BackgroundAnaplasma 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.MethodsWe 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.ResultsAmong 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.ConclusionsThis 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.

The aim of this study was to detect Theileria equi (Laveran 1901) DNA in horses and ticks using real-time PCR and to list the factors associated with infection in animals located in the Seropedica and Petropolis municipalities of the state of Rio de Janeiro. We tested blood samples from 314 horses and samples from 300 ticks, including 191 Amblyomma cajennense, 104 Dermacentor nitens, and 5 Ixodida larvae. Factors inherent to the horse, the ownership, and animal management were obtained from an epidemiological questionnaire and were evaluated in association with the presence of T. equi DNA in the animals. Among the horses in the study, 81 % (n = 253/314) presented T. equi DNA, and the animals of the Seropedica municipality had the highest infection frequency (91 %, n = 128/141, p < 0.001). The factors that had significantly different infection frequencies by chi-squared or Fisher’s exact tests (p < 0.2) were included in a logistic regression model using the R programming package. Work and walking activity (odds ratio [OR] = 5.7, CI = 2.3–14.4), reproductive activity (OR = 3.8, CI = 1.3–11.5), and tick infestation (OR = 2.6, CI = 1.1–6.2) were factors that favored the presence of T. equi DNA in the animals (p < 0.05). Among the tick samples, A. cajennense and D. nitens were the identified species. The presence of T. equi DNA was observed in 9.9 % (n = 19/191) of the A. cajennense samples and 3.8 % (n = 4/104) of the D. nitens samples. A multivariate analysis revealed that the presence of A. cajennense on the animals (OR = 4.1, CI = 1.8–9.1) was associated with the presence of T. equi DNA in the horses. In the studied municipalities, activities related to work, walking, and reproduction and the presence of ticks on the horses, particularly an intense infestation of A. cajennense, are factors that lead to infection with T. equi in the horses.

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.

Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80) for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR) calculated. The overall prevalence of T. equi infection was 81.09% (n = 579), with higher prevalence (p < 0.05) in the Itaguaí (85.43%) when compared to Serrana microregion (76.92%). The geographic area, altitude, farming condition and area of origin of equids were associated (p < 0.05) with seropositivity for T. equi. Equids reared in the Itaguaí microregion (PR = 1.11, p = 0.003) and at altitudes below 500 m (PR = 1.10; p = 0,014) were more likely to be seropositive for T. equi. Furthermore, when equids were born in the farm (PR = 1.10, p = 0.008) and reared with poor farming conditions (PR = 1.13, p = 0.018) they were more likely to be exposed to T. equi. The main ticks found on equids were Amblyomma cajennense and Dermacentor (Anocentor) nitens. The microregions studied are endemic areas for equine theileriosis and there exists enzootic stability for T. equi. Only factors related to the collection area of serum samples influenced the seropositivity of equids for T. equi in that region.Keywords: horses, equine theileriosis, IFAT, prevalence, epidemiology. ResumoAmostras de soro de 714 equídeos das microrregiões de Itaguaí e Serrana, Rio de Janeiro, Brasil, foram submetidas ao teste de imunofluorescência indireta (título 1:80) para Theileria equi. A prevalência entre as microrregiões e os fatores associados à soropositividade foram avaliados e a razão de prevalência (RP) calculada. A prevalência geral para T. equi foi de 81,09% (n = 579), com maior prevalência (p < 0,05) para microrregião de Itaguaí (85,43%), quando comparado a Serrana (76,92%). A região, altitude, nível da propriedade e origem dos equídeos foram associados (p < 0,05) com a soropositividade para T. equi. Equídeos criados na microrregião de Itaguaí (RP = 1,11; p = 0,003) e em altitudes abaixo de 500 m (RP = 1.10; p = 0,014) apresentaram maior chance de se tornarem soropositivos para T. equi. Além disso, quando são criados em propriedades de nível ruim (RP = 1,13; p = 0,018) e nascidos na propriedade (RP = 1,10; p = 0,008) apresentaram mais chance de terem contato com T. equi. As principais espécies de carrapatos encontradas parasitando os equídeos foram Amblyomma cajennense e Dermacentor (Anocentor) nitens. As microrregiões estudadas são endêmicas para theileriose equina e de estabilidade enzoótica para T. equi. Apenas os fatores relacionados à área de coleta das amostras de soro influenciaram a soropositividade dos equídeos para T. equi naquela região.Palavras-chave: equinos, theileriose equina, RIFI, prevalência, epidemiologia.

IVCapacidade tamponante, pH e consistência das fezes em equinos submetidos à sobrecarga dietética com amido oito, 12, 16, 20, 24, 28, 32 4, 8, 12, 16, 20, 24, 28, 32 and 36 hours post-overload, and immediately

-The present study was carried out to evaluate the in situ degradation of dry matter (DM), neutral detergent fiber (NDF) and crude protein (CP) in roughages by the in situ caecal digestion technique in horses. The roughages evaluated were: Lucerne hay (Medicago sativa), peanut (Arachis pintoi cv. Amarillo), desmodio(Desmodium ovalifolium), stylo (Stylosanthes guianensis cv. Mineirão), pigeon pea (Cajanus cajan), lime-yellow pea (Macrotyloma axillare) and coastcross hay (Cynodon dactylon cv. coastcross). The assay was conducted in a complete randomized design with seven roughages and three replications. One mare with a cannula fitted in the caecum was used, fed diet consisting of coastcross hay (80%) and concentrate (20%) at 2.0% BW, four times a day. Nylon 6.5 × 20 cm bags were used with 45 μ/pore, containing 5. Key Words: cannula, digestion, fiber, forage, legume Degradação cecal in situ de alimentos volumosos em equinos RESUMO -Objetivou-se avaliar a degradação da matéria seca (MS), fibra em detergente neutro (FDN) e proteína bruta de alimentos volumosos pela técnica da digestão cecal in situ em equinos. Avaliaram-se as forrageiras: feno de alfafa (Medicago sativa), amendoim forrageiro (Arachis pintoi cv. Amarillo), desmódio (Desmodium ovalifolium), estilosantes (Stylosanthes guianensis cv. Mineirão), guandu (Cajanus cajan), macrotiloma (Macrotyloma axillare) e feno de coastcross (Cynodon dactylon cv. coastcross). O ensaio teve duração de 35 dias e foi realizado em delineamento inteiramente casualizado com sete alimentos e três repetições. Utilizou-se uma égua fistulada no ceco, alimentada com dieta composta por feno de coastcross (80%) e concentrado (20%), fornecida quatro vezes ao dia em quantidade equivalente a 2% do peso vivo. No ceco, foram inseridos 3 a 4 sacos de náilon de porosidade 45 μ de 6,5 × 20 cm, contendo 5,2 g de MS/saco, nos tempos de 2, 4, 6, 8, 12, 24 e 48 horas de incubação. Os parâmetros de degradação da MS de todos os volumosos foram significativos. Não houve ajuste no modelo de degradação da PB e FDN do guandu. O amendoim, estilosantes e macrotiloma apresentaram maior fração potencialmente degradável da MS, com valores de 53, 46,5 e 40%, respectivamente, e os maiores valores da fração solúvel de 20, 21, 28,6%, além de elevadas taxas de degradação (10,36, 20,26 e 14,8%h -1 ). Nestes alimentos, também foram observadas as maiores taxas de degradação da FDN (9,1 e 11,3, 11,2%h -1 ), das frações potencialmente degradáveis (55, 51,8 e 47,2%) e as mais altas taxas de degradação da PB em 48 horas de incubação (87, 95, 94,8%, respectivamente). O amendoim forrageiro, o estilosantes e o macrotiloma apresentam potencial de uso nas dietas para equinos.Palavras-chave: cânula, digestão, fibra, forrageiras, leguminosa R. Bras.

-The objective of this study was to the evaluate physiological, hematological and biochemical parameters of eventing horses fed diets containing soybean oil and submitted to physical effort tests. Twelve horses were used in a completely randomized design with two diets and six replications. The diets used were: diet without soybean oil inclusion (control) and diet with inclusion of 10% soybean oil. The experiment lasted 82 days, with three exercise tests: at the beginning, 60 th and 82 th day of the experiment. There were five data collections for each test: before the test with the horses at rest, immediately after the test, and at 10, 20 and 120 minutes after each test. Body temperature and rate heart were taken and blood samples were collected to evaluate hematological and biochemical variables. The mean values of the physiological, metalogical and biochemical parameters were submitted to non-parametric analysis. The inclusion of soybean oil only altered the γ-glutamyltransferase (GGT) and creatinine concentration in function time of intake of the diet. There was effect of trainings time on the horses conditioning that improved with reduction in the lactate concentration while the glucose concentration increased at the last exercise test.Key Words: horses, nutrition, riding, training Desempenho de equinos de concurso completo de equitação alimentados com dieta contendo óleo de soja RESUMO -Objetivou-se avaliar os parâmetros fisiológicos, hematológicos e bioquímicos de equinos de concurso completo de equitação consumindo dieta contendo óleo de soja e submetidos a testes de esforço físico. Foram utilizados 12 equinos em delineamento experimental inteiramente casualizado, com duas dietas e seis repetições. As dietas utilizadas foram: dieta sem inclusão de óleo de soja (controle); dieta com inclusão de 10% de óleo de soja. O ensaio teve duração de 82 dias e incluiu três testes de esforço físico: ao início, no 60 o e no 82 o dia. Em cada teste, foram realizadas cinco coletas de dados: antes do teste com os animais em repouso; imediatamente após o teste; e 10, 20 e 120 minutos após os testes. Foram avaliadas a temperatura corporal e a frequência cardíaca e realizadas coletas de amostras de sangue para análises hematológicas e bioquímicas. Os valores médios dos parâmetros fisiológicos, hematológicos e bioquímicos foram submetidos à análise não-paramétrica. Somente ao longo do tempo de consumo das dietas, a inclusão do óleo de soja somente alterou a concentração de γ-glutamil transferase (GGT) e creatinina. O tempo de treinamento físico tem efeito no condicionamento dos equinos, que melhora com a redução da concentração de lactato, e na concentração de glicose, que aumentou no teste de esforço físico.

The study aimed to evaluate the risk factors associated with the frequency of IgG antibodies against Babesia bovis and B. bigemina in cattle in southern Mozambique. Eight hundred and nine serum samples were collected from cattle in three provinces namely Maputo, Gaza and Inhambane, and tested by indirect enzyme-linked immunosorbent assay (i-ELISA) to assess the humoral immune response towards B. bovis and B. bigemina. The chi-square test at 5% significance was used to determine whether there was an association between gender, age and geographic origin of seropositive animals. The overall prevalence was 78.8% (548/695) for B. bovis and 76.0% (528/695) for B. bigemina. The origin of the animals showed a significant association (p<0.05) with seropositivity to both agents, while gender and age was not associated (p>0.05). Maputo province had the highest rate of positive animals, with 93.7% (118/126) for B. bovis and 97.6% (123/126) for B. bigemina. In Gaza province 77.3% (321/415) of the animals were positive for B. bovis and 67.5% (280/415) for B. bigemina, while in the province of Inhambane the levels of seropositivity were 70.8% (109/154) and 81.2% (125/154) for B. bovis and B. bigemina respectively. In the present study, the frequency of cattle positive for B. bovis and B. bigemina was shown to increase among older age groups, suggesting that infection and re-infection persisted even after the primary infection. Thus, this region is considered to be in a state of enzootic stability with regards to B. bovis and B. bigemina.