Piroplasm Infection in Domestic Cats in the Mountainous Region of Rio de Janeiro, Brazil.

Piroplasm infections in domestic cats have been reported with increasing frequency in numerous countries. However, in some states of Brazil, little information is available about the occurrence of these parasites. Blood samples were collected from 250 cats treated at a private clinic in the mountainous region of Rio de Janeiro. The samples were each subjected to a blood count, microscopic examination, and molecular research on piroplasms. The animals' clinical histories and epidemiological information were analyzed to identify the risk factors associated with infection. Ticks were recovered during the clinical care and were identified and subjected to molecular analyses to determine the presence of piroplasm DNA. Piroplasms were detected in 2.7% (7/250) of the cats. Nucleotide sequences of Babesia vogeli were identified in six cats, while the Cytauxzoon sp. was identified in one cat. Cats displaying apathy/weakness/prostration and hemorrhage/bleeding were more likely to be infected. In addition, Amblyomma aureolatum was recovered from a cat PCR-negative for piroplasms. This is the first study in Rio de Janeiro that has detected Babesia vogeli in cats. The results obtained here underscore the need for further studies in Rio de Janeiro to investigate the dynamics of such infections and the vectors involved.

Occurrence and Molecular Identification of Hemoparasites in Wild Mammals Kept in Rehabilitation Centers in Brazil.

PURPOSE: Hepatozoonosis and piroplasmosis are diseases caused by apicomplexan protozoa that affect different types of animals, including mammals. The present study aimed to evaluate the occurrence of Hepatozoon spp. and piroplasms in wild mammals kept in captivity in rehabilitation centers in the states of Minas Gerais and Goiás, Brazil. METHODS: For this, blood samples from 152 animals were collected and analyzed by conventional optical microscopy and polymerase chain reaction (PCR). In addition, positive PCR samples were submitted to sequencing for molecular characterization of the specimens found. RESULTS: Microscopic analysis revealed 53 of the 152 animals (28.3%) parasitized by piroplasms. No Hepatozoon sp. was observed. On the other hand, using the primers HepF300/HepR900 and Piro1F/Piro5R, both amplifying fragments of the 18S rDNA gene, eight animals (5.2%) were positive for Hepatozoon spp. and 40 (26.3%) for piroplasms. From the sequencing of the positive samples Hepatozoon canis, Hepatozoon felis, Theileria cervi, Theileria equi and Cytauxzoon felis were identified. In addition to the aforementioned hemoparasites, some animals were found parasitized by microfilaria. Such data ratify the presence of hemoparasites in captive wild animals, and are unprecedented in the two geographical regions covered by the present study. 19.7% of mammals harbored ectoparasites of the genera Amblyomma and Rhipicephalus. CONCLUSION: Wild mammals are infected by several pathogens that can also infect domestic animals, some of them potentially zoonotic which can directly contribute to mortality and species reduction. Therefore, a deep understanding of the parasites, the hosts and the diseases is extremely necessary so that prevention, control and treatment measures are effectively applied.

An atypical case of Babesia bigemina parasitising a dog from a rural area of eastern Mexico / Um caso atípico de Babesia bigemina parasitando um cão de uma área rural do leste do México

A dog that shared habitat with domestic animals in a cattle farm and that was exposed to wildlife was taken to a private practitioner for clinical examination. The analyses conducted on the patient revealed the presence of Babesia bigemina by a molecular test. Clinical signs such as lethargy, anorexia and hyperthermia > 39 °C, pale mucous membranes and blood urine were observed in the patient. The animal was treated with imidocarb dipropionate (two doses each 0.5 ml/10 kg b.w. at an interval of 14 days). On treatment day 7, the clinical signs were mostly reduced. On day 30, PCR was carried out to assess the efficacy of the treatment, with a negative result. This case represents the first report of babesiosis due to B. bigemina in a dog living on a cattle farm in Mexico. It indicates the lower host specify of these pathogens and that dogs can play a role as sentinels of vector-borne parasites in livestock animals.(AU)

Um cão que compartilhava hábitat com animais domésticos em uma fazenda de gado e que foi exposto à vida selvagem foi levado a um clínico particular para que fosse examinado. As análises realizadas no paciente revelaram a presença de Babesia bigemina por um teste molecular. Sinais clínicos, como letargia, anorexia e hipertermia > 39°C, mucosas pálidas e sangue na urina foram observados no paciente. O animal foi tratado com dipropionato de imidocarb (duas doses cada 0,5 ml/10 kg de peso corporal em um intervalo de 14 dias). No dia de tratamento 7, os sinais clínicos foram reduzidos. No dia 30, foi realizada PCR para avaliar a eficácia do tratamento, com resultado negativo. Esse caso representa o primeiro relato de babesiose por B. bigemina em um cão que vive em uma fazenda de gado no México. Isso indica que o hospedeiro inferior especifica esses patógenos, e que os cães podem desempenhar um papel como sentinelas de parasitas transmitidos por vetores em animais de criação.(AU)

Fatal infection caused by Cytauxzoon felis in a captive-reared jaguar (Panthera onca).

Fatal infections with Cytauxzoon felis are common in domestic cats, and jaguars (Panthera onca) are believed carriers of this protozoan. Fatal C. felis infections have never been described in jaguars before. Thus, this study describes such an infection in a 5-month-old captive-reared jaguar that presented hyporexia and died at 26 days after the first clinical signs. During necropsy, samples were taken from several tissues, some were fixed in 10% formalin and process for histopathological diagnosis, and some samples were used for DNA extraction, amplification via PCR and sequencing of the potential pathogens. Grossly, marked splenomegaly and icterus were observed. Histologically, numerous structures that are morphologically compatible with Cytauxzoon spp. schizonts obliterated multiple blood vessels in the brain, leptomeninges, spinal cord, lungs, heart, skeletal muscle, adrenal gland, kidneys, spleen, small intestine and pancreas. C. felis was identified by PCR in many organs. Thus, C. felis infection in jaguars can be fatal, and the clinicopathological findings are similar to those of cytauxzoonosis in other wild and domestic felid species.

CYTAUXZOON FELIS DNA DETECTION IN HEALTHY CATS FROM RIO DE JANEIRO, BRAZIL.

Feline cytauxzoonosis is a disease caused by Cytauxzoon felis, a protozoan that infects the red blood cells and macrophages. It is responsible for an acute and often fatal disease in domestic cats. The purpose of this study was to investigate the occurrence of C. felis infections in healthy cats. Piroplasm forms were seen in the erythrocytes of 2 cats, and C. felis DNA was identified by polymerase chain reaction (PCR) in one of them. The results demonstrate that erythrocytic piroplasmids associated with tick-borne parasitic protozoa may be found circulating in the blood of healthy cats in Rio de Janeiro. These can be differentiated from the morphologically similar forms of species such as Babesia by analysis of DNA, thereby demonstrating the potential for further studies of feline populations in Brazil.

Use of molecular tools for the diagnosis of rangeliosis by Rangelia vitalii in Argentina: A case report.

Vector-borne pathogens are responsible for serious emerging diseases and Rangelia vitalii, the etiologic agent of canine rangeliosis, is one of the most pathogenic tick-borne pathogens for dogs in South America. This protozoan is transmitted by the Amblyomma aureolatum tick bite and the clinical features associated to the disease are fever, hemolytic anemia, jaundice, hepatosplenomegaly and bleeding from natural orifices, mainly from the ear egde. The reports of canine rangeliosis in Argentina are scarce. In the present study we report the detection of Rangelia vitalii in a naturally infected dog from Gualeguay, Entre Ríos, Argentina with history of tick infestation and clinical signs compatible with rangeliosis. An initial blood sample was positive to piroplasmids by blood smear examination and the molecular amplification of a fragment of the 18SrRNA gene. Sequencing of the fragment confirmed the pathogen identity. After treatment with imidocarb dipropionate, the clinical signs remitted and the blood smear tested negative.

Theileria sp. in water buffaloes from Maranhão State, northeastern Brazil / Theileria sp. em búfalos do Estado do Maranhão, nordeste do Brasil

Anaplasma marginale and piroplasm species are widespread among Brazilian cattle herds. Both of these tick-borne pathogens hamper livestock production and cause a significant economic impact. Although buffaloes have demonstrated a high level of adaptability, data on tick-borne pathogens are scarcely reported in Brazil. Thus, the aim of this study was to screen water buffaloes from the state of Maranhão for piroplasm and A. marginale occurrence using PCR assays. All samples were negative for A. marginale. One of the 287 (0.35%) water buffaloes tested was positive for Theileria sp. Sequencing of the 18S rDNA fragment (356 bp) showed that the Theileria sp. identified was closely related to the T. buffeli /orientalis group. Future studies on the clinical signs of infection and the main vector in this country are needed.(AU)

Anaplasma marginale e espécies de piroplasma são amplamente distribuídas no rebanho bovino brasileiro. Ambos os patógenos transmitidos por carrapatos dificultam a produção pecuária e causam um impacto econômico significativo. Embora os búfalos tenham demonstrado um alto nível de adaptabilidade, dados sobre patógenos transmitidos por carrapatos são raramente relatados no Brasil. Assim, o objetivo deste estudo foi investigar búfalos do estado do Maranhão para piroplasmas e A. marginale utilizando-se a técnica da PCR. Todas as amostras foram negativas para A. marginale . Um dos 287 (0,35%) búfalos testados foi positivo para Theileria sp. O sequenciamento de um fragmento do gene 18S rDNA (356 pb) demonstrou que Theileria sp. identificado estava relacionada ao grupo T. buffeli/orientalis . Estudos futuros sobre os sinais clínicos de infecção e o principal vetor neste país são necessários.(AU)

Theileria sp. in water buffaloes from Maranhão State, northeastern Brazil / Theileria sp. em búfalos do Estado do Maranhão, nordeste do Brasil

Abstract Anaplasma marginale and piroplasm species are widespread among Brazilian cattle herds. Both of these tick-borne pathogens hamper livestock production and cause a significant economic impact. Although buffaloes have demonstrated a high level of adaptability, data on tick-borne pathogens are scarcely reported in Brazil. Thus, the aim of this study was to screen water buffaloes from the state of Maranhão for piroplasm and A. marginale occurrence using PCR assays. All samples were negative for A. marginale. One of the 287 (0.35%) water buffaloes tested was positive for Theileria sp. Sequencing of the 18S rDNA fragment (356 bp) showed that the Theileria sp. identified was closely related to the T. buffeli /orientalis group. Future studies on the clinical signs of infection and the main vector in this country are needed.

Resumo Anaplasma marginale e espécies de piroplasma são amplamente distribuídas no rebanho bovino brasileiro. Ambos os patógenos transmitidos por carrapatos dificultam a produção pecuária e causam um impacto econômico significativo. Embora os búfalos tenham demonstrado um alto nível de adaptabilidade, dados sobre patógenos transmitidos por carrapatos são raramente relatados no Brasil. Assim, o objetivo deste estudo foi investigar búfalos do estado do Maranhão para piroplasmas e A. marginale utilizando-se a técnica da PCR. Todas as amostras foram negativas para A. marginale . Um dos 287 (0,35%) búfalos testados foi positivo para Theileria sp. O sequenciamento de um fragmento do gene 18S rDNA (356 pb) demonstrou que Theileria sp. identificado estava relacionada ao grupo T. buffeli/orientalis . Estudos futuros sobre os sinais clínicos de infecção e o principal vetor neste país são necessários.

Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil / Rangelia vitalii, Babesia spp. e Ehrlichia spp. em cães de Passo Fundo, estado do Rio Grande do Sul, Brasil

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.(AU)

Patógenos transmitidos por carrapatos são um problema emergente em todo o mundo, o trabalho objetivou diagnosticar os agentes causais da infecção em cães com suspeita de hemoparasitoses. Cinquenta e oito caninos com sinais clínicos como depressão, diáteses hemorrágicas e febre foram avaliados quanto à apresentação clínica, hemograma, esfregaço sanguíneo, sorologia pelo método de Imunofluorescência Indireta para os agentes Babesia vogeli e Ehrlichia canis e na PCR convencional para Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) e Ehrlichia spp. (gene dsb). Cinco (8,6%) dos 58 cães apresentaram sorologia positiva para Babesia spp. e três (5,1%) para E. canis. Quatro (6,8%) animais mostraram-se positivos para R. vitalii no diagnóstico molecular. Os produtos da PCR foram sequenciados e o DNA encontrado de R. vitalii mostrou 99% de identidade genética com amostras de R. vitalii isoladas no Brasil. Não foi observada a presença de Babesia spp. e E. canis na PCR dos cães avaliados. Os resultados indicaram a presença de R. vitalii e exposição a Babesia spp. e Ehrlichia spp. entre os cães analisados.(AU)

Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil / Rangelia vitalii, Babesia spp. e Ehrlichia spp. em cães de Passo Fundo, estado do Rio Grande do Sul, Brasil

Abstract 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.

Resumo Patógenos transmitidos por carrapatos são um problema emergente em todo o mundo, o trabalho objetivou diagnosticar os agentes causais da infecção em cães com suspeita de hemoparasitoses. Cinquenta e oito caninos com sinais clínicos como depressão, diáteses hemorrágicas e febre foram avaliados quanto à apresentação clínica, hemograma, esfregaço sanguíneo, sorologia pelo método de Imunofluorescência Indireta para os agentes Babesia vogeli e Ehrlichia canis e na PCR convencional para Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) e Ehrlichia spp. (gene dsb). Cinco (8,6%) dos 58 cães apresentaram sorologia positiva para Babesia spp. e três (5,1%) para E. canis. Quatro (6,8%) animais mostraram-se positivos para R. vitalii no diagnóstico molecular. Os produtos da PCR foram sequenciados e o DNA encontrado de R. vitalii mostrou 99% de identidade genética com amostras de R. vitalii isoladas no Brasil. Não foi observada a presença de Babesia spp. e E. canis na PCR dos cães avaliados. Os resultados indicaram a presença de R. vitalii e exposição a Babesia spp. e Ehrlichia spp. entre os cães analisados.

Arthropod-borne pathogens circulating in free-roaming domestic cats in a zoo environment in Brazil.

Recently, tick and flea-borne pathogens have been detected in wild carnivores maintained in captivity in Brazilian zoos. Since free-roaming cats are frequently found in Brazilian zoos, they could act as reservoirs for arthropod-borne pathogens, which could be transmitted to endangered wild carnivores maintained in captivity in these institutions. On the other hand, stray cats in zoos may play a role as sentinels to pathogens that circulate among wild animals in captivity. The present work aimed to detect the presence of Anaplasmataceae agents, hemoplasmas, Bartonella species, piroplasmas, and Hepatozoon sp. DNA in blood samples of 37 free-roaming cats in a Brazilian zoo. Three (8%) cats were positive for Anaplasma spp. closed related to Anaplasma phagocytophilum; 12 (32%) cats were positive for hemoplasmas [two (5%) for Mycoplasma haemofelis, five (13.5%) for Candidatus Mycoplasma haemominutum, and five (13.5%) for Candidatus Mycoplasma turicensis]; 11 (30%) were positive for Bartonella spp., six (16%) were positive Babesia vogeli and one (3%) for Theileria sp. Coinfection with multiple arthropod-borne agentes was observed in sampled cats. None of sampled cats were positive for Ehrlichia spp., Cytauxzoon spp., or Hepatozoon spp. in PCR. This is the first molecular detection of Babesia vogeli and Theileria sp. in domestic cats in Brazil. The control of the population of free-roaming cats in these conservation institutions is much needed aiming to prevent the potential transmission to endangered wild animals maintained in captivity, such as wild neotropical wild felids, as well as to human beings visiting zoos.

Tick-borne pathogens and disease in dogs on St. Kitts, West Indies.

Between 2009 and 2011, we conducted a case-control study of ticks and tick-associated pathogens affecting dogs on the island of St. Kitts, eastern Caribbean, including 55 cases of clinically suspected tick-borne disease (TBD) and 110 presumably healthy animals presented for elective surgeries. Rhipicephalus sanguineus caused year-round infestations of dogs, and 36% of the dogs in the study were infested at the time of examination. Overall, 62% of suspected TBD cases and 24% of presumably healthy dogs tested positive by PCR for infections with: Anaplasma platys (0% and 4%), Babesia canis vogeli (20% and 6%), Babesia gibsoni (18% and 5%), Ehrlichia canis (35% and 7%), and Hepatozoon canis (5% and 2%). Co-infections were documented in 15% of these PCR-positive dogs. Antibodies against A. platys or E. canis were noted in 36% of the dogs. Thrombocytopenia was the most common sign of infection, followed by anemia. This is the first detection of A. platys, B. canis vogeli, or H. canis on St. Kitts and the first detection of B. gibsoni in the Caribbean. We conclude that tick-borne pathogens of dogs are highly prevalent in this region and may present in dogs that appear healthy, in spite of hematologic abnormalities that may increase surgical risk.

Detection and molecular characterization of piroplasms species from naturally infected dogs in southeast Brazil / Detecção e caracterização molecular de piroplasmas em cães naturalmente infectados no Sudeste do Brasil

Rangelia vitalii is a protozoon described from dogs in the south and southeast regions of Brazil. It is phylogenetically related to Babesia spp. that infects dogs, but data on this enigmatic parasite is still limited. The aim of this work was to detect piroplasm species in dogs in the state of Rio de Janeiro, Brazil, by 18S rRNA gene-based PCR assay, restriction fragment length polymorphism (RFLP) and sequence analyses. Of 103 dogs examined, seven (6.8%) were positive for Babesia spp. by PCR. The amplified products were digested by restriction enzymes to differentiate the Babesia species, and one sample was identified as Babesia vogeli. The pattern observed for the other six amplification products did not match with pattern described for large Babesia infecting dogs. Sequencing analysis confirmed these six samples as R. vitalii, with high homologies (99-100%) with a sequence from south Brazil. This study confirms the presence of Babesia vogeli and Rangelia vitalii circulate in domestic dogs in Teresópolis, Rio de Janeiro, Brazil.

Rangelia vitalii é um protozoário que infecta cães e foi descrito nas regiões Sul e Sudeste do Brasil. R. vitalii é filogeneticamente próxima à Babesia spp., mas dados deste misterioso parasito ainda são escassos. O objetivo deste trabalho foi detectar a presença de piroplasmas em cães naturalmente infectados no estado do Rio de Janeiro, através da amplificação do gene 18S rRNA pela PCR, clivagem com enzimas de restrição (RFLP) e caracterização genética através do sequenciamento. De 103 cães, sete (6,8%) foram positivos para Babesia spp. pela PCR. Os produtos amplificados foram digeridos por enzimas de restrição para a diferenciação das espécies de Babesia e uma amostra foi identificada como Babesia vogeli. O padrão de amplificação observado nas outras seis amostras não correspondeu ao padrão descrito para babesias que infectam cães. O sequenciamento das seis amostras confirmou ser uma espécie geneticamente idêntica a R. vitalii apresentando grande homologia (99-100%) com a sequência do sul do Brasil. Este estudo confirma a presença de Babesia vogeli e Rangelia vitalii infectando cães em Teresópolis, Rio de Janeiro, Brasil.

Detection and molecular characterization of piroplasms species from naturally infected dogs in southeast Brazil / Detecção e caracterização molecular de piroplasmas em cães naturalmente infectados no Sudeste do Brasil

Rangelia vitalii is a protozoon described from dogs in the south and southeast regions of Brazil. It is phylogenetically related to Babesia spp. that infects dogs, but data on this enigmatic parasite is still limited. The aim of this work was to detect piroplasm species in dogs in the state of Rio de Janeiro, Brazil, by 18S rRNA gene-based PCR assay, restriction fragment length polymorphism (RFLP) and sequence analyses. Of 103 dogs examined, seven (6.8%) were positive for Babesia spp. by PCR. The amplified products were digested by restriction enzymes to differentiate the Babesia species, and one sample was identified as Babesia vogeli. The pattern observed for the other six amplification products did not match with pattern described for large Babesia infecting dogs. Sequencing analysis confirmed these six samples as R. vitalii, with high homologies (99-100%) with a sequence from south Brazil. This study confirms the presence of Babesia vogeli and Rangelia vitalii circulate in domestic dogs in Teresópolis, Rio de Janeiro, Brazil.(AU)

Rangelia vitalii é um protozoário que infecta cães e foi descrito nas regiões Sul e Sudeste do Brasil. R. vitalii é filogeneticamente próxima à Babesia spp., mas dados deste misterioso parasito ainda são escassos. O objetivo deste trabalho foi detectar a presença de piroplasmas em cães naturalmente infectados no estado do Rio de Janeiro, através da amplificação do gene 18S rRNA pela PCR, clivagem com enzimas de restrição (RFLP) e caracterização genética através do sequenciamento. De 103 cães, sete (6,8%) foram positivos para Babesia spp. pela PCR. Os produtos amplificados foram digeridos por enzimas de restrição para a diferenciação das espécies de Babesia e uma amostra foi identificada como Babesia vogeli. O padrão de amplificação observado nas outras seis amostras não correspondeu ao padrão descrito para babesias que infectam cães. O sequenciamento das seis amostras confirmou ser uma espécie geneticamente idêntica a R. vitalii apresentando grande homologia (99-100%) com a sequência do sul do Brasil. Este estudo confirma a presença de Babesia vogeli e Rangelia vitalii infectando cães em Teresópolis, Rio de Janeiro, Brasil.(AU)