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The present study aimed to investigate the occurrence of Leishmania spp., hemotropic Mycoplasma spp., tick-borne pathogens (TBP), and co-infection in dogs with clinical signs suggestive of visceral leishmaniasis (VL). It also aimed to determine the factors associated with infection and to map the distribution of co-infected dogs in an endemic area in the Northeast region of Brazil. Blood samples from 168 dogs were evaluated for serological analysis to Leishmania spp., Anaplasma spp., Ehrlichia spp., Babesia spp., and molecular assays to Leishmania spp., Anaplasma platys, Ehrlichia canis, Babesia spp., and hemotropic Mycoplasma spp. In serological and molecular analysis, 29.8% and 5.9% of dogs were co-infected. In the regression analysis, seropositivity for Ehrlichia spp., Babesia spp., and Leishmania spp. was significantly associated with the presence of petechiae, young dogs, and weight loss. Serology revealed that co-exposure with Babesia spp. and Ehrlichia spp. was associated with fever and thrombocytopenia, and there was an association between seropositivity for Ehrlichia spp. and Babesia spp. in dogs seropositive for Leishmania spp. The presence of hemotropic Mycoplasma spp. DNA was associated with anorexia. Thus, dogs with clinical VL have co-infection with other pathogens, reinforcing the importance of this study for a better understanding of these co-infections in dogs from endemic areas.

O presente estudo objetivou investigar a prevalência de Leishmania spp., Mycoplasma spp. hemotrópico, patógenos transmitidos por carrapatos (PTC), e coinfecção em cães com sinais clínicos sugestivos de leishmaniose visceral (LV), determinar os fatores associados à infecção, e mapear a distribuição de cães coinfectados em uma área endêmica no Nordeste do Brasil. Amostras de sangue de 168 cães foram avaliadas por análises sorológicas para Leishmania spp., Anaplasma spp., Ehrlichia spp., Babesia spp., e ensaio molecular para Leishmania spp., Anaplasma platys, Ehrlichia canis, Babesia spp., e Mycoplasma hemotrópico. Pelas análises sorológicas e moleculares, 29,8% e 5,9% dos cães apresentaram coinfecção, respectivamente. Na análise de regressão, a soropositividade para Ehrlichia spp., Babesia spp., e Leishmania spp. foram significantemente associadas com a presença de petéquias, cães jovens, e perda de peso. O diagnóstico sorológico revelou que a coexposição à Babesia spp. e Ehrlichia spp. está associada com febre e trombocitopenia, havendo associação entre a soropositividade para Ehrlichia spp. e Babesia spp. em cães soropositivos para Leishmania spp. A presença de DNA de Mycoplasma foi associada à anorexia. Desta forma, cães com sinais de LV possuem coinfecção com outros patógenos, reforçando a importância deste estudo para um melhor entendimento dessas coinfecções em cães de áreas endêmicas.

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Hemoparasite infections are a substantial challenge to agriculture,worldwide. Infected cattle suffer malnutrition, stunting, decreased milk production, and reproductive loss, which can result in great economic loss. The aim of this study was to characterize the predominant hemoparasites affecting cattle in Nicaragua, whose economy is largely dependent on agriculture, and define associated epidemiological factors. Blood from 68 bovines in the municipality of León were analyzed by microscopy and molecular techniques. PCR revealed Anaplasma marginalein 33.82% (IC 95%: 21.84-45.80), 19.11% (IC 95%: 9.03-29.19) of animals were infected with Babesiaspp, and 8.82% (95% CI: 1.34-16.30) were simultaneously co-infected with Babesiaspp and A. marginale. However, A. phagocytophilum, Leishmaniaspp., or Trypanosomaspp were not detected (0%, 95% CI: 0.0-5.20). A. marginale was more frequently detected in males (p=0.041) and calves (p=0.041). This is the first study to report the prevalence of hemoparasites in cattle using molecular diagnosis in western Nicaragua.

Agentes infecciosos chamados hemoparasitas destroem as hemácias em bovinos. O objetivo deste estudo foi detectar os principais hemoparasitas em bovinos no município de León, Nicarágua, e os fatores epidemiológicos associados à sua presença. Foram coletadas 68 amostras de sangue bovino em EDTA e realizada PCR a partir de uma amostra conglomerada selecionada para representar os bovinos do município de León. A análise de PCR revelou que 33,82% (IC 95%: 21,84-45,80) dos animais foram positivos para A. marginale, 19,11% (IC 95%: 9.03-29,19) dos animais estavam infectados com Babesia spp, 8,82% (95% de IC: 1,34-16,30) estavam infectados com Babesia spp. e A. marginale, no entanto, nenhum animal foi considerado positivo para A. phagocytophilum, Leishmania spp. ou Trypanosoma spp (0%, IC 95%: 0,0-5,20). A. marginale foi detectada com maior frequência em machos (p=0,041) e bezerros (p=0,041). Este é o primeiro estudo que relata a prevalência de hemoparasitas em bovinos, aplicando diagnóstico molecular no oeste da Nicarágua.

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Babesiosis and Anaplasmosis are diseases associated with economic losses; ticks and blood-sucking flies are important zoonotic vectors and reservoirs. This study aimed to investigate the presence of anti-Babesia spp. and anti-Anaplasma marginale antibodies using enzyme-linked immunosorbent assay (ELISA), in ruminants at the Catimbau National Park. Blood samples were collected from 119 sheep, 119 goats, and 47 cattle. Rhipicephalus microplus ticks were collected from cattle. ELISA showed seropositivity of 34% (16/47), 20.3% (24/119), and 16% (19/119) for anti-Babesia bovis; 34% (16/47), 15.2% (18/119), and 9% (7/119) for anti-Babesia bigemina; and 34% (16/47), 35.6% (42/119), and 17% (20/119) for anti-A. marginale antibodies in cattle, goats, and sheep, respectively. The information collected using an epidemiological questionnaire showed that mostly are breed in a semi-intensive system, with access to Caatinga vegetation. The circulation of B. bovis, B. bigemina, and A. marginale was confirmed. Thus, based on the prevalence, this suggests this is an enzootic instability area and is prone to outbreaks.(AU)

A Babesiose e a Anaplasmose são doenças associadas a grandes perdas econômicas, sendo carrapatos e moscas sugadoras de sangue importantes vetores e reservatórios zoonóticos dessas enfermidades. Este estudo teve como objetivo investigar a circulação de anticorpos anti-Babesia spp. e anti-Anaplasma marginale usando Ensaio de Imunoabsorção Enzimática (ELISA), e Babesia spp. em ruminantes no Parque Nacional do Catimbau. Foram coletados ectoparasitos e amostras de sangue de 119 ovinos, 119 caprinos e 47 bovinos. Carrapatos Rhipicephalus microplus foram coletados em bovinos. O teste ELISA mostrou soropositividade de 34% (16/47), 20,3% (24/119) e 16% (19/119) para anti-Babesia bovis; 34% (16/47), 15,2% (18/119) e 9% (7/119) para anti-Babesia bigemina; e 34% (16/47), 35,6% (42/119) e 17% (20/119) para anti-A. marginale em bovinos, caprinos e ovinos, respectivamente. As informações coletadas por meio de questionário epidemiológico mostraram a maioria com sistema semi-intensivo, com acesso à vegetação de Caatinga. A circulação de B. bovis, B. bigemina e A. marginale foi confirmada. Assim, com base na prevalência, essa área foi sugerida como de instabilidade enzoótica e propensa a surtos.(AU)

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O artigo apresenta uma revisão sobre os efeitos reprodutivos da infecção pelos principais patógenos transmitidos por vetores que acometem cães, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. e Trypanosoma spp. São discutidas a patologia genital, fertilidade masculina e feminina, transmissão venérea, transmissão vertical, transmissão horizontal mãe-filho e sua relevância na epidemiologia das doenças.(AU)

The article presents a review of the reproductive effects of infection by main vector-borne pathogens of dogs, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. and Trypanosoma spp. Genital pathology, male and female fertility, venereal transmission, vertical transmission, horizontal mother-offspring transmission and their relevance to the epidemiology of the diseases are discussed.(AU)

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The aim of this study was to investigate the association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of pregnant and non-pregnant taurine heifers. Blood samples from 94 females were collected on the first day (D-10) of timed artificial insemination (TAI) protocol and on pregnancy diagnosis (D+34). Hematological parameters were determined and compared between pregnant (PG) and non-pregnant (NPG) heifers, and within group at different sampling days. Real-time PCR (qPCR) was used to determine A. marginale and Babesia bovis infection, and for absolute quantification of Babesia spp. between PG and NPG groups. Correlation analysis was performed between the number of gDNA copies (CN) of Babesia spp. and hematological parameters. On D-10, mean hemoglobin concentration was higher for NPG, and hematocrit and total plasma protein were higher on D+34 for both groups. There was no difference in Babesia spp. CN between groups. In the first qPCR, all heifers were positive for A. marginale and B. bovis. Significant correlations were found between hemoglobin and erythrocyte and between hemoglobin and hematocrit (r = 0.8082 and r = 0.3009, respectively). Low levels of A. marginale and Babesia spp. did not affect hematological parameters of chronically infected pregnant and non-pregnant taurine heifers.(AU)

O objetivo deste estudo foi investigar a associação entre infecção crônica por Anaplasma marginale e Babesia spp. e parâmetros hematológicos de novilhas taurinas prenhes e não prenhes. Sangue de 94 fêmeas foi coletado no primeiro dia (D-10) do protocolo de inseminação artificial em tempo fixo (IATF) e no diagnóstico de gestação (D+34). Parâmetros hematológicos foram comparados entre novilhas prenhes (PG) e não prenhes (NPG) e dentro dos grupos entre dias de coleta. Usando-se PCR em tempo real (qPCR), determinou-se a infecção por A. marginale e Babesia bovis e quantificação absoluta de Babesia spp. Entre os grupos PG e NPG. A análise de correlação foi realizada entre o número de cópias (CN) de Babesia spp. e parâmetros hematológicos. No D-10, a concentração de hemoglobina foi maior para NPG e hematócrito, e proteína plasmática total foram maiores em D+34 para ambos os grupos. Não houve diferença para CN de Babesia spp. entre os grupos. Na primeira qPCR, todas as novilhas foram positivas para A. marginale e B. bovis. Correlações significativas foram encontradas entre hemoglobina/eritrócito e hemoglobina/hematócrito (r=0,8082 e r=0,3009, respectivamente). Baixos níveis de A. marginale e Babesia spp. não afetaram os parâmetros hematológicos de novilhas taurinas prenhes e não prenhes cronicamente infectadas.(AU)

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The present work reviews the epidemiologic situation of Anaplasma marginale and Babesia spp. infections and the occurrence of cattle tick fever outbreaks in Brazil. In areas of tick fever enzootic instability, environmental conditions interfere with the development of Rhipicephalus (Boophilus) microplus: chilly winter in the southern region, floods in the Pantanal, and low humidity in the Caatinga. In contrast, the climatic conditions of stable zones (Cerrado, Amazon and Atlantic Forest biomes) favor tick development. In enzootic areas, tick fever is uncommon because the animals are in frequent contact with the parasite, acquiring immunity naturally during the period of innate resistance; however, outbreaks may occur when calves become infested by considerable numbers of infected ticks during this period or in adults raised in tick-free environments that become infested for the first time when transporting to stable areas. It is necessary to better understand the disease's risk factors under stable conditions and the implications of the mechanical and other vector transmission of A. marginale. To prevent tick fever outbreaks in Brazil, it is important to develop and use anaplasmosis and babesiosis vaccines in cattle from enzootic unstable regions, especially when animals are moved to stable areas.(AU)

O presente trabalho revisa a situação epidemiológica das infecções por Anaplasma marginale e Babesia spp. e a ocorrência de surtos de Tristeza parasitária bovina (TPB) no Brasil. Em áreas de instabilidade enzoótica, as condições ambientais interferem no desenvolvimento do Rhipicephalus (Boophilus) microplus: o frio do inverno na região Sul; as inundações no Pantanal; e a baixa umidade na Caatinga. Por outra parte, as condições climáticas das zonas de estabilidade (os biomas Cerrado, Amazônia e Mata Atlântica) favorecem o desenvolvimento do carrapato. A TPB não é comum nas áreas estáveis, porque os animais estão em contato frequente com os parasitas, adquirindo imunidade naturalmente. Podem, porém, ocorrer surtos quando um número considerável de carrapatos infectados infesta bezerros, durante o período de resistência inata, ou quando adultos que foram criados em ambientes livres de carrapatos infestam-se, pela primeira vez, ao serem transportados para áreas estáveis. É necessário entender melhor os fatores de risco da doença em condições de estabilidade e a implicação da transmissão de A. marginale de forma mecânica, ou por meio de vetores diferentes do carrapato. Para prevenir surtos de TPB, no Brasil, é necessário desenvolver e usar vacinas em bovinos de regiões de instabilidade, especialmente antes de transportá-los para áreas estáveis.(AU)

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Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

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O presente estudo teve como objetivo detectar por meio da Reação em cadeia da Polimerase (PCR) a frequência de Ehrlichiacanis, Babesia spp. e Anaplasma platys em cães, relacionando a prevalência dos achados hematológicos aos resultados positivos pela PCR. Foram avaliadas 209 amostras de sangue de cães atendidos em clínica veterinária particular do município de Queimados, RJ, Brasil, no período de julho a outubro de 2014. Foram realizados hemograma completo e extração de DNA para técnica de PCR. Do total de 209 animais, 19,1% (40/209) animais apresentaram resultado positivo para hemoparasitos pela técnica de PCR. Destes, 52,5% (21/40) foram positivos para E. canis, 27,5% (11/40) positivos para Babesia spp. e 10% (4/40) positivos para A. platys. Quatro animais (1,91%), dos 209 testados, foram positivos para pelo menos dois agentes, caracterizando assim coinfecção. Dos 40 cães positivos para algum dos agentes testados, 25 (62,5%) estavam trombocitopênicos. Ou seja, 15 cães (37,5%) reagiram positivos para hemoparasitos, mas não apresentavam trombocitopenia. A anemia foi um achado comum, sobretudo nas infecções por Babesia spp., 100% (11/11) e E.canis, 90,5% (19/21). A técnica de PCR foi um importante método diferencial na detecção das principais hemoparasitoses caninas, juntamente com os achados clínicos e hematológicos para o diagnóstico preciso da infecção em questão.

The present study aimed to detect, by means of Polimerase chain reaction (PCR), the frequency of Ehrlichia canis, Babesia spp. and Anaplasma platys in dogs, relating the prevalence of hematological findings to positive PCR results. A total of 209 blood samples from dogs treated at a private veterinary clinic in the city of Queimados, RJ, Brazil, from July to October 2014 were evaluated. Complete blood count and DNA extraction were performed for the PCR technique. Of the total of 209 animals, 19.1% (40/209) animals were positive for hemoparasites by the PCR technique. Of these, 52.5% (21/40) were positive for E. canis, 27.5% (11/40) were positive for Babesia spp. and 10% (4/40) positive for A. platys. Four animals (1.91%) of the 209 tested were positive for at least two agents, thus characterizing coinfection. Of the 40 dogs positive for any of the agents tested, 25 (62.5%) were thrombocytopenic. That is, 15 dogs (37.5%) were positive for hemoparasites, but did not have thrombocytopenia. Anemia was a common finding, especially in infections by Babesia spp., 100% (11/11) and E. canis, 90.5% (19/21). The PCR technique was an important differential method in the detection of the main canine hemoparasitoses, together with the clinical and hematological findings for the accurate diagnosis of the infection in question.

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Background: Cerebral babesiosis is a significant cause of mortality in calves, as it can be transmitted through the placenta during gestation and presents with hemolytic and/or neurological symptoms. However, the role of this agent as a cause of abortion is still poorly understood. In endemic areas, the disease primarily affects cattle between the ages of 1 and 12 months and occasionally neonates. This study aimed to describe the epidemiological, clinical, anatomopathological, molecular, and differential diagnostic aspects of a naturally infected newborn Nelore calf. Case: A newborn Nelore calf from Mato Grosso do Sul was discovered dead 1 h after birth and was subjected to necropsy, cytopathological and histopathological examinations by the Pathological Anatomy Laboratory (LAP) at the Federal University of Mato Grosso do Sul (UFMS), as well as complementary in situ hybridization (ISH) and polymerase chain reaction (PCR) tests. Hemoparasite research yielded negative results in all cows within the herd, including the mother of the necropsied calf, which were all clinically healthy. The clinical and hematological examination results of the cows were normal. At necropsy, the calf appeared thin, with a moderately jaundiced carcass and multiple petechiae on the pleural and pericardial surfaces. The lung was inflated until the opening of the thoracic cavity and floated when immersed in formalin, indicating that the calf was born alive and breathing. The spleen and liver were moderately enlarged with rounded edges, and the liver appeared slightly orange. The gray matter of the brain was significantly cherry red. Histologically, the central nervous system capillaries were slightly congested, with numerous punctiform and basophilic structures observed in the erythrocytes obliterating these vessels, either alone or in pairs, measuring 1 to 2 µm in diameter, which were morphologically consistent with Babesia bovis. In the cytological evaluation of the imprint slides of the brain cortex, blood capillaries filled with erythrocytes parasitized by solitary or paired punctiform basophilic structures, approximately 1 µm in diameter, morphologically compatible with B. bovis, were detected. Positive labeling for Babesia spp. was observed in the ISH ex-amination, while the PCR identified B. bovis and B. bigemina in were identified in the brain fragments. The macroscopic findings, including pale or icteric mucous membranes, yellow and enlarged liver, and splenomegaly observed in cases of anaplasmosis, may be confused with those of cerebral babesiosis when the latter does not present with hemoglobinuria. Brain congestion indicates B. bovis infection but does not rule out co-infection with Anaplasma marginale in this case, although this rickettsia was not identified in the blood smear. Discussion: The diagnosis of Babesia bovis infection was based on epidemiological and histological data, as well as the identification of the protozoan in spleen and brain smears and through PCR and ISH exams, which are highly sensitive and can aid in the diagnosis of B. bovis in cases of perinatal deaths. In situ hybridization is effective in cases where the material has autolysis, as the technique allows the genetic material of the agent to be associated with the lesion, even with tissue alterations caused by fixation in formalin. The findings of this study highlight the importance of considering this disease as a differential diagnosis among those that cause abortions or neonatal losses in cattle and emphasize the importance of conducting anatomopathological exams for definitive diagnosis.

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The present study aimed to use polymerase chain reaction (PCR) to detect species of the order Piroplasmida, such as Anaplasma spp., Borrelia spp., and Ehrlichia spp., circulating in the blood of Didelphis aurita in a peridomiciliary environment. Blood samples collected from big-eared opossum (Didelphis aurita) were screened for hemoparasites using PCR. The extracted DNA was tested for tick-borne hemoparasites. We were unable to detect hemoparasites, such as Ehrlichia spp., Babesia spp., Anaplasma spp., and Borrelia spp. Theileria DNA was detected in only one sample screened using PCR for an approximately 650-base pair fragment of the 18S rRNA gene. Sequencing and BLAST analysis of a subset of the PCR amplicons revealed 97% (535/553 bp) identity with Theileria bicornis. The detection of Theileria sp. in D. aurita challenges us to pursue more in-depth studies of marsupial piroplasmosids and to evaluate the morphological aspects of the findings and their possible involvement in zoonoses.

O objetivo do presente estudo foi utilizar a reação em cadeia da polimerase (PCR) para detectar espécies da ordem Piroplasmida, como Anaplasma spp., Borrelia spp. e Ehrlichia spp., circulando no sangue de Didelphis aurita em ambiente peridomiciliar. Amostras de sangue coletadas de gambá da orelha preta (Didelphis aurita) foram triadas para hemoparasitas por meio da PCR. O DNA extraído foi testado para alguns hemoparasitas transmitidos por carrapatos. Não conseguimos detectar hemoparasitos, como Ehrlichia spp., Babesia spp., Anaplasma spp. e Borrelia spp. O DNA de Theileria foi detectado apenas em uma amostra. Um fragmento de ~650 pares de bases do gene 18S rRNA foi sequenciado. e a análise pelo BLAST (um subconjunto de amplicons de PCR) revelou uma identidade de 97% (535/553 pb) com Theileria bicornis. A detecção de Theileria sp. in D. aurita nos desafia a aprofundar estudos sobre piroplasmosídeos marsupiais e avaliar os aspectos morfológicos dos achados e seu possível envolvimento com zoonoses.

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The tick species and tick-borne pathogens present in a group of questing ticks collected from forest fragments in a rural area in the municipality of Divino, Minas Gerais state, Brazil were evaluated. The collected ticks were divided into two groups those collected from around the edges of the fragments and those collected from the interior of the forest. In all the fragments, the ticks were collected using a dragging and flagging technique and by harvesting them from white fabric gaiters. The larvae, nymphs, and adults were all morphologically identified using specific taxonomic keys. The larvae were identified to the genus level. DNA was extracted from the ticks and tested for the presence of Rickettsia spp., Borrelia spp., Ehrlichia spp., Anaplasma spp., Babesia spp., and Theileria spp. using a conventional polymerase chain reaction (PCR). In total, 1,122 questing ticks (750 larvae, 367 nymphs, and five adults) and 18 larval clusters were evaluated. The main species found in the collected tick population were Amblyomma sculptum, A. auricularium, A. aureolatum, and A. pseudoconcolor, along with the larvae of Amblyomma spp. and Dermacentor spp. None of the tick samples gave a positive result when tested by PCR for the presence of DNA from Rickettsia spp., Borrelia spp., Ehrlichia spp., Anaplasma spp., Babesia spp., or Theileria spp.(AU)

Neste estudo avaliou-se a presença de espécies de carrapatos e a detecção de agentes patogênicos a eles associados. Os carrapatos de vida livre foram coletados em fragmentos florestais da área rural do Município de Divino, Minas Gerais, Brasil. Os carrapatos coletados foram divididos em dois grupos, aqueles que foram coletados na área da margem do fragmento (Borda) e os coletados no interior da floresta (Mata). Em todos os fragmentos, os carrapatos foram coletados de acordo com a técnica de arraste aéreo ou no chão e com o uso de perneiras e de flanela. Tanto, as larvas, quanto as ninfas e os adultos foram morfologicamente identificados usando chaves taxonômicas específicas. No caso das larvas, estas foram identificadas até o nível de gênero. Foi realizada extração de DNA dos carrapatos e o DNA extraído foi testado para a presença de Rickettsia spp., Borrelia spp., Ehrlichia spp., Anaplasma spp., Babesia spp., e Theileria spp. por meio de uma estratégia de reação em cadeia da polimerase convencional. No total, 1.122 carrapatos em fase de vida livre (750 larvas, 367 ninfas e 5 adultos) e 18 clusters de larvas foram usados no estudo. As principais espécies identificadas na população de carrapatos coletada foram: Amblyomma sculptum, Amblyomma auricularium, Amblyomma aureolatum e Amblyomma pseudoconcolor e larvas de Amblyomma spp. e Dermacentor spp. Como resultado da detecção de patógenos não foi possível achar DNA de nenhum dos agentes analisados, assim todas as amostras de DNA dos carrapatos testados foram negativas tanto para Rickettsia spp. quanto para Borrelia spp., Ehrlichia spp., Anaplasma spp., Babesia spp. e Theileria spp.(AU)

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The objective was to report an outbreak of tick-borne disease (TBD) on riverside property in the Western Amazon. The death of 25 Nellore cattle was reported on a rural property on the banks of the Purus River, state of Acre. The producer observed animals with staggering walking, drop in productivity, weight loss and evolution to death in approximately 30 days. Fifteen animals from the same batch were selected for clinical evaluation and the ear tip was punctured for hemoparasite research, in addition to blood collection for hematological, biochemical and molecular evaluation. The main laboratory findings were leukocytosis, thrombocytopenia, hypoproteinemia, elevated creatine kinase and reduced urea, creatinine and albumin, as well as visualization of forms suggestive of Anaplasma spp. in 13.33% of the samples. Through PCR, 20% positivity was observed for Anaplasmamarginale and 53.33% for Babesia sp. Hematological and biochemical changes, although highly suggestive, may suffer changes from other factors not related to TBD. Therefore, the presumptive identification of the etiological agent in the blood or confirmatory by molecular methods is essential in the diagnosis. Depending on the stage of the disease, low parasitemia occurs, making it difficult to see hemoparasites in blood smears. The Babesia sp. was the main agent of the outbreak of TBD in the population evaluated, which, when associated with early clinical and laboratory diagnosis, results in adequate therapeutic direction and prophylactic measures, promoting a balance between host, agent and vector.

Objetivou-se relatar um surto de tristeza parasitária bovina (TPB) em propriedade ribeirinha na Amazônia Ocidental. Foi notificado o óbito de 25 bovinos da raça Nelore, em uma propriedade rural às margens do rio Purus, estado do Acre. O produtor observou animais com andar cambaleante, queda na produtividade, perda de peso e evolução ao óbito em aproximadamente 30 dias. Quinze animais do mesmo lote foram selecionados para avaliação clínica e foi procedida a punção a ponta de orelha para pesquisa de hemoparasitos, além da coleta de sangue para avaliação hematológica, bioquímica e molecular. Os principais achados laboratoriais foram anemia, leucocitose, trombocitopenia, hipoproteinemia, elevação da creatina quinase e redução de ureia, creatinina e albumina, além da visualização de formas sugestivas de Anaplasma spp. em 13,33% das amostras. Por meio da PCR, foi observado 20% de positividade para Anaplasma marginale e 53,33% para Babesia sp. As alterações hematológicas e bioquímicas, embora bastante sugestivas, podem sofrer alterações de outros fatores não relacionados à TPB. Por isso, a identificação presuntiva do agente etiológico no sangue ou confirmatória por métodos moleculares é essencial no diagnóstico. A depender da fase da doença, ocorre baixa parasitemia, dificultando a visualização de hemoparasitos em esfregaços sanguíneos. A Babesia sp. foi o principal agente do surto de TPB na população avaliada, que, quando associado ao diagnóstico clínico e laboratorial precoce, resulta no direcionamento terapêutico adequado e medidas profiláticas, promovendo uma relação de equilíbrio entre hospedeiro, agente e vetor.

Resumo

Hemoparasitoses vêm se tornando cada vez mais importantes na clínica médica de pequenos animais. Dentre os agentes causadores encontramos Ehrlichiacanis, Anaplasmaplatys., e Mycoplasma spp., torna-se de grande importância conhecer a epidemiologia nos gatos domésticos. Objetivou-se com esta pesquisa fazer um levantamento retrospectivo de fichas de gatos advindos de consultas no Hospital Veterinário Mário Dias Teixeira (HOVET) que realizaram exame de Reação de Cadeia da polimerase (PCR) no laboratório de biologia molecular, na Universidade Federal Rural da Amazônia, no ano de 2018 e 2019. No total foram 72 amostras de gatos domésticos processadas, sendo 33 machos e 39 fêmeas, 70 animais SRD e 2 Siameses, todos com trombocitopenia, além de outros sinais clínicos que os levaram a precisar de atendimento veterinário, foram categorizados os meses de entrada e processamento das amostras, bairros dos animais e grupos etários. De todos os animais testados, 34,7% obtiveram diagnóstico positivo para uma das enfermidades, sendo o gênero Mycoplasma spp. o que mais prevaleceu em amostras positivas, com maior frequência em fêmeas adultas, bem como foi descrita ocorrência de E. canis apenas nesse sexo, já A. platysfoi descrito com maior frequência em machos, além de achados de infecções concomitantes observado entre os agentes Anaplasmae Mycoplasma. Concluímos que os gatos atendidos no HOVET possuíam parasitismo por diferentes agentes infecciosos.

Hemoparasitosis have become increasingly important in the small animals' internal medicine. Among the causal agents, there are Ehrlichiacanis, Anaplasmaplatys. and Mycoplasma spp., which give the understanding of the epidemiology in domestic cats a great significance. This research aimed to make a retrospective survey of records from cats that came from appointments at the Veterinary Hospital Mário Dias Teixeira (HOVET) and underwent the Polymerase Chain Reaction (PCR) test at the molecular biology laboratory, at the Amazônia Federal Rural University (UFRA), in the years of 2018 and 2019. In total, 72 samples of domestic cats were processed, from which 33 were males and 39 females, 70 of them were mongrel cats and 2 siamese, all of them showed thrombocytopenia amongst other clinical signs that led them to need a veterinary appointment, the months of admission, processing of the samples, districts the animals came from and age group were categorized. 34,7% of all the animals tested showed positive results for one of the diseases, with the genus Mycoplasma spp. being the most prevalent in positive samples, showing a higher rate in adult females, as the occurrence of E. canis was reported only in females, while A. platys was reported with a higher rate in males, as well as concomitant infections following the observation of the agents Anaplasma and Mycoplasma. In conclusion, the cats admitted at HOVET showed parasitism by different infectious agents.

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Objetivou-se neste estudo relatar a frequência e a identidade de patógenos transmitidos por carrapatos em cães residentes de uma área caracterizada por brejo de alta altitude. Amostras sanguíneas (n=203) foram coletadas e molecularmente analisadas via PCR (Babesia spp., Hepatozoon spp., Anaplasma spp. e Ehrlichia spp.) e sequenciamento de DNA. De todas as amostras analisadas, 8,87% (18/203) foram positivas a algum patógeno transmitido por carrapato. Especificamente, 5,42% (11/203) e 3,45% (7/203) foram positivos a Anaplasma platys e Ehrlichia canis, respectivamente. Este estudo fornece, pela primeira vez, evidência científica de infecção de cães por esses patógenos nessa área de alta altitude e reforça o provável papel de R. sanguineus s.l. como vetor de A. platys, principalmente considerando.se que muitos animais positivos eram infestados por essa espécie de carrapato.(AU)

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ABSTRACT: The prevalence of trypanosomiasis in sheep is of worldwide concern. It is a hemoparasitic disease caused by the protozoan Trypanosoma (T) spp., andthe biological cycle of transmission involves susceptible hosts and certain hematophagous flies, such as Tabanids and Stomoxys. The objective of this study was to determine the presence of Trypanosomasp. in sheep (Ovisaries). Through an applied, descriptive-prospective-cross-sectional qualitative study conducted between October 1, 2018, and January 29, 2019, blood samples extracted from the jugular vein were analyzed by the blood smear method using 2 Romanowsky staining techniques (Giemsa and Diff-Quick). Animals studied were between ages of 3 and 10 years, coming from 3 farms in the city of Colimes (Ecuador). Of 100 sampled and processed animals, 2 (2%) were positive for Trypanosoma sp., constituting the first report of this hemoparasite in sheep in Ecuador, and 1 case of Babesia spp. (1%) and 4 cases of Anaplasma marginale (4%) were also identified. However, none of these cases presented symptomatology of any hemotropic-parasitic disease. Subsequently, the positive animals were dewormed.The presence of these parasites wasconfirmedat the study site and,thus, could become a serious animal health problem.

RESUMO: A tripanossomíase em ovelhas é uma hemoparasitose globalmente prevalente causada pelo protozoário chamado Trypanosoma (T) sp. O ciclo de vida de transmissão envolve hospedeiros suscetíveis e alguns dípteros hematófagos como Tabanids e Stomoxys. O objetivo destetrabalho foi determinar a presença de Trypanosoma sp. em ovelhas (Ovisaries). Através de um estudo aplicado, com abordagem qualitativa, do tipo descritivo prospectivo-transversal, realizado entre o dia primeiro de outubro de 2018 e 29 de janeiro de 2019, foram analisadas amostras de sangue extraídas da veía jugular pelo método do esfregaço de sangue, utilizando duas técnicas de coloração Romanowski (Giensa e Diff-Quick). Os animais estudados tinham entre três e dez anos de idade, de três fazendas no municipio de Colimes (Equador). Dos 100 animais amostrados e processados, dois casos (2%) foram positivos para Trypanosoma sp., constituindo o primeiro relato deste hemoparasita em ovelhas no Equador. Além disso, um caso de Babesia sp. (1%) e quatro casos de Anaplasma marginale (4%); entretanto, em nenhum desses casos houve sintomas de qualquer doença hemotropicoparasitária. Animais positivos foram posteriormente desparasitados. A presença desses parasitas foi confirmada no local do estudo e, portanto, pode se tornar um sério problema de saúde animal.

Resumo

Abstract Hemoplasmas are epierythrocytic bacteria that infect mammals. 'Candidatus Mycoplasma haemoalbiventris' was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by 'Ca. M. haemoalbiventris'. Although 'Ca. M. haemoalbiventris' is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

Resumo Hemoplasmas são bactérias epieritrocíticas que infectam mamíferos. 'Candidatus Mycoplasma haemoalbiventris' foi detectado previamente em gambás-de-orelha-branca (Didelphis albiventris) das regiões sul e centro-oeste do Brasil. O presente estudo objetivou: i) triar os gambás para as doenças transmitidas por carrapatos (Piroplasmida e Anaplasmataceae); e ii) detectar e caracterizar as espécies de hemoplasma que infectam gambás nas cidades de Curitiba e Foz do Iguaçu, no Estado do Paraná, sul do Brasil. Trinta amostras de sangue de gambás-de-orelha-branca foram analisadas por PCR. Os animais não estavam infestados por ectoparasitos. O gene endógeno de mamífero gapdh foi amplificado em todas as amostras. Todos os gambás testaram negativos para Theileria/Babesia spp. e Ehrlichia/Anaplasma spp. por PCR, respectivamente, para os genes 18S rRNA e 16S rRNA. Uma PCR gene-específica, baseada no gene 16S rRNA de hemoplasmas, mostrou que três/13 (23,08%; CI 95%: 8,18-50,26%) gambás de Foz do Iguaçu foram positivos para Mycoplasma sp. hemotrópico. Todos os gambás de Curitiba testaram negativos para hemoplasmas. O sequenciamento de fragmentos dos genes 16S e 23S rRNA revelou que os animais estavam infectados pelo 'Ca. M. haemoalbiventris'. Embora 'Ca. M. haemoalbiventris' seja prevalente em gambás no Brasil, os sinais clínicos associados à infecção e os prováveis vetores permanecem desconhecidos.

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Ehrlichia canis, Anaplasma platys, and Babesia vogeli are frequently detected in the veterinary routine with varied pathogenesis that can lead to death, mainly in co-infections. Although canine hemotropic mycoplasmas are considered of low clinical importance, they have recently gained prominence in molecular research. The present study aimed to analyze two hospital populations of dogs, randomly selected from the year 2013 and 2015, from the municipalities of Toledo (n=68) and Cascavel (n=107). Direct examination of blood smears, serology, and PCR were used to detect the presence of E. canis, A. platys, B. vogeli and Mycoplasma sp. Direct blood smear examination was negative for the investigated agents in all samples. Serum immunochromatography (SensPERT ™, VetAll Laboratories, Korea) in 175 animals showed that only 4% (n = 7) had anti-E. canis antibodies, while 60% (n = 105) were positive for Anaplasma sp. All PCR samples were negative for E. canis, while 18.28% (n = 32) were positive for A. platys, 3.42% (n = 6) positive for Babesia vogeli, and 7.41% (n = 13) positive for Mycoplasma sp. The study revealed the presence of A. platys in Toledo and Cascavel by PCR, which accentuates the need to monitor vector populations and usual hosts and to evaluate the potential risk of infection in humans.

Ehrlichia canis, Anaplasma platys e Babesia vogeli são frequentemente detectados na rotina veterinária com patogênese variada que pode levar à morte, principalmente em coinfecções. Embora os micoplasmas hemotrópicos caninos sejam considerados de baixa importância clínica, recentemente ganharam destaque na pesquisa molecular. O presente estudo teve como objetivo analisar duas populações hospitalares de cães, selecionadas aleatoriamente no ano de 2013 e 2015, dos municípios de Toledo (n = 68) e Cascavel (n = 107). O exame direto de esfregaços de sangue, sorologia e PCR foram usados para detectar a presença de E. canis, A. platys, B. vogeli e Mycoplasma sp. O esfregaço sanguíneo direto foi negativo para os agentes investigados em todas as amostras. A imunocromatografia sérica (SensPERT ™, VetAll Laboratories, Coréia) em 175 animais mostrou que apenas 4% (n = 7) tinham anticorpos contra E.canis, enquanto 60% (n = 105) foram positivos para Anaplasma sp. Todas as amostras de PCR foram negativas para E. canis, enquanto 18,28% (n = 32) foram positivas para A. platys, 3,42% (n = 6) positivas para Babesia vogeli e 7,41% (n = 13) positivas para Mycoplasma sp. O estudo revelou a presença de A. platys em Toledo e Cascavel por PCR, o que acentua a necessidade de monitorar populações de vetores e hospedeiros usuais e avaliar o risco potencial de infecção em humanos.

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Abstract The aim of the present study was to detect Cercopithifilaria bainae and other tick-borne pathogens and to perform molecular characterization of the tick Rhipicephalus sanguineus s.l. collected from dogs. Ticks (n = 432, including 8 larvae, 59 nymphs, and 365 adults) were sampled from domiciled dogs (n = 73) living in Campo Grande, Mato Grosso do Sul (Midwest Brazil). All ticks were morphologically identified as R. sanguineus. Genomic DNA was extracted in pools (three to five ticks per animal) and was used for definition of R. sanguineus haplotypes (based on 16S rRNA analysis) and pathogen identification (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli and Rickettsia spp.). Rhipicephal us sanguineus specimens were identified as haplotypes A and B. DNA of Cercopithifilaria bainae (43.83%; 32/73), Ehrlichia canis (24.65%; 18/73), Anaplasma platys (19.17%; 14/73), and Hepatozoon canis (5.47%; 4/73) was detected. The identity of pathogens was confirmed by DNA sequence analysis. The present study confirms the presence of haplotypes A and B of R. sanguineus in the state of Mato Grosso do Sul and its importance as a vector of several pathogens of veterinary concern. Finally, this is the first report to identify C. bainae in ticks in the Midwestern region of Brazil.

Resumo O objetivo do presente estudo foi detectar Cercopithifilaria bainae e outros patógenos transmitidos por carrapatos e realizar a caracterização molecular do carrapato Rhipicephalus sanguineus s.l. coletado em cães. Carrapatos (n = 432, incluindo 8 larvas, 59 ninfas e 365 adultos) foram amostrados de cães domiciliados (n = 73) residentes no município de Campo Grande, Mato Grosso do Sul (centro-oeste do Brasil). Todos os carrapatos foram identificados morfologicamente como R. sanguineus. O DNA genômico foi extraído em pools (três a cinco carrapatos por animal), seguido pela definição de haplótipos (com base no gene 16S rRNA) e pela investigação de patógenos (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli e Rickettsia spp.). Os espécimes coletados foram identificados como haplótipos A e B de R. sanguineus. Foram detectados DNA de Cercopithifilaria bainae (43,83%; 32/73), Ehrlichia canis (24,65%; 18/73), Anaplasma platys (19,17%; 14/73) e Hepatozoon canis (5,47%; 4/73). A identidade dos patógenos foi confirmada por análise de sequência de DNA. O presente estudo confirma a circulação dos haplótipos A e B de R. sanguineus no estado de Mato Grosso do Sul e sua importância como vetor de vários patógenos de interesse veterinário. Finalmente, este é o primeiro relato de C. bainae em carrapatos na região centro-oeste do Brasil.

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Abstract Tick-borne rickettsial pathogens (TBRP) are important causes of infections in both dogs and humans. Dogs play an important role as a biological host for several tick species and can serve as sentinels for rickettsial infections. Our aim was to determine the presence of TBRP in dogs and in dog-associated ticks and their potential risk to human diseases in Medellin, Colombia. DNA for E. canis (16S rRNA and dsb) and A. platys (groEl) was detected in 17.6% (53/300) and 2.6% (8/300) of dogs, respectively. Antibodies against Ehrlichia spp. 82 (27.3%) and Anaplasma spp. 8 (2.6%) were detected in dogs. Antibody reactivity against both agents were found in 16 dogs (5.3%). Eight dogs showed antibody for Rickettsia spp. with titers that suggest 3 of them had a probable exposure to R. parkeri. Rhipicephalus sanguineus s.l. (178/193) was the main tick in dogs, followed by R. microplus (15/193). The minimum infection rates (MIR) in R. sanguineus were 11.8% for E. canis and 3.4% for A. platys. E. canis and A. platys are the main TBRP infecting dogs and ticks and R. sanguineus s.l. is likely involved in the transmission of both agents. Interestingly, we found serological evidence of exposure in dogs for spotted fever group rickettsiae.

Resumo As riquétsias transmitidas por carrapatos (RTC) são causas importantes de infecção em cães e humanos. Os cães exercem um papel essencial como hospedeiros biológicos para diversas espécies de carrapatos, assim como podem ser úteis como sentinelas de infecções por riquétsias. O intuito deste estudo foi determinar a presença de RTC em cães, assim como em seus carrapatos, para determinar o risco potencial de doença humana em Medellín, Colômbia. DNA de Ehrlichia canis (16S rRNA e dsb) e Anaplasma platys (groEl) foi detectado em 17,6% (53/300) e 2,6% (8/300) dos cães, respectivamente. Anticorpos contra Ehrlichia spp. (82; 27,3%) e Anaplasma spp. (8; 2,6%) foram detectados nos cães. Reatividade de anticorpos contra ambos patógenos (Ehrlichia e Anaplasma) foi detectada em 16 cães (5,3%). Oito animais apresentaram anticorpos contra Rickettsia spp., e 3 deles sugerem uma provável exposição a Rickettsia parkeri. Rhipicephalus sanguineus s.l. (178/193) foi a principal espécie de carrapatos, seguida de R. microplus (15/193). A taxa de infecção mínima em R. sanguineus foi 11,8% para E. canis e 3,4% para A. platys. E. canis e A. platys são as principais RTC que infectam cãese R. sanguineus s.l. provavelmente está envolvido na transmissão de ambos os agentes. É evidente, porém, a exposição sorológica dos cães a riquétsias do grupo da febre maculosa.

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Abstract Although anemia has been historically linked to Haemonchus contortus infection, other infectious agents, such as hemotropic mycoplasmas and tick-borne disease pathogens, may also lead to anemic crisis in sheep. This study has aimed to investigate infections related to anemia in a sheep herd from Bandeirantes City, Paraná State, southern Brazil. Seven out of forty-two (16.6%; 95% CI: 8.32-30.6%) sheep were positive for hemoplasmas by a PCR targeting the 16S rRNA gene and all tested negative for A. marginale/A. ovis and Babesia/Theileria spp. by PCR based on msp4 and 18S rRNA genes, respectively. Two (4.7%; 95% CI: 1.32-15.79%) animals were infested with Rhipicephalus microplus ticks. Fecal egg counting was performed in 38 sheep and 24 (63.15%; 95% CI: 47.2-76.6%) presented > 500 eggs per gram. Phylogenetic analysis of partial sequences of the detected hemotropic Mycoplasma sp. 16S and 23S rRNA genes confirmed that the animals were infected with Mycoplasma ovis. Polymorphism analysis of partial 16S rRNA sequences showed three different genotypes of M. ovis infecting sheep assessed in the present study. Mycoplasma ovis and gastrointestinal nematodes occurs in sheep from the northern region of Paraná State.

Resumo Embora a principal causa de anemia seja historicamente relacionada à infecção por Haemonchus contortus, outros agentes infecciosos, como micoplasmas hemotrópicos e patógenos transmitidos por carrapatos, também podem causar quadros anêmicos em ovinos. O presente estudo objetivou investigar infecções relacionadas à anemia em um rebanho de ovinos, na cidade de Bandeirantes, Estado do Paraná, sul do Brasil. Sete (16,6%; 95% CI: 8,32-30,6%) de 42 ovinos foram positivos para hemoplasmas pela PCR do gene 16S rRNA, enquanto todos foram negativos para A. marginale/A. ovis e Babesia/Theileria spp. por ensaios da PCR baseados nos genes msp4 e 18S rRNA, respectivamente. Dois (4,7%; 95% CI: 1,32-15,79%) animais estavam infestados por carrapatos Rhipicephalus microplus. Dos 38 animais nos quais foi realizada a contagem de ovos por grama de fezes (OPG), 24 (63,15%; 95% CI: 47,2-76,6%) apresentaram valores >500 para OPG. A análise filogenética das sequências parciais dos genes 16S rRNA e 23S rRNA de hemoplasmas confirmou a infecção por Mycoplasma ovis. A análise de polimorfismos de um fragmento do gene 16S rRNA mostrou a ocorrência de três genótipos diferentes de M. ovis nos animais. Mycoplasma ovis e nematódeos gastrointestinais ocorrem em ovinos da região nordeste do Estado do Paraná.