P. Braunig

Seroprevalence and risk factors for Neospora caninum in goats in Santa Catarina state, Brazil

Neosporosis is an infectious disease caused by the parasite Neospora caninum. Knowledge regarding neosporosis in goats is still quite limited, especially in the state of Santa Catarina (SC), southern Brazil. Therefore, this study aimed to assess the seroprevalence and risk factors for N. caninum in goats in the western and mountain regions of SC. Blood samples were collected from 654 goats in 57 municipalities. The indirect immunofluorescence test was used for antibody detection against N. caninum. Thirty samples (4.58%) were seropositive, with titers ranging from 1:50 to 1:6400. An epidemiological survey was also conducted in order to identify risk factors for neosporosis in goats. It was found that reproductive problems on the farms, as well as the diet and direct contact with dogs were casual risks for neosporosis. These results indicate that N. caninum infects goats in these regions, which may lead to reproductive problems.

The importance of vertical transmission of Neospora sp. in naturally infected horses

Neospora spp. is a intracellular protozoan phylogenetically closely related to Toxoplasma gondii and Sarcocystis neurona, and it can infect horses leading to the development of reproductive or neurological diseases. We determined the presence of antibodies to Neospora sp. in mares at their parturition time and determine the frequency of vertical transmission in healthy foals to verify the importance of transplacental transmission. The samples were analyzed by indirect immunofluorescence antibody test, showing that seroprevalence in mares is higher than in foals and seropositive mares are likely to transmit the neosporosis to their offspring. This shows that endogenous challenge occurs in horses, and it suggests that this protozoan can be disseminated by means of transplacental transmission in horse species.

Neospora caninum DNA detection by TaqMan real-time PCR assay in experimentally infected pregnant heifers.

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n=29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n=9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1+TAI, n=9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI+NC-1, n=11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Students t-test. At 35 days of gestation the pregnancy rates in the group NC-1+TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P<0.05). At 60 days, the pregnancy rates in the NC-1+TAI group (0/4, 0%) was lower compared to TAI+NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P<0.05). Animals from the group NC-1+TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1+TAI) and 5 animals (TAI+NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI+NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle.

Relation between Neospora caninum and abortion in dairy cows: Risk factors and pathogenesis of disease.

Neosporosis is a parasitic disease cause by Neospora caninum, a parasite of great importance in livestock. This study aimed to evaluate the presence of antibody against N. caninum in dairy cattle with history of abortion, as well as to identify associated risk factors for neosporosis. Animals suspected of neosporosis (n = 130) after clinical examination were randomly selected. Sera samples from 29 farms were submitted to indirect immunofluorescence technique (IFA) in order to detect antibodies against N. caninum, and animals were considered positive if ≥ IFA 1:200. An epidemiological questionnaire was used to verify probable risk factors for neosporosis and their cause-effect relation. Serological results showed that 43.8% of the animals were seropositives for N. caninum. The univariate statistical analysis found a significant relation between neoporosis and age. The number of pregnancies and the number of years that the farms had been producing milk were found as associated risk factors for the disease either by univariate or by multivariate analyses. The cause-effect model found a possible relation between reproductive problems and positive serology for neosporosis (P = 0.06). Therefore, it was concluded that approximately 44% of dairy cows with history of abortion were seropositives for N. caninum and that age and the number of years that the farms had been producing milk are risk factors for parasite infection in dairy cattle.

Anticorpos anti-Neospora caninum em bovinos de leite do sudoeste do estado do Paraná

Since November 2007 until May 2009, 1,778 serum samples of cattle from dairy herds of the Southwest of Parana State, Brazil, were used for search of anti-Neospora caninum antibodies. The frequency of seropositive animals, assessed by IFAT, was 24.2% (431/1,178), showing a relatively high occurrence in the studied population. These results show that Neospora caninum is widely distributed in the dairy cattle in the Southwest of Parana State. The presence of Neospora caninum in a herd is worrisome, since this protozoan is closely related with reproductive disorders and low milk production in cattle.

Intra-uterine exposure of horses to Sarcocystis spp. antigens

The aim of this study was to examine the intra-uterine exposure to Sarcocystis spp. antigens, determining the number of foals with detectable concentrations of antibodies against these agents in the serum, before colostrum ingestion and collect data about exposure of horses to the parasite. Serum samples were collected from 195 thoroughbred mares and their newborns in two farms from southern Brazil. Parasite specific antibody responses to Sarcocystis antigens were detected using the indirect immunofluorescent antibody test (IFAT) and immunoblot analysis. In 84.1% (159/189) of the pregnant mares and in 7.4% (14/189) of foals we detected antibodies anti-Sarcocystis spp. by IFAT. All samples seropositive from foals were also positive in their respective mares. Serum samples of seropositive foals by IFAT, showed no reactivity on the immunoblot, having as antigens S. neurona merozoites. In conclusion, the intra-uterine exposure to Sarcocystis spp. antigens in horses was demonstrated, with occurrence not only in mares, but also in their foals, before colostrum ingestion these occurrences were reduced.

Molecular detection of Cryptosporidium spp. and the occurrence of intestinal parasites in fecal samples of naturally infected dogs and cats

Cats and dogs are hosts of a large number of gastrointestinal parasites and can shed helminth eggs and protozoan oocysts in their feces. The close relationship between companion animals and humans intensifies human exposure to zoonosis caused by parasites. In this study, 177 fecal samples were collected: 128 from dogs and 49 from cats of both sexes and varied ages. One or more intestinal parasites were observed in 56.2% (72/128) of the dog fecal samples and in 53.0% (26/49) of the cat fecal samples. Parasitic monoinfection was present in 70.8% (51/72) of dog samples and in 46.1% (12/26) of cat samples, whereas multi-infection was observed in 29.2% (21/72) and 53.8% (14/26) of dog and cat samples, respectively. The detection frequency of Cryptosporidium spp. was 22.6% (40/177) using Ziehl-Neelsen staining. DNA was extracted from all samples and the Cryptosporidium small subunit ribosomal ribonucleic acid (SSU rRNA) gene was amplified from 5.6% (10/177) of the fecal samples using nested polymerase chain reaction (PCR). Amplification was achieved in 4.6% (6/128) of the dog samples and in 8.2% (4/49) of the cat samples. DNA sequencing of the nested PCR positive samples identified Cryptosporidium canis in 66.6% (4/6) and Cryptosporidium parvum in 33.3% (2/6) of the dog samples and Cryptosporidium felis in 75% (3/4) and Cryptosporidium parvum in 25% (1/4) in the cat samples. The present study thus demonstrated significant levels of gastrointestinal parasite infection in companion animals and highlighted the presence of zoonosis agents.

zCoinfections by Leishmania infantum, Neospora caninum and Toxoplasma gondii in necropsied dogs from the central region of Rio Grande do Sul, Brazil

The present paper is aimed to determine the presence of antibodies for Leishmania infantum, Neospora caninum and Toxoplasma gondii by indirect immunofluorescence (IIF) in dogs (n=78) from the central region in the state of Rio Grande do Sul necropsied in the Veterinary Hospital from Universidade Federal de Santa Maria (UFSM). The data was evaluated regarding epidemiological, anatomic, and histopathologic findings. Of the total animals evaluated, 67.9% (53/78) showed seropositivity for at least one agent. The occurrence of antibodies to L. infantum, N. caninum and T. gondii was 33.3% (26/78) 37.1% (29/78) and 43.5% (34/78), respectively. The mono infections were detected in 9.4% (5/53) of L. infantum, 18.8% (10/53) for N. caninum and 20.7% (11/53) T. gondii. The coinfections occurred in 50.9% (27/53) of animals. There were not anatomical and histopathological lesions regarding these surveyed agents, characterizing them as subclinical animals. The results confirmed the exposition of dogs to these protozoa in the central region of the RS, highlighting the circulation of the causer agent of leishmaniasis in an area considered harmless for the disease.

Investigation of antibodies against Sarcocystis neurona and Sarcocystis cruzi in equines.

ABSTRACTSarcocystis neurona is the primary agent for Equine Protozoal Myeloencephalitis (EPM), important neurological disease characterized by behavior or muscular changes, that impairs animal performance and husbandry. Sarcocystis cruzi is a pathogen related to myositis in cattle. Although related the life cycles of the parasites are distinct. S. neurona has opossums (Didelphis spp.) and S. cruzi, dogs as definitive hosts. However, S. neurona and S. cruzi may undergo cross-reactivity in serological tests, interfering on results of EPM ante-mortem diagnostic tests. In the present study, serology of 189 mares was performed by indirect immunofluorescence antibody test, using antigens of S. neurona and S. cruzi in order to assess the exposure degree of animals to antigens. Analyzing the results, it was observed that most of the animals (84.13%) reacted with at least one protozoal species and the number of animals which showed antibodies against S. cruzi was greater than S. neurona (80.42% and 33.86%, respectively) and a third of seropositive animals reacted to antigens of both species.

Investigation of antibodies againstSarcocystis neurona andSarcocystis cruzi in equines

ABSTRACTSarcocystis neurona is the primary agent for Equine Protozoal Myeloencephalitis (EPM), important neurological disease characterized by behavior or muscular changes, that impairs animal performance and husbandry. Sarcocystis cruzi is a pathogen related to myositis in cattle. Although related the life cycles of the parasites are distinct. S. neurona has opossums (Didelphis spp.) and S. cruzi, dogs as definitive hosts. However, S. neurona and S. cruzi may undergo cross-reactivity in serological tests, interfering on results of EPM ante-mortem diagnostic tests. In the present study, serology of 189 mares was performed by indirect immunofluorescence antibody test, using antigens of S. neurona and S. cruzi in order to assess the exposure degree of animals to antigens. Analyzing the results, it was observed that most of the animals (84.13%) reacted with at least one protozoal species and the number of animals which showed antibodies against S. cruzi was greater than S. neurona (80.42% and 33.86%, respectively) and a third of seropositive animals reacted to antigens of both species.