Occurrence of zoonotic enteric parasites in fecal samples from dogs in shelters, parks, squares and public roads, and the dog guardians' perception of zoonoses as for the risk to public health in the city of Guarapuava, Paraná, Brazil.

The aim of this study was to determine the occurrence of zoonotic enteroparasites in the feces of dogs from public shelters, squares, parks, and public roads in the city of Guarapuava, Paraná, Brazil, and to evaluate the perception of dog guardians regarding zoonoses and their risk to public health. Fecal samples were collected, coproparasitological examinations were performed to detect zoonotic enteroparasites, and polymerase chain reaction (PCR) was used to identify Giardia spp. and Cryptosporidium spp. Questionnaires were given to guardians who walked their dogs in parks, squares, and public roads, as to assess their perception of zoonoses. A total of 333 samples were collected, of these 75, 123, and 135 of them were from public shelters, squares and parks, and public roads, respectively. One or more parasites were identified in 166 (50 %) samples, of which 58/75 (77 %) were from public shelters, 50/123 (41 %) from squares and parks, and 58/135 (43 %) from public roads. The parasites detected included Ancylostoma spp., Giardia spp., Trichuris spp., Toxocara spp., and Cystoisospora spp., with Ancylostoma spp. having the highest occurrence. PCR was performed on 161 samples for convenience due to financial limitations, because only a portion of the study was funded by the municipal government, of which 15.6 % were positive for Giardia spp., and all were negative for Cryptosporidium spp. In total, 246 guardians were interviewed, of which 36 % said they did not collect their animals' feces during walks, 20 % did not use anti-helminthics on their dogs, and 23 % did not know which diseases could be transmitted by feces. Therefore, we conclude that there is a high infection rate of parasites with zoonotic potential in public places, showing the need to raise awareness among guardians about the diseases transmitted by dog feces, correct vermifugation and the importance of collecting feces in public places.

Health evaluation of street dogs in Guarapuava, Brazil.

Free-roaming dogs are removed from the street by the Municipal Animal Control division/shelter and NGOs to safeguard public health as well as to prevent any suffering dogs may face and to find them homes. However, adoption rates are low and there is a lack of public programs aimed at humane control causing increased numbers of street dogs and overcrowding in shelters, compromising health and welfare. We aimed to evaluate the health of dogs from municipal shelters and independent shelters, and community dogs in the city of Guarapuava through clinical examination and complete blood cell count tests. The main changes found in the clinical examination of 297 dogs were hyperthermia in 4.0%, tachypnea in 20.5%, ocular and oral mucosa hypocorada in 6.4%, and ectoparasite infestation in 42.7%. Changes found in the erythrogram were anemia in 15.1% and polycythemia in 3.4%. In the platelet count, thrombocytopenia was found in 10.1% and thrombocytosis in 10.4%. On the leukogram, leukocytosis was observed in 14.8% and leukopenia in 3%. The results reveal the compromised health of some animals, requiring better diagnostic investigation, since certain clinical and complete blood cell count alterations may be caused by zoonotic agents that can endanger human and animal health.

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.

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.

First detection of Feline morbillivirus infection in white-eared opossums (Didelphis albiventris, Lund, 1840), a non-feline host.

Feline Morbillivirus (FeMV) was first detected in 2012 in domestic cats from Hong Kong and was found to be associated with tubulointerstitial nephritis and chronic kidney disease. In subsequent studies in other countries, FeMV was detected in asymptomatic cats. However, it is not clear whether FeMV plays a role as a pathogen in the kidney diseases of cats, and other epidemiological data are still unknown. To date, studies have reported the presence of FeMV exclusively in domestic cats. This study is the first molecular detection of the FeMV RNA associated with pathological and immunohistochemical findings in a synanthropic marsupial, the white-eared opossum (Didelphis albiventris), inhabiting peri-urban areas of north-central Parana, Southern Brazil. Molecular techniques identified the viral RNA in the lungs and kidneys. Histopathologic evaluation of these tissues revealed interstitial pneumonia in the lungs with lymphocytic nephritis and tubular necrosis in the kidneys. Immunohistochemistry assays detected positive intralesional immunoreactivity to N protein of FeMV within the lungs and kidneys. A FeMV opossum strain was isolated in Crandell Rees feline kidney lineage cells, resulting in syncytia formation and cell death. Therefore, these results support the ability of FeMV to infect other mammal species and reinforce the possibility of the opossum to be a disseminator of this virus among domestic and wild animals.

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.

Estudo retrospectivo da leptospirose canina em hospital veterinário escola no município de Londrina, Pr / Retrospective study of canine leptospirosis in a veterinary school hospital in the municipality of Londrina, Pr / Estudio retrospectivo de leptospirosis canina en un hospital veterinario escolar del municipio de Londrina, Pr

O objetivo do trabalho foi relatar a ocorrência de casos de cães com Leptospira spp. no Hospital Veterinário de Londrina. Foram avaliados 5.454 prontuários, durante os meses de janeiro a dezembro de 2016, e selecionados os que apresentaram solicitação de exame para Lepstopira por meio da técnica de campo escuro (CE). Nos positivos pela técnica CE dados sobre o sexo, sinais clínicos, endereço, data do atendimento e resultado na SAM foram coletados. A análise dos dados foi realizada pelo programa EpiInfo (7.2.3.1), dos 5.454 prontuários nos quais foi solicitado exame CE em 7,06% (385/5.454), e nesses, 13,76% (53/385) foi identificada a presença da espiroqueta. Dos animais positivos na técnica CE, 20,75% (11/53) foram reagentes na SAM. O sorovar Canicola foi o mais frequente em 90,91% (10/11) dos animais, apresentando títulos variados de 100 a 12.800. Para o sorovar Pomona 9,09% (1/11) dos animais apresentaram títulação de 100. Os sinais clínicos mais observados foram apatia em 58,49% (31/53), azotemia em 49,06% (26/53) e êmese em 49,06% (26/53) dos casos. Quanto ao sexo, os machos constituíram 71,70% dos casos e as fêmeas 28,30% (p=0,0015). Os resultados demonstraram a presença de Leptospira spp. entre os animais atendidos no hospital veterinário de Universidade Estadual de Londrina e reforçam a importância do sorovar Canicola entre os cães, visto que esse foi o mais frequente entre os sorovares e que apresentou os maiores títulos.(AU)

The purpose of this work is to report the occurrence of cases of dogs with Leptospira spp. at the Veterinary Hospital of Londrina. A total of 5,454 medical records were evaluated from January to December 2016, and data obtained from the examination for Lepstopira using the dark field technique (DF) were selected. In cases which were positive by the DF technique, data on gender, clinical signs, address, care data, and SAM results were collected. Data analysis was performed using the EpiInfo program (7.2.3.1). From the total 5,454 medical records in which DF examination was requested, in 7.06% (385/5.454), and from those, 13.76% (53/385) were positive for Leptospira. From the animals positive in the DF technique, 20.75% (11/53) were reactive in SAM. The Canicola serovar was the most prevalent, being present in 90.91% (10/11) of the animals, with titers ranging from 100 to 12,800. For the Pomona serovar, 9.09% (1/11) of the animals presented a title of 100. The most frequent clinical signs observed were apathy, in 58.49% (31/53); azotemia in 49.06% (26/53); and emesis in 49.06% (26/53) of the cases. As for gender, male animals constituted 71.70% of the cases and female 28.30% (p = 0.0015). The results demonstrated the presence of Leptospira spp. among animals treated at the Veterinary Hospital of the State University of Londrina which reinforces the importance of the Canicola serovar among dogs, since it was the serovar with the highest prevalence and highest titer found.(AU)

El objetivo de este trabajo fue reportar la ocurrencia de casos de perros con Leptospira spp. en el Hospital Veterinario de Londrina. Se evaluaron un total de 5.454 historias clínicas de enero a diciembre de 2016, y se seleccionó los que presentaron solicitud de examen de Lepstopira mediante la técnica de campo oscuro (CE). En los positivos por la técnica de CE se recogieron datos de sexo, signos clínicos, domicilio, fecha de atención y resultado en la SAM. El análisis de los datos se realizó mediante el programa EpiInfo (7.2.3.1), de las 5.454 historias clínicas en las que se solicitó examen CE en el 7,06% (385/5.454), y en estas, el 13,76% (53/385) se identificaron la de espiroqueta. De los animales positivos en la técnica CE, el 20,75% (11/53) fueron reactivos en la SAM. El serovar Canicola fue el más frecuente en el 90,91% (10/11) de los animales, con títulos que oscilaron entre 100 y 12.800. Para el serovar Pomona, el 9.09% (1/11) de los animales tuvo un título de 100. Los signos clínicos más observados fueron apatía en 58.49% (31/53), azotemia en 49.06% (26/53) y emesis en 49.06% (26/53) de los casos. Cuanto al sexo, los machos constituyeron el 71,70% de los casos y las hembras el 28,30% (p = 0,0015). Los resultados demostraron la presencia de Leptospira spp. entre los animales tratados en el Hospital Veterinario de la Universidad Estatal de Londrina y refuerzan la importancia del serovar Canicola entre los perros, ya que este fue el más frecuente entre los serovares y tuvo los títulos más altos.(AU)

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.

Phenotypical and genotypical differences among Leishmania (Leishmania) amazonensis isolates that caused different clinical frames in humans and dogs: A systematic review.

Leishmania (Leishmania) amazonensis is an important etiological agent of American cutaneous leishmaniasis (ACL) in Brazil. The species causes a large spectrum of clinical manifestations in humans and dogs, ranging from cutaneous, cutaneous diffuse, mucocutaneous, and visceral involvement, however, the factors that drive the development of different disease forms by the same species are not yet fully known. In the present work, it was systematically reviewed the studies addressing phenotypic and genotypic characteristics of Leishmania (L.) amazonensis isolates causing cutaneous and visceral clinical frames in humans and dogs, comparing the results observed. For this, four research databases were searched for the following keywords: (Leishmania amazonensis AND visceral leishmaniasis) AND (tropism OR virulence OR visceralization OR adaptations OR mutation OR clinical presentation OR resistance OR survival OR wide spectrum). The results revealed that the complexity disease seems to involve the combination of genetic factors of the parasite (as modifications in molecules related to the virulence and metabolism) and also of the host's immune background and status. Nonetheless, the exact mechanism that leads to different clinical manifestations between strains of the same species is still uncertain and future studies must be developed to better elucidate this phenomenon.

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.

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.

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.

American cutaneous leishmaniasis associated with degradation of native forest, regardless of economic, social and infrastructure vulnerability.

American cutaneous leishmaniasis (ACL) is a notifiable dermatozoonosis with relevant morbidity. The present study aimed to evaluate the epidemiological aspects of cases of ACL reported in Northern Paraná (2007-2016), as well as to georeference and analyse the influence of economic, social and environmental variables. Data from ACL notification forms were obtained and the patients' houses were georeferenced. Descriptive statistics, calculations of disease incidence and proportion of vegetation cover by municipality, spatial analysis, multiple linear regression and vulnerability analysis by census sector (CS) were performed. One thousand four hundred fifty-one cases of ACL were reported in the 89 municipalities of four regional health (RH). The average incidence of cases per 10,000 inhabitants was as follows: 11.58, 8.79, 4.92 and 4.03 in 18thRH, 15thRH, 16thRH and 17thRH. Peaks of incident cases were observed in 2008, 2012 and 2015. There was a statistically significant difference between the HR when comparing the proportions of the variables gender, age, education level, area of residence, clinical form, diagnostic criteria, response to treatment and drug used in case of failure. It was observed that the lower the proportion of remaining Atlantic Forest, the greater the incidence of ACL in the municipality. With regard to clusters analysis, treatment abandonment clusters were observed in the 15thRH and mucous form clusters were observed in the 15thRH, 16thRH and 17thRH. The vulnerability analysis by CS allowed us to observe a statistically significant difference in all vulnerability indicators: economic and social in 15thRH and 17thRH; home infrastructure in 15thRH and 18thRH, urban infrastructure in 15thRH and 18thRH and compound vulnerability index in 15thRH and 18thRH. However, the most vulnerable areas did not always have the highest number of cases. The data presented demonstrate that preventive and health education measures must be mainly directed to areas of greater degradation of native forest; regardless of the vulnerability situation.

Anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state, South Brazil: prevalence and associated factors / Anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em ovinos do estado do Paraná, sul do Brasil: prevalência e fatores associados

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.

Resumo O objetivo deste estudo foi avaliar a soroprevalência e os fatores de risco associados à presença de anticorpos contra Toxoplasma gondii e Neospora caninum em ovinos do Paraná. A detecção dos anticorpos para T. gondii e N. caninum foi realizada, usando-se testes de ELISA indireto caseiro e comercial, respectivamente. A análise de regressão logística múltipla foi usada para verificar os fatores de risco associados à soroprevalência. Anticorpos anti-T. gondii e anti-N. caninum foram observados em 42,7% e em 17,6% dos animais, respectivamente. Os fatores de proteção associados à soropositividade foram "algum nível de confinamento (confinamento ou semiextensivo)" (OR=0,53) para T. gondii e "presença de mão de obra especializada" para N. caminum (OR=0,64). Os fatores de risco foram "presença de gatos" para T. gondii (OR=1,75) e "alimentação de cães com resíduos placentários" (OR=1,79) para N. caninum. Além de apresentar uma significante e simultânea soroprevalência para ambos os agentes (9,9% dos animais), os resultados também indicam que as deficiências em gestão e saneamento ambiental, a presença de reservatórios e os tipos de exploração favorecem a soropositividade. Dessa forma, estudos como estes podem auxiliar em programas sanitários e políticas públicas para a prevenção de T. gondii e N. caninum nos rebanhos ovinos do Paraná.

Evaluation of an active and early surveillance methodology for visceral leishmaniasis by molecular detection in road-killed wild fauna / Avaliação de metodologia de vigilância ativa e precoce da leishmaniose visceral por detecção molecular em fauna silvestre atropelada

Abstract 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.

Resumo O objetivo do presente estudo foi avaliar uma metodologia de vigilância ativa da leishmaniose visceral por meio da detecção de DNA de Leishmania em órgãos de animais silvestres atropelados, de novembro de 2016 a outubro de 2018, no Norte do Paraná, Brasil. Os pontos de coleta dos animais silvestres atropelados foram georreferenciados. Os animais foram autopsiados e amostras de medula óssea, linfonodo, fígado, baço, e pele de orelha foram coletados. DNA genômico foi extraído e submetido à PCR para amplificação de quatro diferentes regiões de Leishmania spp.: 18S, kDNA, HSP70 e ITS1, sequenciamento de DNA foi realizado. Os iniciadores utilizados para a amplificação dos genes kDNA, ITS1 e HSP70 apresentaram resultados inespecíficos. Dos 66 mamíferos coletados em 24 diferentes municípios, um tamanduá-mirim (Tamandua tetradactyla) apresentou DNA de Leishmania spp. em linfonodo na PCR, que amplificou o gene 18S. O sequenciamento de DNA confirmou o resultado e demonstrou a presença do subgênero Viannia. Sugere-se o uso da metodologia apresentada no presente estudo na vigilância ativa e precoce da leishmaniose visceral em área não endêmica.

Techniques for inactivating Toxoplasma gondii oocysts: a systematic review / Técnicas para inativar oocistos de Toxoplasma gondii: uma revisão sistemática

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 <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.

Resumo O oocisto, forma resistente do Toxoplasma gondii, desempenha um papel importante na transmissão desse protozoário. O objetivo desta revisão foi relatar os métodos capazes de inativar oocistos por meio de uma revisão sistemática da literatura realizada nas bases de dados Scientific Electronic Library Online, Web of Science, Science Direct, PubMed e Scopus. As palavras-chave pesquisadas foram (((efeitos OR infectividade OR resistência) AND Toxoplasma) AND oocyst). Foram selecionados 16 artigos que descreveram 309 tratamentos diferentes. Dentre todos os protocolos avaliados, 35,60% (110/309) foram eficazes na inativação de oocistos. Os métodos físicos foram mais eficazes do que outros métodos (p <0,05). Oocistos esporulados e a cepa VEG de T. gondii foram mais resistentes (p <0,05) aos tratamentos. Embora seja eficaz contra vírus e bactérias, o uso de desinfetantes na água tem pouco ou nenhum efeito sobre os oocistos de T. gondii. O uso de radiação e pressão foram eficazes na inativação de oocistos. Como esses tratamentos não incluem mudança de temperatura, podem ser utilizados em alimentos de consumo cru, tais como vegetais, pois não acarretarão substancialmente alterações nas suas características físico-químicas. Portanto, esses métodos podem ser alternativas viáveis ​​para o controle de T. gondii.

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.

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.

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

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