Semi-nested PCR for the specific detection of Habronema microstoma or Habronema muscae DNA in horse faeces.
Abstract: Habronema microstoma and Habronema muscae (Spirurida: Habronematidae) are parasitic nematodes which infect the stomach and/or skin of equids. The accurate diagnosis of gastric habronemosis is central to studying its epidemiology, but data on its distribution and prevalence are lacking, mainly due to the limitations of clinical and coprological diagnosis in live horses. To overcome this constraint, a two-step, semi-nested PCR-based assay was validated (utilizing genetic markers in the nuclear ribosomal DNA) for the specific amplification of H. microstoma or H. muscae DNA from the faeces from horses (n = 46) whose gastrointestinal parasite status had been determined at autopsy and whose faeces were examined previously using a conventional parasitological approach. Of these horses examined at autopsy, some harboured adults of either H. microstoma (n= 19) or H. muscae (n =4), and others (n = 7) harboured both species. Most of them were also infected with other parasites, including strongylid nematodes (subfamilies Cyathostominae and Strongylinae), bots and/or cestodes; there was no evidence of metazoan parasites in 2 horses. Larvated spirurid eggs were detected in the faeces of 1 of the 30 horses (3.3 %) shown to be infected with Habronema at autopsy. For this set of 46 samples, the PCR assay achieved a diagnostic specificity of 100 % and a sensitivity of approximately 97 % (being able to specifically detect as little as approximately 0.02 fg of Habronema DNA). The specificity of the assay was also tested using a panel of control DNA samples representing horse, the gastric spirurid Draschia megastoma and 26 other species of parasites from the alimentary tract of the horse. H. microstoma, H. muscae and D. megastoma could be readily differentiated from one another based on the sizes of their specific amplicons in the PCR. The results of this study showed that the performance of the PCR for the diagnosis of gastric habronemosis was similar to that of autopsy but substantially better than the traditional coprological examination procedure used. The ability to specifically diagnose gastric habronemosis in equids should have important implications for investigating the epidemiology and ecology of H. microstoma and H. muscae.
Publication Date: 2005-01-15 PubMed ID: 15648696DOI: 10.1017/s0031182004006122Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article presents a study that developed and validated a semi-nested PCR-based assay, a molecular genetic testing method, for diagnosing parasitic infections in horses caused by Habronema microstoma and Habronema muscae. The new method demonstrated high accuracy and outperformed traditional approaches, potentially improving epidemiological studies on these parasitic infections.
Objective of the study
- The objective of the study was to overcome limitations of existing diagnosis methods for parasitic nematode infections in horses, and thus improve the understanding of the epidemiology of infections caused by Habronema microstoma and Habronema muscae, nematodes that infect the stomach and/or skin of equids (the horse family).
Methodology
- The study uses a semi-nested PCR-based assay, which is a specialized form of Polymerase Chain Reaction (PCR), a method used to amplify certain pieces of DNA. This method was designed to detect the DNA of either H. microstoma or H. muscae specifically.
- The researchers applied this method to faecal samples obtained from 46 horses whose gastrointestinal parasite statuses had been previously determined at autopsy and via a conventional parasitological approach.
Results
- The PCR assay achieved a diagnostic specificity of 100%, meaning it correctly identified all non-infected horses as non-infected. It also had a sensitivity of approximately 97%, suggesting it was able to correctly detect infected horses almost every time.
- The PCR assay was able to identify as little as 0.02 femtograms (a femtogram is a quadrillionth of a gram) of Habronema DNA, underlining the impressive sensitivity of the method.
- The PCR assay findings corresponded closely to the diagnoses made at autopsy, thus, validating its efficacy and accuracy. In contrast with conventional methods, the PCR assay exhibited superior performance in diagnosing gastric habronemosis, the disease caused by these nematodes.
Implications
- The research demonstrated that the PCR-based method is superior to traditional diagnostic methods for detecting Habronema infections in horses.
- This new diagnostic approach could have significant implications for studying the epidemiology and ecology of H. microstoma and H. muscae.
- By applying this method for the accurate diagnosis of gastric habronemosis, improved prevention, control, and treatment interventions might be developed.
Cite This Article
APA
Traversa D, Giangaspero A, Iorio R, Otranto D, Paoletti B, Gasser RB.
(2005).
Semi-nested PCR for the specific detection of Habronema microstoma or Habronema muscae DNA in horse faeces.
Parasitology, 129(Pt 6), 733-739.
https://doi.org/10.1017/s0031182004006122 Publication
Researcher Affiliations
- Department of Biomedical Comparative Sciences, Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy. traversa@unite.it
MeSH Terms
- Animals
- DNA, Helminth / analysis
- DNA, Ribosomal Spacer / chemistry
- Feces / chemistry
- Female
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Male
- Polymerase Chain Reaction / methods
- Species Specificity
- Spirurida Infections / diagnosis
- Spirurida Infections / veterinary
- Spiruroidea / genetics
Citations
This article has been cited 16 times.- Palozzo A, Traversa D, Marruchella G, Celani G, Morelli S, Petrizzi L. Summer Sores Secondary to a Hoof Crack in an Andalusian Stallion. Pathogens 2021 Aug 16;10(8).
- Barlaam A, Traversa D, Papini R, Giangaspero A. Habronematidosis in Equids: Current Status, Advances, Future Challenges. Front Vet Sci 2020;7:358.
- Marcer F, Negrisolo E, Franzo G, Tessarin C, Pietrobelli M, Marchiori E. Morphological and molecular characterization of adults and larvae of Crassicauda spp. (Nematoda: Spirurida) from Mediterranean fin whales Balaenoptera physalus (Linnaeus, 1758). Int J Parasitol Parasites Wildl 2019 Aug;9:258-265.
- Schuster RK, Sivakumar S. The larval development of Habronema muscae (Nematoda: Habronematidae) affects its intermediate host, Musca domestica (Diptera: Muscidae). Parasitol Res 2017 Feb;116(2):503-509.
- 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.
- Di Cesare A, Castagna G, Otranto D, Meloni S, Milillo P, Latrofa MS, Paoletti B, Bartolini R, Traversa D. Molecular detection of Capillaria aerophila, an agent of canine and feline pulmonary capillariosis. J Clin Microbiol 2012 Jun;50(6):1958-63.
- Lepri E, Veronesi F, Traversa D, Conti MB, Marchesi MC, Miglio A, Mandara MT. Disseminated angiostrongylosis with massive cardiac and cerebral involvement in a dog from Italy. Parasitol Res 2011 Aug;109(2):505-8.
- Buzzell GR, Tariq S, Traversa D, Schuster R. Morphology of the infective larval stage of the equid parasite Habronema muscae (Spirurida: Habronematidae), from houseflies (Musca domestica). Parasitol Res 2011 Mar;108(3):629-32.
- 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.
- Traversa D, Iorio R, Otranto D. Diagnostic and clinical implications of a nested PCR specific for ribosomal DNA of the feline lungworm Aelurostrongylus abstrusus (Nematoda, Strongylida). J Clin Microbiol 2008 May;46(5):1811-7.
- 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.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
