ABSTRACT The aim of this study was to determine the seroprevalence of Neospora caninum in goats from eight provinces of the Piura region, Peru. In total, 361 sera were analysed with the commercial ID Screen Neospora caninum Competition ELISA kit for the detection of anti-N. caninum IgG antibodies. A general seroprevalence of 3.32% (95% CI 1.47-5.17) was found. The province of Morropón presented the highest frequency with 8.11% (95% CI 0-16.91) and the goats of Huancabamba, Paita, Sechura and Talara were seronegative. The animals seropositive to N. caninum were adults (>1 year) and were living at an altitude d»500 meters above sea level.

RESUMEN El objetivo del estudio fue determinar la seroprevalencia de Neospora caninum en caprinos de ocho provincias de la región Piura, Perú. Se analizaron 361 sueros con el kit comercial de ELISA de competencia ID Screen® Neospora caninum Competition para la detección de anticuerpos IgG anti N. caninum. Se halló una seroprevalencia general de 3.32% (IC 95% 1.47-5.17). La provincia de Morropón presentó la mayor frecuencia con 8.11% (IC 95% 0-16.91) y los caprinos de Huancabamba, Paita, Sechura y Talara fueron seronegativos. Los animales seroreactivos a N. caninum fueron adultos (>1 año) y se encontraban habitando una altitud d»500 msnm.

ABSTRACT Neospora caninum is an important worldwide parasite responsible for causing abortion in animals. Due to limited information on the occurrence of infection by this parasite in the state of Rondônia, Brazil, this study aimed to determine the seroprevalence and identify the risk factors associated with the infection in slaughtered cattle, from 19 municipalities distributed in seven microregions of the state. A total of 494 samples were obtained and subjected to anti-N. caninum antibodies, using the Indirect Immunofluorescence Reaction technique. Antibodies were detected in 5.06% (25/494) of the samples, in 30.30% (10/33) of farms, in nine municipalities located in four microregions of Rondônia. Of all the animals analyzed, 4.81% of the females (20/416) and 6.41% of the males (05/78) were seropositive for the parasite, with “abortion in the last 12 months” being considered an important risk factor for the occurrence of infection (OR = 9.54; p = 0.01). The present study points out the prevalence of anti-N. caninum antibodies in 5.06% of slaughtered animals and abortion as the main risk factor associated with infection by N. caninum, thus contributing to the elucidation of the epidemiology of this protozoan in Rondônia, Brazil.

RESUMO Neospora caninum é um importante parasito, responsável por causar aborto em animais e distribuído mundialmente. Devido às informações limitadas sobre a ocorrência da infecção por esse parasito no estado de Rondônia, Brasil, este estudo teve como objetivo determinar a soroprevalência e identificar os fatores de risco associados à infecção em bovinos abatidos, oriundos de 19 municípios, distribuídos em sete microrregiões do estado. Um total de 494 amostras foi obtido e submetido à pesquisa de anticorpos anti-N. caninum, por meio da técnica de reação de imunofluorescência indireta. Os anticorpos foram detectados em 5,06% (25/494) das amostras, em 30,30% (10/33) das propriedades rurais, de nove municípios, localizados em quatro microrregiões de Rondônia. De todos os animais analisados, 4,81% das fêmeas (20/416) e 6,41% dos machos (05/78) foram soropositivos para o parasito, sendo o “aborto nos últimos 12 meses” considerado um importante fator de risco para a ocorrência da infecção (OR = 9,54; P = 0,01). O presente estudo aponta a prevalência de anticorpos anti-N. caninum em 5,06% dos animais abatidos e o aborto como o principal fator de risco associado à infecção por N. caninum, contribuindo assim para a elucidação da epidemiologia desse protozoário em Rondônia, Brasil.

Neospora caninum is a coccidian parasite that occurs worldwide and is one of the most important causes of abortion, especially in cattle. However, no studies have been performed in Namibia to determine the N. caninum status in livestock. Therefore, this study aimed to determine the seroprevalence of N. caninum in cattle and the associated risk factors in the Khomas region of Namibia. A total of 736 sera were collected from cows in 32 farming establishments. These comprised 698 beef and 38 dairy cattle sera and were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Questionnaires were concurrently administered to determine possible risk factors associated with N. caninum seropositivity. A total of 42 sera were positive (all beef), giving an animal-level seroprevalence rate of 5.7%. Eight of the 32 establishments had at least one positive animal, giving a herd-level seroprevalence of 25%. There was no significant association between seropositivity and the presence of dogs, jackals, history of abortions, farm size, number of cattle or average annual rainfall. The establishments with moderate to high numbers of Feliformia were 9.8 times more likely to be seropositive to N. caninum than those with none to low levels of the former (p = 0.0245). The authors concluded that the seroprevalence level of N. caninum in the Khomas region was relatively low compared with other parts of the world and that the role of Feliformia in the epidemiology of bovine neosporosis needed to be further investigated. CONTRIBUTION: Serological evidence of bovine neosporosis and the associated risk factors are reported in Namibia for the first time. This study contributes to the scientific body of knowledge on N. caninum in Africa, which is currently limited.

ABSTRACT: This study investigated the association between neurological signs as well as plausible risk factors and the seroprevalence of Toxoplasma gondii and Neospora caninum infection in dogs of the Campo Grande region of Mato Grosso do Sul, Brazil. In this study, the dogs were divided into two groups based on the presence and the absence of neurological signs (n=30 in each group). Serological diagnosis was performed using the indirect fluorescent antibody test. In the group with neurological disorders, 23.3% and 30% of the dogs had anti-T. gondii and anti-N. caninum antibodies, respectively. Moreover, three dogs from this group showed co-infection with both protozoa. In the group without neurological signs, 16.7% and 13.3% of the dogs were seropositive for T. gondii and N. caninum, respectively. Although presence of neurological signs was not associated with T. gondii and N. caninum infections (P = 0.747 and P = 0.21, respectively), there was a statistical association between T. gondii seropositivity and peripheral neurological alteration (P = 0.016) among dogs with neurological signs. Raw meat ingestion was the only risk factor associated with the presence of anti-N. caninum antibodies (P = 0.041). Results revealed evidence of exposure to N. caninum and T. gondii in dogs irrespective of the presence of neurological signs. Moreover, this study highlighted the need for serological investigation of T. gondii in dogs with disturbances in peripheral nervous systems and not offering raw meat to animals to avoid the risk of N. caninum infection.

RESUMO: Este estudo teve como objetivo investigar a associação entre sinais neurológicos, assim como possíveis fatores de risco e soroprevalência de Toxoplasma gondii e Neospora caninum em cães de Campo Grande, região de Mato Grosso do Sul, Brasil. Nesse estudo, os cães foram divididos em dois grupos, baseado na presença ou ausência de sinais neurológicos (n = 30 em cada grupo). O diagnóstico sorológico foi realizado por meio do teste imunofluorescência indireta (RIFI). No grupo com distúrbios neurológicos, 23.3% e 30% dos cães tinham anticorpos anti-T. gondii e anti-N. caninum, respectivamente. Além disso, três cães deste grupo estavam coinfectados com ambos os protozoários. No grupo sem sinais neurológicos, 16.7% e 13.3% dos cães foram soropositivos para T. gondii e N. caninum, respectivamente. Apesar da presença de sinais neurológicos não ter sido associado a infecção por T. gondii ou N. caninum ( P = 0.747 e P = 0.21, respectivamente), houve associação estatística entre soropositividade para T. gondii e alteração neurológica periférica (P = 0.016) entre os cães com sinais neurológicos. A ingestão de carne crua foi o único fator de risco que apresentou associação com a presença de anticorpos para N. caninum (P = 0.041). Estes resultados mostraram evidência de que a exposição a N. caninum e T. gondii em cães independe da presença de sinais neurológicos. Além disso, este estudo destaca a necessidade de investigação sorológica para T. gondii em cães com distúrbios neurológicos periféricos, e o oferecimento de carne crua aos animais deve ser evitada devido ao risco de infecção por neosporose.

Abstract This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.

Resumo Objetivou-se determinar a ocorrência de anticorpos anti-Toxoplasma gondii, Neospora caninum e Leptospira spp., em ovinos e caprinos criados em aldeias da comunidade indígena Xukuru do Ororubá, Pernambuco, Brasil. Foram analisadas 180 amostras de soro de ovinos e 108 amostras de soro de caprinos de ambos os sexos e diferentes idades. Para a pesquisa de anticorpos foi utilizada a técnica de Reação de Imunofluorescência indireta (RIFI), para os protozoários T. gondii e N. caninum e Aglutinação Microscópica (MAT) para Leptospira spp., com ponto de corte de 1:64, 1:50 e 1:100, respectivamente. A frequência de anticorpos anti-T gondii foi de 16,6% (30/180) em ovinos e 11,1% (12/108) em caprinos. A frequência de anticorpos anti-N. caninum foi de 10,55% (19/180) para ovinos e 20,37% (22/108) para caprinos, enquanto para Leptospira spp., 2,2% (4/180) dos ovinos e 1,85% (2/108) dos caprinos reagiram positivamente. Os resultados obtidos neste estudo são inéditos em comunidades indígenas do país e alertam para o monitoramento de caprinos e ovinos da aldeia indígena Xukuru do Ororubá, quanto à ocorrência e impacto produtivo de infecções por T. gondii, N. caninum e Leptospira spp., além da ocorrência de zoonoses como a toxoplasmose e leptospirose na comunidade indígena.

Abstract The purpose of the study is to explore the epidemiological information concerning the spread of neosporosis in the Moscow region (Russian Federation) and Almaty region (Republic of Kazakhstan). The study is conducted in 2019 in the Moscow region (Russian Federation) and Almaty region (Republic of Kazakhstan). The study sample includes 800 cows (400 animals at each of the two locations), 100 at each of the 4 cattle farms in the Moscow region and 100 at each of the 4 cattle farms in the Almaty region. There were more seropositive cows on the rest of farms as compared to farm No. 1: 1.9 times more on farm No. 2 (p ≤ 0.01), 2.4 times more on farm No. 3 (p ≤ 0.001), and almost 4 times more on farm No. 4 (p ≤ 0.0001). In terms of the abortion rates, the maximum difference between the farms was 5-fold in the Moscow region (p ≤ 0.0001) and 3-fold in the Almaty region (p ≤ 0.001). Positive correlations exist between all the studied parameters (the proportion of seropositive animals, the proportion of seroprevalent animals, the abortion rate, and the stillbirth rate). The results of the study are mainly extremely valuable for the global economy, because the Republic of Kazakhstan and the Russian Federation occupy an important place in the export of meat and dairy products.

Abstract The aim of this study was to determine the presence of deoxyribonucleic acid (DNA) from Toxoplasma gondii, Sarcocystis spp. and Neospora caninum, in tissues of wild boars slaughtered in southern Brazil. A total of 156 samples were collected from different organs of 25 wild boars, and DNA from at least one of the protozoa investigated was detected in 79 samples. To differentiate between infectious agents, restriction fragment length polymorphism was performed using the restriction enzymes DdeI and HpaII. For N. caninum, conventional PCR was performed with specific primers. The DNA of at least one of the studied pathogens was detected in each animal: 26.58% for T. gondii, 68.36% for Sarcocystis spp. and 5.06% for N. caninum. Coinfection between T. gondii and Sarcocystis spp. occurred in 14 animals, between T. gondii and N. caninum in only one male animal, between Sarcocystis spp. and N. caninum in a female, while co-infection with the three agents was equally observed in only one male animal. Considering the high frequency of detection and its zoonotic risk, especially T. gondii, it appears that wild boars can be potential sources of transmission of infectious agents and the adoption of monitoring measures in these populations should be prioritized.

Resumo O objetivo deste estudo foi determinar a presença de ácido desoxirribonucléico (DNA) de Toxoplasma gondii, Sarcocystis spp. e Neospora caninum, em tecidos de javalis abatidos no sul do Brasil. Foram coletadas 156 amostras de diferentes órgãos de 25 javalis, sendo detectado o DNA de pelo menos um dos protozoários pesquisados em 79 amostras. Para diferenciar entre os agentes infecciosos, o polimorfismo do comprimento do fragmento de restrição, foi realizado usando-se as enzimas de restrição DdeI e HpaII. Para N. caninum, a PCR convencional foi realizada com "primers" específicos. O DNA de pelo menos um dos patógenos estudados foi detectado em cada animal: 26,58% para T. gondii, 68,36% para Sarcocystis spp. e 5,06% para N. caninum. Coinfecção entre T. gondii e Sarcocystis spp. ocorreu em 14 animais; entre T. gondii e N. caninum em apenas um animal macho; entre Sarcocystis spp. e N. caninum em uma fêmea, enquanto a coinfecção com os três agentes foi observada igualmente em apenas um animal macho. Considerando-se a alta frequência de detecção e seu risco zoonótico, especialmente T. gondii, constata-se que os javalis podem ser potenciais fontes de transmissão de agentes infecciosos, e a adoção de medidas de monitoramento nessas populações devem ser priorizadas.

Abstract This study establishes the serological frequency against Neospora caninum on day zero and the presence of N. caninum DNA surveyed throughout the gestation of Bubalus bubalis females in a bovine buffalo system in the central zone of the state of Veracruz, Mexico. Blood samples were taken from 11 females in 6 different sampling periods and analyzed for N. caninum antibodies detection on day zero. DNA detection by PCR was performed on all sampling periods. The gestation months of the females were recorded for five trimesters by ultrasonography, as well as births and pregnancy losses. Recorded seropositivity and positivity for agent DNA were 90.9% (95% CI 58.7-9.7) and 36.3% (95% CI 10.9-69.2), respectively, on day zero. N. caninum DNA was detected between 18.1% (95% CI 2.3-51.7) and 45.4% (95% CI 16.7-76.6) over the five trimesters of observation, with three births and three abortions recorded. The studied water buffalo population had a high presence of N. caninum antibodies; however, the detection of N. caninum DNA remained below 47% in the females. The association was only observed in the detection of DNA with pregnant females (P 0.007). Our results support the hypothesis of the resistance of water buffaloes to infection and the onset of clinical signs against infection by N. caninum even upon a high possibility of infection and reinfection described in this production system in Mexico.

Abstract The objective was to describe the seroprevalence of anti-Neospora caninum antibodies in goats and sheep slaughtered in the state of Paraíba and to identify possible associated factors with the infections. Two hundred twenty-nine samples from goats and two hundred five from sheep were analyzed by Indirect Immunofluorescence Reaction (IFAT) using a cutoff point of 1:50. The presence of anti-N. caninum antibodies was identified in 28.4% (65/ 229; 95% Confidence Interval: 22.6-34.2) of the goat samples and in 12.7% (26/ 205; 95% CI: 8.2 – 17.2) of the sheep samples. Contact between goats and dogs (Odds ratio 4.81; CI 1.13 – 2.67; p = 0.041) and cattle (OR. 1.87; CI 1.13 – 2.67; p = 0.002) was identified as a risk factor for goats and contact between sheep and dogs (OR 2.32; CI 1.58 – 3.14; p = 0.026) and history of abortion (OR 1.94; CI 1.28 – 2.90; p = 0.001) was considered a risk factor for sheep. The study revealed a high seroprevalence of anti-N. caninum antibodies in slaughtered goats and sheep in Paraíba. Risk factors such as contact with dogs/cattle and abortion history underscore the need for preventive measures to control infection and enhance animal health management.

Resumo O objetivo deste estudo foi descrever a soroprevalência de anticorpos anti-Neospora caninum em caprinos e ovinos abatidos em abatedouros públicos do Estado da Paraíba, e identificar os possíveis fatores associados às infecções. Foram analisadas 229 amostras de soro de caprinos e 205 de ovinos pela Reação de Imunofluorescência Indireta (RIFI), utilizando-se ponto de corte de 1:50. A presença de anticorpos anti-Neospora caninum foi identificada em 28,4% (65/229; Intervalo de confiança 95%: 22,6-34,2) das amostras de caprinos e em 12,7% (26/205; IC 95%: 8,2-17,2%) das amostras de ovinos. O contato de caprinos com cães (Odds ratio 4,81; IC 2,07-8,87; p = 0,041) e bovinos (OR. 1,87; IC 1,13 – 2,67; p = 0,002) foram identificados como fatores de risco para caprinos; e o contato de ovinos com cães (OR. 2,32; IC 1,58-3,14; p = 0,026) e o histórico de aborto (OR. 1,94; IC 1,28-2,90; p = 0,001) foram considerados fatores de risco para ovinos. O estudo revelou alta soroprevalência de anticorpos anti-N. caninum em caprinos e ovinos abatidos na Paraíba. Fatores de risco, como contato com cães/bovinos e histórico de aborto, destacam a necessidade de medidas preventivas para conter a infecção e melhorar o manejo da saúde animal.

Abstract Dogs can be infected by Toxoplasma gondii and Neospora caninum, for which they function, respectively, as intermediate, and definitive hosts. In the present study seroprevalence against T. gondii and N. caninum antibodies, were determined by indirect fluorescent antibody test (cut off of 16 and 50, respectively), in dogs that were treated at public veterinary hospitals in the metropolitan region of São Paulo and risk factors were identified. Out of the 1,194 samples 125 (10.5%; 95% CI: 8.8-12.3%) were positive for T. gondii and 9 (0.75%, 95% CI: 0.34-1.4%) for N. caninum. For T. gondii, statistical differences were observed between the proportions of positive dogs and different zones of the municipality (p = 0.025), and age (p = 0.02), higher among older dogs. The keepers were invited to answer an epidemiological questionnaire to analyze risk factors, and 471 (39.4%) agreed to be interviewed, and among their dogs 65 (13.8%) were T. gondii seropositive. Age group above 8 years (OR = 3.63; 95% CI: 1.08-12.23) was a risk factor and having a defined breed (OR = 0.49; 95% CI: 0.25-0.96) was a protective factor for T. gondii infection. Because of the low number of dogs positive for N. caninum, risk factors for this coccidium were not determined.

Resumo Cães podem ser infectados por Toxoplasma gondii e Neospora caninum, os quais funcionam, respectivamente, como hospedeiros intermediários e definitivos. Neste estudo, a soroprevalência contra anticorpos anti-T. gondii e N. caninum foi determinada pelo teste de imunofluorescência indireta (ponto de corte de 16 e 50, respectivamente), em cães atendidos em hospitais públicos veterinários da região metropolitana de São Paulo e fatores de risco foram identificados. Das 1.194 amostras, 125 (10,5%; IC 95%: 8,8-12,3%) foram positivas para T. gondii e 9 (0,75%, IC 95%: 0,34-1,4%) para N. caninum. Para T. gondii foram observadas diferenças entre as proporções de cães positivos e diferentes zonas do município (p = 0,025) e idade (p = 0,02), prevalência maior entre os mais velhos. Os tutores foram convidados a responder um questionário epidemiológico para análise de fatores de risco, e 471 (39,4%) concordaram em ser entrevistados, destes 65 cães (13,8%) eram soropositivos para T. gondii. Faixa etária acima de 8 anos (OR = 3,63; IC 95%: 1,08-12,23) foi fator de risco e raça definida (OR = 0,49; IC 95%: 0,25-0,96) foi fator de proteção para a infecção por T. gondii. Devido ao baixo número de positivos para anticorpos anti-N. caninum, fatores de risco para este coccídio não foram determinados.

RESUMEN Neospora caninum es el agente etiológico de la neosporosis en bovinos y perros, se considera como una de las principales causas de aborto en vacas; en perros infectados intrauterinamente e inmunodeprimidos, ocasiona lesiones multisistémicas y son considerados los hospederos definitivos. El objetivo de este estudio fue determinar la seroprevalencia contra Neospora caninum en perros urbanos y rurales del sur oriente del Estado de México y su asociación a factores de riesgo para esta especie. Mediante un muestreo no probabilístico por conveniencia se obtuvieron 70 muestras de suero de perros de una zona urbana y 76 de zona rural. Información sobre edad, género, alimentación y medio ambiente se obtuvo mediante un cuestionario. El análisis estadístico fue por medio de Ji cuadrada, la determinación de la razón de probabilidades (OR) y regresión lineal múltiple discriminante. La seropositividad general a Neospora caninum mediante IFI, fue 51.3% (75/146), en perros urbanos de 48.6% y en rurales de 53.9%, (p=0.314). Para el factor edad, la seropositividad fue mayor a medida que incrementó la misma; el consumo de placentas se reconoció como factor de riesgo. Se identifica una amplia distribución de Neospora caninum en perros, así como una constante exposición al mismo en el área de estudio

ABSTRACT Neospora caninumç is the etiological agent of neosporosis in cattle and dogs. It is considered one of the leading causes of abortion in cows. Dogs are considered the definitive host; in intrauterine infected and immunosuppressed dogs, it causes multisystemic lesions. The aim of this study was to determine the seroprevalence against Neospora caninum in urban and rural dogs from the southeast of Mexico State and its association with risk factors for these species. Through a non-probabilistic convenience sampling, 70 serum samples were obtained from dogs from an urban area and 76 from a rural one. Information on age, gender, diet and environment was obtained through a questionnaire. The statistical analysis was applied by means of Chi square, as well as the determination of odds ratio (OR) and discriminant multiple linear regression. The general seropositivity to Neospora caninum by IFAT was 51.3% (75/146), in urban dogs, it was 48.6%, and 53.9% in rural dogs (p=0.314). As for the age risk factor, seropositivity was higher as it increased; the consumption of placentas was recognized as a risk factor. A wide distribution of Neospora caninum was identified in dogs as well as constant exposure to it in the area studied.

Abstract Bovine Abortion Syndrome (BAS) has a significant impact on commercial dairies. Abortion rates vary between 2 and 20 % and there is little information regarding non-infectious factors affecting fetal stages. We analyzed the joint relationship of infectious and non-infectious factors with the probability of abortion in dairy cows. We used dairy cows from the Ñuble province of Chile (n = 148) at 42 days of pregnancy. We then took monthly blood samples until abortion or calving. We determined plasma concentrations for the following: amino transferase, albumin, cholesterol, globulin, fibrinogen, β-hydroxybutyrate, retinol, α-tocopherol, calcium, phosphorus, glutathione peroxidase, total proteins and urea. Serology was performed for infectious diseases and body conditions were recorded. Our study used a multivariable logistic regression model and proportional hazard regression. There was an (P < 0.05) association with abortion, (OR = 4.27) for Neospora caninum and (HR = 94.35) for Leptospira interrogans, with a low cholesterol concentration (P < 0.05) in the fourth month of pregnancy (OR = 0.61), and a decrease in body condition from month three to four (OR = 0.32). BAS is associated with negative energy balances, protective effects of high cholesterol plasma concentrations in the first trimester of gestation and incidences of Neospora caninum, Leptospira interrogans, and a conjunction of the two. It is important to provide epidemiological surveillance tools that help anticipate herd reproductive losses through management measures, infectious disease control and proper nutritional management.

RESUMO: Toxoplasma gondii, Neospora caninum e Sarcocystis spp. são parasitas detectados em tecidos de animais domésticos e selvagens. As aves são relevantes no ciclo de vida e epidemiologia dos protozoários devido à grande variedade de espécies de aves, hábitos alimentares e migratórios. O objetivo deste estudo foi a detecção molecular de T. gondii, N. caninum e Sarcocystis spp. em diversas espécies de aves naturalmente infectadas. Portanto, amostras de tecido de cérebro e coração foram coletados de aves recebidas e necropsiadas no Laboratório Central de Diagnóstico de Patologias Aviárias (LCDPA), sendo submetidas a extração de DNA e amplificação pela reação em cadeia da polimerase (PCR) do gene 18S rRNA para Sarcocystis spp., gene NC5 para N. caninum e gene repetitivo 529 pares de bases para T. gondii. N. caninum foi detectado em duas aves (02/65; 3,07%), em amostra de cérebro de Rupornis magnisrostris (número acesso: ON182081, 267pb) e em amostras de cérebro e coração de Dendrocygna bicolor (número acesso: ON211312, 267pb). DNA do genero Sarcocystis spp. foi detectado em três aves (03/65; 4,62%), sendo que no sequenciamento genético foram identificados Sarcocystis spp. (número acesso: MW463929) em cérebro de Nymphicus hollandicus e Sarcocystis speeri (número acesso: MW464125) em cérebro e coração de Amazona aestiva. A análise filogenética revelou que Sarcocystis spp. formou um clado com Sarcocystis spp. que utilizam gambá (Didelphis aurita) como hospedeiro definitivo e S. falcatula que utilizam Lóris-molucano (Trichoglossus moluccanus) como hospedeiro intermediário. S. speeri formou um clado com S. speeri que utilizou Mus musculus como hospedeiro intermediário experimental e formou um clado com Sarcocystis columbae, Sarcocystis corvusi, Sarcocystis halieti e Sarcocystis sp. que afetam espécies de aves. O DNA de T. gondii não foi detectado em nenhum tecido. Este é o primeiro relato de detecção de DNA de N. caninum, Sarcocystis spp. e S. speeri em amostras de tecido para essas espécies de aves estendendo a lista de hospedeiros intermediários.

ABSTRACT: Toxoplasma gondii, Neospora caninum and Sarcocystis spp. are parasites detected in tissues of domestic and wild animals. Birds are relevant in the life cycle and epidemiology of protozoa due to the wide variety of bird species, feeding and migratory habits. The aim of this study was the molecular detection of T. gondii, N. caninum and Sarcocystis spp. in several species of naturally infected birds. Therefore, samples of brain and heart tissue were collected from birds received and necropsied at the Central Laboratory for the Diagnosis of Avian Pathologies (LCDPA), undergoing DNA extraction and amplification by the polymerase chain reaction (PCR) of the 18S rRNA gene to Sarcocystis spp., NC5 gene for N. caninum and repetitive gene 529 base pairs for T. gondii. N. caninum was detected in two birds (02/65, 3.07%), in a brain sample of Rupornis magnisrostris (accession number: ON182081, 267pb) and in a brain and heart sample of Dendrocygna bicolor (accession number: ON211312, 267pb). DNA of the genus Sarcocystis was detected in three birds (03/65, 4.62%), and in the genetic sequencing Sarcocystis spp. (accession number: MW463929) in brain of Nymphicus hollandicus and Sarcocystis speeri (accession number: MW464125) in brain and heart of Amazona aestiva. Phylogenetic analysis revealed that Sarcocystis spp. formed a clade with Sarcocystis spp. that use skunk (Didelphis aurita) as definitive host and Sarcocystis falcatula that use Moluccan loris (Trichoglossus moluccanus) as intermediate host. S. speeri formed a clade with S. speeri that used Mus musculus as an experimental intermediate host and formed a clade with Sarcocystis columbae, Sarcocystis corvusi, Sarcocystis halieti and Sarcocystis sp. that affect bird species. T. gondii DNA was not detected in any tissue. This is the first report of DNA detection of N. caninum, Sarcocystis spp. and S. speeri in tissue samples for these bird species extending the list of intermediate hosts.

Resumo A ocorrência de anticorpos anti-Toxoplasma gondii e anti-Neospora caninum já foram descritos em cães de praticamente todos os estados brasileiros, entretanto, no estado do Amazonas, há poucos estudos sobre esses coccídios. Neste estudo, a ocorrência de cães domiciliados de Manaus, reagentes a T. gondii e N. caninum, e os fatores de risco para a infecção foram avaliados. Amostras de sangue de 154 cães foram obtidas, e um questionário foi aplicado aos tutores com informações sobre os animais. As amostras foram analisadas quanto à presença de anticorpos anti-T. gondii e N. caninum, pela reação de imunofluorescência indireta, ponto de corte 16 e 50, respectivamente. Associações entre as variáveis estudadas e a presença de anticorpos contra os coccídios foram feitas pelo teste de qui-quadrado, exato de Fisher ou G (p<0,05). Das 154 amostras, 19 (12,3%; IC 95% = 7,1% - 17,5%) foram reagentes a T. gondii, e a associação (p <0,05) foi observada entre a presença de anticorpos e contato com outros cães. A ocorrência de cães reagentes a N. caninum foi de 1,9% (IC 95% = 0,4% - 5,6%) com 3 dos 154 cães positivos. Nenhuma associação (p>0,05) foi observada entre a presença de anticorpos anti-N. caninum e as variáveis estudadas.

Abstract The presence of antibodies anti-Toxoplasma gondii and Neospora caninum have been described in dogs from virtually all Brazilian states, however in the state of Amazonas, there are few studies on these coccidia. In this study the occurrence of antibodies against T. gondii and N. caninum and risk factors were determined in domiciliated dogs of Manaus, AM. Blood samples were collected from 154 dogs and, during the harvest, a questionnaire was applied with questions related to the animals. The samples were analyzed, for the presence of anti-T. gondii and N. caninum antibodies, by indirect fluorescence antibody test, with cutoff of 16 and 50, respectively. Associations between the variables studied and the presence of antibodies were made by chi-square test, fisher's exact test or G test (p<0.05). Of the 154 samples, 19 (12.3% 95% CI = 7.1% - 17.5%) were reagents to T. gondii, and association (p <0.05) was observed between the presence of antibodies and contact with other dogs. The occurrence of dogs reactive to N. caninum was 1.9% (95% CI = 0.4% - 5.6%) with 3 of the 154 dogs positives, and no association (p>0.05) was observed between the presence of N. caninum antibodies, and the variables studied.

Resumo A bartonelose é uma zoonose transmitida por vetores, com distribuição mundial, que infecta várias espécies de mamíferos. Apesar dos estudos conduzidos no Brasil, informações sobre Bartonella em cães são escassas. Portanto, foi realizado um estudo retrospectivo para investigar a exposição a Bartonella sp. em cães cães, utilizando-se o ensaio de imunofluorescência indireta (IFA). Trezentas e trinta e cinco amostras de soro de cães, previamente testadas para patógenos transmitidos pelos vetores, Toxoplasma gondii e Neospora caninum foram avaliadas quanto à presença de anticorpos IgG contra Bartonella sp. Todos os cães eram oriundos da região metropolitana de Ribeirão Preto, Nordeste do Estado de São Paulo. Vinte e oito amostras (8,3%) foram positivas para Bartonella sp. no ponto de corte de 64. Entre as 28 amostras positivas, 16 (57,1%) também foram positivas para Ehrlichia canis, 12 (42,8%) para Babesia vogeli, cinco (17,8%) para T. gondii e três (10,7%) para L. infantum e N. caninum. Os resultados demonstraram que os cães amostrados foram expostos a Bartonella sp. Como os animais eram de proprietários particulares, nossos achados demonstram que as pessoas também podem ter sido expostas a Bartonella sp. São necessários estudos para avaliar se a infecção por B. canis ou E. canis constitui fator de risco para infecção por Bartonella em cães.

Abstract Bartonellosis is a vector-borne zoonotic disease with worldwide distribution that infect a broad spectrum of mammalian species. Despite the recent studies carried out in Brazil, information regarding Bartonella in dogs are scarce. Therefore, we performed a retrospective study to investigate the exposure to Bartonella sp. in dogs by indirect immunofluorescence assay (IFA). Three hundred and thirty-five archived serum samples from dogs previously tested for vector-borne pathogens, Toxoplasma gondii, and Neospora caninum were screened for the presence of IgG antibodies to Bartonella sp. All dogs originated from the Metropolitan region of Ribeirão Preto, northeast of the State of São Paulo. Twenty-eight samples (8.3%) were positive for Bartonella sp. at the cut-off of 64. Among the 28 seropositive samples for Bartonella sp., 16 (57.1%) were also seropositive for Ehrlichia canis, 12 (42.8%) for Babesia vogeli, five (17.8%) for T. gondii and three (10.7%) for L. infantum and N. caninum. Our results demonstrated that dogs sampled were exposed to Bartonella sp. Since all the animals sampled in the present study were from private owners, our findings demonstrate that these people may also be exposed to Bartonella sp. Further studies designed to assess whether the infection by other arthropod-borne pathogens such as B. vogeli and E. canis are risk factors for Bartonella infection are needed.