Molecular Characterization of the First Internal Transcribed Spacer of Ribosomal DNA of the Most Common Species of Eyeworms (Thelazioidea: Thelazia)

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Abstract

Spirurid nematodes of the genus Thelazia cause ocular infection in several mammals and are transmitted by dipteran flies. Of the 10 described species, T. gulosa, T. rhodesi, T. skrjabini (affecting cattle), T. lacrymalis (affecting horses), and T. callipaeda (affecting carnivores and humans) are the most common. The aim of this study was to characterize the first internal transcribed spacer (ITS1) ribosomal DNA sequences of these 5 species. The length of the ITS1 sequences ranged from 357 bp (T. lacrymalis) to 905 bp (T. callipaeda). Interspecific differences ranged from 35 to 77%. Polymorphic sites were detected in each species. Intraspecific variation varied from 0.3 to 2.5%. The characteristics of the ITS1 of Thelazia spp. show similarities to those of other Spirurida. The findings of this present study show that the ITS1 represent a powerful genetic marker for the molecular identification of eyeworms and a useful tool for developing molecular epidemiological techniques to study Thelazia spp. transmission patterns and prevalence in definitive and intermediate hosts.

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Most cited references 27

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Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer.

Michael Blouin (2002)

DNA sequence divergence at internal transcribed spacer regions (ITS-1 and ITS-2) was compared with divergence at mitochondrial cox1 or nad4 loci in pairs of congeneric nematode species. Mitochondrial sequences accumulate substitutions much more quickly than internal transcribed spacer, the difference being most striking in the most closely related species pairs. Thus, mitochondrial DNA may be the best choice for applications in which one is using sequence data on small numbers of individuals to search for potential cryptic species. On the other hand, internal transcribed spacer remains an excellent tool for DNA diagnostics (quickly distinguishing between known species) owing to its lower level of intraspecific polymorphism.

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Current status and epidemiological observation of Thelazia callipaeda (Spirurida, Thelaziidae) in dogs, cats and foxes in Italy: a "coincidence" or a parasitic disease of the Old Continent?

Donato Traversa, Ezio Ferroglio, Riccardo P Lia … (2003)

Thelazia callipaeda is a spirurid nematode which causes ocular infections in dogs and man and, occasionally, in cats, foxes and rabbits. The intermediate host and vector of T. callipaeda is unknown. For a long time T. callipaeda incidence was reported only from the Russian Federation and the Far East, but recently it has also been found in Italy. In order to investigate the spread of T. callipaeda in Italy, a survey was carried out in two sites, site A in the Piedmont region (North West Italy), and site B in the Basilicata region (Southern Italy). Dogs, cats and foxes in site A and dogs in site B were examined for eyeworms, using different procedures and timing. From January 1995 to August 2002, 91 dogs, 4 cats and 903 fox carcasses were examined in site A, and from October 1999 to January 2003, 443 dogs were examined in site B, and the eyeworms collected were identified using morphological keys. Twenty-one (23.07%) and 185 (41.76%) of the dogs from sites A and B, respectively, were found to be infected by eyeworms; furthermore, all the cats examined and 46 fox carcasses (5.1%) were positive for eyeworms. All the nematodes collected were identified as T. callipaeda. These results indicate that T. callipaeda is not confined to Eastern Europe and Asia, but that it has spread to the Old Continent, and to both Northern and Southern Italy. Considering the high prevalence of infected dogs reported in some municipalities (e.g. 60.14% of 138 dogs examined in a municipality from site B), it is assumed that one or more vectors are significantly present in the areas under investigation. Furthermore, there is good reason to believe that T. callipaeda is also present in other European countries. Speculation as to the origins of this parasitic infestation in Europe and the biology of T. callipaeda and its vector/s is also discussed.

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Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea).

Neil B Chilton, Robin Gasser, Ian Beveridge (1995)

The nucleotide sequence of the second internal transcribed spacer (ITS-2) from ribosomal DNA has been determined for 3 members of the Hypodontus macropi species complex. Sequences were compared from nematodes collected from 3 species of Australian macropodid marsupial, Petrogale persephone, Macropus robustus robustus and Thylogale billardierii. The ITS-2 of each operational taxonomic unit ranged from 287 to 292 bases in length, and had a GC content of 36.6-40.1%. Differences in nucleotide sequence between nematodes from the different host species ranged from 25.0% to 28.3%. The data suggest that H. macropi from P. persephone represents a different species to those in M. r. robustus and T. billardierii. The unique feature of this study is that it represents a comparison of the ribosomal DNA sequences of nematode species which are morphologically indistinguishable but which have been demonstrated to be genetically distinct (i.e. cryptic) species based on electrophoretic data. The results also demonstrate further that morphological characters alone are often not adequate for species recognition. Differences between these 3 species of H. macropi in their recognition sites for restriction endonucleases, indicates that a PCR-RFLP approach could be used, in conjunction with allozyme electrophoresis, to establish how many species are present within the H. macropi complex.

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Author and article information

Journal

Title: Journal of Parasitology

Abbreviated Title: Journal of Parasitology

Publisher: American Society of Parasitologists

ISSN (Print): 0022-3395

ISSN (Electronic): 1937-2345

Publication date Created: February 2004

Publication date (Print): February 2004

Volume: 90

Issue: 1

Pages: 185-188

Article

DOI: 10.1645/GE-115R1

PubMed ID: 15040692

SO-VID: 0c8185fc-9893-4fff-96f3-ed9798c910c4

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Molecular Characterization of the First Internal Transcribed Spacer of Ribosomal DNA of the Most Common Species of Eyeworms (Thelazioidea: Thelazia)

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Most cited references 27

Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer.

Current status and epidemiological observation of Thelazia callipaeda (Spirurida, Thelaziidae) in dogs, cats and foxes in Italy: a "coincidence" or a parasitic disease of the Old Continent?

Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea).

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