Seropositivity for Toxocara spp. in Individuals with Animal Hoarding Disorder in Southern Brazil: An Alarm for Public Health.

PURPOSE: Animal hoarding has been associated with unhealthy human, animal and environmental conditions that predispose such individuals to serious life-threatening risks such as arson, malnutrition, cruelty and zoonosis. The study aimed to evaluate the presence of anti-Toxocara spp. antibodies among individuals with animal hoarding disorder in Curitiba, Brazil. METHODS: 65 residences with register of animal hoarder behavior were visited and 11 residences were included in the study, with a total of 19 individuals consenting participation. A short questionnaire was applied to gather information regarding hoarders and their dogs/cats, and serum samples were screened to detect antibodies (IgG) against antigens of Toxocara spp. RESULTS: Overall, 14/19 individuals (73.7%) presented anti-Toxocara spp. antibodies. In 8/11 (72.7%) households at least one person was seropositive. Seropositivity was higher among women (10/13; 76.9%) than men (4/6; 66.7%). A total of 442 dogs (14-30 dogs; average = 23.3 per household) and 31 cats (1-20 cats; average = 4.8 per household) were observed. To the authors' knowledge, this was the first study to survey occurrences of toxocariasis among animal hoarders. The high population densities of dogs observed during visits, in conjunction with absence of veterinary care and unsanitary conditions, may indicate that situations of high levels of animal infection and soil contamination were present. CONCLUSION: In summary, the seroprevalence observed in this study indicated that there was a high risk of Toxocara spp. infection among individuals with animal hoarding disorder. Provision of educational programs to reduce the risk of infection in this population is warranted.

Serosurvey of Toxoplasma gondii and Toxocara spp. co-infection in pregnant women in low-income areas of Brazil.

Introduction: Despite human toxoplasmosis and toxocariasis having been listed among the top six most neglected parasitic zoonoses worldwide, presenting similar associated risk factors and transmission routes, few studies have been conducted in pregnant women and the consequences of concurrent infection remain to be fully established. Accordingly, the present study has serologically assessed the co-infection and associated risk factors for Toxoplasma gondii and Toxocara spp. in pregnant women, assisted by the public Unified National Health System (SUS) in southeastern Brazil. Materials and Methods: Blood samples were collected and tested for IgG antibodies against Toxoplasma gondii by chemiluminescence immunoassay and against Toxocara spp. by enzyme-linked immunosorbent assay (ELISA). An epidemiological questionnaire was applied to gather socioeconomic information to assess the risk factors associated with seropositivity to toxocariasis/toxoplasmosis by univariate analysis followed by logistic regression. Results: Overall, seropositivity was 69/280 (24.6, 95% CI: 19.96-30.01) for T. gondii and 56/280 (20.0, 95% CI: 15.73-25.08) for Toxocara spp. Co-infection was observed in 25/280 (8.9, 95% CI: 6.12-12.85) pregnant women, with increased odds (OR: 3.3, CI 95%: 1.77-6.14, p = 0.0002). Logistic regression revealed that a higher educational level (high school or college) significantly reduced the likelihood of co-infection seropositivity, owning cats increased the odds of toxocariasis, and older pregnant women presented significantly higher T. gondii seropositivity. Conclusion: Co-infection herein highlights the importance of educational programs in the prevention of toxocariasis and toxoplasmosis in pregnant women and other high-risk populations.

Risk factors for toxocariasis during incarceration: the One Health intervention approach.

Despite potential exposure to soil-transmitted helminths, especially when stray dogs and cats are present, toxocariasis in inmate populations remains to be established. Accordingly, the present study assessed the seroprevalence and associated risk factors of toxocariasis at the Women's State Penitentiary of Parana, Brazil. A total of 234/370 (63.2%; 95% CI 58.2-68.0) women inmates and 28/87 (32.2%; 95% CI 23.3-42.6) correctional officers were seropositive for anti-Toxocara spp. IgG by ELISA, with inmates 2.62-fold more likely positive (p = 0.00000026). The univariate model has identified that non-white (OR = 1.58, p = 0.047) and older than 39 years (OR = 1.28, p = 0.032) inmates were associated with mild but significant odds for seropositivity. Elementary or higher educational level was considered a protective factor for seropositivity. The presence of Toxocara spp. eggs was observed in 10/15 (66.7%) collected soil samples by centrifuge-flotation in Zinc Sulfate, and molecular analysis by PCR identified only Toxocara cati in these eggs. An intervention program was established with regular trap-neuter-release, with gradual removal for adoption (donation campaigns), treatment, and euthanasia when necessary (particularly due to advanced sporotrichosis). In addition, an educational awareness agenda was proposed, aiming to reduce soil contamination and accidental intake by the incarcerated population. A total of 40 feral cats were trapped, 20 males and 20 females, mostly adults. After trapping, 36 cats were neutered, treated, and microchipped in the Veterinary Teaching Hospital (VTH) at the Federal University of Paraná. Five trapped feral cats were euthanized, four diagnosed with advanced sporotrichosis, and one already neutered cat (not herein) with complications due to feline immunodeficiency virus (FIV). Female inmates presented higher seroprevalence for Toxocara spp. antibodies when compared to correctional officers, significantly associated with age, self-declared ethnicity (non-white), and lack of formal education. Despite the non-natural scenario of a state penitentiary, the One Health approach of Toxocara spp. has highlighted the interdisciplinary nature of the study and its relevance in understanding the complex interactions between human, animal, and environmental factors, particularly impacting female inmates. Further studies should establish the rate of inmate infection over time while deprived of liberty.

One health approach to toxocariasis in quilombola communities of southern Brazil.

BACKGROUND: Toxocariasis has been listed among the most neglected parasitic diseases worldwide, with approximately one fifth of the global population exposed, particularly those living under poverty. In Brazil, communities of descendants of enslaved blacks (quilombola) have historically had some of the highest rates of vulnerability and poverty, characterized by lack of health assistance, poor quality of life, and nutritional insecurity. METHODS: A cross-sectional sampling of quilombola individuals living in four communities of southern Brazil, as well as their dogs and the soil, was carried out from December 2021 to March 2022. Sociodemographic and other information such as water source, alimentary habits, and dog and cat ownership were gathered using a semi-structured questionnaire for assessing toxocariasis risk factors. Human serum samples were tested by ELISA for anti-Toxocara spp. IgG antibody detection was carried out on dog feces and hair, and soil samples were surveyed for presence of Toxocara spp. eggs. RESULTS: Overall, 172/208 individuals (82.7%, 95% CI = 77.0-87.2) were seropositive, the highest seroprevalence rate to date in Brazil. Male gender (P = 0.029), educational level (P = 0.026), and drinking water source (P = 0.043) were associated with seropositivity by univariate analysis. Final logistic regression revealed increased odds (P = 0.017, OR = 7.6, 95% CI = 1.5-42.7) to have seropositivity in individuals > 50 years old (< 10 years old). As expected, individuals with soil contact were more likely seropositive (P = 0.038, OR = 4.4, 95% CI = 1.1-18.8). Although retrieved in only 5/96 (5.2%) dog feces, Toxocara spp. eggs were found in 18/60 (30.0%) soil samples. CONCLUSIONS: The high vulnerability and seroprevalence observed in quilombola communities clearly demand a One Health approach for detection, monitoring, and prevention of infection by Toxocara spp. in both human and dog populations.

One health approach to toxocariasis in Brazilian indigenous populations, their dogs, and soil contamination.

Introduction: Although socioeconomic vulnerability and lifestyle factors may contribute to the transmission of Toxocara spp., no study has investigated indigenous populations in Brazil using the One Health approach. Methods: Accordingly, this study assessed anti-Toxocara spp. antibodies in Brazilian indigenous people and healthcare professionals by enzyme-linked immunosorbent assay. Presence of Toxocara spp. eggs (feces and hair) in dogs as definitive hosts and in soil samples of the indigenous communities were also recovered and molecularly investigated. Results: Overall, 342/463 (73.9%) indigenous individuals and 46/147 (31.3%) non-indigenous healthcare professionals were seropositive for Toxocara spp. In addition, T. canis eggs were retrieved from 9/194 (4.6%) dog fecal samples and 4/204 (2.0%) dog hair samples, mainly from the Paraná State communities (3/42; 7.1%). Soil contamination was observed only in the Paraná State communities (36/90; 40.0%), with the molecular detection of T. canis. River water consumption was also associated with indigenous seropositivity (Odds ratio, 11.4). Discussion: Indigenous individuals in Paraná State communities were 2.72-fold more likely to be seropositive than those in São Paulo State, likely due to a lack of sanitary infrastructure. In this scenario, a primarily soil-transmitted disease may also have become waterborne, with embryonated eggs probably spread to water supplies by rain. Full-time healthcare professionals in daily contact with indigenous communities were 9.2-fold more likely to be seropositive than professionals who visited sporadically, suggesting exposure to Toxocara spp. during their work and raising health concerns. In addition, the findings herein showed a significantly higher seroprevalence in indigenous people than in healthcare workers (χ2 = 85.5; p < 0.0001), likely due to overtime exposure to Toxocara spp. In conclusion, Brazilian indigenous communities are highly exposed to toxocariasis, with poor infrastructure and contact with contaminated river water as associated risk factors and a higher risk of infection in healthcare professionals working full-time in these communities.

Seroprevalence and associated risk factors of strongyloidiasis in indigenous communities and healthcare professionals from Brazil.

Strongyloides stercoralis, a pathogenic roundworm, is considered endemic in several tropical and subtropical areas worldwide. Indigenous populations have the highest soil-transmitted helminthiases-related mortality rates, but the prevalence and risk factors associated with S. stercoralis in Brazilian indigenous populations have not been established. Thus, the present study aimed to assess the seroprevalence and associated risk factors for S. stercoralis in indigenous communities and the healthcare professionals serving them in Brazil. Indigenous populations living in nine communities and healthcare professionals were tested for anti- S. stercoralis antibodies by ELISA. A questionnaire was used to assess socio-epidemiological information. Associated risk factors for seropositivity were tested by chi-square or Fisher's exact tests, using univariate analyses and multivariate logistic regression. Overall, 174/463 (37.6%; CI 95%: 33.3-42.1) indigenous persons and 77/147 (52.4%; 95% CI: 44.3-60.3) healthcare professionals were seropositive for anti- S. stercoralis antibodies. Seropositivity among the two groups was statistically significant (p = 0.0016; OR = 0.547; 95% CI: 0.376-0.796) and revealed that healthcare professionals were 1.83 times more likely to be seropositive. The multivariate analysis showed that being male or being adult were also risk factors, while having a septic tank as a sanitary facility represented a protective factor for S. stercoralis exposure in indigenous persons. None of the variables evaluated were associated with S. stercoralis exposure in the professional group. The study herein has reported a high seroprevalence to Strongyloides stercoralis in indigenous communities of Brazil and healthcare professionals, warning for potential public health concerns of strongyloidiasis in such populations.

Toxocara canis 30-35 kDa excretory-secretory antigen is an important marker in mice challenged by inocula containing different parasite load levels.

The Western-blotting technique was applied to identify antigenic fractions of excretory-secretory Toxocara canis antigen recognized by IgG antibodies throughout an experimental infection in mice challenged by different inocula. Mice were inoculated with 5, 50 and 500 embryonated eggs and serum samples were collected 15, 30, 60, 90 and 120 days post-infection. Serum samples were analyzed using an excretory-secretory Toxocara antigen. Antibodies recognized antigenic fractions from 30 to 90 kDa. The protein fraction of 30-35 kDa was the most frequently recognized regardless of the size of inoculum and the stage of infection represented by the different collection times, but the antigenic recognition was more evident in groups infected with 50 and 500 eggs. This study presents an antigenic panel of the excretory-secretory antigen of T. canis and suggests that the 30-35 kDa antigenic fraction is a promising marker of the infection and should be further explored in future studies on experimental toxocariasis.

Toxocara canis 30-35 kDa excretory-secretory antigen is an important marker in mice challenged by inocula containing different parasite load levels

ABSTRACT The Western-blotting technique was applied to identify antigenic fractions of excretory-secretory Toxocara canis antigen recognized by IgG antibodies throughout an experimental infection in mice challenged by different inocula. Mice were inoculated with 5, 50 and 500 embryonated eggs and serum samples were collected 15, 30, 60, 90 and 120 days post-infection. Serum samples were analyzed using an excretory-secretory Toxocara antigen. Antibodies recognized antigenic fractions from 30 to 90 kDa. The protein fraction of 30-35 kDa was the most frequently recognized regardless of the size of inoculum and the stage of infection represented by the different collection times, but the antigenic recognition was more evident in groups infected with 50 and 500 eggs. This study presents an antigenic panel of the excretory-secretory antigen of T. canis and suggests that the 30-35 kDa antigenic fraction is a promising marker of the infection and should be further explored in future studies on experimental toxocariasis.

One Health approach on human seroprevalence of anti-Toxocara antibodies, Toxocara spp. eggs in dogs and sand samples between seashore mainland and island areas of southern Brazil.

Toxocariasis, caused by Toxocara spp. nematodes, is among the top 5 neglected parasitic diseases worldwide; however, no comprehensive study to date has serologically compared infections in people and their dogs and environmentally contaminated soil or sand of mainland and island locations. Accordingly, this study aimed to assess the seroprevalence of anti-Toxocara antibodies in traditional human seashore populations, the presence of eggs in dogs' feces and hair, and the presence of eggs in environmental samples from islands compared to the adjacent mainland of southern Brazil. Overall, 212/328 (64.6%) people were positive for Toxocara spp. antibodies, including 125/190 (65.8%) island and 87/138 (63.0%) mainland residents. For dog samples, 12/115 (10.43%) were positive for the presence of Toxocara spp. eggs, all from dogs living in islands, and 22/104 (21.15%) dog hair samples contained eggs of Toxocara spp. Environmental contamination with Toxocara spp. eggs was observed in 50/130 (38.46%) samples from all sampled sites. No significant association was found between risk factors (age, sex, educational level, monthly income, owning dogs or cats, ingestion of treated water, and consumption of raw or uncooked meat) and Toxocara spp. seropositivity. The present study is the first concurrent report on people, their dogs, and environmental contamination of Toxocara spp. The high prevalence we observed in the seashore populations of both in island and mainland areas may be caused by exposure to contaminated sand and climatic factors favoring frequent exposure to Toxocara spp. In conclusion, seashore lifestyle and living conditions of both island and mainland areas may have predisposed higher contact with infected pets and contaminated soil, favoring the high prevalence of toxocariasis.

Serosurvey and associated risk factors of anti-Toxocara spp. antibodies in bovines from slaughterhouses of southeastern Brazil.

BACKGROUND: Toxocariasis, caused by a nematode species of the genus Toxocara, has been described as one of the most prevalent zoonotic helminthiases worldwide. Human transmission may occur by ingesting Toxocara spp. larvae from raw or undercooked meat or organs; however, no comprehensive serosurvey study has been conducted to date investigating the role of cattle as paratenic hosts. The aim of the study reported here was to assess the prevalence of anti-Toxocara spp. antibodies and associated risk factors in bovines from two slaughterhouses located in Presidente Prudente, southeastern Brazil. METHODS: Blood samples were collected and tested by indirect enzyme-linked immunosorbent assay (ELISA). Cattle farmers voluntarily responded to an epidemiologic questionnaire. RESULTS: Overall, 213 of the 553 (38.5%) bovine samples were assessed as seropositive for anti-Toxocara spp. antibodies by indirect ELISA. Multivariate analysis revealed that the source of beef cattle and the presence of dogs or cats at the farm were associated with seropositivity. The use of feedlot systems was associated with lower likelihood of seropositivity. CONCLUSIONS: These results indicate a high level of anti-Toxocara seropositivity in slaughterhouse cattle, with potentially contaminated meat posing an infection risk to humans. In addition, the presence of dogs and cats where the slaughtered beef cattle were raised was statistically associated with bovine seropositivity, probably due to the overlapping environment at the farm and the lack of pet deworming. The use of feedlot systems was a protective factor likely due to the absence of dog and cat contact, elevated feeding troughs that avoid contact with contaminated soil or grass, and younger age at slaughter of feedlot cattle. In summary, bovines may be used as environmental sentinels of Toxocara spp. contamination, and high seropositivity of slaughterhouse cattle may indicate a potential risk of human toxocariasis through the ingestion of raw or undercooked contaminated meat.

Culture isolation and molecular identification of Blastocystis sp. in Brazilian human isolates: preliminary results.

Blastocystis sp. is a protist commonly found in stool samples of humans and animals. Biological and genetic factors of this organism remain controversial. The present study aimed to develop and implement the Blastocystis in vitro culture of Brazilian human isolates for routine use. The fecal isolates (n = 20) were maintained in our laboratory by several passages in Pavlova's medium. Cultures were monitored every 72 h by light microscopy. Genomic DNA was extracted to identify the subtypes (STs). In most isolates, the vacuolar form was prevalent. The amoeboid, granular and cystic forms were observed during in vitro cultivation. STs 1, 2, 3, 4 and 7 were identified. Our preliminary results show the generation time and forms present in the in vitro culture of Blastocystis subtypes isolated from Brazilian human isolates. Therefore, we emphasize the use of in vitro culture as a tool in future studies for the better understanding of the biological aspects of Blastocystis sp.

Effect of sex on Toxocara canis larval migration to the cerebellum during experimental infection of Rattus norvegicus

The presence of Toxocara canis third instar larvae in the cerebellum of Rattus norvegicus may alter rodent behavior and movement. In this study, we investigated whether the sex of the rodent affects the migration of larvae to the cerebellum. Thirty-six Rattus norvegicus specimens (18 males and 18 females) were infected with 300 T. canis eggs and were euthanized after 60 days. The cerebellum was removed and treated with 0.5% HCl to recover the T. canis larvae. The number of larvae recovered from male rodents was significantly higher than in females, suggesting that the sex of the animal influences larval migration to the cerebellum

Experimental toxocariasis in BALB/c mice: relationship between parasite inoculum and the IgG immune response

BALB/c mice were inoculated with 5-500 Toxocara canis infective eggs, and bled at 15-120 days post infection (dpi) to evaluate the dynamics of IgG antibody response and larvae distribution. Positive results were observed in all occasions for every inoculum, and a direct proportional relationship between antibody detection and the parasitic load was observed. In samples collected at 60 dpi, detection of IgG was more intense, especially with the 50 and 500 egg doses; also, a correlation between antibody level and egg count was observed with these two inocula. At 120 dpi, a decrease in antibody titer was observed for all groups; and at the end of the experiment, larvae were recovered from carcass, liver and brain. In the liver, larvae were only found in mice inoculated with 500 T. canis eggs. In carcasses, these were recovered in all groups, and the group inoculated with 50 eggs showed the highest percentage of larvae in the brain.

Experimental toxocariasis in BALB/c mice: relationship between parasite inoculum and the IgG immune response.

BALB/c mice were inoculated with 5-500 Toxocara canis infective eggs, and bled at 15-120 days post infection (dpi) to evaluate the dynamics of IgG antibody response and larvae distribution. Positive results were observed in all occasions for every inoculum, and a direct proportional relationship between antibody detection and the parasitic load was observed. In samples collected at 60 dpi, detection of IgG was more intense, especially with the 50 and 500 egg doses; also, a correlation between antibody level and egg count was observed with these two inocula. At 120 dpi, a decrease in antibody titer was observed for all groups; and at the end of the experiment, larvae were recovered from carcass, liver and brain. In the liver, larvae were only found in mice inoculated with 500 T. canis eggs. In carcasses, these were recovered in all groups, and the group inoculated with 50 eggs showed the highest percentage of larvae in the brain.

In vitro action of antiparasitic drugs, especially artesunate, against Toxoplasma gondii.

INTRODUCTION: Toxoplasmosis is usually a benign infection, except in the event of ocular, central nervous system (CNS), or congenital disease and particularly when the patient is immunocompromised. Treatment consists of drugs that frequently cause adverse effects; thus, newer, more effective drugs are needed. In this study, the possible activity of artesunate, a drug successfully being used for the treatment of malaria, on Toxoplasma gondii growth in cell culture is evaluated and compared with the action of drugs that are already being used against this parasite. METHODS: LLC-MK2 cells were cultivated in RPMI medium, kept in disposable plastic bottles, and incubated at 36ºC with 5% CO2. Tachyzoites of the RH strain were used. The following drugs were tested: artesunate, cotrimoxazole, pentamidine, pyrimethamine, quinine, and trimethoprim. The effects of these drugs on tachyzoites and LLC-MK2 cells were analyzed using nonlinear regression analysis with Prism 3.0 software. RESULTS: Artesunate showed a mean tachyzoite inhibitory concentration (IC50) of 0.075µM and an LLC MK2 toxicity of 2.003µM. Pyrimethamine was effective at an IC50 of 0.482µM and a toxicity of 11.178µM. Trimethoprim alone was effective against the in vitro parasite. Cotrimoxazole also was effective against the parasite but at higher concentrations than those observed for artesunate and pyrimethamine. Pentamidine and quinine had no inhibitory effect over tachyzoites. CONCLUSIONS: Artesunate is proven in vitro to be a useful alternative for the treatment of toxoplasmosis, implying a subsequent in vivo effect and suggesting the mechanism of this drug against the parasite.

In vitro action of antiparasitic drugs, especially artesunate, against Toxoplasma gondii / Ação in vitro de drogas antiparasitárias, especialmente artesunato, contra Toxoplasma gondii

INTRODUCTION: Toxoplasmosis is usually a benign infection, except in the event of ocular, central nervous system (CNS), or congenital disease and particularly when the patient is immunocompromised. Treatment consists of drugs that frequently cause adverse effects; thus, newer, more effective drugs are needed. In this study, the possible activity of artesunate, a drug successfully being used for the treatment of malaria, on Toxoplasma gondii growth in cell culture is evaluated and compared with the action of drugs that are already being used against this parasite. METHODS: LLC-MK2 cells were cultivated in RPMI medium, kept in disposable plastic bottles, and incubated at 36ºC with 5% CO2. Tachyzoites of the RH strain were used. The following drugs were tested: artesunate, cotrimoxazole, pentamidine, pyrimethamine, quinine, and trimethoprim. The effects of these drugs on tachyzoites and LLC-MK2 cells were analyzed using nonlinear regression analysis with Prism 3.0 software. RESULTS: Artesunate showed a mean tachyzoite inhibitory concentration (IC50) of 0.075µM and an LLC MK2 toxicity of 2.003µM. Pyrimethamine was effective at an IC50 of 0.482µM and a toxicity of 11.178µM. Trimethoprim alone was effective against the in vitro parasite. Cotrimoxazole also was effective against the parasite but at higher concentrations than those observed for artesunate and pyrimethamine. Pentamidine and quinine had no inhibitory effect over tachyzoites. CONCLUSIONS: Artesunate is proven in vitro to be a useful alternative for the treatment of toxoplasmosis, implying a subsequent in vivo effect and suggesting the mechanism of this drug against the parasite.

INTRODUÇÃO: Toxoplasmose é geralmente uma infecção benigna, exceto nos eventos de doença ocular, congênito e do sistema nervoso central, e particularmente quando o paciente é imunocomprometido. O tratamento consiste de drogas que frequentemente causam efeitos adversos, então novas drogas, mais efetivas são necessárias. Neste estudo, a possível atividade de artesunato, uma droga usada com sucesso no tratamento da malária, sobre o crescimento de Toxoplasma gondii em cultura celular é avaliado e comparado à ação de drogas que já estão sendo utilizadas contra este parasita. MÉTODOS: Células LLC-MK2 foram cultivadas em meio RPMI, mantidas em garrafas plásticas descartáveis e incubados a 36ºC com 5% CO2. Taquizoítos da cepa RH foram usados. As seguintes drogas foram testadas: artesunato, cotrimoxazol, pentamidina, pirimetamina, quinino e trimetoprima. Os efeitos dessas drogas sobre taquizoítos foram analisados por análise regressiva não linear com o software Prism 3.0. RESULTADOS: Artesunato mostrou uma concentração inibitória media (IC50) de 0,075µM e uma toxicidade sobre células LLC MK2 de 2,003µM. Pirimetamina foi efetiva a uma IC50 de 0,482µM e uma toxicidade de 11,178µM. Trimetoprima sozinha foi efetiva contra o parasita in vitro. Cotrimoxazol também foi efetivo contra o parasita, mas a concentrações mais altas que aquelas observadas para artesunato e pirimetamina. Pentamidina e quinino não tiveram efeitos inibitórios sobre os taquizoítos. CONCLUSÕES: Provou-se que artesunato in vitro pode ser uma alternativa útil para o tratamento da toxoplasmose, implicando um subsequente efeito in vivo e sugerindo o mecanismo desta droga contra o parasita.