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      Differential Detection of Echinococcus Spp. Copro-DNA by Nested-PCR in Domestic and Wild Definitive Hosts in Moghan Plain, Iran

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          Abstract

          Background

          Despite Echinococcus granulosus, there are merely two old reports of E. multilocularis infection among Iranian canids of Moghan Plain, the only area known endemic for the species. We detected specific DNA markers in fecal samples by PCR (Copro-PCR) for differential diagnosis of Echinococcus species in living canids.

          Methods

          Totally 144 fecal samples from domestic dogs, red foxes and a golden jackal were examined for genus-specific Echinococcus coproantigens using ELISA. Forty two positive or ambiguous samples were further examined for Echinococcus species-specific DNA markers by two different set of nested-PCR.

          Results

          Twenty five out of 144 (17.4%) animals were contaminated with E. granulosus including 14 (23.7%) domestic dogs, 10 (11.9%) red foxes and one (100%) golden jackal. But none of them harboured E. multilocularis species-specific Copro-DNA. The overall prevalence of E. granulosus and E. multilocularis infections in canids of the area was estimated to be 17.4% and 0.0%, respectively. There was a significant relation between the results of Copro-PCR and CA-ELISA.

          Conclusion

          The lack of E. multilocularis infection, compared to previous reports may be due to the differences in used diagnostic methods and/or recently limited territories of wild canids and altered their food resources in this particular area.

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          Most cited references29

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          Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA.

          A multiplex polymerase chain reaction (PCR) was evaluated for the identification of morphologically indistinguishable eggs of the taeniid tapeworms from carnivores using primers targeting mitochondrial genes. The primers for Echinococcus multilocularis (amplicon size 395 bp) were species-specific as assessed by in silico analysis and in the PCR using well-defined control samples. The design of primers that specifically amplify DNA from E. granulosus or Taenia spp. was not possible. The primers designed for E. granulosus also amplified DNA (117 bp) from E. vogeli, and those designed for Taenia spp. amplified products (267 bp) from species of Mesocestoides, Dipylidium and Diphyllobothrium. Nevertheless, as our diagnostic approach includes the concentration of taeniid eggs by sequential sieving and flotation, followed by their morphological detection, this non-specificity has limited practical importance. Sequence analysis of the corresponding amplicon can identify most of the described E. granulosus genotypes. Taenia spp. can be identified by direct sequencing of the 267 bp amplicon, or, for most species, by restriction fragment length polymorphism (RFLP) analysis. The multiplex PCR was readily able to detect 1 egg (estimated to contain 7000 targets, as determined by quantitative PCR). Having been validated using a panel of well-defined samples from carnivores with known infection status, this approach proved to be useful for the identification of taeniid eggs from both individual animals and for epidemiological studies.
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            The present status of human helminthic diseases in Iran.

            M Rokni (2008)
            Over the last few decades there have been several marked changes in the human helminthiases found in Iran. Fascioliasis is emerging as an important chronic disease of humans, especially in the northern province of Gilan (where outbreaks in 1989 and 1999 involved >7000 and >10,000 cases, respectively) and, more recently, in the western province of Kermanshah. In contrast, no cases of urinary schistosomiasis, a disease that once affected thousands of individuals in south-western Khuzestan province, have been reported in Iran in recent years, and no cases of dracunculiasis have been seen in the country since the mid-1970s. Approximately 1% of all admissions to surgical wards are attributable to cystic echinococcosis, which is still considered endemic, but only a few cases of alveolar echinococcosis have been recorded. Over the last decade, there appears to have been a generally downward trend in the incidence of intestinal helminthiases in Iran. Recent estimates of the prevalences of ascariasis and strongyloidiasis, for example, lie between just 0.1% and 0.3%, and <1% of the population now appears to be infected with hookworm. In contrast, human infection with Hymenolepis and Enterobius remains relatively common. There have been a few case reports of toxocariasis and a few sero-epidemiological investigations of this disease but problems in accurate diagnosis have prevented good estimates of the general prevalence of this nematode infection. Just nine cases of pentastomiasis (all caused by Linguatula), 12 of dirofilariasis, one of gongylonemiasis, and three of moniliformiasis have been formally recorded in Iran.
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              Present situation of echinococcosis in the Middle East and Arabic North Africa.

              Echinococcosis is one of the major zoonotic parasitic diseases in the Middle East and Arabic North Africa from Morocco to Egypt. Both cystic and alveolar echinococcosis has been reported from these areas. However, cystic echinococcosis is more prevalent and has been reported from all countries in the Middle East and Arabic North Africa. Alveolar echinococcosis is less prevalent and has been reported only from Iran, Turkey, Iraq and Tunisia. Present situation of echinococcosis in dogs and other definitive hosts, animal intermediate hosts and humans in the Middle East and Arabic North Africa has been reviewed. Echinococcus granulosus is highly prevalent in Iran, Turkey, Iraq, Morocco, Tunisia, and Libya. In the Levant countries, the cystic echinococcosis is also highly endemic. In Oman, it is endemic with low prevalence and a very low level in Cyprus. Various surveys have indicated that hydatid cysts are commonly found in sheep, cattle, goats and camels throughout the Middle East and Arabic North Africa. Sheep are the most infected animals of these regions. Most of studies on human have been focused on surgical reports although several population studies have been performed using serological and imaging techniques. Human cystic echinococcosis (CE) is prevalent in the Middle East and Arabic North Africa. It is hyper endemic in Iran, Turkey, Iraq, Jordan, Morocco, Libya, Tunisia, and Algeria, and endemic in Egypt. Studies on the strain specificities of E. granulosus in the Middle East revealed sheep strain (G1) present in sheep, goats, cattle, camels and humans, and the camel strain (G6) in camels, sheep, cattle as well as humans. Dog/sheep strain seems to be more prevalent in the foregoing regions in documented reports from Iran and Jordan. However, a strain of E. granulosus, which resembles the horse strain (G4) strain, has been reported from Jordan. Strain specifications of E. granulosus in Arabic North Africa showed that sheep/dog strain (G1) have been reported from Tunisia and Libya both from humans and animals. However, in Egypt the human cases reported are of camel/dog strain.
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                Author and article information

                Journal
                Iran J Parasitol
                Iran J Parasitol
                IJPA
                Iranian Journal of Parasitology
                Tehran University of Medical Sciences
                1735-7020
                2008-238X
                Jan-Mar 2013
                : 8
                : 1
                : 107-113
                Affiliations
                [1 ]Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
                [2 ]Immunology of Infectious Diseases Research Center (IIDRC), Rafsanjan University of Medical Sciences, Rafsanjan, Iran
                [3 ]Department of Medical Parasitology, Pasteur Institute of Iran, Tehran, Iran
                [4 ]Dept. of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
                Author notes
                [* ] Corresponding author: Tel.: +98 913 1936267, Email: mzarebidaki@ 123456gmail.com
                Article
                IJPA-8-107
                3655248
                23682268
                502c4135-d250-4b01-a583-1fd308b5d8f5
                © 2013 Iranian Society of Parasitology & Tehran University of Medical Sciences

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 21 May 2012
                : 14 December 2012
                Categories
                Original Article

                Parasitology
                echinococcus,copro-dna,canids,iran
                Parasitology
                echinococcus, copro-dna, canids, iran

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