225 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Neotropical Helminthology Neotropical Helminthology, 2024, vol. 18 (2), 225-239 ORIGINAL ARTICLE / ARTÍCULO ORIGINAL ZOONOTIC FOCI OF TOXOCARA SP.: COMPARISON OF TWO TOURIST REGIONS IN THE STATE OF SUCRE, VENEZUELAFOCOS ZOONÓTICOS DE TOXOCARA SP.: COMPARACIÓN DE DOS REGIONES TURÍSTICAS EN EL ESTADO SUCRE, VENEZUELA Erika Gomez-Martinez 1,2 , Márcia Viviana Gonçalves-Vallejos 1 , María Betania Bravo- Rodríguez 1 , Marcos Tulio-Díaz 2 , Zulay Simoni-González 2 , Del Valle Guilarte 2 , Elizangela Farias-da Silva 2 , ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403 DOI: https://dx.doi.org/10.62429/rnh20242181825 Este artículo es publicado por la revista Neotropical Helminthology de la Facultad de Ciencias Naturales y Matemática, Universidad Nacional Federico Villarreal, Lima, Perú auspiciado por la Asociación Peruana de Helmintología e Invertebrados Af nes (APHIA). Este es un artículo de acceso abierto, distribuido bajo los términos de la licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0) [https:// creativecommons.org/licenses/by/4.0/deed.es] que permite el uso, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citada de su fuente original. ABSTRACT Toxocariasis is a human infection caused by the larvae of the nematodes Toxocara canis (Werner, 1782) or Toxocara cati (Schrank, 1788), which are parasitic in animals and represents a serious public health problem. T e objective of this study was to investigate the information and prevalence of parasitic toxocariasis in the state of Sucre, Venezuela, particularly in the municipalities of Montes and Sucre, in communities considered vulnerable due to social and Zeca Manuel-Salimo 1,1 , Luziana de Sousa-Xavier 5 , Grafe Oliveira-Pontes 5 1 Programa de Pós-Graduação em Medicina Tropical, Universidade do Estado do Amazonas, Av. Pedro Teixeira, s/n, Dom Pedro, 69040-000 Manaus-AM, Brasil. 2 Instituto de Investigaciones em Biomedicina y Ciencias Aplicadas Dra. Susan Tai, Universidad de Oriente IIBCAUDO. Av. Universidad, CEP: 6101 Cumaná-Sucre, Venezuela. 3 Centro de Estudos Superiores de Tabatinga, Universidade do Estado do Amazonas, Av. da Amizade, 74 - Centro, CEP 69640-000, Tabatinga – AM, Brasil. 4 Faculdade de Ciências de Saúde, Universidade Lúrio, Nampula 364, CEP: 3100 Moçambique. 5 Fundação de Medicina Tropical Doutor. Heitor Vieira Dourado, Av. Pedro Teixeira, s/n, Dom Pedro, 69040-000, Manaus--AM, Brasil.* Corresponding author: grafeoliveira@gmail.com Er ika Gomez-Martinez: https://orcid.org/0000-0002-2467-8831Márcia Viviana Gonçalves-Vallejos: https://orcid.org/0000-0003-0657-4120María Betania Bravo-Rodríguez: https://orcid.org/0009-0005-2368-362XMarcos Tulio-Díaz: https://orcid.org/0009-0007-6392-3004Zulay Simoni-Gonzalez: https://orcid.org/0000-0002-5237-8238Del Valle Guilarte: https://orcid.org/0009-0006-3000-2463Elizangela Farias da-Silva: https://orcid.org/0000-0002-3386-6842Zeca Manuel-Salimo: https://orcid.org/0000-0002-0834-8728Luziana de Sousa-Xavier: https://orcid.org/0009-0000-6515-691XGrafe Oliveira Pontes: https://orcid.org/0000-0003-0755-1086
226 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. economic defciencies within the epidemiological/sanitary monitoring. In these municipalities, many stray dogs are living near homes and maintaining contact with other domestic dogs. Te seroprevalence of T. canis was evaluated through the ELISA method. A seroprevalence of IgG anti- T. canis was obtained in the parish of San Juan of 90.12% and the parish of Cumanacoa of 82.14%. In the parish of San Juan, when evaluating the prevalence of parasites by species, T. canis was present in 18.37% of the feces of the canines evaluated. On the other hand, in the parish of Cumanacoa, T. canis was present in 9.90% of the canine samples analyzed; similarly, the presence of Toxocara sp. in soils in both parishes. In summary, the determination of two zoonotic foci of Toxocara sp. in the state of Sucre, Venezuela, since all the factors necessary for the transmission of toxocariasis are present. Keywords : Dogs – Epidemiology – Larva migrans – Parasitic disease – Toxocara canis – Zoonosis RESUMEN La toxocariasis es una infección humana causada por las larvas de los nematodos Toxocara canis (Werner, 1782) o Toxocara cati (Schrank, 1788), que son parásitos en los animales, y representa un grave problema de salud pública. El objetivo de este trabajo fue investigar la información y prevalencia de toxocariasis parasitaria en el estado Sucre, Venezuela, particularmente en los municipios de Montes y Sucre, en comunidades consideradas vulnerables por defciencias sociales y económicas en el monitoreo epidemiológico/sanitario. En estos municipios hay muchos perros callejeros que viven cerca de las casas y mantienen contacto con otros perros domésticos. La seroprevalencia de T. canis se evaluó mediante el método ELISA. Se obtuvo una seroprevalencia de IgG anti- T. canis en la parroquia San Juan de 90,12% y en la parroquia Cumanacoa de 82,14%. En la parroquia San Juan, al evaluar la prevalencia de parásitos por especie, T. canis estuvo presente en el 18,37% de las heces de los caninos evaluados. Por otra parte, en la parroquia Cumanacoa, T. canis estuvo presente en el 9,90% de las muestras caninas analizadas; Asimismo, la presencia de Toxocara sp. en suelos de ambas parroquias. En resumen, la determinación de dos focos zoonóticos de Toxocara sp. en el estado Sucre, Venezuela, ya que están presentes todos los factores necesarios para la transmisión de la toxocariasis. Palabras clave: Epidemiología – Enfermedad parasitaria – Larva migrans – Perros – Toxocara canis – Zoonosis INTRODUCTION Toxocariasis is a human infection caused by the larvae of the nematodes Toxocara canis (Werner, 1782) or Toxocara cati (Schrank, 1788), which are parasitic in animals. Te clinical manifestations of the disease are usually asymptomatic, which contributes to the disease being neglected. And yet, toxocariasis has been shown to be especially prevalent among children from socioeconomically disadvantaged populations, both in the tropics and subtropics and in industrialized nations (Macpherson, 2013; Rostami et al., 2019; Ma et al., 2020).In humans, the infection is acquired accidentally, as there is no normal development of the parasite in humans, only the larval stage survives as it is a paratenic host in this parasitosis (Taylor et al ., 2017). Infection occurs by ingestion of infective eggs found in poorly washed vegetables and fruit originating from a contaminated environment, geophagy, ingestion of undercooked paratenic hosts (chicken, beef or lamb), putting contaminated objects in the mouth, as well as not washing hands after playing with or petting infected dogs (Holland et al., 1995; Vázquez et al., 1997; Ulloque-Badaracco et al., 2023). Children are the most infected due to their poor hygienic habits and are often in contact with dogs and puppies, as well as exposed to the pets’ living environment (Cabral-Monica et al., 2022). Toxocariasis is a widely distributed zoonosis, in which the human need to keep pets and companion animals such as dogs and cats around ensures the persistence of the parasite and infection in humans over time. with rates ranging from 1% in Spain to 86% in Santa Lucia; from 3.6% to 24.7% in Brazil; 47.5% in Colombia; from 34.9% to 66.6% in Venezuela and from 37.9% (Roldán et al., 2010a). In Argentina, a study carried out in children showed a 67.7% seroprevalence of anti- Toxocara IgG antibodies (López et al., 2005), while in Paraguay a 78% seroprevalence for toxokaryosis was reported (Rivarola et al., 2009). In Peru, a few reports have informed of frequencies from 7.8% to 44.92% in rural populations from diferent places (Roldán et al., 2010a).
227 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Toxocara canis infection in dogs has worldwide distribution rates ranging from 0 to 99.4% (Barriga, 1988; Heymann & American Public Health Association, 2004; Borges & Ferreira, 2020). In Venezuela, studies on T. canis infection were carried out in urban areas such as Caracas, reporting prevalences of 66.6%, in rural and suburban communities of the Capital District and the State of Miranda 20% and 70% (Incani, 1996; Delgado & Rodríguez-Morales, 2009). In the Brazilian Amazon, the prevalence in urban children < 5 years of age has been reported to be 21.5%,increasing to 26.7% in the rural general population, and to 26.8%or 52% in the riverine population (Oliart-Guzmán et al., 2014). Furthermore, studies on canine toxocariasis carried out in Maracaibo reported a prevalence of 11.4% (Ramírez-Barrios et al., 2004) and 63,16% in soil samples from leisure parks (Cazorla-Perfetti et al., 2007). In eastern Venezuela, particularly in the state of Anzoátegui, (Fernández et al., 2009) carried out a study on children aged 6 to 8 years, obtaining a prevalence of 16%, and (Gómez, 2004) carried out a study in the Nueva Toledo neighborhood of El Peñón, state of Sucre, on children aged 2 to 15 years, with prevalences of 77.14%. T us, the present study evaluated the seroprevalence of toxocariasis in the municipalities of Montes and Sucre, which are regional tourist destinations in Venezuela. T ese regions were selected to be considered vulnerable due to social and economic deprivation, the large number of stray dogs found in the area, and the fact that most family groups have dogs as pets. MATERIAL AND METHODSStudy area T e present study was conducted in the municipalities of Sucre and Montes, in the state of Sucre, Venezuela (Fig. 1). T e municipality of Sucre is located in a mountainous area with a hot, semi-arid climate, with average temperatures of 27°C and 440 mm of precipitation with a rainy season from July to November (Pérez, 2006). T e second site was Cumanacoa in the municipality of Montes. It is characterized by a hot sub-humid climate with temperatures ranging from 21°C to 27°C and annual precipitation between 900 and 1500 mm. Figure 1 . Map of the state of Sucre, Venezuela with the respective municipalities evaluated: Sucre (10,24198° N, 64,27980° O), Cumanacoa (10,24720° N, 63,92019° O), Google Maps. Population sample To calculate the representative sample size, a formula proposed by Martínez (2003) for f nite populations, with a known sampling frame, was used. T e prevalence for toxocariosis of 74.14% was considered (Gómez, 2004). In the parish of San Juan, 172 blood and faeces samples were taken from people who voluntarily attended the survey. T e same occurred in the parish of Cumanacoa, where 100 blood and stool samples were taken, respectively. A clinical-epidemiological survey was administered to each individual, regardless of gender and age, in each parish, with prior informed consent (OPAS, 1993).
228 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. Human faecal samples All the people who atended and participated in the collection were asked to provide a stool sample for the evaluation of the diferent helminths including Ascaris lumbricoides (Linnaeus, 1758). As an ascaridium, A. lumbricoides cross-reacts with T. canis when using immunological techniques using sera to evaluate for anti- T. canis antibodies. Serum samples of individuals presenting with Ascaris lumbricoides eggs in their stool was excluded from the study. Human blood samples Blood samples were obtained from the participants by venous puncture and transferred to the parasitology laboratory at the Instituto de Investigaciones en Biomedicina y Ciencias Aplicadas “Dr. Susan Tai”, Universidad de Oriente (IIBCA-UDO), Venezuela, where they were processed for serum collection. Te collected sera were frozen at -20°C until analysis. Determination of anti-T . canis IgG antibodies by ELISA method A commercial Toxocara -IgG ELISA kit (RIDASCREEN Toxocara -IgG), which uses excretion-secretion antigens, was used for serological diagnosis. No absorption of sera with Ascaris suum (Goeze, 1782) antigen was performed. However, all individuals with A. suum eggs in their faeces were excluded from analysis. For the evaluation and interpretation of the results, the sample was considered as positive for anti- T. canis antibodies when its index was higher than 1.10, as a negative sample, that with an index lower than 0.90 and as an indeterminate sample that with an index between 0.90 – 1.10 (Fernández et al., 2009). Dog Fecal Samples In both parishes, with the consent of the owners the dog fecal samples were collected in the morning, at midday and in the afternoon, and placed in airtight containers and transported to the laboratory. If not analyzed on the day of collection, they were stored at 4°C. Soil samples Soil samples were collected from the courtyards of homes in communities from the San Juan and Cumanacoa parishes. Tese samples were obtained by superfcially scraping the courtyards with a spatula, 5 cm from any faecal matter after a random selection of sites with bare soil. Soil samples were collected upto 10 cm deep and stored in labeled polyethylene bags at room temperature until processing (Leventhal & Cheadle, 1992). Parasitological diagnosis Small portions of all the stool samples were examined by direct microscopic examination after mixing with 0.85% physiological saline and Lugol’s solution (Botero & Restrepo, 1998). In addition, concentration and enrichment techniques such as: spontaneous tube sedimentation and fotation by Willis-Malloy were applied (Quinn et al., 1980; Camacho et al., 2006). It was also carried out with soil samples taken in each parish. Tese examinations were also done on soil samples taken in each parish. Statistical analysis Te results obtained were expressed as percentages and represented in tables and graphs. Te seroprevalence of T. canis infection was determined by applying the formula described by Gordis (2004). Te Chi-square test ( χ ²) was used to establish the association between the clinical-epidemiological aspects evaluated and the seroprevalence of T. canis in the inhabitants of San Juan and Cumanacoa parishes (Martínez, 2003). Data was analyzed using SPSS 18 software. Ethics aspects: Tis research was approved by the IIBCAUDO Bioethics and Biosafety Committee, (CBBIIBCAUDO-2009-09-3) year 2009, September, project 3. RESULTS The seroprevalence of T. canis was evaluated using the Toxocara IgG commercial kit. Of the 172 serum samples analyzed from the parish of San Juan, 10 samples were discarded because they belonged to people who had A. lumbricoides eggs in their faeces. Of the remaining 162 sera, 146 (90.12%) were positive, 9 (5.56%) indeterminate and 7 (4.32%) negatives. The average sample index of the positive cases was 3.03 of the indeterminate 0.99 and of the negative 0.55 (table 1). From the 100 serum samples analyzed from the Cumanacoa parish, Montes municipality, 16 sera were discarded because they belonged to people who had A. lumbricoides eggs in their stool samples. Of the 84 remaining sera, 69 (82.14%) were positive, 6 (7.14%) indeterminate and 9 (10.17%) negatives, the average sample rate for positive cases was 2.18, for indeterminate cases 0.96 and for negative cases 0.60 (table 1).
229 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Table 1 . Seroprevalence and summary statistics of sample rates for IgG anti- T. canis antibodies in inhabitases of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. Inhabitants of San Juan / Cumanacoa Samplesn.% χ SDLLULPositive146/6990,12/82,143,03/2,181,22/0,941,20/1,137,74/5,04Indeterminate9/65,56/7,140,99/0,960,08/0,040,91/0,921,10/1,02Negative7/94,32/10,170,55/0,600,14/0,210,37/0,340,73/0,89 n: number of cases, χ : mean, SD: standard deviation, LL: lower limit, UL: upper bound %: percentage. To understand the epidemiology of toxocariasis in both parishes of Sucre municipality, Sucre state, Venezuela, the seroprevalence of anti- T. canis IgG was analyzed according to gender; it was found that females were more afected in both San Juan and Cumanacoa parishes. However, the chi-square test did not show a statistically signifcant association (table 2). Table 2 . Association between genders of inhabitants and seroprevalence of anti- Toxocara canis IgG in San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. IgG Anti-Toxocara canis in Inhabitants of San Juan / CumanacoaPositivesIndeterminateNegativesTotalGendern%n%n%n% Females102/4569.86/65,227/377.80/502/628.57/66.67111/5468.52/,64.29Males44/2430.14/34,782/322.20/505/371.43/33.3351/3031.48/35.71Total 146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 5,66 ns p= 0,06 / χ 2 = 0,58 ns p= 0,75 n: number of cases %: percentage χ 2: chi-square ns: not signifcant p: probability Table 3 show the seroprevalence of anti- T. canis IgG and its distribution according to age group, with signifcant results in the Cumanacoa and San Juan parishes where the 0-10 age group was the most afected. Curiously, the chi-square test showed no signifcant association within this age group from the San Juan parish, which was also the most afected. Table 3 . Association between age and seroprevalence of anti-Toxocara canis IgG in inhabitants of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. IgG Anti-Toxocara canis in Inhabitants of San Juan / CumanacoaPositivesIndeterminateNegativesTotalAge (years)n%n%n%n% 0-10 55/3837,67/55,075/455,56/66,676/485,71/44,44 66/4640,74/54,7611-20 31/1321,23/18,841/011,11/00/0 0/0 32/1319,75/15,4821-30 21/914,38/13,041/011,11/00/3 0/33,33 22/1213,59/14,2931-40 13/1 8,90/1,450/0 0/01/114,29/11,11 14/2 8,64/2,3841-50 4/1 2,74/1,450/2 0/33,330/1 0/11,11 4/4 2,47/4,7651-60 11/4 7,53/5,802/022,22/00/0 0/0 13/4 8,02/4,7661-70 7/3 4,80/4,350/0 0/00/0 0/0 7/3 4,32/3,57(Continúa Tabla 3)
230 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. 71-80 3/0 2,05/00 0/00/0 0/0 3/0 1,85/081-90 1/0 0,70/00 0/00/0 0/0 1/0 0,62/0Total146/69 100/1009/6 100/1007/9 100/100162/84 100/100 χ 2 = 12.80 ns p= 0.69 n: number of cases %: percentage χ 2: chi-square ns: not signifcant *s: signifcant p: probability T able 4 show the seroprevalence of anti- T. canis IgG and its distribution according to handwashing practices before eating, food preparations and contact with soil by individuals in both parishes. Tere was no statistically signifcant association with any of the variables assessed by the chi-square test. Tis was contrary to the self-declared food handling and handwashing practices. Majority of those testing positive for anti- T. canis IgG antibodies individuals declared washing their hands before eating (84.93% San Juan parish) and (92.75% Cumanacoa parish) or cleaning their food together with handwashing before consumptio (75.34% for San Juan parish, and 85.51% for Cumanacoa parish). We also disclose that at the time of the surveys many of the people, particularly children, had dirty fngernails. In addition, if they washed their hands or wash their food (fruits and vegetables), they did so using water from the rivers that transversed these parishes. Te water used was neither treated nor boiled, further compounding to the environmental contamination of these uncooked foodstufs with T. canis eggs and other parasites. Te results from both parishes were comparable. Table 4 . Association between hand washing practices before eating, foodstuf handling parctices, contact with soil and seroprevalence of anti-Toxocara canis IgG in inhabitants of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela IgG ANTI-Toxocara canis in Inhabitants of San Juan / CumanacoaPositivesIndeterminateNegativesTotaln%n%n%n% Hand washing before eating Yes124/6484,93/92,757/577,78/83,337/8100/88,89138/7785,19/91,67No22/515,07/71,252/122,22/16,670/10/11,1124/714,81/8,33Total146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 1,62 ns p= 0,44 / χ 2 = 0,74 ns p= 0,69Foodstuf washing and cleaningYes110/5975,34/85,517/477,78/66,675/771,43/77,78122/7075,31/83,33No36/1024,66/14,492/222,22/66,672/228,57/22,2240/1424,69/16,67Total146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 0,09 ns p= 0,96 / χ 2 = 1,63 ns p= 0,44Contact with soilYes92/4763,01/68,127/377,78/506/485,71/44,44105/5464,81/64,29No54/2236,99/31,882/322,22/501/514,29/55,5657/3035,19/35,71Total146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 2,21 ns p= 0,33 / χ 2 = 2,52 ns p= 0,28 n: number of cases %: percentage χ 2: chi-square ns: not signifcant p: probability An analysis of anti- T. canis IgG seroprevalences and their distribution with regard to presence and contact with dogs by the participants from the two parishes is shown in table 5. Te sero-conversion status was not associated with the contact with dogs. (Continúa Tabla 3)
231 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Table 5 . Association between the presence of dogs, contact with dogs and the seroprevalence of anti- Toxocara canis IgG in inhabitants of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. IgG ANTI-Toxocara canis in Inhabitants of San Juan / CumanacoaPositivesIndeterminateNegativesTotaln%n%n%n% Dog presentYes99/2367,81/33,338/366,89/504/657,14/66,67111/3268,52/38,10No47/4632,19/66,671/311,11/503/342,86/33,3351/5231,48/61,90Total146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 2,19 ns p= 0,33 / χ 2 = 4,14 ns p= 0,13Contact with dogsYes60/4841,10/69,577/677,78/1003/442,86/44,4470/5843,20/69,04No86/2158,90/30,432/022,22/04/557,14/55,5692/2656,80/30,96Total146/69100/1009/6100/1007/9100/100162/84100/100 χ 2 = 4,659 ns p= 0,09 / χ 2 = 5,25 ns p= 0,07 n: number of cases %: percentage χ 2: chi-square ns: not signifcant p: probability Tese tables show the seroprevalence of anti- T. canis IgG and its distribution according to age, deworming and time of deworming of pet dogs of the participants from in both parishes. However, there were no signifcant associations established (Table 6). Table 6 . Role of age, deworming status, and time of deworming of pet dogs in the anti-Toxocara canis IgG seroprevalence of inhabitants of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. IgG ANTI-Toxocara canis in Inhabitants of San Juan / CumanacoaVariablesPositivesIndeterminateNegativesTotaln%n%n%n% Age of dog6 months-1 year41/1341,41/56,524/050/04/3100/5049/1644,14/50Older than 1 year58/1058,59/43,484/350/1000/30/5062/1655,86/50Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 5,47 ns p= 0,06 / χ 2 = 3,39 ns p= 0,18Received dewormerYes39/1439,40/60,873/237,60/33,331/525/83,3343/2138,74/65,63No60/960.60/39,135/162,50/66,673/175/16,6768/1161,26/34,37Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 0,34 ns p= 0,84 / χ 2 = 1,07 ns p= 0,59Deworming timeAt birth10/010,10/01/012,50/00/00/011/09,91/0Monthly11/011,11/00/00/00/00/011/09,91/0Every 6 months11/611,11/26,091/012,50/01/325/5013/911,71/28,13Annually7/87,07/34,781/212,50/33,330/20/33,338/127,21/37,50Never60/960,61/39,135/162,50/66,673/175/16,6768/1161,26/34,37Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 3,16 ns p= 0,92 / χ 2 = 3,35 ns p= 0,50 n: number of cases %: percentage χ 2: chi-square ns: not signifcant p: probability
232 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. It was observed that puppy owners do not provide adequate deworming treatment, of which after 3- or 4-weeks puppies begin to shed T. canis eggs freely into the soil, where they become infective. Consulted databases did not produce any published work on this parasitosis that considered these epidemiological variables. Both parishes had diferent patterns of behavior in relation to these items, but no statistically signifcant association was found in either parish. Tere was a statistically signifcant association with the collection of canine faeces by their owners in the parish of Cumanacoa, where 91.30% of the animals defecated in the yard and 78.26% of the owners did not collect the dog fecal matter. Tus, these faeces, together with the evolutionary forms of parasites in the contaminated fecal material can develop and be ready for infection (Table 7). Table 7 . Association between location of dog defecation, collection of dog faeces by dog owners, the frequency of dog faeces collection and disposal, and the seroprevalence of anti- Toxocara canis IgG in inhabitants of San Juan and Cumanacoa (Sucre and Montes municipality), Sucre state, Venezuela. IgG ANTI-Toxocara canis in Inhabitants of San Juan / CumanacoaVariablesPositivesIndeterminateNegativesTotaln%n%n%n% Location of dog defecationCourtyard81/2181,82/91,307/287,50/66,674/5100/83,3392/2882,88/87,50Ticket17/017,17/00/00/00/00/17/015,32/0House1/21,01/8,701/112,50/33,330/10/16,672/41,80/12,50Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 7,73 ns p= 0,10 / χ 2 = 1,59 ns p= 0,45Collection of faecesYes51/551,52/21,746/175/33,334/5100/66,6761/2654,96/81,26No48/1848,48/78,262/225/66,670/10/33,3350/645,05/18,75Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 5,05 ns p= 0,08 / χ 2 = 8,01 ns p= 0,02Contact with the siteYes51/2051,52/86,966/275/66,674/4100/83,3361/1154,96/34,37No48/348,48/13,042/125/33,330/20/16,6750/2145,05/65,63Total99/23100/1008/3100/1004/6100/100111/32100/100 χ 2 = 5,05 ns p= 0,08 / χ 2 = 8,01*s p= 0,02 n: number of cases %: percentage χ 2: chi-square ns: not signifcant *s: signifcant p: probability In summary, we have established prevalence of the Toxocara infection in human subjects, dogs (pets and strays) and in soil samples obtained from the vicinity of the homes and playgrounds. Tus, we have identifed two zoonotic foci of potential Toxocara sp. transmission and maintenance parasite life cycle in Sucre state. An epidemiological map was charted comparing the situation in Sucre state with other states in Venezuela (Fig. 2).
233 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Figure 2 . Prevalence of Toxocara sp. in Sucre state compared to states in Venezuela with existing reports of Toxocara sp. studies. DISCUSSION Te parish of San Juan, Sucre municipality, Sucre state, Venezuela, is a rural area dedicated to agriculture and animal husbandry (pigs, cattle, and poultry) usage. Tese activities are both a source of income and food, as well as an area of regional tourism. Te municipality of Montes carries out similar economic activities, especially agriculture and livestock farming with a special emphasis on sugar cane, cofee, vegetable crop production and cattle, poultry and pig rearing.We found high seroprevalence of T. canis (Table 1), nonetheless, globally, the prevalences reported are lower than those of this study: in Colombia, a prevalence of 39% was reported, in Argentina 37% and in Brazil 28% (Ulloque-Badaracco et al., 2023). However, a study in Paraguay (78%), but not in Turkey (7.6%) and Italy (31.87%) reports prevalences similar to those obtained in this study (Rivarola et al ., 2009; Akdemir, 2010; Qualizza et al ., 2011). In Venezuela, most studies on T. canis infection in humans have been carried out in the central north, west and south of the country. In Amazonas, a seroprevalence of 19.66% was found (Oliart-Guzmán et al. , 2014), in preschool children in Aragua state, Venezuela, were detected in 29% (65/224) of children. Te seroprevalence in the diferent preschools studied ranged between 4.2% and 60.6% (Martínez et al., 2015). In Zulia state, García et al. (2004) found a prevalence of 9.72% while a study conducted in Barquisimeto, a seroprevalence of 44.60% was reported (Delgado et al ., 2009).Few studies have been conducted on T. canis infection in eastern Venezuela. Gómez (2004), for instance, obtained a 77.14% prevalence in Sucre State, using the ELISA method, while Fernández et al. (2009), using the RIDASCREEN ELISA technique, reported a prevalence of 19.0% in Anzoátegui State. It is noteworthy that participants evaluated in this study, from both parishes, have high prevalence rates, a refection of zoonotic foci present in these communities.
234 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. Te seroprevalence of anti- T. canis IgG according to gender, was found that females were more afected in both San Juan and Cumanacoa parishes (Table 2). Comparable results were obtained by Espinoza et al. (2003) and Gómez (2004), who found a higher seroprevalence in females with 57.80% and 61.11%, respectively. However, these fndings difer from those by Martín et al. (2008), Delgado et al . (2009), Akdemir (2010) and Wisniewska et al. (2011) who report that 57.63%, 51.86%, 77.80% and 62.14%, respectively, were in males. Nevertheless, Roldán et al. (2010b) mentioned that the gender of the individual seems not to be an important predisposing factor for toxocariasis in human populations.Te results found in the age group of both municipalities coincide with those reported by various authors such as Aguiar-Santos et al. (2004) who reported a prevalence of 60% in children under 10 years of age and Gómez (2004) who reported a prevalence of 40.74% in children aged 2-4 years. Similarly, López et al . (2005) reported 67.70% in children aged 1-14 years while Fernández et al. (2009) recorded a prevalence of 22.50% in children aged 6 years and Rivarola et al. (2009) found a prevalence of 64.15% in children aged 3 to 9 years. However, none of these researchers found age-group specifc diferences in the occurrence of toxocariasis. García et al . (2004) suggested that the reduced risk of infection in older individuals might be due to improved hygienic habits and spending more time away from home thus distancing them from sources of infection. It was proposed that this parasitic infection was most likely to occur at home more than at school or at work, where sanitary and infrastructural conditions were better.It is important to highlight that in the two parishes; the schools are close to rivers and surrounded by felds, where children carry out their recreational activities, and where dogs defecate openly consequently depositing parasite eggs. Additionally, many of these communities and schools lack piped water and thus draw water from the rivers for consumption by children and residents. Subsequently, residents of these communities, including school children, are exposed to contracting this parasitosis both at home and in their schools.Tese results difer from those obtained by Acero et al. (2000), who found that 46.15% of children never washed their hands before eating and by Gómez (2004) who found that 62.96% of the Toxocara seropositive children did not wash their hands before eating, but 100% of them declared washing their food before consumption, particularly those to be eaten raw. Most participants from San Juan (63,01%) and Cumanacoa (68,12%), had contact with T. canis soil contaminated, had parasitized dogs and did not wash their hands well before eating or ingesting food. Tese participants were predisposed to high risk of easily getting infected. Tese rates were comparable to results found by Acero et al. (2000) who reported that 53.85% of toxocara-infected children consumed soil, had poor hygiene practices, López et al. (2003) found that 30% of toxocariasis positive children had a history of geophagy whilst Delgado et al. (2009) noted that 96.3% of the infected children had a history of geophagy.In both evaluated parishes, there were large tracts of land, contaminated with canine faeces, and consequently T. canis eggs. At the same time, these felds served as playgrounds for the children or fruit and vegetable gardens.Concerning the presence of dogs in the homesteads, 67.81% of the seropositive individuals in the San Juan parish stated that they had dogs as pets, which difered from those of Cumanacoa parish (33.33%). However, it should be noted that majority of the inhabitants from both parishes declared coming in contact with other canines in the area due to the large and uncontrolled population of stray dogs found in both parishes. Gómez (2004) found that 52.26% of seropositive participants had dogs while Martín et al. (2008) reported that 74.57% of those seropositive for IgG anti- T. canis antibodies owned dogs. However, in a study evaluating playgrounds in Turkey, Akdemir (2010) found that only 7.60% of those seropositive had dogs. We note that having dogs as pets was not signifcantly associated with testing positive for the IgG anti- T. canis antibodies.Fan et al . (2004) argued that toxocariasis is not related to dog breeding, but dog breeding is a condition that in most cases contributed to the occurrence of infection, especially in low socio-economic societies. Similarly, Alonso et al . (2004) pointed out that neither direct contact with dogs, even puppies, nor the presence of the animals in the home are sufcient conditions for acquiring T. canis infection; other epidemiological circumstances had to coincide to favor the risk of an infection being acquired from a contaminated environment.Some studies suggest that the likelihood of infection had more to do with individual susceptibility, and behavioral and hygienic habits rather than with the environmental factors present in the population. Considering this, we sought to understand if epidemiological variables such as age of the canines present, deworming and time of deworming of the individuals who indicated that they owned dogs and the seroprevalence of anti- T. canis IgG.
235 Zoonotic foci of Toxocara sp.: in state of Sucre, Venezuela Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Te majority of the seropositive individuals in the parish of San Juan stated that their dogs defecated in the yard (81.82%), 51.52% reported collecting dog faeces and frequenting the place of disposal, 27.27% disposed of these faeces in the thicket and 15.15% accumulated them until the waste collection day. Tis was indicative that these individuals also had frequent and direct contact with T. canis contaminated dog faeces in addition to being surrounded by animal waste subsequently increasing the possibility of parasite transmission and reinfection. In the databases consulted, no published studies on this parasitosis were found that took these epidemiological variables into account.One of the main sources of T. canis infection found within the homesteads are gardens and yards heavily contaminated with dog faeces. Additionally, the ease with which unwormed dogs access homes greately contributes to this contamination and high likelihood of infection. It is estimated that one gram of faeces from an infected animal can harbour about 10.000 T. canis eggs, while a female parasite can shed up to 200.000 eggs daily into the environment. Larval eggs can survive up to 10 years in the environment if conditions are right, thanks to their high resistance (García et al ., 2004).Taking into account the observations by García et al . (2004) and Alonso et al . (2004), geophagia, the high dog population, the manner in which dogs are kept without restrictions of space or specifc places to defecate, and the lack of collection of excrement from stray dogs, the parishes of San Juan and Cumanacoa have become a favorable place for the transmission of this zoonosis. In these localities, the prevalent conditions expose the inhabitants, especially children, to the parasite-contaminated excrement.Te population sample studied corresponded to individuals from two diferent socio-economic and cultural backgrounds, but with the same social defciencies. Tis could explain why no major diferences in risk factors association were found between serology-positive and serology-negative patients. Additionally, a representative sample of canine faeces was also taken and evaluated for parasitological contamination in these communities. In the parish of San Juan, 93.87% (46 of 49) of the canine faecal samples were parasitized of which the prevalence of T. canis was 18.37%. Similarly, in the parish of Cumanacoa, 65.40% (53 of 81) of the samples were parasitized, of which 9.90% were the T. canis species. Tis is indicative of which shows the risk of infection with T, canis in both communities. Tses fndings are similar to those of Ramírez-Barrios et al. (2004) and Devera et al. (2008) that showed a 11.4% and 16.7% prevalence of canine intestinal parasites in Venezuela, specifcally in the city of Maracaibo and Ciudad Bolívar.In addition to the faecal samples, soil samples were also analyzed. Of the 35 and 28 soil samples from San Juan and Cumanacoa parishes analyzed, 5.0% and 10.71% were positive for Toxocara sp. Eggs, respectively. We however declare that at the time of soil collection, the weather condition in the localities was unpredictable, with constant and torrential rains, such that the parasite eggs in the soil could have been washed by Surface run of into the rivers that fow through the communities. Despite the weather conditions, the presence of Toxocara sp. eggs in the soil of both parishes in Sucre municipality, Sucre State, Venezuela, demonstrates the high risk of contact and possible accidental ingestion by children in these communities. Tis subsequently predisposes them to acquiring the Visceral or Ocular Larva Migrans syndrome, in addition to advancing high infection rates in dogs. Very heterogeneous result was reported by Laiño et al. (2024), (1.5%) and Antonio et al. (2020) (32.5%).Comparing with other studies around Venezuela, from the two zoonotic foci evaluated in Sucre State demonstrates that this region has one of the highest seroprevalences of toxocariasis in humans. However, Anzoátegui State reports the highest prevalence of Toxocara sp. in canines while Zulia State has the highest rate of soil infected with Toxocara sp. eggs (Cazorla-Perfetti et al ., 2007; Tortolero et al ., 2008; Devera et al ., 2008; Fernández et al ., 2009; Nieves et al. , 2012; Apóstol et al., 2013). We are of the opinion that the high seroprevalence is attributed to the rural condition of both parishes evaluated in Sucre State. It is further compounded by the lack of a Zoonosis department in the local government to manage the large number of stray dogs in the communities and promote civic education on disease, dynamics of transmission, consequences of infection. As a zoonosis and geohelminthiasis, this parasitosis represents a public health problem for both parishes in Sucre State. Its epidemiology depends closely on maintaining the chain of infected domestic and/or stray dogs and cats, on human contact with soil contaminated with the infected and on the general habits and attitudes that tend to acquire the infection. It is crucial that the public health authorities implement a control and health education plan to raise awareness of the forms of contagion of this parasitosis and the most appropriate way to prevent it. Primary prevention should focus on veterinarial control of pets, culling of stray dog populations and improving habits and attitudes that tend to maintain the infection in both municipalities of Sucre State, Venezuela.
236 Neotropical Helminthology (Lima). Vol. 18, Nº2, jul - dec 2024 Gomez-Martinez et al. Author contributions : CRediT (Contributor Roles Taxonomy) EGM = Erika Gomez-Martinez MVGV = Márcia Viviana Gonçalves-Vallejos MBBR = María Betania Bravo-Rodríguez MTD = Marcos Tulio-Díaz ZSG = Zulay Simoni-Gonzalez DVG = Del Valle Guilarte EFS = Elizangela Farias-da Silva ZMS = Zeca Manuel-Salimo LSX = Luziana de Sousa-Xavier GOP = Grafe Oliveira-Pontes Conceptualization : EGM, GOP, ZSG, DVG Data curation : GOP, LSX, ZMS Formal Analysis : EGM, DVG, ZSG, GOP Funding acquisition : EGM, GOP, LSX, MBBR, MVGV, MTD, ZMS, ZSG, DVG, EFS Investigation : EGM, DVG, ZSG, MTD, MBBR, EFS Methodology : EGM, DVG, ZSG Project administration : EGM, DVG, ZSG Resources : EGM, DVG, ZSG Software : EGM, DVG, ZSG Supervision : EGM, ZMS, GOP Validation : EGM, GOP Visualization : EGM, GOP Writing – original draft : EGM, GOP, LSX, MVGV, ZMS Writing – review & editing : EGM, GOP, LSX, MBBR, MVGV, MTD, ZMS, ZSG, DVG, EFS ACKNOWLEDGMENTS Tis work was fnanced by Instituto de Investigaciones Cientifcas en Biomedicina y Ciencias Aplicadas, “Susan Tai” IIBCAUDO, the Universidad de Oriente, estate Sucre, Venezuela. Te authors would like to thank all the participants and their families who accepted to participate in the research and contributed to the study. BIBLIOGRAPHIC REFERENCES Acero, M., Muñoz, M., Flores, A., & Nicholis, R. (2000). Seroprevalencia de anticuerpos contra Toxocara canis y factores de riesgo en niños, Ciudad Bolívar, Bogotá. Biomédica (Bogotá), 21, 256-263. Aguiar-Santos, A., Andrade, L., & Medeiros, Z. (2004). Human toxocariasis: Frequency of anti Toxocara antibodies in children and adolescents from an outpatient clinic for lymphatic flariasis in Recife, Northcast, Brasil. Revista do Instituto de Medicina Tropical de São Paulo, 46 , 81-85.Akdemir, C. (2010). Visceral larva migrans among children in Kutahya (Turkey) and an evaluation of playgrounds for T. canis eggs. Te Turkish Journal of Pediatrics, 53, 158-162.Antonio, D.R., Daniel, A.L.V., José, V.R.F., Dario, A.R.I., & Yanitza, B.M.Y. (2020). Toxocara spp. y otros helmintos en muestras de suelo de plazas y parques de Puerto Ordaz, Municipio Caroní, Estado Bolívar, Venezuela. Kasmera , 48 , e48231618Apóstol, P., Pasceri, P., & Javitt-Jiménez, M. (2013). Detección de huevos de Toxocara sp. en suelos de tres parques públicos de la zona centro de Barquisimeto, estado Lara. Revista del Colegio de Médicos Veterinarios, 5, 38-47.Alonso, J., López, M., Bojanich, M., & Marull, J. (2004). Infección por Toxocara canis en población adulta sana de un área subtropical de Argentina. Parasitología latinoamericana, 59, 61-64.Barriga, O. (1988). A critical look at the Importance, prevalence and control of Toxocariasis and the possibilities of immunological control. Veterinary Parasitology, 29 , 195-234.Borge, H., & Ferreira, J. M. (2020). Leucocorias e seus diagnósticos diferenciais: Um relato de caso. Revista da Faculdade de Medicina de Teresópolis , 4, 42-50.
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