Open Access
Issue
Parasite
Volume 31, 2024
Article Number 25
Number of page(s) 6
DOI https://doi.org/10.1051/parasite/2024017
Published online 17 May 2024

© A.L.P. Fernandes et al., published by EDP Sciences, 2024

Licence Creative CommonsThis is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Introduction

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, an obligate intracellular coccidian protozoan. Felids are the only definitive hosts, since the T. gondii biological cycle is complete in these animals [2, 6]. Cats can excrete millions of oocysts and a single animal is capable of spreading infection to many hosts [6].

This multisystemic disease has three infective parasite forms (oocysts, tachyzoites and bradyzoites) and can be transmitted to cats mainly through ingestion of raw or undercooked meat containing cysts with bradyzoites. In humans, besides ingestion of cysts, transmission can also occur through sporulated oocysts from feces of infected cats in the environment and transplacentally (tachyzoites) [7, 23]. Risk factors associated with feline infection are sex, age, eating habits, coexistence with other species, and consumption of raw and undercooked meat and contaminated water [25].

Toxoplasma gondii seroprevalence in domestic cats in the world is 30–40%, and Brazil is highlighted as one of the countries with the most reports [20]. In the semi-arid region of Paraíba, a significant seroprevalence of 43.8% was observed in cats with IgG anti-T. gondii antibodies that presented age and hunting habits as risk factors [12]. By contrast, a seroprevalence of 26% was observed in cats from Rolim de Moura, state of Rondônia, North of Brazil, with no risk factors identified [26].

In Brazil, toxoplasmosis has a seroreactivity between 56.4% and 91.6% in women during pregnancy. Therefore, it is an important disease to One Health, especially in this group and in immunocompromized people. This disease has a great impact as it can result in miscarriage and serious neurodevelopment malformations, such as microcephaly and hydrocephalus. It is a major factor in global causes of infant morbidity and mortality [21, 28]. Furthermore, ocular toxoplasmosis is one of the relevant causes of uveitis in several countries and can explain up to 60% of cases of chorioretinitis [4].

Toxoplasmosis is an emerging, neglected zoonosis that is growing exponentially in Brazil. In urban areas, stray and domestic cats play a crucial role in transmission and maintenance of this agent as they are the only definitive hosts in direct contact with humans. Thus, health surveillance actions that assess seroprevalence and risk factors associated with feline toxoplasmosis are essential in order to obtain early diagnosis, reduce the risk of transmission, provide guidance for owners and develop prophylactic actions. For this reason, we selected a domestic cat population due to intense contact with humans and then aimed to characterize the epidemiological situation of toxoplasmosis in these cats in João Pessoa, capital of the state of Paraíba, Northeast region of Brazil.

Material and methods

Ethics

The present work met the standards for research involving animals in accordance with Regulation No. 38/18, which establishes restrictions on use of animals in research. The research was started only after we received a letter of approval from the Ethics Committee on the Use of Animals (CEUA) and CEP (research ethics committee) of the Federal University of Paraíba (UFPB), authorizing the research under Protocol number 3304170821. For an animal’s participation in the study, prior consent from the owner was required.

Area, sampling and blood collection procedures

The research was carried out in João Pessoa, state of Paraíba, Brazil and was performed at clinics and owners’ houses from October 2021 to February 2022. Domestic cats of both sexes aged between 1 and 15 years were selected. Cats aged under 12 months were not included in the study due to possible interference with serological tests due to antibody levels [1].

Number of blood samples collected for the experiment was determined by simple random sampling, as follows:

Meaning:

N = number of individuals sampled,

Z = normal distribution value for the 95% confidence level,

p = expected prevalence of 50%,

d = absolute error of 7%.

In this way, 267 feline blood samples were selected for collection. Blood collections were performed by puncturing the external jugular vein or cephalic vein using a disposable syringe and scalp with a maximum blood volume of 2 mL per cat at a clinic or the owner’s home with containment and collection according to cat friendly practices using bags, blankets and offering sachets and snacks.

The samples were stored in tubes without anticoagulant. Sera were separated from whole blood by centrifugation for 3,000 RPM at room temperature for 10 min with 80-2b Laboratory Centrifuge equipment with fixed rotor, divided into aliquots, transferred to 1.5 mL flat-bottom plastic microtubes, identified and stored at −20 °C.

Diagnosis by indirect fluorescent antibody test

Indirect fluorescent antibody test (IFAT) was performed to diagnose toxoplasmosis. Tests were performed to detect anti-T. gondii (IgG) according to the technique described by [3]. For this purpose, the RH strain of T. gondii (tachyzoites) was fixed on glass slides. Positive and negative controls were used on each slide for monitoring. Samples that demonstrated a reaction at a 1:16 dilution were classified as positive and then diluted sequentially, in multiples of four, until maximum reactive dilution for titration with the aim of measuring the amount of antibodies in each sample [5, 11].

Statistical analysis

In addition, a handwritten epidemiological questionnaire about the animal’s habits and health was administered orally by one interviewer to cat owners. It contained information regarding the following items and their respective categories: age (1 year/1–5 years/over 5 years), sex (female/male), street access (no/yes), castration (no/yes), type of food (pet food/homemade food/both), place where animals defecate (sandbox/yard/newspaper) and hunting habits (no/yes). These data were used to define factors associated with infection.

Analysis of factors associated with infection was divided into two moments. Firstly, a bivariate analysis was carried out where information obtained in epidemiological questionnaires (independent variables) was crossed with results of diagnostic tests (dependent variables) using chi-square or Fisher’s exact test at a confidence level of 20%. Secondly, independent variables that were significant were subjected to a multivariate analysis using Poisson regression with robust variance at a significance level of 5%. All procedures were performed in SPSS 25 for MAC.

Georeferencing

It was decided to carry out a spatial analysis in order to verify occurrence of patterns in geographic areas by checking distribution of georeferenced points, which were obtained through addresses of owners’ homes plotted in the Google Maps app, with a level of accuracy of 20 m. After tabulating georeferenced points, a non-parametric interpolation method based on Kernel density was carried out, which made it possible to estimate distribution intensities of heat points. Kernel width (1,944 m), interpolations, spatial resolution (X: 79.21; Y: 79.21), number of rows (268) and columns (250) were automatically adjusted by ArcGis 10.4 app. Digital maps of João Pessoa are available online by the city hall and the state and federal network by IBGE.

The city’s geographic limits were plotted on a digital georeferenced map of João Pessoa, based on a map of Brazilian municipalities from 2001, obtained from the João Pessoa city hall website. Plotting and processing digital maps were carried out using ArcGIS version 9.1.

Results and discussion

Seroprevalence of antibodies against T. gondii in cats was 17.22% (46/267; 95% CI = [12.7–21.8]). The titers ranged from 1:16 to 1:16,384, and the most frequent titers were 1:1,024 (9/46 animals; 19.5%) and 1:8,192 (9/46 animals; 19.5%) (Table 1).

Table 1

Antibody titers of anti-Toxoplasma gondii antibodies by IFAT of seropositive cats in the state of Paraíba, Northeast region of Brazil, in different dilutions.

The seropositivity of 17.22% demonstrates that domestic cats still have contact with T. gondii and produce antibodies against the protozoan. Thus, these animals play a role in toxoplasmosis epidemiology [6] since, in periods of immunosuppression, parasites that are in tissue cyst format can be reactivated and the feline can eliminate oocysts in feces [1].

Unlike other studies carried out in Northeast region of Brazil that detected prevalences of positive cats for Toxoplasma gondii varying between 47.7%, and 71.2%, Patos – Paraíba, and Fernando de Noronha – Pernambuco, respectively [12, 17], this research showed low seroprevalence. This possibly occurred because the studied population was made up only of domestic cats that had owners, unlike other studies where the majority of the population was stray animals. In these conditions, where owners provide care regarding nutrition, hygiene and veterinary care, there is a reduction in risk of infection [7].

Seroprevalence studies can be influenced by other factors such as the diagnostic technique used, cut-off point and where the target population lives. There are experiments that applied modified agglutination tests and indirect hemagglutination to analyze titer of anti-T. gondii in cats [12, 13]. In this research, ITAF was adopted as a diagnostic method since it has high sensitivity and specificity, in addition to being low cost and having specific conjugates for each species [15].

Table 2 presents the analysis of factors associated with T. gondii infection in cats from João Pessoa, where it was observed that the variables age (more than 60 months: RP = 5.744, CI = [1.368–24.121], p = 0.017) and region where they live (urban area: RP = 8.515, CI = [4.424–16.390], p < 0.0001) represented a significant risk.

Table 2

Factors associated with risk of Toxoplasma gondii infection in cats resulting from a univariate and multivariate analysis, from João Pessoa, state of Paraíba, Brazil, from October 2021 to February 2022.

Animals over 60 months were 5,744 times more likely to have anti-T. gondii. This factor associated with risk of protozoan infection is described as classic, since older animals have a greater chance of becoming infected due to the possibility of longer exposure [24, 27].

Another factor associated with Toxoplasma gondii infection was that cats lived in urban areas, which were 8,515 times more likely to be positive compared to those that did not live solely in that region. Felines in urban areas possibly acquire T. gondii by ingesting leftover food from humans, due to low hunting availability [10, 16, 19]. Although this study targeted domestic animals, the vast majority of which tend to eat pet food, some owners reported offering both pet food and homemade food to their animals. The practice of offering raw meat without heat treatment to cats is still adopted, which may favor occurrence of infected animals, as in the environment oocysts undergo sporulation, becoming infectious and are subsequently ingested by intermediate hosts through contaminated water and food. This is the most common way for cats and humans to acquire toxoplasmosis [6].

When analyzing Figure 1 where cases of T. gondii are distributed on Kernel map it is observed that areas with more intense colors represent the largest clusters of seropositive animals per km2. Most cases were concentrated in neighborhoods in the east and south of the city, which also have the largest number of inhabitants [14]. It is important to highlight that seropositive animals will not necessarily eliminate oocysts in their feces because in any given period of time only 1% of cats are found actively excreting oocysts [9], being felines in this location unlikely to transmit toxoplasmosis to humans [8]. We believe that in João Pessoa, the environmental characteristics of high rainfall can help oocysts to survive, spread and remain accessible to potential hosts.

thumbnail Figure 1

Map of the city of João Pessoa with estimated Kernel density for feline toxoplasmosis from October 2021 to February 2022.

In all cases, handling cat feces when collecting it from a litter box should be avoided by immunocompromized people and pregnant women, given the risks and possibilities of contamination [22]. It is also necessary to pay attention to more socioeconomically vulnerable populations, which in João Pessoa are more concentrated in south and west neighborhoods [18]. In these locations, campaigns are needed to educate the population about transmission of T. gondii, its impacts, effects and prevention, in addition to educating about the role of domestic cats in the epidemiological chain of the disease.

Conclusion

In conclusion, the seroprevalence of T. gondii infection in domestic cats in João Pessoa is considered low. However, it is important to emphasize that felines play an important role in the epidemiology of this disease. It is nonetheless suggested that cats in this research play a secondary role in transmission of toxoplasmosis. Therefore, it is crucial to reinforce information about prophylaxis and to monitor the epidemiological profile of the animal population.

Acknowledgments

The authors would like to thank all owners and their cats for kindly participating in the research.

Conflicts of interest

The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this article.

References

  1. Afonso E, Thulliez P, Gilot-Fromont E. 2006. Transmission of Toxoplasma gondii in an urban population of domestic cats (Felis catus). International Journal for Parasitology, 36(13), 1373–1382. [Google Scholar]
  2. Calero-Bernal R, Gennari SM. 2019. Clinical toxoplasmosis in dogs and cats: an update. Frontiers in Veterinary Science, 6, 54. [Google Scholar]
  3. Camargo ME. 1974. Introdução às técnicas de imunofluorescência. Revista Brasileira de Patologia Clínica, 10, 87–107. [Google Scholar]
  4. Cortés JÁ, Roncancio Á, Uribe LG, Cortés-Luna CF, Montoya JG. 2019. Approach to ocular toxoplasmosis including pregnant women. Current Opinion in Infectious Diseases, 32(5), 426–434. [Google Scholar]
  5. De Moura AB, Trevisani N, De Quadros RM, Ledo G, De Souza AP, Sartor AA. 2015. Anticorpos contra Toxoplasma gondii em gatos apreendidos pelo centro de controle de zoonoses de Lages, SC. Archives of Veterinary Science, 20(1), 1–7. [Google Scholar]
  6. Dubey JP, Cerqueira-cézar CK, Murata FHA, Kwok OCH, Yang Y, Su C. 2020. All about toxoplasmosis in cats: the last decade. Veterinary Parasitology, 283, 109145. [Google Scholar]
  7. Dubey JP, Lago EG, Gennari SM, Su C, Jones JL. 2012. Toxoplasmosis in humans and animals in Brazil: high prevalence, high burden of disease, and epidemiology. Parasitology, 139(11), 1375–1424. [Google Scholar]
  8. Dubey JP. 2010. Toxoplasmose de Animais e Humanos. CRC Press: Boca Raton, EUA. p. 1–30. [Google Scholar]
  9. Elmore SA, Jones JL, Conrad PA, Patton S, Lindsay DS, Dubey JP. 2010. Toxoplasma gondii: epidemiology, feline clinical aspects, and prevention. Trends in Parasitology, 26(4), 190–196. [Google Scholar]
  10. Feitosa TF, Costa FTR, Ferreira LC, Silva SS, Santos A, Silva WI, Brasil AWL, Vilela VLR. 2021. High rate of feline immunodeficiency virus infection in cats in the Brazilian semiarid region: Occurrence, associated factors and coinfection with Toxoplasma gondii and feline leukemia virus. Comparative Immunology, Microbiology and Infectious Diseases, 79, 101718. [Google Scholar]
  11. Feitosa TF, Vilela VLR, Dantas ES, Souto DVO, Pena HFJ, Athayde ACR, Azevêdo SS. 2014. Toxoplasma gondii and Neospora caninum in domestic cats from the Brazilian semi-arid: seroprevalence and risk factors. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 66(4), 1060–1066. [Google Scholar]
  12. Gonzales C, Vargas-Calla A, Gomez-Puerta LA, Robles K, Lopez-Urbina MT, Gonzalez AE. 2022. Seroprevalence of Toxoplasma gondii and associated risk factors in cats from Lima, Peru. Veterinary Parasitology: Regional Studies and Reports, 31, 100733. [Google Scholar]
  13. Huertas-López A, Sukhumavasi W, Álvarez-García G, Martínez-Subiela S, Cano-Terriza D, Almería S, Dubey JP, García-Bocanegra I, Cerón JJ, Martínez-Carrasco C. 2021. Seroprevalence of Toxoplasma gondii in outdoor dogs and cats in Bangkok, Thailand. Parasitology, 148(7), 843–849. [Google Scholar]
  14. IBGE. 2010. Instituto Brasileiro de Geografia e Estatística. Brasília: IBGE. Disponível em: https://www.ibge.gov.br/cidades-e-estados.html? [Google Scholar]
  15. Liu Q, Wang ZD, Huang SY, Zhu XQ. 2015. Diagnosis of toxoplasmosis and typing of Toxoplasma gondii. Parasites & Vectors, 8, 292. [Google Scholar]
  16. Lugoch G, Noro M, De Andrade J. 2018. Metanálise da prevalência de toxoplasmose em gatos e ovinos no Brasil. Revista de Ciência Veterinária e Saúde Pública, 6, 41–70. [Google Scholar]
  17. Magalhães FJR, Ribeiro-Andrade M, Souza FM, Lima Filho CDF, Biondo AW, Vidotto O, Navarro IT, Mota RA. 2017. Seroprevalence and spatial distribution of Toxoplasma gondii infection in cats, dogs, pigs and equines of the Fernando de Noronha Island, Brazil. Parasitology International, 66, 43–46. [Google Scholar]
  18. Maior MMS, Cândido GA. 2015. Vulnerabilidade socioeconômica: um estudo transversal para o município de João Pessoa – PB. Revista Principia – Divulgação Científica e Tecnológica do IFPB, João Pessoa, 24, 72–87. [Google Scholar]
  19. Meireles LR, Galisteo AJ Jr, Pompeu E, Andrade HF Jr. 2004. Toxoplasma gondii spreading in an urban area evaluated by seroprevalence in free-living cats and dogs. Tropical Medicine International Health, 9(8), 876–881. [Google Scholar]
  20. Montazeri M, Mikaeili Galeh T, Moosazadeh M, Sarvi S, Dodangeh S, Javidnia J, Daryani A. 2020. The global serogical prevalence of Toxoplasma gondii in felids during the last five decades (1967–2017): a systematic review and meta-analysis. Parasites & Vectors, 13, 82. [Google Scholar]
  21. Nascimento TL, Pacheco CM, De Sousa FF. 2017. Prevalência de Toxoplasma gondii em gestantes atendidas pelo Sistema Único de Saúde. Ciência & Saúde, 10, 96–101. [Google Scholar]
  22. Oliveira GMS, Simões JM, Schaer RE, Freire SM, Nascimento RJM, Pinheiro AMCM, Carvalho SMS, Mariano APM, Carvalho RC, Munhoz AD. 2019. Frequency and factors associated with Toxoplasma gondii infection in pregnant women and their pets in Ilhéus, Bahia, Brazil. Revista da Sociedade Brasileira de Medicina Tropical, 52, e20190250. [Google Scholar]
  23. Opsteegh M, Haveman R, Swart AN, Mensink-Beerepoot ME, Hofhuis A, Langelaar MF, van der Giessen JW. 2012. Seroprevalence and risk factors for Toxoplasma gondii infection in domestic cats in The Netherlands. Preventive Veterinary Medicine, 104(3–4), 317–326. [Google Scholar]
  24. Retmanasari A, Widartono BS, Wijayanti MA, Artama WT. 2017. Prevalence and risk factors for toxoplasmosis in Middle Java, Indonesia. EcoHealth, 14, 162–170. [Google Scholar]
  25. Rocha KS, Lima MS, Monteiro TRM, Honorio BET, Pinho APVB, Paz GSD, Scofield A, Cavalcante GG, Magalhães-Matos PC, Sampaio Junior FD, Abel I, Langoni H, Moraes CCG. 2020. Serological prevalence of Toxoplasma gondii infection in cats (Belém, Pará, Brazil). Revista Brasileira de Parasitologia Veterinária, 26, 29. [Google Scholar]
  26. Silva ALP, Lima EF, Silva Filho GM, Ferreira LC, Campos BA, Bison I, Brasil AWL, Parentoni RN, Feitosa TF, Vilela VLR. 2023. Seroepidemiological survey of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in domestic cats (Felis catus) in Rolim de Moura, State of Rondônia, North Brazil. Tropical Medicine and Infectious Disease, 2023(8), 220. [Google Scholar]
  27. Souza LZ, Rodrigues RGA, Oliveira DADD, Roman JL, Pinto SB, Bittencourt LHFB, Oyafuso MK. 2017. Soroprevalência de Toxoplasma gondii em gatos domiciliados em Palotina, Paraná, Brasil. Arquivos Ciência Veterinária e Zoologia UNIPAR, 20, 123–126. [Google Scholar]
  28. Teimouri A, Nassrullah OJ, Hedayati P, Bahreini MS, Alimi R, Mohtasebi S, Salemi AM, Asgari Q. 2022. Prevalence and predictors of Toxoplasma gondii infection in psychiatric inpatients in Fars Province, Southern Iran. Frontiers in Psychiatry, 13, 891603. [Google Scholar]

Cite this article as: Fernandes ALP, de Melo Alves M, Silva JO, Bison I, de Castro Tavares Silva A, Parentoni RN, dos Santos JRS, Feitosa TF, Vilela VLR & de Lima Brasil AW. 2024. Geoepidemiology, seroprevalence and factors associated with Toxoplasma gondii infection in domicilied cats from Paraíba (Brazil). Parasite 31, 25.

All Tables

Table 1

Antibody titers of anti-Toxoplasma gondii antibodies by IFAT of seropositive cats in the state of Paraíba, Northeast region of Brazil, in different dilutions.

Table 2

Factors associated with risk of Toxoplasma gondii infection in cats resulting from a univariate and multivariate analysis, from João Pessoa, state of Paraíba, Brazil, from October 2021 to February 2022.

All Figures

thumbnail Figure 1

Map of the city of João Pessoa with estimated Kernel density for feline toxoplasmosis from October 2021 to February 2022.

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