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Cryptosporidium spp. is one of the most important pathogens infecting nursing calves worldwide. This study aimed to investigate the intestinal microbiota of dairy calves during the first month of life and the impact of diarrhea caused by Cryptosporidium on a Brazilian farm. Fecal samples from 30 calves were collected during the first month of life, and fecal scores were recorded. Samples from the second, third, and fourth days of life were analyzed by DNA sequencing of the 16S rRNA gene. In addition, samples of sixteen calves positive for Cryptosporidium spp. were retrospectively chosen according to the development of diarrhea: four and two days before diarrhea, at the onset of diarrhea, after four days of diarrhea, at the end of diarrhea, and after six days of diarrhea resolution. Diarrhea was observed in all calves (100%), starting at day 5 of life, and all calves tested positive for Cryptosporidium in at least one sample. The microbiota richness increased with age but was retarded by diarrhea. Compositional changes associated with Cryptosporidium infection included increases in Fusobacterium, Prevotella, and Peptostreptococcus, as well as decreases in Collinsella and Lachnospiraceae. In conclusion, Cryptosporidium infection has the potential to decrease richness and change the composition of the intestinal microbiota of dairy calves.

The purpose of this investigation was to verify the effectiveness of composting in the sanitization of sludge from urban sewage (SS). The treatments (T) used consisted of SS mixed with sugarcane bagasse (SB), tree pruning residues (TP), poultry litter (PL) or grass clipping (GC) at a carbon/nitrogen ratio of approximately 30/1 and ratios of T1 (SS+SB), T2 (SS+SB+TP), T3 (SS+SB+GC), T4 (SS+SB+PL) or T5 (SS+SB+TP+GC). Temperature was measured daily for over 280 days of composting. Every two weeks, the compounds were analyzed for thermotolerant coliforms and viable helminth eggs using the multiple-tube and Yanko techniques; and once each month to identify Cryptosporidium spp. and Giardia spp. (oo)cysts using the sucrose and zinc sulfate centrifugal flotation technique and nested PCR followed by sequencing. The maximum temperatures measured in the thermophilic phase of the compounds ranged from 55°C to 64.8°C, and at day 280, weight reductions of 27% (T1), 48% (T2), 63% (T3), 66% (T4) and 64% (T5) were observed. The absence of fecal coliforms was observed from day 98 (T4), day 126 (T1), day 196 (T3 and T5) and day 210 (T2). All treatments resulted in helminth egg inviability and the absence of protozoan (oo)cysts. Only T4 and T5 were positive for G. duodenalis according to the sequencing analysis. In conclusion, high temperatures during the composting thermophilic phase made the use of pathogens required by legislation unfeasible; therefore, it is important to investigate the viability of protozoa in compounds to ensure a safe final product for human and animal health.

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. HIGHLIGHTS Viable oocysts and cysts persist after the stabilization pond treatment system.; High concentration of viable Cryptosporidium was quantified in treated sewage.; An accurate methodology to concentrate and purify (oo)cysts in wastewater was described.; PMA-PCR is a cheap and efficient methodology to detect (oo)cysts viability.; Agro-industrial effluents might influence protozoa concentration in domestic sewage.;

Esta revisão sistemática reuniu informações sobre a distribuição espacial no sul do Brasil do flebotomíneo Migonemyia migonei, um possível vetor de espécies de Leishmania causadoras da leishmaniose visceral (LV). Os artigos foram pesquisados nas bases de dados PubMed, Scielo, Web of Science e Scopus usando as palavras-chave: “Migonemyia migonei AND Paraná”, “Migonemyia migonei AND Santa Catarina”, “Migonemyia migonei AND Rio Grande do Sul”, “phlebotomine AND Parana” , “flebotomíneo AND Paraná” e “flebotomíneo AND Paraná”, “flebotomíneo AND Santa Catarina;”, “flebotomíneo AND Santa Catarina” e “flebotomíneo AND Santa Catarina”, “flebotomíneo AND Rio Grande do Sul;”, “flebotomíneo AND Rio Grande do Sul” e “sandfly AND Rio Grande do Sul”. A busca inicial identificou 322 artigos que atenderam aos critérios de seleção. Em seguida, arquivos vazios ou duplicados foram excluídos. Os títulos foram triados e os textos completos foram obtidos. Esta revisão incluiu 36 artigos, abrangendo 72 dos 399 (18,04%) municípios do Paraná, um dos 295 (0,33%) de Santa Catarina, dois dos 497 (0,40%) do Rio Grande do Sul. Mg. migonei foi encontrado em 54 municípios do Paraná estado, em um município de Santa Catarina e em um do Rio Gr ande do Sul. Com base na ampla distribuição de Mg. migonei nos municípios paranaenses, é necessária maior atenção quanto aos casos de LV em humanos e animais, além de investigações epidemiológicas desses casos de suspeita de autoctonia, bem como maiores esforços de prevenção e controle. Mais estudos são necessários em Santa Catarina e Rio Grande do Sul.

Abstract Cattle are an important source of zoonotic species of Cryptosporidium for humans. The aim of this study was to investigate the presence of Cryptosporidium, identify the species and determine the risk factors relating to environment, animals and management among dairy calves in eight Brazilian states. A total of 408 fecal samples from calves aged 1-60 days were analyzed. An epidemiological questionnaire was completed. Sample screening was performed using Ziehl-Neelsen technique and the positive samples were subjected to nested PCR. Cryptosporidium species were identified by means of the PCR-RFLP technique, using SSPI, ASEI and MBOII enzymes. The Ziehl-Neelsen technique showed that 89.7% (35/39) of the farms and 52.9% (216/408) of the samples were positive. Through nested PCR, these protozoa were detected in 54.6% of the samples. The 56 samples subjected to PCR-RFLP presented Cryptosporidium parvum. There was higher prevalence of the parasite in animals aged 7 to 28 days (62.6%). Diarrhea, ages between seven and 28 days and a spring water source were factors associated with the risk of infection. The calf hutch-type management system was associated with reduced infection. These findings demonstrate the high level of Cryptosporidium spp. circulation in cattle herds and the predominance of the species C. parvum.

We report on apparent temporal progression of probable sources of infection and transmission routes for global human toxoplasmosis outbreaks as described in published articles. We searched the Scientific Electronic Library Online, Web of Science, PubMed, and Scopus databases for articles on Toxoplasma, toxoplasmosis, and outbreaks. We found that transmission routes for Toxoplasma gondii varied by decade. In the 1960s and 1990s, toxoplasmosis outbreaks mainly occurred through ingestion of cysts in meat and meat derivatives; in the 1980s, through milk contaminated with tachyzoites; in 2000, due to the presence of oocysts in water, sand, and soil; and in 2010, due to oocysts in raw fruits and vegetables. Our study suggests a possible change in the epidemiology of reported toxoplasmosis outbreaks. Because of this change, we suggest that greater attention be paid to the disinfection of vegetables, as well as to the quality of water used for drinking and irrigation.

O estudo foi realizado entre julho de 2014 e maio de 2016, em 21 propriedades hortícolas do estado do Paraná, Brasil. Foram coletadas duas amostras de vegetais folhosos e uma amostra de água de irrigação por propriedade. As amostras de água foram analisadas pela técnica do substrato cromogênico para avaliar a contaminação por coliformes totais e Escherichia coli, e os vegetais foram avaliados pelas técnicas de Willis (1921), Hoffman et al. (1934), Faust et al. (1939) e quanto à contaminação por parasitas. Observou-se presença de E. coli em 80,95% (17/21) das amostras de água; com relação aos vegetais, 19 (45,23%) continham pelo menos uma espécie de parasita, tais como: ancilostomatídeos, Chilomatix spp., Dipillidium spp., Entamoeba spp., Strongyloides spp., Trichuris spp., larva de vida livre, larva de nematódeo, oocisto não-esporulado. Houve associação estatística entre o destino do esgoto (fossa seca) e a positividade aos parasitas. Os dados mostram contaminação fecal em número significativo de amostras e confirmam a necessidade de maiores exigências sanitárias durante o cultivo de hortaliças folhosas, que, na maioria das vezes, são consumidas cruas.

The Brazilian agricultural model is based on the intensive use of pesticides. Once in the environment, these compounds might target hydric sources, mainly through soil lixiviation or during crop application, exposing water biota as well as humans to contaminants. The objective of this research was identify the presence of 14 pesticides (ametrine, atrazine, azoxystrobin, carbendazim, carbofuran, clomazone, chlorpirifos, diuron, hexazinone, imidacloprid, malathion, simazine, tebuconazole and tebuthiuron) in source water samples and drinking water samples from the city of Londrina, Paraná State, Brazil. 24 water samples were collected biweekly from December 2014 to October 2015. The sample preparation was done using Solid Phase Extraction (SPE) with 3 mL/500 mg C-18 cartridges. Pesticide determination was performed applying Liquid Chromatography tandem Mass Spectrometry (LC-MS/MS). The frequencies of detection of some compounds in source water were: atrazine (85%), azoxystrobin (95%), carbendazim (83%), diuron (75%), imidacloprid (95%), simazine (12%), hexazinone (4%), tebutiuron (4%) and tebuconazole (33%). In the water supply, the following were detected: atrazine (79%), azoxystrobin (95%), carbendazim (8%), diuron (83%), imidacloprid (95%), simazine (12%), hexazinone (4%), tebuthiuron (8%) and tebuconazole (33%). Some of these compounds are included in Brazilian legislation, and in these cases, the concentrations determined in this study varied between 241 and 7 ng L?¹, which were lower than the maximum allowable concentrations for each of the compounds.

Abstract The oocyst, a resistant form of Toxoplasma gondii, plays an important role in the transmission of this protozoan. The objective of this review was to report the methods capable of inactivating oocysts through a systematic review of the literature carried out in the Scientific Electronic Library Online, Web of Science, Science Direct, PubMed and Scopus databases. The keywords searched were (((effects OR infectivity OR resistance) AND Toxoplasma) AND oocyst). We selected 16 articles that described 309 different treatments. Among all the protocols evaluated, 35.60% (110/309) were effective in inactivating oocysts. Physical methods were more effective than other methods (p

Neosporosis, caused by the protozoan Neospora caninum, has been widely reported and discussed as a major disease associated with reproductive problems in herds of various animal species. This study aimed to perform a systematic review and meta-analysis on the prevalence and risk factors associated with ovine neosporosis worldwide. This is the first systematic review, which was performed according to the established preferred reporting items in systematic reviews and meta-analyses (PRISMA) guidelines to address ovine neosporosis. Four databases were used in this study: PubMed, SciELO, Web of Science, and Scopus. The selected keywords were “neospora AND sheep,” “neosporosis AND sheep,” “neospora AND lamb,” “neosporosis AND lamb,” “neospora AND ewe,” and “neosporosis AND ewe.” The meta-analysis calculations were performed with random-effects models for proportions, and heterogeneity was tested using the I2 test. This study identified 24 studies on ovine neosporosis, nine of which evaluated the factors associated with the risk of seropositivity. After meta-analysis, the pooled estimated prevalence was 12% (95% CI, 8-16%; I² p-value < 0.01). The evaluation of the included studies allowed the identification of ovine neosporosis distribution in continents according to demonstrated prevalence rates. We confirmed via meta-analysis that the use of indirect immunofluorescence assay (IFAT) or enzyme-linked immunosorbent assay (ELISA) in the diagnosis of ovine neosporosis was not a source of data heterogeneity. The presence of dogs on the properties was reported as the main risk factor associated with the occurrence of ovine neosporosis; however, this was not confirmed by the meta-analysis.

Abstract The aim of this study was to evaluate the seroprevalence and factors associated with the presence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state. The detection of antibodies for T. gondii and N. caninum was performed by homemade and commercial indirect ELISA, respectively. Multiple logistic regression analysis was used to verify the factors associated with the seroprevalence. Antibodies anti-T. gondii and anti-N. caninum were observed in 42.7% and in 17.6% of the animals, respectively. The protective factors associated to seropositive were “some level of confinement” (full or semi-extensive confinement) (OR=0.53) for T. gondii and “use of skilled labor” (OR=0.64) for N. caninum. The risk factors were “presence of cats” (OR=1.75) for T. gondii and “feeding of dogs with sheep placental remains” (OR=1.79) for N. caninum. In addition, to presenting a significant and simultaneous seroprevalence for both agents (9.9% of the animals), the results also indicate that deficiencies in management and environmental sanitation, the presence of reservoirs, and types of exploitation enhance the seropositivity. Thus, studies like this might support sanitary programs and public policies for the prevention of T. gondii and N. caninum in the sheep herds of Paraná state.

Although leptospirosis has been considered a major concern in urban areas, no study to date has spatially and simultaneously compared both owner and dog serology in households of major cities. Accordingly, the aim of the present study was to assess the seroprevalence of Leptospira antibodies, evaluate associated risk factors and conduct spatial analyses in 565 randomly selected households, which included 597 dog owners and 729 dogs in Londrina, Southern Brazil. Seropositivity by MAT were detected in in 11/597 (1.84%) owners and in 155/729 (21.26%) dogs. The risk factors were evaluated with logistic regression analysis and spatial factors and case distribution were evaluated with kernel density analyses. The sera of 14/155 (9.03%) dogs reacted for more than one serovar with the same titer. Canicola was the most frequent serogroup, detected in 3/11 (27.27%) owners and 76/155 (49.03%) dogs. The highest titer among the owners was 1:3,200 and was detected in the same household with a titer of 1:800 in the dog. Simultaneous owner-dog seropositivity was found in 7/565 (1.23%) households, with three reacted against serogroup Canicola. Positive owners were detected in 4/565 (0.70%) households and positive dogs were detected in 141/565 (24.95%) households. The associated risks of infection for dogs were different from those associated with infection in owners. Risk analyses for Canicola also identified specific factors of infection. Regardless of owner and dog cases were not statistically clustered, the kernel map has shown dog positivity occurrence in the same hot locations and near positive owners. The dependent variable analysis and logit model suggested a greater likelihood of peri-domiciliary contact with Leptospira. In conclusion, exposure to Leptospira infection was significantly higher in dogs than in their owners and human cases spatially overlapped dog cases, implicating dogs as potential environmental sentinels for this disease. In addition, the associated risk may vary according to serogroup, and the observed simultaneous Canicola seropositivity of owner and dog has suggested intradomicile-transmitted infection.

Abstract The aim of this study was to determine the occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and their associated factors among goats in farms from the Paraná state, South Brazil. The serological analysis was performed by indirect ELISA on 629 goat serum samples collected from 32 farms distributed in five mesorregions from the state. Seropositivity was observed in 30.7% of the animals for T. gondii, 6.3% for N. caninum, and 3.0% for both agents. Final multiple regression models showed that the use of assisted reproduction decreased the chance of seropositivity for T. gondii (PR=0.70) and the slaughtering site on the farm (PR=2.03) increased it. To N. caninum, the supplying concentrated feed to animals (OR=4.80) and animal pre-weaning death (OR=9.96) increased the chance of seropositivity. The variables associated with these seropositivities suggest deficiencies in sanitation and reproductive management. In addition, it recognizes critical points of infection for animal production management.

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.

Abstract The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.

Abstract The objective of this study was to determine factors associated with vegetable contamination with zoonotic protozoan. Samples of water, soil and vegetables were collected from July/2014 to May/2016, totaling 83 samples, 21 properties of Londrina region, Paraná, Brazil. DNA amplification of Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis in the samples was conducted using polymerase chain reaction (PCR). The PCR results were positive for T. gondii in 12.9% (8/62), Cryptosporidium spp. in 11.3% (7/62) and G. intestinalis in 25.8% (16/62) of the samples. DNA sequencing identified C. parvum in five samples and G. intestinalis Assemblage E in three. The statistical associations demonstrated greater probability of positive samples for T. gondii and for at least one of the three protozoa when the source of irrigation water was the river; a greater chance of positive samples for Cryptosporidium spp. when deer were present on the property; and a smaller chance of positive samples for at least one of the three etiologic agents when soil was supplemented with limestone. The results expose some critical contamination points, providing support for training farmers on good management practices during the production process.

Abstract Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

The objective of this study was to investigate the presence of anti-Leishmania spp. antibodies using an indirect immunofluorescence technique in the serum of equidae slaughtered in two slaughterhouses registered with the Federal Inspection Service. Titers greater than 40 were considered reagents. Blood samples were collected from 398 animals of both sexes, with different ages, and coming from six Brazilian states, of which 46.0% (183/398) were reactive, indicating that these animals were exposed to the leishmaniasis agent that is widely distributed in Brazil.

Water is an important vehicle for pathogens such as Cryptosporidium spp. and Giardia duodenalis. In fact, these organisms are responsible for almost 58% and 38% of the waterborne outbreaks reported in ~60 years. Characteristics related to the environmental phase of these protozoa and the different types of hosts are important factors related to environmental contamination. In cryptosporidiosis and giardiasis, outbreaks caused by waterborne pathogens are identified as major risk factors for contamination of water resources by untreated sewage and the entrainment of cattle feces in rainwater. Further, this review covers taxonomic, biologic, and epidemiologic aspects such as prevalence, risk factors, and molecular characterization of these protozoa observed in humans and cattle in order to elucidate the role of these hosts in environmental pollution, and consequently, as a source of infection for susceptible humans, especially in rural areas. Alternative water resources such as wells and springs are used without water treatment and potability monitoring. Epidemiological data are useful, but insufficient to characterize the source of infection, thereby requiring the use of appropriate molecular methods for subtyping the protozoa detected in environmental and feces samples as well as for assessing public health risk.

ABSTRACT The aim of this study was to perform a systematic review of scientific papers that used spatial analysis tools in cases of leishmaniasis, in Brazil. The search for articles was carried out in PubMed, SciELO, Scopus and Web of Science databases. The keywords used in the identification of the articles were Thematic map AND Leishmaniasis, Spatial analysis AND Leishmaniasis, and Geoprocessing AND Leishmaniasis, in English language. A total of 360 articles were found, and 11 of them were analyzed after screening by title and abstract as well as reading of the full articles. The States studied were Sao Paulo, Acre, Maranhao, Piaui, Minas Gerais, Parana and Tocantins. Cutaneous leishmaniasis occurred predominantly in rural areas, with clusters in forest reserve regions or modified forest areas. Conversely, visceral leishmaniasis mainly occurred in peripheral and central urban areas associated with poorer environments and urban infrastructure, including worse sanitation. We conclude that the spatial distribution of leishmaniasis is closely related to the living environment of the risk population. The analyzed articles associated geospatial data with some risk factors for the disease, pointing out the locations where most cases occur, creating a relevant source to define control strategies.

Abstract The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.

Human toxoplasmosis, a protozoonosis caused by Toxoplasma gondii, has been described as a worldwide foodborne disease with important public health impact. Despite infection has reportedly varied due to differences in alimentary, cultural and hygienic habits and geographic region, social vulnerability influence on toxoplasmosis distribution remains to be fully established. Accordingly, the present study has aimed to assess T. gondii seroprevalence and factors associated to social vulnerability for infection in households of Ivaiporã, southern Brazil, with 33.6% population making half minimum wage or less, ranked 1,055th in population (31,816 habitants), 1,406th in per capita income (U$ 211.80 per month) and 1,021st in HDI (0.764) out of 5,570 Brazilian cities. Serum samples and epidemiological questionnaires were obtained from citizen volunteers with official City Secretary of Health assistance in 2015 and 2016. In overall, serosurvey has revealed 526/715 (73.57%) positive samples for anti-T. gondii antibodies by Indirect Fluorescent Antibody Test. Logistic regression has shown a significant increase associated to adults (p = 0.021) and elderly (p = 0.014) people, illiterates (p = 0.025), unemployment (p

O objetivo deste trabalho foi detectar anticorposcontra T. gondii emcãescom sinais clínicoscompatíveis com toxoplasmose atendidos em um Hospital Veterinário. Foram submetidos a sorologia 598 cães no período de 1993 a 2013. A pesquisa de anticorpos da classe IgG contra T. gondii foi realizada pela Reação de Imunofluorescência Indireta (RIFI); amostras com título ≥ 16 foram consideradas positivas. Entre os cães com sinais clínicos, 259 (43,31%) foram soropositivos para T. gondii com títulos variando de 16 a 4096; já entre os animais hígidos, 79 (22,25%) foram reagentes. Os sinais mais evidentes foram os neurológicos (30,43%) e uveíte (8,03%). Entre as variáveis raça, sexo e idade, apenas a última exerceu influência sobre os resultados observados, com maior frequência de soropositivos em cães idosos. Palavras-chave: caninos; RIFI; sinais clínicos; sorologia; Toxoplasma gondii; zoonose.

The leishmaniasis are a group of globally widespread parasitic diseases caused by a protozoa of the genus Leishmania, the state of Paraná accounts for 98% of leishmaniasis human cases in southern Brazil and the dogs might serve as a sentinel of the risk of infection to humans in a given area. The aim of this work was to study the epidemiology of leishmaniasis in dogs residents in two different environments, they are: recycling of solid waste centers (Group 1) and a neighborhood adjacent to a forest (Group 2), both in Londrina, Paraná State, Brazil and to determine epidemiological variables associated to infection in these different realities. Serum samples, randomly collected, from 61 dogs from Group 1 and from 74 dogs from Group 2 were submitted to Indirect Immunofluorescence Antibody Test (IFAT) to research for anti-Leishmania IgG antibodies. Of the 135 dogs analyzed, 13 animals (9.6%) were positive; nine (14.7%) in Group 1 and four (5.4%) in Group 2, the difference between the groups is not statistically significant. These results suggest the presence of the protozoan Leishmania spp. in both environments studied which indicate potential for infection in the human population.

ABSTRACT: The present study aimed to investigate an abortion outbreak in a dairy goat herd in the municipality of Arapoti, Parana, Brazil. At the beginning of the outbreak, blood samples were collected from 33 goats with clinical signs; later, of the whole goat herd, two cats and two dogs. Milk samples were collected from 78 lactating goats. Four environmental soil samples and four samples of feed residue from goat feeders were collected too. Immunofluorescence antibody test (IFA) was used for serodiagnosis, the molecular analysis was conducted by means of the polymerase chain reaction (PCR), for the isolation of the etiological agent the bioassay was used. The results of the IFA revealed that 76.53% (137/179) of the goats, two dogs and two cats were seropositive for Toxoplasma gondii. Bioassay revealed one buffy coat and two milk sample having viable T. gondii. In the PCR, 11 whole blood samples, eight milk, three feeder troughs, and all soil samples were positive. The findings of the present study confirmed an outbreak caused by environmental contamination (of soil and feed) with T. gondii oocysts that could have been shed by kittens that lived on the farm and had access to the stock of goat food, facilitating this contamination, which reinforces the need for veterinary assistance and good management practices on farms.

Abstract The aim of this study were to detect antibodies anti-Leishmania spp. and anti-Trypanosoma cruzi in two different populations of domestic cats (Felis catus domesticus) from North Paraná referred for surgical castration and to determine which characteristics of the animals studied may be associated with seropositivity. Serum samples from 679 cats were analyzed using enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT) in series. Associations between age, sex, race, year of care and animal group were verified using the simple logistic regression. Percentage of 8.5% (58/679) of cats were positive for Leishmania spp. and 7.6% (51/673) for T. cruzi by the tests ELISA and IFAT. Animals collected by non-governmental animal protection organizations presented more seropositivity for Leishmania spp. (p

[This corrects the article DOI: 10.1371/journal.pone.0180906.].

The effects of managerial systems on the occurrence of gastrointestinal helminths in Free-Range Chickens (FRCs) from northern Paraná, Brazil were investigated. The most predominant (23.3%; 61/262) cestode observed was Raillietina cesticillus; Heterakis gallinarum (71.4%; 187/262) and Ascaridia galli (45%; 118/262) were the predominant nematodes; Postharmostomum commutatum was the only trematode observed in 2.7% (7/262) of FRCs. The most elevated parasitic burdens were associated with Heterakis gallinarum, Ascaridia galli, and Raillietina cesticillus. Significant (p ? 0.05) associations were observed when the effects of the types of bedding, soil type, and fence restriction of FRCs were considered relative to the possibility of helminthiasis. The type of bedding, the length of the sanitary break, and the presence of shading significantly (p ? 0.05) influenced the possibility of FRCs being infected by H. gallinarum. Most parameters evaluated were significantly associated with infection of FRCs by A. galli. These findings suggest that FRCs from northern Paraná are infected by a wide-range of gastrointestinal helminths, but more frequently by R. cesticillus, H. gallinarum, and A. galli. Moreover, the type of floor bedding, the soil type, and the usage of fences in the management of FRCs is directly related to gastrointestinal helminthiasis.

Este estudo avaliou a dinâmica da população de cães e gatos e o perfil de seus respectivos guardiões na cidade de Jaguapitã, sul do Brasil. A razão homem:cão e homem:gato foi de 4,6 e 21,5, respectivamente. Em comparação, mais cães (n=2,460) do que gatos (n=571) eram domiciliados havendo significativamente mais cães machos (55.8%; 679/1,217) em relação as fêmeas (44.2%; 538/1217). A maioria dos gatos (69.2%; 180/260) não recebeu qualquer medicação antiparasitária, não foi vacinada contra raiva (91.2%; 237/260) ou qualquer doença infecciosa específica (91.5%; 238/260). Menos da metade destes (40.8%; 106/260) tinha idade inferior a um ano e, um número significante de gatos não apresentava definição racial (81.2%; 211/260) e não era castrado (93.5%; 243/260). A maior parte dos cães não apresentava raça definida (69.5%; 846/1217), tinha entre um e quatro anos de idade (42.6%; 519/1217) e não era castrada (96.3%; 1172/1,217). Um grande número de cães recebeu medicação antiparasitária (71%; 865/1,217), mas a maioria desses não foi imunizada contra raiva (63.8% 777/1,217) ou outra doença infecciosa (58.6%; 713/1,217). Cães e/ou gatos estavam presentes na maioria dos domicílios visitados (68.7%; 770/1,120), nos quais 54.4% (610/1,120) possuíam somente cães, 4.9% (55/1,120) possuíam somente gatos, enquanto 9.4% (105/1,120) possuíam cães e gatos. Os resultados obtidos são similares aos descritos em outras cidades do Brasil e outros países. O livre acesso à rua associado ao reduzido nível de imunização contra as doenças infecciosas de cães e gatos e à conduta de posse responsável adotada pelos guardiões torna esses animais altamente susceptíveis às doenças infecciosas e zoonóticas.

This study aimed to determine the prevalence of Toxoplasma gondii in swine slaughtered in Iporã, Northwest Paraná state. Blood samples were obtained from 500 finishing swine. All animals were raised under intensive farming; the blood samples were analyzed using the immunofluorescence antibody test (IFAT). Animals with titers of ?64 were considered positive for T. gondii infection. Of the tested samples, 63 (12.6%) were positive by IFAT; 58 (92.06%) of these showed titers of 64 (4.7%), with titers 256 and two (3.1%) titers of 1024. These pigs might be considered a source of T. gondii infection for humans

Increasing human-animal proximity has also increased risks of zoonotic disease transmission. This study measured the prevalence of anti-Toxoplasma gondii, anti-Leishmania spp. and anti-Leptospira spp. IgG antibodies in dogs and cats attended by the Birth Control Project (BCP) from 2004 to 2012 to determine which environmental variables were significantly associated with infectious agents in the study population. Blood samples from 271 dogs and 415 cats were collected by jugular or cephalic venipuncture for serological analysis. Indirect immunofluorescence assays were used to detect anti-T. gondii and anti-Leishmania spp. IgG antibodies and microscopic serum-agglutination tests were used to detect anti- Leptospira spp. antibodies. Data tabulation and statistical analyses were performed using EpiInfo (3.4.3). Among canine sera samples, 20.6% (56/271), 1.3% (3/236), and 21.6% (51/236) were positive for T. gondii, Leishmania spp., and Leptospira spp., respectively. Higher T. gondii prevalence was found in dogs of unknown breeding history (p=0.0009), those fed raw or undercooked meat (p=0.0101), and those that did not receive the polyvalent vaccine (p=0.0147). There was no statistically significant association between leishmaniosis and leptospirosis and variables evaluated in the epidemiological questionnaire. Among feline sera samples, 20.9% (87/415) were positive for T. gondii. The highest prevalence rates were observed in felines that hunted rodents (p=0.0388) and that consumed untreated water (p=0.0294). These results confirmed that ingestion of raw or undercooked meat, drinking untreated water, and predation of rodents were important risk factors for T. gondii infection. A low Leishmania spp. serodiagnostic prevalence was observed. The most frequent Leptospira spp. serovars were Copenhageni, Canicola, and Butembo.

Toxoplasmosis is a zoonosis caused by the Toxoplasma gondii protozoa. The congenital form happens when the parasite reaches the fetus through the placenta and causes damages of different intensities, which can be consequence of the strain virulence, of the immune response capability of the pregnant woman or even of the pregnancy stage, and can result in fetal death or severe clinical symptomatology. The aim of this paper was to determine the prevalence of antibodies to Toxoplasma gondii antibodies in pregnant woman seen at the Basic Health Units (BHU) in the city of Umuarama (PR) and check for possible toxoplasmosis associations with social-demographic characteristics, behavior and environmental variables. In this study, a total of 813 pregnant women who had their prenatal exams at the BHU in the city of Umuarama (PR) were included. Blood samples collected from these subjects were submitted to IgG and IgM anti-T. gondii antibody survey. Each pregnant woman answered an epidemiological questionnaire for the detection of variables, which were later analyzed by the EpiInfo program. The prevalence detected for IgG and IgM anti-T. gondii antibodies were 56.8% and 1.0%, respectively. The variables age group, education level, per capita income, number of pregnancies, ingestion of fresh sausage, presence of peridomiciliary cats in the backyard and the habit of ingesting farm milk showed association to the infection by T. gondii.

A água é um importante veículo de transmissão de patógenos, entre eles Cryptosporidium spp e Giardia duodenalis, respectivamente responsáveis por 58% e 38% dos surtos de veiculação hídrica registrados em cerca de 60 anos. Características relacionadas às fases ambientais destes protozoários e a grande quantidade de hospedeiros são importantes fatores relacionados à contaminação ambiental. Em surtos de criptosporidiose e giardíase por veiculação hídrica são apontados como principais fatores de risco a contaminação dos mananciais, utilizados para a dessedentação humana, por esgoto não tratado e o desague de fezes de animais devido ao carreamento por água da chuva. Além destes fatores, esta revisão aborda aspectos taxonômicos, biológicos e epidemiológicos, como prevalência, fatores de risco e caracterização molecular destes protozoários em humanos e bovinos como forma de elucidar a importância destes hospedeiros na contaminação ambiental e, consequentemente, como fonte de infecção para humanos suscetíveis, especialmente em áreas rurais. Nestas áreas são frequentemente utilizadas fontes alternativas de abastecimento de água, como poços e nascentes, sem tratamento e monitoramento de potabilidade. Dados epidemiológico auxiliam, porém não são suficientes para caracterizar fontes de infecção, sendo necessário o uso de adequadas ferramentas de caracterização molecular para subtipagem de protozoários identificados em fezes e amostras ambientais e avaliação do risco de saúde pública.

Toxoplasmosis is a worldwide zoonosis with important public health implications. The aims of this study were to determine and identify factors associated to the seroprevalence of anti-Toxoplasma gondii antibodies in humans and dogs from Jataizinho, Parana State, Brazil. Serum samples from 280 humans and 766 dogs, from rural and urban areas, were tested by indirect immunofluorescent antibody test. Positive dilutions were ≥ 1:16. Epidemiological informations were obtained through an individual questionnaire and data tabulation was performed using EpiInfo 3.5.3 (CDC, Atlanta). Statistical significance was evaluated by chi-square (ᵪ2) or Fisher’s exact tests with a 5.0 % significance level. The seroprevalence in humans was 70.00% (196/280), as follows: 69.08% (143/207) in rural and 72.60% (53/73) in a slum urban community. The seroprevalence in dogs was 52.35% (401/766), as follows: 71.74% (99/138) in rural and 48.08% (302/628) in urban area. In rural areas, there was no factor associated to risk of infection. In urban slum community, visualizing more than four rats at home was a factor statistically associated to seropositivity for human toxoplamosis. Infection of urban dogs was associated to age, contact with stray dogs or rodents, and free access to street. The results indicated a similar possibility of contact with Toxoplasma gondii between dogs and humans living in rural or slum urban areas of small municipalities. Keywords: IFAT; prevalence; Toxoplasma gondii; zoonosis.

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

Toxoplasmosis, caused by Toxoplasma gondii, has traditionally been considered an important water and foodborne protozoonosis with important public health considerations. Although felids play a well-established role as definitive hosts, canine epidemiological involvement in the parasite's life cycle remains questionable and controversial. The increasing closeness of the human-dog bond, particularly seen in urban settings, has been recognized as a historically unprecedented worldwide movement. Sharing daily lives in the same households, dogs may be exposed to similar associated risks of T. gondii infection as their owners. Thus, epidemiological assessment of the intra-domiciled environment, especially among socio-economically different human populations, may provide novel information regarding the actual role of dogs in animal and human toxoplasmosis. Despite spatial approaches being recently used for other water and foodborne diseases, no study has been conducted on the simultaneous spatial seroprevalence of both human and animal IgG anti-T. gondii antibodies in urban areas of major cities. Accordingly, the aim of the present study was to assess the seroprevalence and associated variables of Toxoplasma infection in owners and their domiciled dogs in Londrina, southern Brazil. Human and canine seroprevalence rates and variables associated with seroprevalence were investigated through representative random sampling among 564 households, which included 597 owners and 729 dogs. Overall, statistically significant differences between the seroprevalence of human and dog anti-T. gondii antibodies were found by Immunofluorescence Antibody Testing in 248/597 (41.54%) owners and 119/729 (16.32%) dogs. Through multiple analysis, significant concomitant variables for seropositivity of household individuals (people and dogs) were determined, including public sewer service, yard cleaning frequency, and having a dirty yard. Although no statistically significant multiple logistic model was observed among owners, univariate analysis detected associations with monthly income, soil contact, and occupation. Among dogs, the absence of other dogs and the absence of a dirty yard were concomitant significantly protective associated factors. Age differences between seropositive and seronegative individuals was significant only for human beings, with the median age of negative individuals significantly higher than positive individuals. Although no spatial clusters were identified for humans or residences, a significant cluster was identified for dogs. In conclusion, characteristics of urban toxoplasmosis may include significantly higher owner seroprevalence than their owned dogs, with canine seroprevalence directly associated with having more dogs and a dirty backyard, and spatial differences in both human and dog exposures. Although not a good indicator for human foodborne diseases, dogs may be a reliable sentinel for environmental infection. Moreover, such a holistic approach may provide crucial information for more focused prevention and monitoring programs, particularly in households with multiple pets and trash-filled backyards.

Leishmaniasis is an important metazoonosis caused by protozoa of the genus Leishmania and has a heteroxenic life cycle involving invertebrate and vertebrate hosts. Transmission occurs during the blood meal of infected phlebotomine sand flies in wild species, domestic animals, and humans. The dog is a reservoir for the parasite causing visceral leishmaniasis (VL), whereas in American tegumentary leishmaniasis (ATL), dogs are erratic hosts that are accidentally infected, as in humans. Dogs are considered an important indicator of the parasite and its vectors in the environment, thus highlighting the importance of diagnosis in these animals. This study aimed to assess the seroepidemiology of Leishmania spp. in dogs in the municipality of Telêmaco Borba that were part of a castration campaign. Blood samples from 191 dogs were collected, and their owners were surveyed on various epidemiological variables. Serological analysis was performed using indirect immunofluorescence (IIF) and rapid immunochromatography (DPP®). Screening by IIF identified 13 (6.81%) positive animals, none of which were positive for the DPP® test, which is specific for VL. Statistical analysis of the questionnaire responses indicated a significant association between seropositivity and the presence of stacked or composting leaves in the backyard (p = 0.0498), forest areas (squares, woods, parks) near the residence (p = 0.0015), and poorly healing ulcerated or nodular epidermal lesions in the dog (p = 0.0138). This study revealed the presence of anti-Leishmania spp. IgG antibodies in dogs residing in Telêmaco Borba, suggesting the presence of the parasite and vector in the environment. In addition, the existence of stacked or composting leaves in the backyard, forest areas near the residence, and epidermal lesions in dogs are factors associated with Leishmania spp. infection in pet dogs.

The aim of this study was to determine the frequency of gastrointestinal parasites in dogs and cats of Londrina, Paraná. A survey of the results of fecal examinations, the technique of Faust, Willis, Hoffmann and / or direct examination, performed in routine Laboratory of Parasitology, Department of Veterinary Preventive Medicine, State University of Londrina in the period January 2000 to December 2011 and the Clinical Laboratory Veterinary January 2008 to December 2011. 2668 fecal samples were analyzed, of which 2290 (85.83%) and 378 dogs (14.17%) of cats. Of the total, 851 (37,16%) dogs and 166 (43,91%) feline samples were positive for at least one parasite. Isospora spp. was the most frequent in both species with 8.82% and 11.64% of dogs infected cats. Regarding the form of infection, 740 (86.96%) and 139 dogs (83.73%) cats had single infection, while 111 (13.04%) dogs and 27 (16.27%) cats had multiple infection gastrointestinal parasites. The ocurrence of a significant number of parasites and the close contact between animals and humans demonstrates the need for a more effective control and specific, whereas the reduction of the parasitic load of animals and thus decreases environmental exposure of humans to important zoonosis.

The aim of the present study was to verify the association between seropositivity for IgG anti-Toxoplasma gondii antibodies and social, economic and environmental variables of pregnant women attending the public health centers of Paraná, Brazil. From January 2007 to July 2010, 2226 pregnant women were interviewed and detection of anti-T. gondii specific IgG and IgM antibodies was performed by chemiluminescence test. Seropositivity for anti-T. gondii IgG was observed in 1151 (51.7%) pregnant women, 29 of which (1.3%) presented IgM reagent with IgG of high avidity. The variables associated with the presence of IgG were residency in the rural area, more than one pregnancy, less than or equal to eight years schooling, low per capita income, age group, raw or poorly cooked meat ingestion, and contact with the soil. There was neither association with raw fruit and vegetable ingestion nor with the presence of cats in the residencies. Keywords: Seroepidemiology, Prevalence, Toxoplasmosis, Pregnancy

The pathogenic fungi of the species Cryptococcus neoformans and Cryptococcus gattii could be found in the environment on pigeon guano, soil with pigeon and bird excreta and vegetable material from trees. The aim of this study was analyze the presence of the Cryptococcus pathogenic species on soil with pigeon excreta, soil with vegetable material and decayed wood of trunk hollows of trees from public places in the city of Londrina, PR. Thirty public places were selected from squares, parks and streets in the city centre where samples were collected from: soil with pigeon excreta (n=120), soil with vegetable material (n=120) and decayed wood of trunk hollows of trees (n=120), during June 2009 until March 2010. These collect comprised the seasons: winter, spring and summer, during this period 360 samples were obtained and analyzed in four different times. The species were identified by means of microscopic analysis from culture and their rising on selective medium. A total of five samples were positive to Cryptococcus spp. (1,38%) three were positive to Cryptococcus neoformans (0,84%) in two samples of soil with pigeon excreta, the other one on soil with vegetable material. The samples were originated from a park, a street and a square, placed in the city centre area of Londrina. Cryptococcus gattii was isolated from two samples (0,54%), one in soil with pigeon excreta and another in vegetable material obtained from the soil of two parks that were not protected against sunshine and rain. Cryptococcus spp. was not isolated in none of the 120 swabs samples of decayed wood trunk hollows from the 12 species of trees sampled. The results of the analyses from the environment confirmed the occurrence of the pathogenic species Cryptococcus neoformans e Cryptococcus gattii in places were the human and animal susceptible populations could be exposure to infectious fungi propagators. Os fungos patogênicos das espécies Cryptococcus neoformans e Cryptococcus gattii podem ser encontrados no meio ambiente em guano de pombos, solo com excrementos de pombos e pássaros, além de matéria vegetal de árvores. O objetivo deste estudo foi investigar a presença das espécies patogênicas de Cryptococcus em solo com excrementos de pombos, solo com matéria vegetal e em madeira em decomposição de partes ocas de árvores existentes em lugares públicos da cidade de Londrina, PR. Trinta locais foram selecionados dentre praças, parques e ruas da região central da cidade onde amostras foram coletadas de: solo com excretas de pombos (n=120), solo com matéria vegetal da base das árvores (n=120) e swab de matéria vegetal em decomposição de partes ocas de árvores, no período de junho de 2009 a março de 2010. As coletas abrangeram três estações do ano: inverno, primavera e verão, nesse período 360 amostras foram obtidas e analisadas em quatro diferentes etapas. As espécies foram identificadas por meio de análise microscópica da cultura e crescimento em meios seletivos. Um total de cinco amostras foi positivo para Cryptococcus spp. (1,38%), três amostras foram identificadas como C. neoformans (0,84%), duas amostras de solo com excrementos de pombos e uma amostra vegetal do solo. As amostras eram provenientes de um parque, uma rua e uma praça localizados na área central de Londrina. C. gattii foi isolado das outras duas amostras (0,54%), uma em solo com excrementos de pombos e outra em amostra de matéria vegetal do solo obtida de dois parques da cidade localizados em área desprotegida contra insolação e chuvas. Não foi obtido isolamento de Cryptococcus spp. das 120 amostras de swab de madeira em decomposição de partes ocas das 12 espécies de árvores amostradas. Os resultados das análises do meio ambiente confirmaram a ocorrência das espécies patogênicas Cryptococcus neoformans e Cryptococcus gattii em locais onde a população suscetível de humanos e animais podem ser expostos aos propágulos infecciosos dos fungos.

This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16), T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15%) of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.O objetivo do estudo foi investigar anticorpos anti-Toxoplasma gondii e isolar o parasita do cérebro de equídeos abatidos em matadouros-frigoríficos no Brasil. Colheram-se amostras de 398 cérebros e sangue de equídeos machos e fêmeas de idades variadas, provenientes de seis estados brasileiros. As amostras de soro foram avaliadas pelo teste de imunofluorescência indireta (IFI) para T. gondii (ponto de corte ≥ 64), e os fragmentos de cérebros foram submetidos ao bioensaio em camundongos. Por meio da IFI, 46 (11,6%) equídeos foram soropositivos. Pelo bioensaio em camundongos, 14 (3,5%) cérebros de equídeos testados foram positivos. Em doze dos bioensaios, os camundongos foram positivos somente pela IFI (ponto de corte ≥ 16) e T. gondii foi isolado nos outros dois bioensaios. A PCR-RFLP com base em 18S rDNA para diferenciar entre T. gondii, Neospora caninum, e Sarcocystis neurona foram feitas em todos os 14 cérebros e dois foram positivos apenas para T. gondii. De dois isolados positivos para T. gondii e do cérebro positivo à PCR em que realizou-se a PCR-RFLP, com base em 13 marcadores e SAG2, a genotipagem mostrou ser o T. gondii tipo I para todas as amostras. A IFI de soros de equídeos e do bioensaio em camundongos identificaram positividade em 60 (15%) amostras testadas. Os resultados mostram que alguns cavalos enviados para abate foram expostos ao T. gondii.

The role of rodents in the epidemiology of toxoplasmosis was investigated in Londrina, Paraná State, Brazil. One hundred and eighty-one Rattus rattus and one Mus musculus were caught in 37 places. Blood and tissues were collected and submitted to the indirect fluorescence antibody test (IFAT) and the bioassay. Serum samples from 61 contacting dogs were also collected. Sixteen rats (8.8%) were positive for Toxoplasma gondii, but just two of them were positive by serology and bioassay test. Antibodies were found in nine (4.9%) rats. Tissues of nine rats bioassayed were positive and four isolates were obtained. Restriction fragment length polymorphism (RFLP) analysis was performed using 12 markers (SAG1, SAG2, SAG2-alt, C22-8, C29-2, L358, PK1, BTUB, GRA6, SAG3, Apico, CS3). Genotyping revealed that the four strains isolated from this study have been isolated before in cats and chickens from Brazil. None of the isolates was identified like clonal archetypal T-types I, II, and III. The rats presented lower serologic Toxoplasma gondii prevalence (8.8%) compared to contacting dogs (70.5%).

OBJETIVO: Avaliar a suscetibilidade das gestantes à toxoplasmose em serviço público de saúde de dois municípios da região oeste do Paraná. MÉTODOS: Foram avaliadas 422 gestantes por meio da pesquisa sorológica de anticorpos IgG e IgM anti-Toxoplasma gondii (ELISA e MEIA). As soronegativas repetiram a sorologia no segundo e terceiro trimestre de gestação. Em um dos municípios, também foi realizada a triagem neonatal em 27 recém-nascidos para detecção de IgM anti-Toxoplasma gondii pelo teste de fluorometria. Todas as gestantes responderam a um questionário epidemiológico, para análise dos fatores associados ao risco da infecção pelo Toxoplasma gondii. Para análise estatística, foram consideradas a variável dependente da presença de IgG anti-Toxoplasma gondii e as variáveis independentes contidas no questionário epidemiológico. RESULTADOS: A prevalência de anticorpos IgG anti-Toxoplasma gondii nas gestantes foi de 59,8 e 60,6%. Em um dos municípios, as variáveis associadas à presença de anticorpos IgG foram baixo nível de escolaridade e mais de uma gestação. Não houve associação com os outros fatores investigados, como a ingestão de carnes cruas ou mal cozidas, vegetais crus, salames coloniais, manipulação de terra ou areia, horta em casa e gatos em casa. No outro município, não foi observada associação estatística com nenhuma das variáveis estudadas. Não foi confirmado nenhum caso de infecção aguda nem de soroconversão em ambos os municípios. Nenhuns dos recém-nascidos avaliados apresentou positividade para toxoplasmose. CONCLUSÃO: A toxoplasmose é comum nas gestantes atendidas pelo serviço público de saúde da região estudada e há 40% delas suscetíveis à infecção. Esse dado reforça a necessidade de manter o programa implantado nesses municípios.PURPOSE: To evaluate the susceptibility to toxoplasmosis in pregnant women in the public health service from two cities in the western region of Paraná, Brazil. METHODS: Four thousand twenty-two pregnant women were evaluated for anti-Toxoplasma gondii IgG and IgM by ELISA and MEIA. Seronegative pregnant women repeated the serology in the second and third trimester of pregnancy. Neonatal screening of 27 newborns was also performed in one of the cities to detect IgM anti- Toxoplasma gondii by fluorometry. All pregnant women answered an epidemiological questionnaire to analyze the factors associated with the risk of infection by Toxoplasma gondii. For statistical analysis, the presence of IgG anti-Toxoplasma gondii was considered as the dependent variable and the variables contained in the epidemiological questionnaire as the independent ones. RESULTS: The prevalence of anti-Toxoplasma gondii IgG in pregnant women was 59.8 and 60.6%. In one of the cities, the variables associated with the presence of IgG antibodies were low educational level and more than one pregnancy. There was no association with other factors studied such as consumption of raw or undercooked meat, consumption of raw vegetables, consumption of colonial salami, handling soil or sand, the presence of a home vegetable garden and cats in the household. In the other city there was no statistical association with the variables studied. No case of acute infection and no seroconversion were confirmed in either city. None of the infants evaluated were positive for toxoplasmosis. CONCLUSION: Toxoplasmosis is common in pregnant women attended by the public health service in the region studied and 40% of them are susceptible to the infection. These data reinforce the need to keep the screening program in these cities.

The study of the prevalence of anti-leptospiral antibodies in pet dogs in urban areas and the variables associated with the risk of infection can aid in the identification of the serovars present in the region and indicated the reservoirs involved in this important zoonosis. The aim of the present study was to determine the prevalence of anti-leptospiral antibodies in 653 dogs from 369 households distributed throughout the urban area of the municipality of Jataizinho, Paraná (PR), and to investigate the variables associated with the risk of leptospiral infection in these animals. Serum samples were subjected to microscopic agglutination tests (MATs) against 23 serovars of pathogenic leptospires. Among the 132 (20.21%) reactive samples, 15 (11.36%) were reactive for more than one serovar, and the most likely serovar could not be determined. Among the 117 (88.73%) samples that reacted to a single serovar, the serovar Canicola was considered to be the most likely in 89 cases (76.07%). Variables associated with the risk of disease were investigated using an epidemiological questionnaire administered to each owner, and the data obtained were analyzed using the statistical software EpiInfo. The significant variables (p ? 0.05) associated with seropositivity for leptospirosis in pet dogs in the urban area of the municipality of Jataizinho, PR, were the presence of rats in the household, adult or elderly dogs, free access of the dog to the street and contact with stray dogs. The results indicate a high prevalence of pet dogs in the urban area of Jataizinho, PR, with anti-leptospiral antibodies and suggest the need to educate the population of the municipality about the vaccination of dogs, as well as, the epidemiological role of the dog in urban leptospirosis. O estudo da prevalência de cães domiciliados em áreas urbanas com anticorpos antileptospiras e de variáveis associadas ao risco de infecção, pode auxiliar na identificação dos sorovares presentes na região e sugerir os reservatórios envolvidos nesta importante zoonose. O objetivo deste trabalho foi determinar a prevalência de anticorpos antileptospiras em 653 cães de 369 domicílios distribuídos na área urbana do município de Jataizinho, PR, e estudar variáveis associadas ao risco de infecção por leptospiras nesses animais. As amostras de soro foram submetida a prova de soroaglutinação microscópica (SAM) frente a 23 sorovares de leptospiras patogênicas. Das 132 (20,21%) amostras de cães reagentes, 15 (11,36%) reagiram para mais de um sorovar, não sendo possível determinar o sorovar mais provável, e das 117 (88,73%) que reagiram para um sorovar somente, o sorovar Canicola foi considerado o mais provável em 89 (76,07%). Variáveis associadas ao risco da doença foram investigadas através de um questionário epidemiológico aplicado a cada proprietário e os dados obtidos foram analisados pelo programa estatístico EpiInfo. As variáveis significativas (p ? 0,05) associadas à soropositividade para leptospirose nos cães domiciliados em área urbana do município de Jataizinho, PR, foram: visualizar ratos no ambiente familiar, animal adulto ou idoso, livre acesso do cão à rua e contato com cães errantes. Os resultados indicam uma alta prevalência de cães domiciliados em área urbana de Jataizinho, PR, com anticorpos antileptospiras e sugerem a necessidade de conscientizar a população do município sobre a vacinação dos cães, assim como, o papel epidemiológico do cão na leptospirose urbana.

The objective of this study was to investigate the presence of Trichinella spp. in the musculature and anti-Toxoplasma gondii and anti-Leptospira spp. antibodies in the serum of equidae slaughtered at two abattoir-cold storage facilities licensed by the brazilian Federal Inspection Service. Masseter muscle and blood samples were collected from 398 animals of both sexes and different ages originating from six different Brazilian states. The serum samples were subjected to the Indirect Fluorescence Antibody Test for T. gondii (IFAT ³ 64) obtaining 46 (11.6%) samples reagent. For Leptospira spp. Antibody quantification, the serum samples were tested by microscopic seroagglutination test (MAT ³ 100), and 123 (30.9%) reagent results were obtained. It was possible to identify the most likely infecting serovar in 95 samples (77.2%): Hardjo (26.3%) and Autumnalis (12.6%) were the ones most prevalent. None of the 398 masseter muscle samples analyzed revealed any presence of Trichinella spp. larvae. The slaughtered equidae that were investigated were probably exposed to the etiologic agents of toxoplasmosis and leptospirosis, but they did not present Trichinella infection.O objetivo deste estudo foi investigar a presença de Trichinella spp. na musculatura e anticorpos anti-Toxoplasma gondii e anti-Leptospira spp. em soro de equídeos abatidos em dois matadourosfrigoríficos com serviço de Inspeção Federal. Amostras de músculo masséter e sangue foram coletados de 398 animais de ambos os sexos e várias idades, provenientes de seis diferentes estados brasileiros. As amostras de soro foram submetidas à Reação de Imunofluorescência Indireta para T. gondii (IFI ³ 64) obtendo 46 (11,6%) amostras reagentes. Para Leptospira spp. as amostras de soro foram testadas pelo teste de soroaglutinação microscópica (MAT ³ 100) e 123 (30,9%) amostras foram reagentes. Foi possível identificar o sorovar mais provável em 95 amostras (77,2%) sendo Hardjo (26,3%) e Autumnalis (12,6%) os mais encontrados. Nenhuma das 398 amostras de músculo masséter analisados revelou qualquer presença de larvas de Trichinella spp. Os equídeos destinados ao abate que foram investigados provavelmente foram expostos aos agentes da toxoplasmose e leptospirose, porém não apresentaram infecção por Trichinella.

The purpose of this study was to determine the seroprevalence of antibodies against Toxoplasma gondii and Leishmania spp. in humans and dogs living in two rural settlements in northern Paraná State. An epidemiological questionnaire was applied to obtain socio-demographic information and possible associations with the infections, and the data were analyzed using EpiInfo®. Blood samples were collected from 216 humans and 169 dogs, and tested by indirect immunofluorescence assay. The prevalence of toxoplasmosis in humans was 79.1% (171/216) and in dogs was 82.2% (139/169). Among the variables analyzed for toxoplasmosis in humans the presence of young cats in the household (p = 0.031) and higher frequency with individuals > 18 years showed a significant association. A higher frequency of seropositive was observed in dogs aged > 1 year. The prevalence of leishmaniasis in humans was 7.4% (16/216) and in dogs was 8.2% (14/169). The variable presence of forest less than 200 meters from the residence had a significant association among both humans and dogs. Also for dogs, there was association with the presence of organic matter (leaves) around the household. In conclusion, it can be stated that there is a high spread of T. gondii in both species and the occurrence of anti-Leishmania spp. antibodies in humans and dogs indicates that there is transmission of Leishmania spp. in these localities.O objetivo deste estudo foi determinar a prevalência de anticorpos contra Toxoplasma gondii e Leishmania spp. em humanos e cães que vivem em dois assentamentos rurais no norte do Paraná. Um questionário epidemiológico foi aplicado para obter informações sociodemográficas e possíveis associações com as infecções, e os dados foram analisados pelo EpiInfo®. Amostras de sangue foram coletadas de 216 pessoas e 169 cães, e testados por imunofluorescência indireta. A prevalência de toxoplasmose em humanos foi de 79,1% (171/216) e em cães foi de 82,2% (139/169). Entre as variáveis analisadas para toxoplasmose em humanos a presença de gatos jovens no domicílio (p = 0,031) e maior freqüência de indivíduos >18 anos mostraram uma associação significativa. Uma maior freqüência de soropositivos foi observada em cães com idade >1 ano. A prevalência de leishmaniose em seres humanos foi de 7,4% (16/216) e em cães foi de 8,2% (14/169). A variável presença de floresta a menos de 200 metros da residência apresentou uma associação significativa entre os seres humanos e cães. Também para os cães, houve associação com a presença de matéria orgânica (folhas) em torno da casa. Em conclusão, pode afirmar-se que existe uma grande propagação de T. gondii em ambas as espécies e a ocorrência de anticorpos anti-Leishmania spp. em humanos e cães, indica que há transmissão de Leishmania spp. nessas localidades.

The aim of this study was to determine the seroprevalence of Cutaneous Leishmaniasis (CL) in dogs of Bela Vista do Paraiso, Parana state, compare the IFA and ELISA techniques and identify the vectors possibly involved in the cycle of the parasite. Were collected blood samples from 489 dogs that were subjected to detection of anti-Leishmania sp. by IFA and by ELISA. Were considered positive samples ³40 titers in IFA and for ELISA ³ 0.174 optical density. Among the samples analyzed, 222 (45.4%) were positive by IFA and 189 (38.7%) by ELISA. Comparing the tests were found 176 positive samples (36.0%) and 254 negative (51.9%) for both techniques. The sensitivity of ELISA was 79.3% and specificity was 95.1%. The global coefficient of the test was 87.0% with kappa coefficient of 0.75. Analysis of variables for dogs with positive serology by IFA showed significant differences regarding the absence of forests and lack of contact with other animal species. Positive sera by ELISA in dogs variables that showed significant differences were the type of riparian vegetation existing in the environment, the lack of garbage, open sewers and released directly into rivers or streams and garbage thrown on wasteland, burned or buried. The result obtained with the capture of sandflies was the predominance of Lutzomyia whitmani with 79.9% of the species collected. The result obtained with the capture of sandflies was the predominance of Lutzomyia whitmani with 79.9% of the species collected. The results showed that the LTA is widespread in the canine population of Bela Vista do Paraiso, and both the IFI and ELISA can be used for diagnosis. So the dog appears as a link between wild and peridomestic cycle of CL may become an amplifier of disease in this ecosystem.O trabalho teve como objetivo determinar a soroprevalência da Leishmaniose Tegumentar Americana (LTA) canina no município de Bela Vista do Paraíso, Paraná, comparar as técnicas de imunofluorescência indireta (IFI) e ensaio imunoenzimático (ELISA) e identificar as espécies de flebotomíneos presentes, possivelmente envolvidas no ciclo do parasito. Amostras de sangue de 489 cães foram submetidas à pesquisa de anticorpos anti-Leishmania sp. pela IFI e ELISA. Foram consideradas positivas as amostras que apresentaram título ³ 40 na IFI e densidade ótica ³ 0.174 no ELISA. Entre as amostras analisadas, 222 (45,4%) foram reagentes pela IFI e 189 (38,7%) pelo ELISA. Comparando-se os testes foram encontradas 176 amostras positivas (36,0%) e 254 negativas (51,9%) para ambas as técnicas. A sensibilidade do ELISA foi de 79,3% e a especificidade foi de 95,1%. O coeficiente global do teste foi de 87,0% com coeficiente Kappa de 0,75. A análise das variáveis para cães com sorologia positiva pela IFI demonstrou diferença significativa em relação à ausência de matas e ausência de convívio com outras espécies animais. Em cães sororeagentes pelo ELISA as variáveis que apresentaram diferença significativa foram o tipo de mata ciliar existente no ambiente, a ausência de lixo, o esgoto a céu aberto e lançado diretamente em rios ou córregos e do lixo lançado em terreno baldio, queimado ou enterrado. O resultado obtido com a captura dos flebotomíneos foi a predominância do Lutzomyia whitmani com 79,9% das espécies coletadas. Os resultados demonstraram que a LTA está amplamente disseminada na população canina do município de Bela Vista do Paraíso, e que tanto a IFI como o ELISA podem ser utilizados para o diagnóstico. Deste modo o cão apresenta-se como um elo entre o ciclo silvestre e o peridomiciliar da LTA, podendo tornar-se um sinalizador do agente no ecossistema da doença nesse ecossistema.

The aim of the present work was to determine the prevalence of IgG and IgM anti-Toxoplasma gondii antibodies and the factors associated to the infection in pregnant women attended in Basic Health Units in Rolândia, Paraná, Brazil. The sample was divided in two groups: group I (320 pregnant women who were analyzed from July 2007 to February 2008) and group II (287 pregnant women who were analyzed from March to October 2008). In group I, it was found 53.1% of pregnant women with IgG reactive and IgM non-reactive, 1.9% with IgG and IgM reactive, 0.3% with IgG non-reactive and IgM reactive and 44.7% with IgG and IgM non-reactive. In group II, it was found 55.1% with IgG reactive and IgM non-reactive and 44.9% with IgG and IgM non-reactive. The variables associated to the presence of IgG antibodies were: residence in rural areas, pregnant women between 35-40 years old, low educational level, low family income, more than one pregnancy, drinking water which does not originate from the public water supply system and the habit of handling soil or sand. Guidance on primary prevention measures and the quarterly serological monitoring of the pregnant women in the risk group are important measures to prevent congenital toxoplasmosis.

The toxoplasmosis is a zoonosis caused by Toxoplasma gondii and affects a lot of species of carnivores and omnivores, including the human. The rodents are important in the transmition cycle because they act as an infection font to felines, the definitive host of this protozoan. The objective of this work was to evaluate the Modified Agglutination Test (MAT) for the serologic diagnosis of toxoplasmosis in rats, comparing with the Indirect Fluorescent Antibody Test (IFAT), which has been considered the golden standard in animal toxoplasmosis diagnosis. Kappa test was used for comparing the serologic tests (IFAT and MAT) and for determination of cutoff appropriate to MAT in this animal species. 182 rats were caught on local recycling of solid waste and solid residue storage in Londrina city, Paraná. Out of the 182 rats, nine (4.94%) were positive to IFAT at a dilution of 1:16, and 17 (9.34%) and five (2.75%) were reactive to MAT in dilutions 1:25 and 1:50, respectively. The comparison of results between the techniques presented kappa coefficients of 0.26 and 0.55, respectively at 1:25 and 1:50 dilutions of MAT. It can be concluded that the dilution 1:50 is the most suitable to be used as cutoff for detecting T. gondii antibodies in rats using MAT, because agreed with IFAT.A toxoplasmose é uma zoonose causada pelo Toxoplasma gondii que acomete várias espécies carnívoras e onívoras, incluindo o ser humano. Os roedores são importantes na cadeia epidemiológica da doença por servirem de fonte de infecção aos felídeos, os hospedeiros definitivos deste protozoário. O objetivo deste trabalho foi avaliar o Teste de Aglutinação Modificada (MAT) na detecção de anticorpos contra T. gondii em ratos, comparando-o à Reação de Imunofluorescência Indireta (RIFI), considerada padrão ouro para o diagnóstico da toxoplasmose animal. Empregou-se o teste kappa para a comparação dos testes sorológicos (RIFI e MAT) e para a determinação do ponto de corte mais apropriado para a utilização do MAT, nesta espécie animal. Foram capturados 182 ratos em locais de reciclagem e armazenagem de resíduos sólidos na cidade de Londrina, PR. Destes, nove (4,94%) foram positivos na RIFI na diluição 1:16; e 17 (9,34%) e cinco (2,75%) foram sororreagentes ao MAT nas diluições 1:25 e 1:50, respectivamente. A comparação dos resultados entre as técnicas apresentou coeficientes kappa de 0,26 e 0,55, respectivamente às diluições 1:25 e 1:50 do MAT. Conclui-se que a diluição 1:50 é a mais indicada para ser o ponto de corte na detecção de anticorpos contra T. gondii em ratos pelo MAT, por apresentar melhor concordância à RIFI.

Paracoccidioidomycosis, caused by the dimorphic fungus Paracoccidioides brasiliensis, is the most prevalent human systemic mycosis in Latin America. Paracoccidioidomycosis affects mainly male rural workers, causing granulomatous lesions in organs such as lungs, liver and spleen. The participation of other animal species in the fungus eco-epidemiology in not well understood. The aim of this study was to evaluate the infection by P. brasiliensis in horses from the Northern Region of Paraná State. The serum samples from 100 horses were assayed by ELISA and Immunodiffusion test, using gp43 and exoantigen as antigens, respectively. A seropositivity of 30% was observed by ELISA test, although all samples were negative by immunodiffusion test. These results suggest that horses can be indicator of fungus presence in the ambient.A paracoccidioidomicose, causada pelo fungo dimórfico Paracoccidioides brasiliensis, é a micose sistêmica de maior prevalência em humanos na América Latina. A paracoccidioidomicose afeta principalmente trabalhadores rurais do sexo masculino, provocando lesões granulomatosas em órgãos como pulmões, fígado e baço. A participação de outras espécies de animais na eco-epidemiologia do fungo não é bem compreendida. Este trabalho teve como objetivo avaliar a infecção por P. brasiliensis em cavalos da Região Norte do Estado do Paraná. Foram analisadas 100 amostras de soros de cavalos por ELISA e por Imunodifusão em gel de agar, utilizando como antígenos a gp43 e o exoantígeno de P. brasiliensis, respectivamente. A soropositividade observada foi de 30% por ELISA enquanto que por imunodifusão todos os soros foram negativos. Os resultados obtidos sugerem que cavalos podem ser indicadores da presença do fungo no meio ambiente.