Ribosómico 18S/genética , Theileria/genética , Theileriosis/parasitología , Animales , Brasil , Secuencia de Consenso , ADN Protozoario/química , ADN Protozoario/aislamiento & purificación , Enfermedades Endémicas/veterinaria , Enfermedades de los Caballos/sangre , Caballos , Funciones de Verosimilitud , ARN Protozoario/sangre , ARN Protozoario/genética , ARN Ribosómico 18S/sangre , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Alineación de Secuencia/veterinaria , Theileria/clasificación , Theileriosis/sangreRESUMEN
The agents of equine piroplasmosis, Theileria equi and Babesia caballi, are endemic in Trinidad, West Indies. While transmission is mainly by ixodid ticks, transplacental transmission of T. equi has also been reported. This disease has contributed to foetal losses as well as morbidity and mortality of neonatal foals and adult horses. Previous 18S rRNA-based phylogenetic studies indicated a noticeable degree of variation within and among B. caballi and T. equi isolates from different geographical regions. The objective of this study was to evaluate the diversity of T. equi and B. caballi obtained from horses in Trinidad by amplifying a region of the 18S rRNA gene. The phylogenetic trees for T. equi sequences obtained from horses in 2006 and 2011-2013 revealed that Trinidad sequences were of genotype A. Additionally, all of the B. caballi sequences from Trinidad were grouped together with other B. caballi sequences of genotype A. However, T. equi sequences from horses in Saint Kitts and Nevis clustered with sequences of genotype C. This study also identified two genotypes of T. equi in the equine population of Brazil. All of the T. equi and B. caballi sequences obtained from horses in Trinidad belong to genotype A and were similar to T. equi and B. caballi sequences of the same genotype that were submitted to GenBank™ databases. Countries in close proximity to Trinidad have T. equi sequences belonging to genotype C; therefore, movement of horses between these countries can introduce a new genotype of T. equi into the equid population of Trinidad.
Asunto(s)
Babesia/genética , Babesiosis/epidemiología , Enfermedades de los Caballos/parasitología , Caballos/parasitología , ARN Ribosómico 18S/genética , Theileria/genética , Theileriosis/epidemiología , Animales , Babesiosis/parasitología , Babesiosis/transmisión , Brasil/epidemiología , ADN Protozoario/genética , Femenino , Genotipo , Ixodidae/parasitología , Masculino , Filogenia , Análisis de Secuencia de ADN , Theileriosis/parasitología , Trinidad y Tobago/epidemiologíaRESUMEN
Abstract We evaluated the distribution of piroplasmids in equids from the Mato Grosso state in Midwestern Brazil using molecular methods and the interspecific genetic diversity. For this, 1,624 blood samples of equids from 973 farms were examined by PCR, using primer pairs that amplify a fragment of the genes rap-1 and ema-1 of Babesia caballi and Theileria equi, respectively. For molecular characterization and phylogenetic studies, 13 and 60 sequences of the rap-1 and ema-1 genes, respectively, were used to build a dendogram using maximum parsimony. B. caballi and T. equi were detected in 4.11% and 28.16% of the farms, respectively, and molecular prevalence was 2.74% for B. caballi and 25.91% for T. equi. The location of the farms and animals raised in the Pantanal ecoregion influence the probability of equids testing positive for B. caballi and T. equi . Moreover, age and herd purpose were variables significantly associated with T . equi infection. The sequences of B. caballi presented 1.95% intraspecific variability, contrasting with 2.99% in T. equi. Dendrograms for both species demonstrated the presence of subgroups with high values of support of branches. However, it is not possible to associate these groups with geographic origin and/or ecoregion.
Resumo Foi avaliada a distribuição de piroplasmídeos em equídeos do Estado de Mato Grosso, no Centro-Oeste do Brasil, utilizando-se métodos moleculares e a diversidade genética interespecífica. Para isso, 1.624 amostras de sangue de equídeos de 973 fazendas foram examinadas pela PCR, usando pares de oligonucleotídeos que amplificam um fragmento dos genes rap-1and ema-1 de Babesia caballi e Theileria equi, respectivamente. Para caracterização molecular e estudos filogenéticos, foram utilizadas 13 e 60 sequências dos genes rap-1 e ema-1, respectivamente, para construção de um dendograma utilizando máxima parcimônia. B. caballi e T . equi foram detectados em 4,11% e 28,16% das fazendas, respectivamente, e a prevalência molecular foi de 2,74% para B. caballi e 25,91% para T. equi. A localização das fazendas e animais criados na ecorregião do Pantanal influenciam a probabilidade de equídeos serem positivos para B. caballi e T. equi. Além disso, idade e propósito do rebanho foram variáveis, significativamente, associadas à infecção por T. equi. As sequências de B . caballi apresentaram variabilidade intraespecífica de 1,95%, contrastando com 2,99% em T. equi. Dendrogramas para ambas as espécies demonstraram a presença de subgrupos com altos valores de sustentação dos ramos. No entanto, não é possível associar esses grupos com origem geográfica e/ou ecorregião.
Asunto(s)
Animales , Theileriosis/epidemiología , Babesia/genética , Babesiosis/epidemiología , Variación Genética/genética , Theileria/genética , Enfermedades de los Caballos/epidemiología , Filogenia , Especificidad de la Especie , Theileriosis/diagnóstico , Theileriosis/parasitología , Babesiosis/diagnóstico , Babesiosis/parasitología , Brasil/epidemiología , Prevalencia , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/parasitología , CaballosRESUMEN
We evaluated the distribution of piroplasmids in equids from the Mato Grosso state in Midwestern Brazil using molecular methods and the interspecific genetic diversity. For this, 1,624 blood samples of equids from 973 farms were examined by PCR, using primer pairs that amplify a fragment of the genes rap-1 and ema-1 of Babesia caballi and Theileria equi, respectively. For molecular characterization and phylogenetic studies, 13 and 60 sequences of the rap-1 and ema-1 genes, respectively, were used to build a dendogram using maximum parsimony. B. caballi and T. equi were detected in 4.11% and 28.16% of the farms, respectively, and molecular prevalence was 2.74% for B. caballi and 25.91% for T. equi. The location of the farms and animals raised in the Pantanal ecoregion influence the probability of equids testing positive for B. caballi and T. equi . Moreover, age and herd purpose were variables significantly associated with T . equi infection. The sequences of B. caballi presented 1.95% intraspecific variability, contrasting with 2.99% in T. equi. Dendrograms for both species demonstrated the presence of subgroups with high values of support of branches. However, it is not possible to associate these groups with geographic origin and/or ecoregion.
Asunto(s)
Babesia/genética , Babesiosis/epidemiología , Variación Genética/genética , Enfermedades de los Caballos/epidemiología , Theileria/genética , Theileriosis/epidemiología , Animales , Babesiosis/diagnóstico , Babesiosis/parasitología , Brasil/epidemiología , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/parasitología , Caballos , Filogenia , Prevalencia , Especificidad de la Especie , Theileriosis/diagnóstico , Theileriosis/parasitologíaRESUMEN
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.
Asunto(s)
Variación Genética , Enfermedades de los Caballos/parasitología , Theileria/genética , Theileriosis/parasitología , Animales , Brasil , Caballos , Filogenia , Reacción en Cadena de la Polimerasa , ARN Protozoario/análisis , ARN Ribosómico 18S/análisis , Análisis de Secuencia de ARNRESUMEN
Theileria equi infection prevalence was calculated from 1000 blood samples obtained from apparently healthy horses in western Mexico. Samples were sent to the Animal Biotechnology Laboratory of the University of Guadalajara (Mexico) for T. equi diagnosis. Nested polymerase chain reaction (nPCR) was used as a diagnostic method to detect pathogen DNA. Using primers for the merozoite antigen-1 (EMA-1) gene, 19.70±2.47% of the horses (95% CI, 17.23-22.17%) tested positive for T. equi. There was no significant association between gender and T. equi infection. However, prevalence was higher among stabled horses (25.81%) than that among grazing horses (15.02%). The positivity rate was also higher among Quarter Horse (24.70%), Lusitano (35.90%), and Costa Rican Saddle Horse (47.37%) breeds than that among the other seven breeds investigated in this study. The percentage of T. equi infection was higher among adult horses (≥ 4years old, 25.05%) than that among colts and fillies (2-4years old, 15.48%), yearlings (1-2years old, 10.49%), and foals (<1year old, 10.34%). This is the first study of T. equi infection prevalence among horses in Mexico by nPCR . The results indicate that the equine piroplasmosis (EP) caused by T. equi is enzootic in western Mexico.
Asunto(s)
Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/epidemiología , Reacción en Cadena de la Polimerasa/veterinaria , Theileria/aislamiento & purificación , Theileriosis/diagnóstico , Theileriosis/epidemiología , Animales , Babesiosis/epidemiología , Babesiosis/parasitología , Femenino , Enfermedades de los Caballos/parasitología , Caballos/parasitología , Masculino , México/epidemiología , Prevalencia , Theileria/genética , Theileriosis/parasitologíaRESUMEN
Theileriosis is a worldwide protozoal tick-borne disease caused by Theileria equi, which may produce a variety of clinical signs and turn infected horses into lifetime carriers. This study has aimed to perform a serological and molecular detection of T. equi and associated factors in sports horses from six areas of northeastern Brazil. In overall, 59.6% horses were positive by indirect immunofluorescence assay and 50.4% by polymerase chain reaction. No significant association was found when presence of ticks, age, gender, anemia or total plasma proteins was analyzed with seropositivity and molecular techniques. Although a significant association of infection was found in two cities. Thus, local risk factors other than presence of ticks, horse age, gender, anemia and total plasmatic proteins may dictate prevalence of T. equi infection in sports horses, even in highly endemic areas with no control of infection prior to horse competitions.
Asunto(s)
Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/parasitología , Theileria/aislamiento & purificación , Theileriosis/diagnóstico , Theileriosis/parasitología , Anemia/parasitología , Anemia/veterinaria , Animales , Brasil/epidemiología , ADN Protozoario/análisis , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente Indirecta , Enfermedades de los Caballos/inmunología , Caballos/parasitología , Reacción en Cadena de la Polimerasa , Proteínas Protozoarias/genética , Factores de Riesgo , Análisis de Secuencia de ADN , Deportes , Theileria/genética , Theileria/inmunología , Theileriosis/epidemiologíaRESUMEN
Equine piroplasmosis caused by Theileria equi and Babesia caballi is endemic in Trinidad and Tobago. Transmission occurs by ticks of the family Ixodidae. T. equi can also be transmitted transplacentally; however transplacental transmission of B. caballi is unknown. This study aims to investigate transplacental transmission of equine piroplasmosis from thoroughbred mares naturally infected via the tick vector. Whole blood and serum samples were collected from 117 mares in the fifth month of pregnancy. Blood samples were also collected from each of their foals (89 in total) within the first 36h of birth. Additionally, all foals were observed for clinical signs within 30days post - partum. All samples were examined microscopically for intra-erythrocytic piroplasms. Serum ELISA tests and PCR analysis on whole blood were performed to determine the presence of T. equi and B. caballi. Thirty-four (30.6%) mares and 14 (15.7%) of their foals were seropositive for T. equi. Twenty-seven (24.3%) mares were positive for T. equi by conventional (c) PCR. Real time (q) PCR analysis based on the ema - 1 gene revealed that seven (8%) foals were positive for T. equi. Eighty-nine (76.1%) mares and 38 (42.7%) foals were seropositive for B. caballi. Four (3.4%) mares were positive for B. caballi by cPCR. Three out of the four cPCR positive mares either had resorptions, or stillbirths for that pregnancy. From this study, there is strong evidence that transplacental transmission of B. caballi can occur leading to foetal losses. Six foals (7%) were positive for B. caballi by qPCR. Of these six, four were born to B. caballi seropositive mares. In this study a foal born of a T. equi seropositive mare was 55.7 times more likely to be serologically positive for T. equi than a foal born to a T. equi seronegative mare. Similarly a foal born of a B. caballi seropositive mare was 39.4 times more likely to be serologically positive for B. caballi than a foal born to a mare that was serologically negative for B. caballi at the fifth month of pregnancy. This is as a result of the ingestion of colostrum containing antibodies to these pathogens. Mares should be screened during pregnancy and their foals closely monitored at parturition for evidence of equine piroplasmosis so that treatment can be implemented earlier for a better prognosis.
Asunto(s)
Babesiosis/transmisión , Enfermedades de los Caballos/transmisión , Complicaciones Parasitarias del Embarazo/veterinaria , Theileriosis/transmisión , Aborto Veterinario/epidemiología , Aborto Veterinario/parasitología , Animales , Anticuerpos Antiprotozoarios/sangre , Babesia/genética , Babesia/inmunología , Babesiosis/epidemiología , Babesiosis/parasitología , ADN Protozoario/sangre , ADN Protozoario/aislamiento & purificación , Eritrocitos/parasitología , Femenino , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/parasitología , Caballos , Modelos Logísticos , Placenta/parasitología , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Embarazo , Complicaciones Parasitarias del Embarazo/epidemiología , Complicaciones Parasitarias del Embarazo/parasitología , Factores de Riesgo , Bazo/parasitología , Encuestas y Cuestionarios , Theileria/genética , Theileria/inmunología , Theileriosis/epidemiología , Theileriosis/parasitología , Trinidad y Tobago/epidemiologíaRESUMEN
Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed.
Asunto(s)
Babesia , Babesiosis/parasitología , Enfermedades de los Perros/parasitología , Ehrlichia canis , Ehrlichiosis/veterinaria , Theileriosis/parasitología , Animales , Brasil , Perros , Ehrlichiosis/parasitologíaRESUMEN
Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from buffaloes to cattle.