High occurrence of viable forms of Cryptosporidium and Giardia in domestic sewage from an agricultural region of Brazil.

Cryptosporidium and Giardia are the main etiologies of waterborne outbreaks caused by protozoa. These parasites are commonly detected in wastewater; however, there is little knowledge about the concentration of viable forms in treated sewage, mainly in small communities. To understand more about the presence of viable oocysts and cysts in domestic sewage, we monitored the affluent and effluent of a wastewater treatment plant (WWTP) in inner-city Brazil. Ten samplings and seven follow-ups were performed in 2020. Samples were concentrated by centrifugation, filtration and purified by fluctuation. Viability was accessed by propidium-monoazide (PMA) associated with nPCR and qPCR. Both viable protozoa were detected in all raw sewage samples (average: 438.5 viable oocysts/L). Regarding treated sewage, Cryptosporidium was detected in all of the samples (average: 92.8 viable oocysts/L) and Giardia was detected in 70% with viable cysts in 30%. Considering the follow-ups, 31.17% of Cryptosporidium viable oocysts remained in the effluent after the treatment. High amounts of Cryptosporidium and a high frequency of Giardia were detected, therefore both arrived at WWTP and were discharged into the river. These alert the presence of agro-industrial effluents into domestic sewage and demonstrated the effectiveness of the concentration technique for monitoring protozoa in wastewater.

Experimental inoculation of Neospora caninum tachyzoites in eared doves (Zenaida auriculata).

Neospora caninum is an apicomplexan parasite distributed worldwide. Although a positive association between the presence of birds and abortions in cattle associated to N. caninum has been reported, the role of the birds in the epidemiologic cycle of the parasite is unknown. To the best knowledge, no experimental studies have evaluated N. caninum in the eared dove, Zenaida auriculata. Therefore, we aimed to determine whether Z. auriculat can act as intermediate host for N. caninum. Eighteen birds were divided into four groups, G1, G2, G3, and G4 (control); G1, G2 and G3 received 2 × 106 tachyzoites of NC-1 strain via different routes: subcutaneous, intramuscular, and intraperitoneal, respectively. G4 composed of three birds. Serum samples were collected weekly, and one bird each from G1, G2 and G3 was euthanized on the 7th and 14th day post-inoculation (dpi). The remaining birds were euthanized after the 28th dpi. Tissues from the doves were evaluated using histopathological analysis, PCR and dog bioassay to detect the parasite. Dogs were fed with tissues from the birds and monitored for 30 days. Serum samples were collected weekly from the dogs for serological analysis, and feces samples were collected daily until the end of the experiment for coproparasitological examinations. No dove showed clinical signs of the infection; however, all of them seroconverted after the inoculation, with stronger immunological response in the G3 birds. The lung tissue of one G3 bird showed positive PCR results; it was euthanized on the 7th dpi, and an inflammatory infiltrate was observed in the lung and kidney from this dove. The dogs did not shed oocysts or seroconverted. Our results indicate that the intraperitoneal route induced infection in the doves; however, the parasite may have been eliminated by the host, and the doves may be resistant to chronic infection.

Vertical transmission of Neospora caninum in bovine fetuses from a slaughterhouse in Brazil.

Neospora caninum is considered as one of the main causes of reproductive failure in cattle. Vertical transmission is the main route of infection in the bovine host and plays an important role in maintaining the parasite in the herd. Molecular detection of N. caninum is important to determine the occurrence of the disease and to evaluate the genetic diversity of the parasite. The present study aimed at assessing the vertical transmission of N. caninum using molecular techniques to detect the parasite in tissue samples from bovine fetuses collected in a slaughterhouse in the state of São Paulo, Brazil. Seventy fetuses and 70 blood samples from pregnant cows were collected in a slaughtering line. Fresh samples of heart and brain tissue from fetuses were analyzed using molecular assays. Serum samples from fetuses and cows were subjected to an indirect fluorescent antibody test (IFAT) to detect antibodies against N. caninum. Nested PCR targeting the internal transcriber 1 (ITS1) region of the protozoan organism was used in the molecular testing. From the total of fetuses examined, 71.42% were positive for N. caninum by PCR. A higher number of heart samples (47.1%) were positive for the parasite using this technique. Antibodies against the protozoa were detected in 12.9% of serum samples of cows; 2.8% of fetuses were seropositive for this pathogen. Our results show that vertical transmission of N. caninum occurs in cattle from this region of Brazil, and that the use of different diagnostic techniques contributes to successful diagnosis of congenital transmission of the parasite in cattle.

Evaluation of a recombinant rhoptry protein 2 enzyme-linked immunoassay for the diagnosis of toxoplasmosis acquired during pregnancy.

The aim of this study was to evaluate an enzyme-linked immunoassay with recombinant rhoptry protein 2 (ELISA-rROP2) for its ability to detect Toxoplasma gondii ROP2-specific IgG in samples from pregnant women. The study included 236 samples that were divided into groups according to serological screening profiles for toxoplasmosis: unexposed (n = 65), probable acute infection (n = 48), possible acute infection (n = 58) and exposed to the parasite (n = 65). When an indirect immunofluorescence assay forT. gondii-specific IgG was considered as a reference test, the ELISA-rROP2 had a sensitivity of 61.8%, specificity of 62.8%, predictive positive value of 76.6% and predictive negative value of 45.4% (p = 0.0002). The ELISA-rROP2 reacted with 62.5% of the samples from pregnant women with probable acute infection and 40% of the samples from pregnant women with previous exposure (p = 0.0180). Seropositivity was observed in 50/57 (87.7%) pregnant women with possible infection. The results underscored that T. gondii rROP2 is recognised by specific IgG antibodies in both the acute and chronic phases of toxoplasmosis acquired during pregnancy. However, the sensitivity of the ELISA-rROP2 was higher in the pregnant women with probable and possible acute infections and IgM reactivity.

Indirect ELISAs with sucrose subcellular fractions of Neospora caninum as antigens for diagnosis of neosporosis in cattle.

Neosporosis is one of the major causes of abortion in cattle, and it is responsible for significant economic losses in those animals. Thus, this study aimed to evaluate indirect ELISA using subcellular fractions of Neospora caninum obtained via sucrose gradient separation. Eighty-five sera from dairy cattle previously tested using indirect immunofluorescence assay (IFA) were used. Three distinct bands were separated at 1.0 M, 1.4 M, 1.6 M, and the pellet at 1.8 M, which were identified as fractions one (F1), two (F2), three (F3), and four (F4), respectively. These fractions showed parasite membranes in the F1, rhoptry and conoids in the F2, mitochondria in the F3, and tachyzoite ghosts remain in F4. Indirect ELISAs for IgM, and IgG were performed. Additionally, sensitivity, specificity, and kappa values were defined considering the IFA as the gold standard. The highest and lowest specificities were observed for F1 (76 %) and F3 (16 %), respectively. F2 and F4 showed the highest sensitivity (93.3 %), kappa agreement (0.46), and Negative Preventive Value (NPV) (73 %) respectively. It was possible to standardize indirect ELISAs using whole soluble antigen and subcellular fractions of N. caninum, and F2 and F4 showed higher sensitivity (93.3 %), kappa (0.41), and NPV values (75 %) than F1, and F3, which could be used for epidemiology studies such as screening.

Seroprevalence and risk factors for Neospora caninum and Toxoplasma gondii in dairy cattle from São Paulo State, Brazil.

Neospora caninum is a major cause of reproductive loss in cattle worldwide as it leads to abortion and animal repositioning. Although Toxoplasma gondii does not cause a reproductive problem in cattle, consuming raw or uncooked beef poses the risk of transmission. This study aimed to evaluate the occurrence of anti-N. caninum and anti-T. gondii antibodies in dairy cattle in the West and Northwest regions of São Paulo State, Brazil. A total of 653 serum samples from dairy cows were analyzed using an indirect immunofluorescence assay (IFA). Epidemiological data from the farms were associated with the serological results of the animals by logistic regression based on the presence of antibodies. The frequencies of the antibodies against N. caninum and T. gondii were 41.6% (272/653) and 11.5% (75/653), respectively. A statistically significant association was observed between: the serum anti-N. caninum antibodies and breed, history of food supplementation for calves, introduction of outside animals that later presented reproductive problems, and history of reproductive problems by the trimester of gestation. The present study highlights the importance of neosporosis in dairy cattle in the study regions and that the inclusion of this parasite in the investigation of animals with reproductive disorders is important.

Seroprevalence of Neospora caninum and risk factors associated with infection in water buffaloes (Bubalus bubalis) from Maranhão State, Brazil.

The aim of this work was to determine the seroprevalence of Neospora caninum in buffaloes (Bubalus bubalis) from Maranhão State, Brazil, and identify risk factors associated with infection. In total, 338 buffaloes of different ages and both sexes from four farms were randomly selected. Information about the study region, animals and management was collected using an epidemiological questionnaire. Sera were subjected to an indirect fluorescent antibody test (IFAT) to detect anti-N caninum antibodies, while N. caninum DNA was detected using the polymerase chain reaction (PCR) assays. The overall seroprevalence of N. caninum in buffaloes was 27.5% (93/338), while DNA was detected in 3.0% (3/101) samples. The proportion of positive animals detected by IFAT and PCR simultaneously was 2.6% (2/77). The risk factors for N. caninum infection were contact with fomites (p = 0.022), management conditions (p = 0.005), calving interval of 20 months (p = 0.010) and deworming management (application 3 times a year in adults and calves, p = 0.020; change of anthelmintic group, p = 0.040). By multivariate analysis, management conditions was a risk factor for N. caninum infection with odds ratio of 2.2 (95% CI 1.0-4.6). This is the first report of the prevalence and risk factors for neosporosis in B. bubalis of Maranhão State, Brazil. Thus, N. caninum is widely distributed in buffalo herds in Maranhão, with management conditions being a risk factor for infection. Further studies should be conducted to elucidate the importance of buffaloes in the epidemiology of neosporosis in Maranhão State.

Socioeconomic factors associated with infection by Toxoplasma gondii and Toxocara canis in children.

The intense contact of children with domestic animals or environments contaminated with faeces of these animals, together with habits related to lack of hygiene, can facilitate infection by zoonoses. The study evaluated the seroprevalence and risk factors associated with toxoplasmosis and toxocariasis in schoolchildren in the city of Jataizinho, Paraná. Of the 412 children aged 4-15 years, 56.8% (234/412) presented antibodies reactive to Toxoplasma gondii, 42.5% (175/412) presented antibodies reactive to Toxocara canis, and 27.4% (113/412) were reactive for the two species. The analysis of risk factors showed that prevalence of toxoplasmosis and toxocariasis was associated with the level of education of the child's mother (less than eight years of schooling), age range (10-15 years) and the presence of cats in the residence. In addition, family income (up to a minimum wage), presence of a dog, the habit of playing in soil/sand and eosinophilia were associated with Toxocara canis infection. There was an association between the two zoonoses (p < .01), indicating the existence of coinfection. The results show high prevalence of these two important zoonoses, alerting to the need of implementing control measures in order to reduce the incidence and risks of sequelae in children.

Epidemiological study of toxoplasmosis outbreaks in Brazil.

In Brazil, notification of toxoplasmosis outbreaks and epidemiological investigation is a mandatory activity of health surveillance. We investigated the risk factors for toxoplasmosis during outbreaks, notifications of outbreaks by the health secretary and reports in the literature. Other factors related to the municipalities were determined through the Institute of Geography and Statistics portal. We found that fruits and vegetables were the most described transmission routes in outbreaks, and oocysts were the most common parasitic form; in recent years; there has been an increase in outbreak notifications. We also found that municipalities with high municipal human development index have a higher number of toxoplasmosis infections during outbreaks. There is a need to raise awareness among the population and producers regarding good water management and quality practices and to facilitate the acquisition of complex data to improve preventive strategies.

Evaluation of quantitative polymerase chain reaction for the detection of Toxoplasma gondii oocysts shed by cats.

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.

Seroprevalence and risk factors for Neospora caninum and Toxoplasma gondii in goats of Maranhão State, Brazil.

We estimated the seroprevalence and possible risk factors for neosporosis and toxoplasmosis in goats in the state of Maranhão, Brazil. In addition, the variables related to these animals and the management of the farm were investigated in terms of the significance of the associations. In total, 383 serum samples from goats, of both sexes and different ages, were collected from 15 farms in four municipalities. The indirect immunofluorescence test was used for antibody detection against Neospora caninum and Toxoplasma gondii. The overall seroprevalence of N. caninum in goats was 26.4% (101/382; IC 95% 22.3-31.1), and 114 out of 383 serum samples were T. gondii-seropositive (29.8%, IC 95% 25.4-34.5). In addition, the seroprevalence of coinfection of T. gondii and N. caninum in goats was 8.6% (33/382; IC 95% 6.2-11.8). The risk factors significantly associated with the seroprevalence of N. caninum were age, type of sheepfold floor, rearing system, feeding, pasture area cultivated, cats having access to the feed deposits, worming, slaughter place of the animals, history of abortion and the presence of dogs and cats. Regarding the seroprevalence of T. gondii infection, age, category, presence of other species and purpose of breeding were the risk factors. To our knowledge, this is the first report of the seroprevalence and risk factors for N. caninum and T. gondii in goats in the state of Maranhão, Brazil, which provides basic data for the implementation of strategies and control measures against neosporosis and toxoplasmosis.

Toxoplasma gondii infection in wild boars (Sus scrofa) from the State of São Paulo, Brazil: Serology, molecular characterization, and hunter's perception on toxoplasmosis.

The consumption of wild boar meat, common in many countries, became popular in Brazil after the hunting of these animals was authorized in 2013. The meat of these animals is often consumed by hunters and their social groups, and their offal is occasionally used as supplemental food in the diet of hunting dogs. Given the high frequency of foodborne diseases related to wild boar meat consumption in other countries, including toxoplasmosis, knowledge on these diseases is essential for risk assessment and elaboration of education campaigns for the exposed public. Thus, this study aimed diagnosing, isolating, and genotyping Toxoplasma gondii in hunted wild boars. For that, we obtained samples of serum and tissues (brain, tongue, diaphragm, and heart) from 26 wild boar hunted in three areas in São Paulo State, Brazil, based on convenience sampling strategy. The serum samples were submitted to the indirect immunofluorescence reaction test (IFAT) test while the tissue samples (n = 22) were used to perform a bioassay in mice to isolate the parasite. The isolated samples were genetically characterized by PCR-RFLP with SAG1, 5' and 3' SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico markers. Questionnaires were also formulated and applied to wildlife hunters to assess knowledge about toxoplasmosis. The seroprevalence of T. gondii was 76.9% (20/26), with titers ranging from 16 to 1024. Viable parasites accounted for 4.5% (1/22) of the samples. The ToxoDB #6 genotype of TgJava1 alone was detected. Most interviewed hunters, 84.2% (16/19) consume game meat and a few of them (15.7%; 3/19) prefer undercooked meat. Also, 15.7% (3/19) of the hunters reported supplementing their hunting dogs' diet with wild boar meat and/or offal. As antibodies to T. gondii were detected in 76.9% (20/26) of the studied wild boars, we concluded that infection by T. gondii is frequent in wild boars used for human and animal consumption in the studied areas. Although genotype #6 is commonly found in Brazil in domestic animals, wild animals, and humans, causing everything from mild clinical symptoms to death, this study found, for the first time, the detection of this genotype in wild boars. These results also reaffirm the importance of these animals as a possible source of T. gondii infection for humans and domestic animals.

Toxoplasma gondii infection impairs the colonic motility of rats due to loss of myenteric neurons.

BACKGROUND: Toxoplasma gondii infection causes intestinal inflammation and diarrhea indicating possible intestinal motor dysfunction. Anatomical studies have shown alterations in the colonic myenteric plexus, but it is unknown whether this impacts motility and therefore whether motility is a target for treatment. We determined whether colonic coordinated movements are compromised by toxoplasmic infection and how it is associated with anatomical changes. METHODS: Male Wistar rats were evaluated at 6, 12, 24, 48, and 72 hours and 30 days postinfection (dpi) and controls. Infected rats received orally 5 × 103 sporulated oocysts of strain ME-49 (genotype II) of T gondii. The colon was collected for anatomical analysis (including the myenteric plexus immunolabeled with HuC/D, nNOS, and ChAT) and motility analysis in vitro (conventional manometry). Fecal output was measured daily. KEY RESULTS: At 12 hours postinfection, T gondii caused hypertrophy of the muscularis externa layer of the distal colon. There was loss of total, nitrergic, and cholinergic myenteric neurons in the proximal colon at 30 day postinfection (dpi); however, only loss of cholinergic neurons was found in the distal colon. Contractile complexes in the middle and distal colon were longer in duration in infected animals, which was associated with slower migration of the colonic motor complex. However, gastrointestinal transit time and fecal pellet output remained unchanged during the T gondii infection. CONCLUSIONS AND INFERENCES: Toxoplasma gondii caused myenteric neuronal loss in the proximal and distal colon and altered the motility pattern in the middle and distal colon to a more propulsive phenotype.

Neospora caninum in free-range chickens (Gallus gallus domesticus) from southern Brazil.

Neospora caninum is an obligate intracellular parasite that can infect many domestic and wild animals, including birds. These animals are important sources for monitoring of environmental contamination, as they could become infected through sporulated oocysts; however, the real role of birds in the biological cycle of N. caninum remains uncertain. This study aimed to determine the prevalence of anti-N. caninum antibodies, evaluate associated factors, detect the parasite by molecular testing of free-range chickens from Brazil, and evaluate different techniques for its serological diagnosis. Blood samples of 366 chickens from 25 farms were collected for serological assays. The indirect fluorescent antibody test (IFAT) and the indirect enzyme-linked immunosorbent assay (ELISA) were used to detect anti-N. caninum antibodies. Chickens that tested seropositive by IFAT had their brain tissues and a pool of organs (heart, lung, and liver) submitted to PCR for molecular detection of the parasite. Out of 366 chickens, 65 (17.8%) and 163 (44.6%) were seropositive by IFAT and ELISA, respectively. Brain tissues (n=60) and the pools of organs (n=65) were negative in the PCR. Our results showed a high prevalence of antibodies in free-range chickens and that IFAT is the more sensitive technique for the detection of anti-N. caninum antibodies.

Epidemiology of a toxoplasmosis outbreak in a research institution in northern Paraná, Brazil.

Toxoplasmosis is a reportable disease in Brazil. The objective of this study was to investigate a toxoplasmosis outbreak at a research institution in Londrina-PR, Brazil. The outbreak was reported in October 2015; however, the first cases occurred in August 2015. Blood samples were collected from 674 persons at the institution. Samples were collected from soil, water (water tank) and food (vegetables) served in the restaurant. Each participant responded to an epidemiological questionnaire. For the blood samples, a chemiluminescent microparticle immunoassay was performed to detect IgM, IgG and specific IgG avidity antibodies; 10.8% (73/674) had evidence of acute toxoplasmosis. Statistical analysis showed a significant association (p < .001) between acute infection and eating lunch in the restaurant of the institution. Regarding the types of food offered in the restaurant during the period, there was a significant association between consuming raw salad (p < .001) and becoming ill. We conclude that the vegetables or raw vegetables served in the restaurant were probably the source of infection; however, the long period between exposure and case reporting made it difficult to identify the source of transmission.

Isolation, genetic and immunohistochemical identification of Toxoplasma gondii from human placenta in a large toxoplasmosis outbreak in southern Brazil, 2018.

The present study aimed to describe a molecular analysis of environmental and pork samples, the isolation, genetic identification and immunohistochemistry (IHC) of Toxoplama gondii from placenta and amniotic fluid from five pregnant women that miscarried during a toxoplasmosis outbreak in 2018, Santa Maria, Rio Grande do Sul. Environmental and pork samples were submitted to polymerase chain reaction (PCR); placenta and amniotic fluid samples to histopathology, IHC, mouse bioassay and PCR. All samples were genotyped by PCR-RFLP with 11 loci. Histopathologic and IHC were compatibles with toxoplasmosis. All pregnants were positive in PCR and bioassay, the genotypes were compared, and all were equal suggesting a same source of infection. Among the environmental and food samples, a sludge sample from a water tank and two porks samples were positive in PCR, and the genotypes were different from the pregnant women isolates. It is concluded that obtain and compare isolates is essential to elucidate outbreak source.

Isolation and molecular characterization of Toxoplasma gondii from placental tissues of pregnant women who received toxoplasmosis treatment during an outbreak in southern Brazil.

Toxoplasma gondii is a protozoan that has great genetic diversity and is prevalent worldwide. In 2018, an outbreak of toxoplasmosis occurred in Santa Maria, Brazil, which was considered the largest outbreak ever described in the world. This paper describes the isolation and molecular characterization of Toxoplasma gondii from the placenta of two pregnant women with acute toxoplasmosis who had live births and were receiving treatment for toxoplasmosis during the outbreak. For this, placental tissue samples from two patients underwent isolation by mice bioassay, conventional PCR and genotyping using PCR-RFLP with twelve markers. Both samples were positive in isolation in mice. The isolate was lethal to mice, suggesting high virulence. In addition, the samples were positive in conventional PCR and isolates submitted to PCR-RFLP genotyping presented an atypical genotype, which had never been described before. This research contributes to the elucidation of this great outbreak in Brazil.

Investigation and environmental analysis of samples from outbreak of toxoplasmosis at research institution in Londrina, Paraná, Brazil, 2016.

The objective of this study was to report an outbreak of human toxoplasmosis at a research institution in Londrina, Paraná, from December 2015 to February 2016. Blood samples from 26 symptomatic individuals were collected and the microparticle chemiluminescence immunoassay was performed to detect IgM, IgG and specific IgG avidity test in the official laboratory. A total of 20 people with symptoms and serology compatible with acute toxoplasmosis (IgM positive and IgG with low avidity) were selected as cases, while 45 asymptomatic employees working in the same teams and during the same shifts were selected as controls. All the participants of the investigation answered an epidemiological questionnaire. Three samples of water and one sludge from the institution's supply cisterns, 10 soil samples, 11 plant samples, three cat fecal samples and one domestic feline cadaver were collected for analysis of the polymerase chain reaction (PCR) for T. gondii. After analyzing the epidemiological data, the consumption of vegetables in the restaurant of the institution was the only variable associated with the occurrence of the disease. In laboratory results, all the samples showed negative results to PCR. The rapid recognition of the outbreak, early notification and investigation could have broken the chain of transmission early, thus preventing the emergence of new cases. In addition, the adoption of good food handling practices could have prevented the occurrence of the outbreak.

Seroprevalence and risk factors for Neospora caninum and Toxoplasma gondii in dairy cattle from São Paulo State, Brazil / Soroprevalência e fatores de risco para Neospora caninum e Toxoplasma gondii em bovinos leiteiros do Estado de São Paulo

Abstract Neospora caninum is a major cause of reproductive loss in cattle worldwide as it leads to abortion and animal repositioning. Although Toxoplasma gondii does not cause a reproductive problem in cattle, consuming raw or uncooked beef poses the risk of transmission. This study aimed to evaluate the occurrence of anti-N. caninum and anti-T. gondii antibodies in dairy cattle in the West and Northwest regions of São Paulo State, Brazil. A total of 653 serum samples from dairy cows were analyzed using an indirect immunofluorescence assay (IFA). Epidemiological data from the farms were associated with the serological results of the animals by logistic regression based on the presence of antibodies. The frequencies of the antibodies against N. caninum and T. gondii were 41.6% (272/653) and 11.5% (75/653), respectively. A statistically significant association was observed between: the serum anti-N. caninum antibodies and breed, history of food supplementation for calves, introduction of outside animals that later presented reproductive problems, and history of reproductive problems by the trimester of gestation. The present study highlights the importance of neosporosis in dairy cattle in the study regions and that the inclusion of this parasite in the investigation of animals with reproductive disorders is important.

Resumo Neospora caninum é uma das principais causas de perda reprodutiva em bovinos em todo o mundo, pois leva ao aborto e ao reposicionamento animal. Embora o Toxoplasma gondii não cause problemas reprodutivos em bovinos, o consumo de carne bovina crua ou não cozida apresenta risco de transmissão. Este estudo teve como objetivo avaliar a ocorrência de anticorpos anti-N. caninum e anti-T. gondii em bovinos leiteiros nas regiões Oeste e Noroeste do Estado de São Paulo, Brasil. Um total de 653 amostras de soro de bovinos leiteiros, foi analisado utilizando-se o ensaio de imunofluorescência indireta (IFI). Os dados epidemiológicos das propriedades foram associados aos resultados sorológicos dos animais por meio de regressão logística. As frequências dos anticorpos contra N. caninum e T. gondii foram 41,6% (272/653) e 11,5% (75/653), respectivamente. Foi observada associação estatisticamente significativa entre: soropositividade para N. caninum e raça, histórico de suplementação alimentar para bezerros, introdução de animais externos que, posteriormente, apresentaram problemas reprodutivos e histórico de problemas reprodutivos no segundo trimestre de gestação. O presente estudo destaca a importância da neosporose em bovinos leiteiros, nas regiões de estudo, e a inclusão deste parasita na investigação de animais com distúrbios reprodutivos é fundamental.

Spatial and simultaneous seroepidemiology of anti-Leishmania spp. antibodies in dog owners and their dogs from randomly selected households in a major city of southern Brazil.

Although leishmaniasis has been described as a classic example of a zoonosis requiring a comprehensive approach for control, to date, no study has been conducted on the spatial distribution of simultaneous Leishmania spp. seroprevalence in dog owners and dogs from randomly selected households in urban settings. Accordingly, the present study aimed to simultaneously identify the seroprevalence, spatial distribution and associated factors of infection with Leishmania spp. in dog owners and their dogs in the city of Londrina, a county seat in southern Brazil with a population of half a million people and ranked 18th in population and 145th in the human development index (HDI) out of 5570 Brazilian cities. Overall, 564 households were surveyed and included 597 homeowners and their 729 dogs. Anti-Leishmania spp. antibodies were detected by ELISA in 9/597 (1.50%) dog owners and in 32/729 (4.38%) dogs, with significantly higher prevalence (p = 0.0042) in dogs. Spatial analysis revealed associations between seropositive dogs and households located up to 500 m from the local railway. No clusters were found for either owner or dog case distributions. In summary, the seroepidemiological and spatial results collectively show a lack of association of the factors for infection, and the results demonstrated higher exposure for dogs than their owners. However, railway areas may provide favorable conditions for the maintenance of infected phlebotomines, thereby causing infection in nearby domiciled dogs. In such an urban scenario, local sanitary barriers should be focused on the terrestrial routes of people and surrounding areas, particularly railways, via continuous vector surveillance and identification of phlebotomines infected by Leishmania spp.