Specific identification of Habronema microstoma and Habronema muscae (Spirurida, Habronematidae) by PCR using markers in ribosomal DNA.
Abstract: Gastric or cutaneous habronemosis caused by Habronema microstoma Creplin, 1849 and Habronema muscae Carter, 1865 is a parasitic disease of equids transmitted by muscid flies. There is a paucity of information on the epidemiology of this disease, which is mainly due to limitations with diagnosis in the live animal and with the identification of the parasites in the intermediate hosts. To overcome such limitations, a molecular approach, based on the use of genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal DNA, was established for the two species of Habronema. Characterisation of the ITS-2 revealed sequence lengths and G+C contents of 296 bp and 29.5% for H. microstoma, and of 334 bp and 35.9% for H. muscae, respectively. Exploiting the sequence difference (approximately 40%) between the two species of nematode, primers were designed and tested by the polymerase chain reaction (PCR) for their specificity using a panel of control DNA samples from common equid endoparasites, and from host tissues, faeces or muscid flies. Effective amplification from each of the two species of Habronema was achieved from as little as 10 pg of genomic DNA. Hence, this molecular approach allows the specific identification and differentiation of the DNA from H. microstoma and H. muscae, and could thus provide a molecular tool for the specific detection of Habronema DNA (irrespective of developmental stage) from faeces, skin and muscid fly samples. The establishment of this tool has important implications for the specific diagnosis of clinical cases of gastric and cutaneous habronemosis in equids, and for studying the ecology and epidemiology of the two species of Habronema.
Publication Date: 2004-07-24 PubMed ID: 15271381DOI: 10.1016/j.mcp.2004.01.006Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper presents a molecular technique for identifying and differentiating between two species of Habronema, a parasite affecting horses. This method, which relies on analyzing specific DNA markers, could be a significant tool for diagnosing parasitic diseases in horses and for studying these parasites’ ecology and epidemiology.
Methodology Used
- The research makes use of genetic markers within the second internal transcribed spacer (ITS-2) of ribosomal DNA to establish a method for identifying and distinguishing between the two species of Habronema.
- The researchers characterised the ITS-2 of the two species, noting variations in sequence lengths and G+C contents for each species.
- Based on the distinct differences in DNA sequences between the two nematode species, the team designed targeted primers for polymerase chain reaction (PCR) tests.
- The primers’ specificity was tested via PCR using control DNA samples from common equine endoparasites and from horse tissues, faeces, and muscid flies.
- The team achieved successful amplification from each Habronema species using as little as 10 picograms of genomic DNA.
Findings and Implications
- The implemented molecular approach allowed for the specific identification and differentiation of DNA from Habronema microstoma and Habronema muscae.
- The promising findings suggest this method could potentially detect Habronema DNA, regardless of the developmental stage, in faeces, skin, and muscid fly samples.
- This breakthrough could be critical in accurately diagnosing clinical cases of gastric and cutaneous habronemosis in equines, diseases caused by these parasites.
- Furthermore, the ability to differentiate between these closely related species has significant implications for studying the ecology and epidemiology of Habronema species, which could lead to more effective prevention strategies and treatments for the diseases they cause.
Cite This Article
APA
Traversa D, Giangaspero A, Galli P, Paoletti B, Otranto D, Gasser RB.
(2004).
Specific identification of Habronema microstoma and Habronema muscae (Spirurida, Habronematidae) by PCR using markers in ribosomal DNA.
Mol Cell Probes, 18(4), 215-221.
https://doi.org/10.1016/j.mcp.2004.01.006 Publication
Researcher Affiliations
- Department of Biomedical Comparative Sciences, Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy.
MeSH Terms
- Animals
- Base Sequence
- DNA, Ribosomal / blood
- DNA, Ribosomal / genetics
- Feces / parasitology
- Genetic Markers
- Horses / parasitology
- Molecular Sequence Data
- Muscidae / genetics
- Polymerase Chain Reaction
- Spirurida Infections / genetics
- Spiruroidea / genetics
- Stomach / parasitology
Citations
This article has been cited 14 times.- Febronio AMB, Boos GS, Batista RLG, Amorim DB, Guimarães JP, Bianchi MV, Mariani DB, Koproski L, Mari C, Parente JEV, Sonne L, Werneck MR, Marques SMT, Driemeier D, Kolesnikovas CKM, Groch KR, Sobotyk C, Verocai GG, Groch KR, Díaz-Delgado J. Crassicaudiasis in three geographically and chronologically distant Cuvier's beaked whales (Ziphius cavirostris) stranded off Brazil.. Int J Parasitol Parasites Wildl 2021 Dec;16:262-269.
- Naseem MN, Raza A, Allavena R, McGowan M, Morgan JAT, Constantinoiu C, Tabor AE, James P. Development and Validation of Novel PCR Assays for the Diagnosis of Bovine Stephanofilariasis and Detection of Stephanofilaria sp. Nematodes in Vector Flies.. Pathogens 2021 Sep 17;10(9).
- Liu Y, Zhao Z, Yang X, Yang L, Yang B, Zheng W, Li W, Luo X, Wang R, Gu W, Wang P. Haematobium irritans and Haematobium titillans as potential vectors of Parabronema skrjabini in camels (Camelus bactrianus) in Inner Mongolia, China.. Parasitology 2020 Nov;147(13):1509-1514.
- Barlaam A, Traversa D, Papini R, Giangaspero A. Habronematidosis in Equids: Current Status, Advances, Future Challenges.. Front Vet Sci 2020;7:358.
- Jian R, Wang SW, Zhang WX, Zhang LP. Morphological and molecular identification of Habronema spp. (Nematoda: Habronematidae) from donkeys in Xinjiang, China, and notes on the taxonomical status of Habronema majus (Creplin, 1849) and H. microstoma (Schneider, 1866).. Syst Parasitol 2017 May;94(4):511-525.
- Rakhshandehroo E, Sharifiyazdi H, Shayegh H, Ahmadi A. Molecular and morphological comparison of two different types of Habronema muscae (Nematoda: Habronematidae) in horse.. Parasitol Res 2014 Dec;113(12):4439-45.
- Traversa D, Otranto D. Biotechnological advances in the diagnosis of little-known parasitoses of pets.. Parasitol Res 2009 Jan;104(2):209-16.
- Iorio R, Slapeta J, Otranto D, Paoletti B, Giangaspero A, Traversa D. Phylogenetic relationships of Habronema microstoma and Habronema muscae (Spirurida: Habronematidae) within the order Spirurida inferred using mitochondrial cytochrome c oxidase subunit 1 (cox1) gene analysis.. Parasitol Res 2009 Apr;104(5):979-84.
- Chen W, Sun HY, Xie MQ, Bai JS, Zhu XQ, Li AX. Development of specific PCR assays for the detection of Cryptocaryon irritans.. Parasitol Res 2008 Jul;103(2):423-7.
- Traversa D, Iorio R, Klei TR, Kharchenko VA, Gawor J, Otranto D, Sparagano OA. New method for simultaneous species-specific identification of equine strongyles (nematoda, strongylida) by reverse line blot hybridization.. J Clin Microbiol 2007 Sep;45(9):2937-42.
- Naem S. The comparative morphology of three equine habronematid nematodes: SEM observations.. Parasitol Res 2007 Oct;101(5):1303-10.
- Naem S. First SEM observations on adult Habronema microstoma (Spirurida: Habronematidae), a parasite of the horse.. Parasitol Res 2007 Aug;101(3):743-9.
- Naem S. Equine stomach worm, Drashia megastoma (Spirurida: Habronematidae): first SEM report.. Parasitol Res 2007 Sep;101(4):913-8.
- Naem S. First description of the horse stomach worm, Habronema muscae (Spirurida: Habronematidae) by scanning electron microscopy.. Parasitol Res 2007 Jul;101(2):427-32.
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