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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.

Abstract The present study aimed to perform an epidemiological and morphological identification of Eimeria infection in sheep in Brazil. Fecal samples from sheep were collected from 20 farms in northern Paraná, Brazil. An epidemiological questionnaire was used to evaluate the risk factors. Fecal samples containing oocysts per gram of feces (OoPG) ≥1000 were subjected to the modified Willis-Mollay method to perform oocyst identification. Sporulated oocysts were observed microscopically for morphological identification. A total of 807 fecal samples were collected. Based on the morphological characteristics of the sporulated oocysts, 10 species of Eimeria were identified, with main species observed: Eimeira ovinoidalis (98.1%), Eimeria crandallis (87.6%), Eimeria parva (79.1%), and Eimeria bakuensis (60.8%). Only 2.6% (7/268) of the sheep were infected with a single species, 4.8% (13/268) contained two different species, and 92.5% (248/268) were infected with three or more species. The analysis of risk factors showed that an intensive rearing, no rotation of pasture, dirt, and slatted floors, and age up to 12 months were associated with infection. This study showed a high prevalence of Eimeria natural infection in sheep from northern Paraná, Brazil. Furthermore, based on the risk factors, good management and hygiene practices must be employed to avoid infection.

Abstract Felines are definitive hosts of Toxoplasma gondii and can shed oocysts in their feces, contaminating the environment. Sporulated oocysts are highly resistant to the environment and have higher infectivity, which are attributed to many toxoplasmosis outbreaks. The aim of the present study was to evaluate a quantitative polymerase chain reaction (qPCR) technique for the detection of T. gondii oocysts shed by cats. Twelve cats from a previous vaccine experiment were challenged orally with 600 cysts of the TgDoveBr8 strain on day 72. Fecal samples were collected daily using the centrifugal flotation technique, with microscopic examination (Sheather technique) and qPCR for 20 days after the challenge. Cats from all groups shed oocysts in their feces. Five negative cats in the Sheather were positive according to qPCR on the 3rd day post-inoculation (dpi). Oocysts were detected on the 4th dpi using the Sheather; however, there was no statistical difference between the two methods (p=0.1116). In addition, there was no statistically significant difference in oocyst shedding between the groups according to the Sheather technique (p=0.6534) and qPCR (p=0.9670). In conclusion, these results demonstrate that qPCR can be used as an alternative to the Sheather to detect and quantify T. gondii oocysts.

In this study, we aimed to investigate an outbreak of abortion in a herd of beef sheep on a property in Ivaiporã, north central Paraná, Brazil. A total of 64 blood samples were collected from the sheep, 33 of which were mothers with reports of abortion or stillbirth, and 7 with weak lamb birth. For epidemiological investigation of the parasite, fourteen samples were collected from different sources on the property, six from water, three from soil, one from trough, and four from feed. Indirect immunofluorescence (IFI) analysis and polymerase chain reaction were performed for detecting anti-Toxoplasma gondii antibodies in the sheep and molecular identification of the protozoan in the environment, respectively. The IFI results showed that 95.3% of the sheep were seropositive for anti-T. gondii antibodies; however, the environmental samples tested negative. During the visit to the property and interview with the owner, free-roaming cats were observed on the property for biological control of rodents that had access to the food deposits. This type of management may have triggered the outbreak, which reinforces the importance of preventive management and veterinary medical monitoring on sheep farms.

ABSTRACT: Toxoplasmosis, neosporosis, and leishmaniasis are important diseases of worldwide distribution and can affect both pets and humans. Hunting dogs have been trained to hunt domestic and wild animals, which makes them more exposed to parasitic infections. The present study aimed to evaluate the seroprevalence of Toxoplasma gondii, Neospora caninum, and Leishmania spp. in hunting dogs from a rural area in Mato Grosso do Sul, Brazil. Serum samples were collected from 39 American Foxhound dogs, and the sex and age variables were recorded. Serum samples were subjected to an indirect fluorescent antibody test (IFAT) to detect antibodies. Seroprevalence was 35.9%, 15.4%, and 2.6% for Toxoplasma gondii, Neospora caninum e Leishmania spp., respectively. There was no statistical difference between genders for these diseases (P>0.05). Results demonstrated a circulation of these protozoa in hunting dogs in a rural area of the state of Mato Grosso do Sul, which can contribute to the epidemiology of these diseases.

Abstract The aim of this study was to verify the presence and identify the species of haemosporidian parasites in eared doves (Zenaida auriculata) in Brazil. Two hundred and eleven male and female eared doves were trap-captured in four different regions of Londrina city, in southern Brazil. Whole blood was collected in EDTA tubes through heart puncture after euthanasia in a CO2 chamber. A nested PCR targeting the mitochondrial cytochrome b gene (cyt b) of Haemoproteus spp./Plasmodium spp. was performed, followed by an enzymatic digestion to identify the genus. Phylogenetic trees were constructed to determine the closely related species. Out of 211 eared doves, 209 (99.05%) were positive for Haemoproteus spp. and/or Plasmodium spp. RFLP analysis showed that 72.72% (152/209) of eared doves were positive only for Haemoproteus spp., 6.22% (13/209) were positive only for Plasmodium spp., and 21.05% (44/209) of eared doves had mixed infections. Genetic analysis found four samples that were homologous with Haemoproteus multipigmentatus and one that was homologous with Plasmodium sp. This is the first molecular study of hemoparasites from eared doves in Brazil, and it is also the first description of H. multipigmentatus and Plasmodium spp. infection in eared doves in Brazil.

The aim of the present study was to detect Eimeria spp. in eared doves (Zenaida auriculata) from Brazil. Two hundred and fourteen birds were trap-capture in different regions of Londrina city, Paraná state, Southern Brazil. Fecal samples were collected, and DNA extraction was performed. A nested PCR based on the subunit I of the cytochrome c oxidase gene of the Eimeria mitochondrial genome was used to detect the DNA presence of this coccidian in eared dove feces. From 214 birds, 171 (79.9%) were positive for Eimeria spp. There was significantly difference of positivity between the site of capture. When analyzing the genders, the numbers of positive males were 84/103 (81.5%) and females 87/111 (78.4%). To the best of the authors’ knowledge, this is the first study with molecular prevalence of Eimeria sp. in Z. auriculata. Further studies should be done to identify the species of Eimeria that infect eared doves Z. auriculata.

Abstract Bovine coccidiosis is a disease of major importance in cattle herds across the world. The disorder mainly affects young calves, and E. bovis and E. zuernii are considered the most pathogenic species of the genus, however, E. alabamensis have been described in grazing calves. In this study, the prevalence of Eimeria spp. was evaluated in calves on dairy farms in the northern region of the state of Paraná, Brazil. Four hundred calves on 44 dairy farms were tested for the presence of coccidian oocysts. The positives were re-examined and the oocysts were morphometrically analyzed for species identification. All the farms were contaminated and 205 animals (51.25%) presented Eimeria spp. oocysts. Among these, 146 animals (71.22%) were co-infected by two or more species of coccidia. Ten species of Eimeria were identified: E. bovis (in 30.25% of the positive samples), E. alabamensis (26.75%), E. zuernii (22.00%), E. ellipsoidalis (18.50%), E. auburnensis (13.75%), E. canadensis (8.00%), E. cylindrica (7.25%), E. subspherica (5.00%), E. bukidnonensis (3.00%) and E. brasiliensis (0.75%). This study demonstrates the high prevalence of Eimeria spp. in the northern region of Paraná, Brazil, and detection for the first time in our region the pathogenic species E. alabamensis.

Abstract Cryptosporidium is a protozoan parasite with a wide range of hosts, including humans. However, only a few Cryptosporidium species have been described in birds (C. meleagridis, C. baileyi, C. galli and C. avium). The aim of this study was to investigate the occurrence of Cryptosporidium spp. in feces of eared doves (Zenaida auriculata), followed by molecular characterization of the parasite. A total of 196 animals of both sexes were trap-captured; the animals were culled and the intestinal contents were collected for DNA extraction. After extraction, a nested-PCR (nPCR), which amplifies a fragment of the 18S rRNA gene of Cryptosporidium spp., was performed. The amplicons obtained were purified and sequenced. PCR analysis revealed that 30 animals (15.3%) were positive for Cryptosporidium spp. There was no significant sex-dependent enrichment of Cryptosporidium occurrence (p > 0.05). Only 15 out of the 30 positive samples were successfully sequenced and their species determined, of which, 13 (86.7%) and 2 (13.3%) were C. meleagridis and C. galli, respectively. Herein, we present for the first time a molecular characterization of Cryptosporidium from feces of eared doves (Z. auriculata) and propose that these birds are a potential source of C. meleagridis infection in humans.

A neosporose é uma das principais causas de abortamentos em bovinos no mundo todo. O objetivo do presente estudo foi avaliar a ocorrência de anticorpos contra Neospora caninum em soros de bovinos de leite e cães provenientes de propriedades da região centro norte do estado do Paraná. Sangue com e sem EDTA foram colhidos de 400 bovinos e de 46 cães provenientes de 20 propriedades. Para detecção de anticorpos contra N. caninum a Reação de Imunofluorescência Indireta (RIFI) foi realizada, e bovinos e cães foram considerados positivos quando apresentaram títulos ? 100 e ? 50, respectivamente. Anticorpos contra-N. caninum foram detectados em 20,1% (80/400) dos bovinos, cujos títulos variaram de 100 a 1600. A n-PCR apresentou apenas dois animais positivos (0,5%) para os bovinos. Dos 46 cães estudados 19,6% (9/46) foram considerados positivos na RIFI, com títulos variando de 50 a 6400. Os resultados permitem concluir que a ocorrência de anticorpos contra N. caninum em bovinos e cães na mesorregião centro norte do Paraná foram semelhantes aos observados em outras mesorregiões do Paraná. A n-PCR de sangue total revelou uma baixa positividade em bovinos e cães, mostrando que a probabilidade de se detectar parasitemia em estudos epidemiológicos é baixa.

A neosporose é uma das principais causas de abortamentos em bovinos no mundo todo. O objetivo do presente estudo foi avaliar a ocorrência de anticorpos contra Neospora caninum em soros de bovinos de leite e cães provenientes de propriedades da região centro norte do estado do Paraná. Sangue com e sem EDTA foram colhidos de 400 bovinos e de 46 cães provenientes de 20 propriedades. Para detecção de anticorpos contra N. caninum a Reação de Imunofluorescência Indireta (RIFI) foi realizada, e bovinos e cães foram considerados positivos quando apresentaram títulos ? 100 e ? 50, respectivamente. Anticorpos contra-N. caninum foram detectados em 20,1% (80/400) dos bovinos, cujos títulos variaram de 100 a 1600. A n-PCR apresentou apenas dois animais positivos (0,5%) para os bovinos. Dos 46 cães estudados 19,6% (9/46) foram considerados positivos na RIFI, com títulos variando de 50 a 6400. Os resultados permitem concluir que a ocorrência de anticorpos contra N. caninum em bovinos e cães na mesorregião centro norte do Paraná foram semelhantes aos observados em outras mesorregiões do Paraná. A n-PCR de sangue total revelou uma baixa positividade em bovinos e cães, mostrando que a probabilidade de se detectar parasitemia em estudos epidemiológicos é baixa.