Urinary capillariosis in a cat from Rio de Janeiro, Brazil - Clinical, morphological and phylogenetic characterization.

Capillaria spp. infections of the urinary tract of domestic carnivores are uncommon worldwide. Infections are rarely diagnosed and are typically asymptomatic. This study aimed to evaluate a case of capillariosis in a cat from the state of Rio de Janeiro, Brazil. A seven-year-old female cat with apathy and reduced appetite was presented. Urine analysis revealed C. plica eggs in urine sediment, and cystitis was evidenced by the presence of bacteria, pyuria, proteinuria and hematuria. The subject was treated with 50 mg/kg fenbendazole for five days. Urine samples were frozen for molecular analysis and species confirmation. Polymerase chain reaction for amplification of the 18S rRNA gene followed by sequencing confirmed the occurrence of Capillaria sp. There has been limited phylogenetic study of Capillaria spp. in cats, so further studies are needed to identify the species present in different locations and associated with feline pathogenesis.

Molecular Survey of Bartonella Species in Shelter Cats in Rio De Janeiro: Clinical, Hematological, and Risk Factors.

The present study aimed to detect Bartonella DNA in cats belonging to shelters, and to evaluate risk factors, clinical signs, and hematological abnormalities associated with infection. Complete blood counts and screening for the presence of Bartonella DNA were performed on cats' ethylenediamine tetraacetic acid anticoagulant-blood samples. Eighty-three cats (39.9%) were positive for Bartonella species. Bartonella DNA was also detected in fleas and in the blood of cats infested by positive flea. Cats that had not been sterilized, had outdoor access, had histories of fights, and had concurrent flea infestation were more likely to be infected by Bartonella species (P < 0.05). Age and sex were not associated with infection. Fifty-one (38.6%) symptomatic cats were positive to Bartonella species (P > 0.05). Clinical conditions most commonly observed were signs of respiratory abnormality and Sporothrix species coinfection (P > 0.05). Regarding hematological changes, eosinophilia was associated with infection (P < 0.05). A high frequency of Bartonella species infection was found in shelter cats and highlights the importance of adequate flea-control programs to prevent infection in cats and consequently in adopters and other animals.

Epidemiological survey of Neorickettsia risticii in equids from the State of Rio de Janeiro, Brazil / Levantamento epidemiológico de Neorickettsia risticii em equídeos do Estado do Rio de Janeiro

Equine neorickettsiosis (EN), also known as Potomac Horse Fever, is a non-contagious disease caused by the bacterium Neorickettsia risticii of the Anaplasmataceae family. The objectives of this study were to detect the presence of anti-N. risticii antibodies by the indirect immunofluorescence assay (IFA) and of its DNA by qPCR in equids at high and low altitude regions in the State of Rio de Janeiro, Brazil, and to identify factors associated with seropositive equids by multiple logistic regression analysis. The frequency of anti-N. risticii antibodies was 16.05% (n=113/704). The animal age and breeding region were the factors that influenced the seropositivity rate for N. risticii in the equids (p<0.05). Equids from the lowland region had higher seropositivity (p<0.05; OR=5.87) compared to those of the mountain region. The presence of snails on the farm was a factor associated with this result (p<0.05; OR=2.88). In the lowland region, age of the animal and site of breeding were protective factors for the detection of antibodies anti-N. risticii in equids, with lower frequency of seropositivity in younger animals (p<0.05; OR=0.06) and in animals raised in dry areas (p<0.05; OR=0.22). The presence of the target DNA of N. risticii by qPCR was not observed in any of the samples tested. The existence of seropositive equids for N. risticii demonstrates a possible circulation of this agent in the studied area, and that the age related characteristics and equids breeding region are important factors regarding seropositivity in the State of Rio de Janeiro.(AU)

A Neorickettisiose equina (NE), também conhecida como Febre do Cavalo de Potomac, é uma doença não contagiosa causada pela bactéria Neorickettsia risticii da família Anaplasmataceae. Os objetivos deste estudo foram detectar a presença de anticorpos anti-N. risticii através da reação de Imunofluorescência Indireta (RIFI) e do DNA dessa bactéria através da qPCR em equídeos de regiões de alta e baixa altitude no Estado do Rio de Janeiro, Brasil; e identificar os fatores associados com a soropositividade dos equídeos através da análise de regressão logística múltipla. A frequência de anticorpos anti-N. risticii foi de 16,05% (n=113/704). Observou-se que a idade e a região de criação foram os fatores que influenciaram a taxa de soropositividade para N. risticii nos equídeos (p<0,05). Equídeos da região de baixada apresentaram maior soropositividade (p<0,05; OR=5,87) quando comparado aos criados em região de montanha. A presença de caramujos na propriedade foi um fator associado a este resultado (p<0,05; OR=2,88). Na região de baixada, animais mais jovens (p<0,05; OR=0,06), criados em áreas secas (p<0,05; OR=0,22) demonstraram serem fatores de proteção na detecção de anticorpos anti-N. risticii. Não foi observada a presença do DNA-alvo de N. risticii através da qPCR em nenhuma das amostras testadas. A existência de equídeos soropositivos para N. risticii demonstra a possível circulação desse agente na área estudada, e as características inerentes a idade e a região de criação dos equídeos são fatores importantes relacionados à soropositividade no estado do Rio de Janeiro.(AU)

Occurrences of anti-toxoplasma gondii and anti-neospora caninum antibodies in sheep from four districts of tocantins state, brazilian legal amazon region

Toxoplasmosis and neosporosis have been recognized as economically important diseases with considerable impact on the livestock industry. Little is known concerning the occurrence of Toxoplasma gondii and Neospora caninum in sheep from Tocantins state, Brazil. Here, we investigated antibodies against these parasites and associated factors in 182 sheep from Araguaína, Santa Terezinha do Tocantins, Arguianópolis and Palmeiras do Tocantins districts, Tocantins. Sheep sera were assayed for T. gondii and N. caninum IgG antibodies by indirect fluorescence antibody test (IFAT), using cut-off point at a dilution of 1:40 and 1:25 respectively. The prevalence of seropositive animal for T. gondii was 13.74% and 13.74% for N. caninum. None of the characteristics studied including reproductive problems, presence of cats, presence of dogs and veterinary care (p>0.05) was associated with occurrence of T. gondii or N. caninum infection. Only breed was identified as associated factor for the occurrence of toxoplasmosis in sheep (p<0.05). The present study is the first report on serum occurrence of T. gondii and N. caninum in sheep from the state of Tocantins, Brazil.

Toxoplasmose e Neosporose são reconhecidas por doenças economicamente importantes com impacto considerável na indústria pecuária. Pouco se sabe sobre a ocorrência de Toxoplasma gondii e Neospora caninum em ovelhas do estado do Tocantins, Brasil. Foram investigados a ocorrência de anticorpos contra estes parasitos e fatores associados em 182 ovelhas das cidades de Araguaína, Santa Terezinha do Tocantins, Arguianópolis e Palmeiras do Tocantins, Tocantins. Os soro das ovelhas foram testados para anticorpos IgG anti-T. gondii e anti-N. caninum pela Reação de Imunofluorescência Indireta (RIFI), usando os pontos de corte na diluição de 1:40 e 1:25, respectivamente. A prevalência de animais soropositivos para T. gondii foi de 13.74% e para N. caninum, 13.74%. Nenhuma das características estudadas incluindo problemas reprodutivos, presença de gatos, presença de cães e cuidados veterinários (p>0.05) foram associadas com a ocorrência infecção por T. gondii ou N. caninum. Somente raça foi identificada como fator associado à ocorrência de toxoplasmose em ovelhas (p<0.05). O presente trabalho é o primeiro relato da ocorrência sérica de T. gondii e N. caninum em ovelhas do estado do Tocantins, Brasil.

Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil.

BACKGROUND: Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. METHODS: We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. RESULTS: Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs. CONCLUSIONS: This is the first report of A. phagocytophilum in ixodid ticks from Brazil. The detection of A. phagocitophylum in A. cajennense, an aggressive feeder on a wide variety of hosts, including humans, is considered a public health concern.

Detection of Anaplasma phagocytophilum in Brazilian dogs by real-time polymerase chain reaction.

Anaplasma phagocytophilum was detected in dogs from Brazil in the municipalities of Seropédica and Itaguaí, Rio de Janeiro state, by real-time polymerase chain reaction (PCR) using SYBR Green to detect the amplification. Of 253 samples, 18 (7.11%) were positive, with a threshold cycle (Ct) ranging from 31 to 35 cycles. The PCR product from a positive sample was cloned and sequenced. The sequence obtained demonstrated 100% identity with other A. phagocytophilum sequences published in the GenBank database. The analytical sensitivity of RT-PCR using SYBR Green system was able to detect 3 plasmid copies when defined numbers of plasmid copies containing 122 base pairs from the msp2 gene were used. The assay was considered specific when DNA from bacteria (Anaplasma platys, Anaplasma marginale, Ehrlichia canis, Neorickettsia risticii, Rickettsia rickettsii) closely related to A. phagocytophilum was placed in the reaction. These results demonstrate that the canine granulocytic anaplasmosis agent is present in regions in which dogs could be a source of infection for tick vectors. The current study reports the detection of A. phagocytophilum, a zoonotic agent responsible for Human granulocytic anaplasmosis, in Brazilian dogs.

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs / Ensaio imunoenziático (ELISA) para detecção de anticorpos IgM contra Leishmania chagasi em cães

Visceral leishmaniasis is an emergent zoonosis with an increasing number of new cases in Brazil where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. An enzyme-linked immunosorbent assay (ELISA), based upon the use of a total soluble antigenic preparation of L. chagasi, was adapted for the detection of IgM antibodies in the serum of infected dogs. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 percent. A total of 110 serum samples were taken from dogs in Belo Horizonte, Minas Gerais, Brazil, and examined for anti-L. chagasi IgM antibody by ELISA and indirect fluorescent antibody test (IFAT). About 25 percent (n=27) of all the dogs tested were found serologically positive for L. chagasi by IFAT, while 89.09 percent (n=98) were seropositive by ELISA. The results obtained by ELISA and IFAT were significantly different (P<0.01). The combined use of ELISA and IFAT is recommended in order to enable veterinary services to more efficiently detect canine visceral leishmaniasis.

A leishmaniose visceral é uma zoonose emergente, com elevado número de novos casos no Brasil, onde o cão doméstico é um importante reservatório do parasito no ciclo infeccioso da Leishmania chagasi. Um ensaio de imunoadsorção enzimática (ELISA) baseado em antígeno bruto de L. chagasi foi adaptado para a detecção de anticorpos IgM em soros de cães infectados. As diluições ótimas do antígeno e dos soros controles positivo e negativo foram determinadas através de titulação em bloco. A sensibilidade e especificidade do ELISA teste foram de 100 por cento. Um total de 110 amostras de soros foram obtidas de cães oriundos de Belo Horizonte, Minas Gerais, Brasil, e avaliadas pelo ELISA e pela reação de imunofluorescência indireta (RIFI) para anticorpos IgM anti-L. chagasi. Aproximadamente 25 por cento (n=27) dos cães testados foram sorologicamente positivos para L. chagasi pela RIFI, enquanto 89.09 por cento foram soropositivos pelo ELISA. Os resultados obtidos pelo ELISA e pela RIFI foram significativamente diferentes (P<0.01). A associação do ELISA e da RIFI deve ser recomendada a fim de permitir a detecção mais eficiente da leishmaniose visceral canina pelos serviços veterinários.

Serodiagnosis of Babesia equi in horses submitted to exercise stress

A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.

A reação de fixação do complemento (RFC), realizada em microplacas, utilizando-se antígeno bruto de Babesia equi, foi adaptada para a detecção de anticorpos em soros de eqüinos infectados. A reação de imunofluorescência indireta (RIFI) e o ensaio imunoenzimático (ELISA) também foram utilizados para o imunodiagnóstico de B. equi. Amostras de soro foram obtidas de 15 eqüinos aparentemente sadios, submetidos a treinamento físico em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Todas as amostras testadas foram positivas para B. equi. Não houve diferença significativa (P<0,01) entre estes 3 testes sorológicos, ou entre a condição de estresse e repouso. A combinação da RFC com a RIFI ou ELISA pode ser recomendada a fim de evitar a entrada de eqüinos portadores em áreas consideradas livres da doença.