Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-18T03:04:01.925Z Has data issue: false hasContentIssue false

Molecular characterization of Toxocara spp. eggs isolated from public parks and playgrounds in Shiraz, Iran

Published online by Cambridge University Press:  07 May 2018

M. Choobineh
Affiliation:
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
F. Mikaeili*
Affiliation:
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
S.M. Sadjjadi*
Affiliation:
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
S. Ebrahimi
Affiliation:
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
S. Iranmanesh
Affiliation:
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
*
Authors for correspondence: F. Mikaeili, Fax.: +98 71 32305291, E-mail: mikaeelf@sums.ac.ir and S.M. Sadjjadi, Fax.: +98 71 32305291, E-mail: smsadjjadi@sums.ac.ir
Authors for correspondence: F. Mikaeili, Fax.: +98 71 32305291, E-mail: mikaeelf@sums.ac.ir and S.M. Sadjjadi, Fax.: +98 71 32305291, E-mail: smsadjjadi@sums.ac.ir

Abstract

Human toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

Type
Research Paper
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Akhlaghi, L, et al. (2006) An investigation on the toxocariasis seroprevalence in children (2–12 years old) from Mahidasht area of Kermanshah Province 2003–2004. Razi Journal of Medical Sciences 13, 4148.Google Scholar
Alavi, SM, et al. (2009) Clinical spectrum and diagnostic tools of fever of unknown origin among hospitalized patients in Razi Hospital (2006–2008), Ahvaz. Jundishapur Journal of Microbiology 2, 152157.Google Scholar
Azizi, S, et al. (2007) Histopathologic changes and larval recovery of Toxocara cati in experimentally infected chickens. Parasitology Research 102, 4752.Google Scholar
Berenji, F, et al. (2015) Soil contamination with Toxocara spp. eggs in public parks of Mashhad and Khaf, north east of Iran. Iranian Journal of Parasitology 10, 286.Google Scholar
Borecka, A (2004) Differentiation of Toxocara spp. eggs isolated from the soil by the PCR-linked RFLP method. Helminthologia 41, 185187.Google Scholar
Borecka, A and Gawor, J (2008) Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs. Journal of Helminthology 82, 119122.Google Scholar
Changizi, E, et al. (2007) Gastrointestinal helminthic parasites in stray cats (from north of Iran). Iranian Journal of Parasitology 2, 2529.Google Scholar
Choi, D, et al. (2012) Transmission of Toxocara canis via ingestion of raw cow liver: a cross-sectional study in healthy adults. Korean Journal of Parasitology 50, 2327.Google Scholar
Cianferoni, A, et al. (2006) Visceral larva migrans associated with earthworm ingestion: clinical evolution in an adolescent patient. Pediatrics 117, e336339.Google Scholar
Dado, D, et al. (2012) Detection of zoonotic intestinal parasites in public parks of Spain. Potential epidemiological role of microsporidia. Zoonoses and Public Health 59, 2328.Google Scholar
Deplazes, P, et al. (2011) Role of pet dogs and cats in the transmission of helminthic zoonoses in Europe, with a focus on echinococcosis and toxocarosis. Veterinary Parasitology 182, 4153.Google Scholar
Despommier, D (2003) Toxocariasis: clinical aspects, epidemiology, medical ecology, and molecular aspects. Clinical Microbiology Reviews 16, 265272.Google Scholar
Dubná, S, et al. (2007) Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic. Veterinary Parasitology 15, 8186.Google Scholar
Emamapour, SR, Borji, H and Nagibi, A (2015) An epidemiological survey on intestinal helminths of stray dogs in Mashhad, north-east of Iran. Journal of Parasitic Disease 39, 266271.Google Scholar
Fahrion, A, et al. (2011) Toxocara eggs shed by dogs and cats and their molecular and morphometric species-specific identification: is the finding of T. cati eggs shed by dogs of epidemiological relevance? Veterinary Parasitology 177, 186189.Google Scholar
Fallah, M, Azimi, A and Taherkhani, H (2007) Seroprevalence of toxocariasis in children aged 1–9 years in western Islamic Republic of Iran, 2003. Eastern Mediterranean Health Journal 13, 10731077.Google Scholar
Fogt-Wyrwas, R, Jarosz, W and Mizgajska-Wiktor, H (2007) Utilizing a polymerase chain reaction method for the detection of Toxocara canis and T. cati eggs in soil. Journal of Helminthology 81, 7578.Google Scholar
Gao, X, et al. (2017) Influence of land use and meteorological factors on the spatial distribution of Toxocara canis and Toxocara cati eggs in soil in urban areas. Veterinary Parasitology 15, 8085.Google Scholar
Garedaghi, Y and Shabestari, AS (2012) Prevalence of Toxocara eggs in public parks of Tabriz city, center of east Azarbaijan Province, Iran. Veterinary Clinical Pathology 5, 357361.Google Scholar
Gasser, RB, et al. (2006) Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. Nature Protocols 1, 31213128.Google Scholar
Gawor, J, et al. (2014) Risk of human toxocarosis in Poland due to Toxocara infection of dogs and cats. Acta Parasitologica 60, 99104.Google Scholar
Ghashghaei, O, et al. (2016) Contamination of soil samples of public parks with Toxocara spp. eggs in Kermanshah, Iran. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 42, 4750.Google Scholar
Hajipour, N, et al. (2016) A survey study on gastrointestinal parasites of stray cats in Azarshahr, (East Azerbaijan province, Iran). Journal of Parasitic Disease 40, 12551260.Google Scholar
Hezarjaribi, HZ, et al. (2016) Parasitic contamination of surface and deep soil in different areas of Sari in north of Iran. Journal of Coastal Life Medicine 4, 861864.Google Scholar
Holland, CV and Smith, HV (2006) Toxocara: the enigmatic parasite. Wallingford, CAB International.Google Scholar
Hosseini-Safa, A, et al. (2015) Seroepidemiology of toxocariasis in children (5–15 yr old) referred to the pediatric clinic of Imam Hossein Hospital, Isfahan, Iran. Iranian Journal of Parasitology 10, 632.Google Scholar
Ito, K, et al. (1986) Three cases of visceral larva migrans due to ingestion of raw chicken or cow liver. Journal of Japanese Society of Internal Medicine 75, 759766.Google Scholar
Khademvatan, S, Abdizadeh, R and Tavalla, M (2014) Molecular characterization of Toxocara spp. from soil of public areas in Ahvaz southwestern Iran. Acta Tropica 135, 5054.Google Scholar
Khazan, H, et al. (2012) Prevalence of Toxocara spp. eggs in public parks in Tehran City, Iran. Iranian Journal of Parasitology 7, 38.Google Scholar
Lescano, SZ, Nakhle, MC and Chieffi, PP (1998) Effect of ‘in vitro’ cultivation time on the infectivity of Toxocara canis eggs. Revista do Instituto de Medicina Tropical de São Paulo 40, 201202.Google Scholar
Li, MW, et al. (2007) PCR tools for the verification of the specific identity of ascaridoid nematodes from dogs and cats. Molecular and Cellular Probes 21, 349354.Google Scholar
Maleki, B, et al. (2016) Soil contamination of public places with Toxocara spp. eggs in Kermanshah, Iran, in 2014. Novelty in Biomedicine 4, 105109.Google Scholar
Maraghi, S, et al. (2012) Seroprevalence of toxocariasis in hypereosinophilic individuals in Ahwaz, south-western Iran. Journal of Helminthology 86, 241244.Google Scholar
Maraghi, S, et al. (2014) Study on the contamination of Abadan public parks soil with Toxocara spp. eggs. Journal of Environmental Health Science and Engineering 12, 86.Google Scholar
Mehrabani, D, Sadjjadi, SM and Oryan, A (2002) Prevalence of gastrointestinal nematode parasites in stray dogs in Shiraz, Southern Iran. Journal of Applied Animal Research 22, 157160.Google Scholar
Mikaeili, F, et al. (2013) Toxocara nematodes in stray cats from Shiraz, southern Iran: intensity of infection and molecular identification of the isolates. Iranian Journal of Parasitology 8, 593600.Google Scholar
Mikaeili, F, et al. (2017) Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions. Acta Parasitologica 26, 549556.Google Scholar
Motazedian, H, et al. (2006) Prevalence of helminth ova in soil samples from public places in Shiraz. Eastern Mediterranean Health Journal 12, 562565.Google Scholar
Nijsse, R, et al. (2015) Environmental contamination with Toxocara eggs: a quantitative approach to estimate the relative contributions of dogs, cats and foxes, and to assess the efficacy of advised interventions in dogs. Parasites and Vectors 28, 397.Google Scholar
Nourian, AA, et al. (2008) Seroepidemiological study for toxocariasis among children in Zanjan-northwest of Iran. Pakistan Journal of Biological Sciences 11, 1844.Google Scholar
Otero, D, et al. (2017) Environmental contamination with Toxocara spp. eggs in public parks and playground sandpits of Greater Lisbon, Portugal. Journal of Infection and Public Health 11, 9498.Google Scholar
Overgaauw, PAM (1997) Aspects of Toxocara epidemiology: toxocarosis in dogs and cats. Critical Reviews in Microbiology 23, 233251.Google Scholar
Ozlati, M, et al. (2016) Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool. Veterinary World 9, 14711477.Google Scholar
Paller, VG and de Chavez, ER (2014) Toxocara (Nematoda: Ascaridida) and other soil-transmitted helminth eggs contaminating soils in selected urban and rural areas in the Philippines. Scientific World Journal 2014, 386232386237.Google Scholar
Paquet-Durand, I, et al. (2007) Prevalence of Toxocara spp., Toxascaris leonina and ancylostomidae in public parks and beaches in different climate zones of Costa Rica. Acta Tropica 104, 3037.Google Scholar
Pawar, RM, et al. (2012) Detection and molecular characterization of ascarid nematode infection (Toxascaris leonina and Toxocara cati) in captive Asiatic lions (Panthera leopersica). Acta Parasitologica 57, 6773.Google Scholar
Pezeshki, A, et al. (2017) Detection of Toxocara spp. eggs in the soil of public places in and around of Ardabil City, northwestern Iran. Iranian Journal of Parasitology 12, 136.Google Scholar
Rubinsky-Elefant, G, et al. (2010) Human toxocariasis: diagnosis, worldwide seroprevalences and clinical expression of the systemic and ocular forms. Annals of Tropical Medicine and Parasitology 104, 323.Google Scholar
Sadjjadi, SM, et al. (2000) Seroprevalence of Toxocara infection in school children in Shiraz, southern Iran. Journal of Tropical Pediatrics 46, 327330.Google Scholar
Sadjjadi, SM, et al. (2001) Prevalence and intensity of infestation with Toxocara cati stray cats in Shiraz, Iran. Veterinarski Archiv 71, 149157.Google Scholar
Saraei, M, et al. (2012) Contamination of soil and grass to Toxocara spp. eggs in public parks of Qazvin, Iran. Asian Pacific Journal of Tropical Biomedicine 2, S1156S1158.Google Scholar
Sardarian, K, et al. (2015) Prevalence of zoonotic intestinal parasites in household and stray dogs in rural areas of Hamadan, Western Iran. Tropical Biomedicine 32, 240246.Google Scholar
Schantz, PM (1989) Toxocara larva migrans now. The American Journal of Tropical Medicine and Hygiene. 41, 2134.Google Scholar
Shahraki, MK, et al. (2017) Epidemiological study of Toxocar canis in children under 14-years-old and dogs in Zabol and Chabahar Districts, southeast of Iran. Iranian Journal of Parasitology 12, 101107.Google Scholar
Sharif, M, et al. (2007) Toxocara cati infections in stray cats in northern Iran. Journal of Helminthology 81, 6366.Google Scholar
Studzińska, MB, et al. (2017) Variations in the rate of infestations of dogs with zoonotic nematodes and the contamination of soil in different environments. International Journal of Environmental Research and Public Health 14, E1003.Google Scholar
Taher, HM (2017) Soil contamination with intestinal parasites eggs in public parks and playgrounds in Kirkuk city. Tikrit Journal of Pure Science 22, 5255.Google Scholar
Tamura, K, et al. (2011) MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28, 27312739.Google Scholar
Tan, JS (1997) Human zoonotic infections transmitted by dogs and cats. Archives of Internal Medicine 157, 19331943.Google Scholar
Tavalla, M, et al. (2012) Prevalence of parasites in soil samples in Tehran public places. African Journal of Biotechnology 11, 45754578.Google Scholar
Tavassoli, M, et al. (2008) Toxocara spp. eggs in public parks of Urmia City, West Azerbaijan Province Iran. Iranian Journal of Parasitology 3, 2429.Google Scholar
Thomas, D and Jeyathilakan, N (2014) Detection of Toxocara eggs in contaminated soil from various public places of Chennai city and detailed correlation with literature. Journal of Parasitic Diseases 38, 174180.Google Scholar
Vafae Eslahi, A, et al. (2017) Road killed carnivores illustrate the status of zoonotic helminthes in Caspian Sea littoral of Iran. Iranian Journal of Parasitology 12, 230235.Google Scholar
Wiwanitkit, V and Waenlor, W (2004) The frequency rate of Toxocara species contamination in soil samples from public yards in a urban area ‘Payathai’, Bangkok, Thailand. Revista do Instituto de Medicina Tropical de São Paulo 46, 113114.Google Scholar
Zibaei, M and Sadjjadi, SM (2010) Prevalence of helminth ova in soil samples from public places in Shiraz. Eastern Mediterranean Health Journal 16, 580.Google Scholar
Zibaei, M and Sadjjadi, SM (2017) Trend of toxocariasis in Iran: a review on human and animal dimensions. Iranian Journal of Veterinary Research 18, 233242.Google Scholar
Zibaei, M and Uga, S (2008) Contamination by Toxocara spp. eggs in sandpits in Kobe, Japan. Journal of Environmental Control Technology 26, 3237.Google Scholar
Zibaei, M, Sadjjadi, SM and Sarkari, B (2007) Prevalence of Toxocara cati and other intestinal helminths in stray cats in Shiraz, Iran. Tropical Biomedicine 24, 3943.Google Scholar
Zibaei, M, et al. (2010) Soil contamination with Toxocara spp. eggs in the public parks from three areas of Khorram Abad, Iran. Nepal Medical College Journal 12, 6365.Google Scholar
Zibaei, M, Sadjjadi, SM and Jahadi-Hosseini, SH (2014) Toxocara cati larvae in the eye of a child: a case report. Asian Pacific Journal of Tropical Biomedicine 4, S5355.Google Scholar
Zibaei, M, Sadjjadi, SM and Maraghi, S (2017) The occurrence of Toxocara species in naturally infected broiler chickens revealed by molecular approaches. Journal of Helminthology 91, 633636.Google Scholar