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      Comparison of Internal Transcribed Spacers and Intergenic Spacer Regions of Five Common Iranian Sheep Bursate Nematodes

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          Abstract

          Background

          Accurate identification of sheep nematodes is a critical point in epidemiological studies and monitoring of drug resistance in flocks. However, due to a close morphological similarity between the eggs and larval stages of many of these nematodes, such identification is not a trivial task. There are a number of studies showing that molecular targets in ribosomal DNA (Internal transcribed spacer 1, 2 and Intergenic spacer) are suitable for accurate identification of sheep bursate nematodes. The objective of present study was to compare the ITS1, ITS2 and IGS regions of Iranian common bursate nematodes in order to choose best target for specific identification methods.

          Methods

          The first and second internal transcribed spacers (ITS1and ITS2) and intergenic spacer (IGS) of the ribosomal DNA (rDNA) of 5 common Iranian bursate nematodes of sheep were sequenced. The sequences of some non–Iranian isolates were used for comparison in order to evaluate the variation in sequence homology between geographically different nematode populations.

          Results

          Comparison of the ITS1 and ITS2 sequences of Iranian nematodes showed greatest similarity among Teladorsagia circumcincta and Marshallagia marshalli of 94% and 88%, respectively. While Trichostrongylus colubriformis and M. marshalli showed the highest homology (99%) in the IGS sequences. Comparison of the spacer sequences of Iranian with non-Iranian isolates showed significantly higher variation in Haemonchus contortus compared to the other species.

          Conclusion

          Both the ITS1 and ITS2 sequences are convenient targets to have species-specific identification of Iranian bursate nematodes. On the other hand the IGS region may be a less suitable molecular target.

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          Most cited references20

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          Helminths, arthropods and protozoa of domesticated animals

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            The use of nuclear ribosomal DNA markers for the identification of bursate nematodes (order Strongylida) and for the diagnosis of infections.

            Many bursate nematodes are of major importance to animal health. Animals are often parasitized by multiple species that differ in their prevalence, relative abundance and/or pathogenicity. Implementation of effective management strategies for these parasites requires reliable methods for their detection in hosts, identification to the species level and measurement of intensity of infection. One major problem is the difficulty of accurately identifying and distinguishing many species of bursate nematode because of the remarkable morphological similarity of their eggs and larvae. The inability to identify, with confidence, individual nematodes (irrespective of their life-cycle stage) to the species level by morphological methods has often led to a search for species-specific genetic markers. Studies over the past 15 years have shown that sequences of the internal transcribed spacers of ribosomal DNA provide useful genetic markers, providing the basis for the development of PCR-based diagnostic tools. Such molecular methods represent powerful tools for studying the systematics, epidemiology and ecology of bursate nematodes and, importantly, for the specific diagnosis of infections in animals and humans, thus contributing to improved control and prevention strategies for these parasites.
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              Genetic markers for strongylid nematodes of livestock defined by PCR-based restriction analysis of spacer rDNA.

              Twenty-four species of parasitic nematode (order Strongylida) from sheep, goats, cattle or pigs were characterised using a polymerase chain reaction-linked restriction fragment length polymorphism technique (PCR-RFLP). The ribosomal (r)DNA region spanning the first internal transcribed spacer (ITS-1), 5.8S rRNA gene and the second internal transcribed spacer (ITS-2) (designated ITS) was amplified from genomic DNA by polymerase chain reaction (PCR), digested separately with four restriction endonucleases (RsaI, HinfI, DraI or NlaIII) and the fragments separated by agarose gel electrophoresis. The PCR products amplified from all species appeared as a single band of approximately 870 bp in size, except for Ostertagia ostertagi whose product was approximately 1250 bp. The PCR-RFLP analysis of ITS revealed characteristic restriction patterns for all species, except for C. surnabada and C. oncophora which had identical patterns. The study demonstrated that ITS contains useful genetic markers for the identification of a range of strongylid nematodes of livestock. These markers should be of use in specific PCR assays for the identification of developmental stages of the parasites where morphological characters are unreliable.
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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
                September 2014
                : 9
                : 3
                : 350-357
                Affiliations
                [1 ]Dept. of Pathobiology, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran
                [2 ]School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
                [3 ]Teagasc, Animal Production Research Centre, Athenry, Co Galway, Ireland
                [4 ]Dept. of Parasitology, Faculty of Veterinary Medicine , University of Tehran, Tehran, Iran
                Author notes
                [* ]Correspondence theo.dewaal@ 123456ucd.ie
                Article
                4316566
                25678919
                c67b4fb5-123d-4215-aa83-bfd68c1953c9
                Copyright: © Iranian Journal of Parasitology & Tehran University of Medical Sciences

                This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.

                History
                : 20 February 2014
                : 12 June 2014
                Categories
                Original Article

                Parasitology
                internal transcribed spacers,intergenic spacer,similarity score,bursate nematodes
                Parasitology
                internal transcribed spacers, intergenic spacer, similarity score, bursate nematodes

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