Patterns of Transmission and Sources of Infection in Outbreaks of Human Toxoplasmosis.

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.

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.

High titers of anti-Leishmania spp. antibodies in apparently healthy dogs in the North Pioneer Mesoregion of the state of Paraná, Brazil.

Leishmaniasis is an anthropozoonosis with vector transmission, and knowledge regarding the occurrence of this parasitosis in sentinels can contribute to infection and disease control measures in humans. The objectives of this study were to evaluate the occurrence of Leishmania exposure and infection in dogs from urban and rural areas in the North Pioneer Mesoregion of the state of Paraná, to evaluate possible risk factors, and to analyze the statistical agreement between the serological techniques that were used. Using a convenience sampling, serum and whole blood samples were collected to perform serological and molecular assays, respectively. The enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT) identified 29/204 (14.2%) and 20/204 (9.8%) seropositive dogs, respectively. Five dogs (2.4%) were seropositive for both serological tests, and four dogs presented high titers in the IFAT. None of the samples tested positive for Leishmania spp. DNA according to polymerase chain reaction analysis. No factors were significantly associated with infection. Leishmania parasites circulate in urban and rural dogs in the North Pioneer Mesoregion of the state of Paraná. Despite the absence of clinical cases, seropositive animals with high antibody titers should serve as a warning to the local population that should be properly informed regarding the prevention.

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 implementation of the primary, secondary and tertiary prevention measures of the Surveillance Program of Gestational and Congenital Toxoplasmosis in the city of Londrina-PR.

Toxoplasmosis acquired during pregnancy is one that can lead to death or malformations of the foetus, and it is a complex disease to diagnose. The objective of the study was to evaluate the Surveillance Program of Gestational and Congenital Toxoplasmosis. To assess primary prevention, 424 pregnant women were interviewed regarding their knowledge of prevention measures in 2019. Secondary prevention measures were assessed, and the results of anti-Toxoplasma gondii serological tests were collected from pregnant women, from 2015 to 2018. In tertiary prevention measures, babies of mothers with a recent suspicion of T. gondii infection were screened to verify forwarding to the reference service. As a result, 45.5% (192/424) reported that they had received guidance from health professionals; 35.4% (68/192) changed their risk habits. The variables of schooling and age, having received prior guidance from health professionals and feline possession, proved to be significant when associated with the notions of preventive measures. 90.2% (17,423/19,319) of pregnant women had undergone serological tests to detect anti-T. gondii antibodies, but there was an excess in requests for tests and medication and only 40.6% (26/64) of the children were referred to the reference hospital. The Program presents positive results about the performance of serological screening in prenatal care; however, the dissemination of knowledge as for the prevention of toxoplasmosis and the request for tests need to be improved.

Urban capuchin monkeys Sapajus nigritus (Goldfuss, 1809) (Primates, Cebidae) as environmental bioindicators of leishmaniasis.

The aim of this study was to investigate the presence of Leishmania sp. DNA and anti-Leishmania spp. antibodies in free-ranging Sapajus nigritus from an urban forest located in a city in the North Central region of the state of Paraná. For the indirect diagnosis, the direct agglutination test was used with promastigote forms of Leishmania (V.) braziliensis, where it was possible to detect the agglutination reaction in 53.33% of the S. nigritus blood samples. For direct diagnosis, the samples were submitted to real-time quantitative polymerase chain reaction, which confirmed the presence of Leishmania spp. DNA in 26.66% of the tested samples. It reinforces the importance of considering the concept of One Health in the face of diseases with high prevalence, such as leishmaniasis and the need for health education measures. This result shows that the animals in the present study have a role as environmental bioindicators for leishmaniasis.

Absence of Trichinella spp. larvae in carcasses of road-killed wild animals in Paraná state, Brazil.

Trichinella spp. are zoonotic parasites that are widely distributed in warm-blooded carnivores and omnivores, including humans. Until the present moment, Brazil has been considered by World Animal Health Organization free from the domestic cycle of trichinellosis, whereas the parasite's sylvatic cycle has the status of infection in limited zones. However, neighboring countries such as Argentina have reports of parasite larvae in the wild fauna. The present study aimed to determine the occurrence of Trichinella spp. in road-killed wild animals in Paraná, Brazil. Biological samples from 71 wild animals-29 Didelphis albiventris, 11 Nasua nasua, ten Cerdocyon thous, seven Dasypus novemcinctus, six Leopardus guttulus, six Sphiggurus spinosus and two Puma concolor-collected from November 2016 to November 2021 were subjected to artificial digestion, following the methodology described in the REGULATION (EC) No. 2075/2005. No Trichinella spp. larvae were detected in the carcasses of the road-killed wild animals. However, considering the wide spectrum of possible reservoirs that could act as a link between the sylvatic and domestic cycles and considering the current Brazilian status of sylvatic trichinellosis in limited zones, frequent monitoring of wild fauna remains necessary.

Techniques for inactivating Toxoplasma gondii oocysts: a systematic review.

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 <0.05). Sporulated oocysts and the T. gondii VEG strain were more resistant (p <0.05) to treatments. Although it is effective against viruses and bacteria, the use of disinfectants in water has little or no effect on T. gondii oocysts. The use of radiation and pressure were effective in inactivating oocysts, as these treatments do not include changes in temperature, they can be used in foods for raw consumption, such as vegetables, as it will not cause substantially changes in their physical and chemical characteristics. Therefore, these methods can be viable alternatives for the control of T. gondii.

Evaluation of an active and early surveillance methodology for visceral leishmaniasis by molecular detection in road-killed wild fauna.

The present study aimed to evaluate a methodology for active surveillance of visceral leishmaniasis by detecting Leishmania DNA in organs of wild road-killed animals from November 2016 to October 2018 in the North of Paraná, Brazil. The collection points of road-killed wild animals were georeferenced. The animals were autopsied and samples of bone marrow, lymph node, liver, spleen, and ear skin were collected. Genomic DNA was extracted and subjected to PCR for amplification of Leishmania spp. 18S, kinetoplastic DNA (kDNA), HSP70, and ITS1 genes, and DNA sequencing was performed. The primers used for the amplification of kDNA, ITS1, and HSP70 genes presented non-specific results. Of the 66 mammals collected from 24 different municipalities, one Southern Tamandua (Tamandua tetradactyla) presented DNA of Leishmania spp. in lymph nodes by 18S PCR. DNA sequencing confirmed the results of the subgenus, Viannia, identification. We suggest using the methodology showed in the present study in the active and early surveillance of visceral leishmaniasis in a non-endemic area.

Urban Horses As Environmental Bioindicators for Leishmaniasis.

The presence of DNA and anti-Leishmania spp. antibodies in the serum of 112 healthy horses was investigated by evaluating the physical examination, from a rural society located in the north central region of Paraná. The antigens of Leishmania amazonensis, Leishmania braziliensis, and Leishmania infantum were used to perform the indirect enzyme-linked immunosorbent assay, where it was possible to detect the reaction in 27.67% of the samples. These were also subjected to the real-time quantitative PCR, which confirmed the presence of Leishmania spp. DNA in 67.34% of the tested samples. The results show that the tested animals were previously exposed to the protozoan. Thus, these animals can be considered environmental bioindicators of the presence of Leishmania spp. at the study site. The material used in this study (serum), although not ideal, proved to be effective and less invasive. Taking into account the importance of the disease, the absence of more in-depth information on the species, the high zootechnical value of these animals, and their strictly close contact with the urban area and the human species, it is essential that further studies are carried out to elucidate the epidemiological profile of them in the face of the disease, as well as the possibility of them acting not only as hosts but also as reservoirs.

Parasites in road-killed wild felines from North of Paraná state, Brazil.

This study aimed to identify the intestinal parasites of road-killed wild felines in the North Central and North, Paraná state, southern Brazil. The animals were monitored by sampling previously established transects. The places where the felines were run over were mapped, the animals were identified, and the gastrointestinal tract was evaluated. The feces were submitted to coproparasitological techniques of spontaneous sedimentation, floating in hypersaturated NaCl solution and centrifugal floating in zinc sulfate. All the parasitic structures detected were photomicrographed. In the coproparasitological analyses were identified oocysts of Cystoisospora spp., eggs of Ancylostomatidae, and Capillaria spp.; eggs of Aelurostrongylus spp., Toxocara spp., Physaloptera spp., Taenia spp., and Spirometra spp.; Aelurostrongylus abstrusus larvae; and eggs and adults of Ancylostoma cati and Taenia spp. One of the cats was parasitized by a flea of Ctenocephalides felis felis. Based on these results, the animals analyzed in this study supplied important samples for the evaluation of parasitic diversity of North of Paraná and suggested that this region may have conditions that allow the maintenance of these parasites life cycles in the environment and among wildlife.

Anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state, South Brazil: prevalence and associated factors.

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.

Molecular diagnosis of the curly lettuce parasitic contamination from hydroponic cultivation from supermarkets.

The consumption of vegetables has increased in recent years due to the search for a healthier diet that is rich in fiber and has fewer calories. To assess the parasitic contamination of lettuce sold in markets, a survey of parasites was carried out from a supermarket chain in the city of Londrina, Paraná. A total of thirty samples of lettuce were purchased in the ten markets visited, three in each, of which ten were conventionally cultivated, ten were hydroponically cultivated, and ten were organically cultivated. All samples were analyzed using the sedimentation methods of Hoffman, Pons and Janer and the fluctuation method of Faust and colleagues and Willis with adaptations. In addition, the samples were subjected to DNA extraction by a commercial kit and polymerase chain reaction to detect Toxoplasma gondii, Cryptosporidium spp. and Giardia spp., which are protozoa that cause food and waterborne parasitic outbreaks. All samples were negative for sedimentation and flotation techniques. One of the hydroponically cultivated samples was positive for T. gondii. The results demonstrate the risk of curly lettuce contamination from hydroponic cultivation and the need for proper cleaning of these foods before consumption.

Characterization of prenatal healthcare for implementation of congenital toxoplasmosis surveillance program: cross-sectional study.

BACKGROUND: Prenatal toxoplasmosis remains a neglected disease worldwide and few government programs focusing on its prevention are available. Success in these programs has been extensively reported in the literature, yet the strategies used for their implementation, as a model for such actions in different communities, have not been described. OBJECTIVE: To describe the aspects of prenatal care strategies in 13 municipalities within the regional healthcare unit of Araçatuba, in the northwestern region of the state of São Paulo in 2017, focusing on congenital toxoplasmosis. DESIGN AND SETTING: Descriptive study on prenatal healthcare within the Brazilian National Health System, in 13 participating municipalities. METHODS: Data on serological tests, referral clinics, notifications, healthcare strategies, health education and drugs for infected children were requested through a questionnaire that was sent via e-mail to people responsible for healthcare services in these municipalities. RESULTS: Major differences relating to diagnoses, reference outpatient clinics, notifications, health education and healthcare and drugs for infected children were reported among the prenatal strategies of these 13 municipal healthcare services. CONCLUSION: The lack of standardized prenatal strategy in the study area may compromise the prevention of infection. However, our identification of each aspect of prenatal care corroborates the need to implement a healthcare surveillance program relating to congenital toxoplasmosis.

Artisan fresh cheese from raw cow's milk as a possible route of transmission in a toxoplasmosis outbreak, in Brazil.

BACKGROUND: The objective of this study was to report an outbreak of human toxoplasmosis occurring in the municipality of Montes Claros de Goiás, Goiás, Brazil, from December 2015 to August 2016. Seven acute cases in June 2016 triggered the subsequent search. METHODS: A total of 251 individuals were selected through an active search, of which 114 (45.4%) agreed to participate in the research and blood collection. For serological diagnosis were used enzyme-linked immunosorbent assay (ELISA) for IgG and IgM and avidity tests. RESULTS: Of the 114 serum samples evaluated, 12.3% (14/114) showed antibodies against Toxoplasma gondii, with a profile indicative of acute infection. Samples of artisan fresh cheese, public water, vegetables and irrigation water were collected. Toxoplasma gondii DNA fragments were amplified using the polymerase chain reaction from two samples of artisan fresh cheese and a sample of irrigation water from the vegetable garden. A control case study was carried out, and the variable cow's artisan fresh cheese consumption was statistically significant (p = .01). CONCLUSIONS: The results showed that cheese analysed and/or irrigated water of vegetable represented an important route of transmission for the disease. This is the first reported outbreak possibly caused by cow's artisan fresh cheese. It is difficult to prove that these routes were the cause of the outbreak; however, the findings allow us to infer that the individuals involved in the outbreak were in contact with these risk factors.

Spatial and Simultaneous Seroprevalence of Anti-Leptospira Antibodies in Owners and Their Domiciled Dogs in a Major City of Southern Brazil.

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.

Toxoplasma gondii and Neospora caninum infections and factors associated in goats in the Parana state, Southern Brazil.

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.

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.

Seroepidemiology of ovine toxoplasmosis and neosporosis in breeding rams from Rio Grande do Sul, Brazil.

This study determined the prevalence of ovine toxoplasmosis and neosporosis and the risk factors associated with the development of these diseases in breeding rams from the State of Rio Grande do Sul (RS), Southern Brazil. Serum samples (n = 1,800) from breeding rams maintained on 705 sheep farms from seven mesoregions were evaluated serologically to detect anti-IgG Toxoplasma gondii by indirect ELISA and anti-IgG Neospora caninum by the indirect immunofluorescence assay (IFA). The prevalence of T. gondii was 33.05% (595/1,800); seropositivity to N. caninum was 18.44% (332/1,800). Additionally, there was simultaneous seropositivity (8.94%;161/1,800) to N. caninum and T. gondii. The variables size of the property (<500 ha) (Prevalence Ratio, PR = 1.36); breeding system (semi-intensive/intensive) (PR = 1.23); and natural mounting without control (PR = 1.50) were considered as risk factors for the occurrence of T. gondii. Size of the property (<500 ha) (PR = 1.58) and natural mounting without control (PR = 2.32) were risk factors associated with the prevalence of N. caninum in rams. Additionally, separation of ewes prior to parturition was considered as a protective factor for the occurrence of T. gondii (PR = 0.82) and N. caninum (PR = 0.74). These results demonstrated that these two parasitic disease agents are endemic in rams throughout all regions of RS.