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Author |
Gasser, R.B.; Hung, G.-C.; Chilton, N.B.; Beveridge, I. |
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Title |
Advances in developing molecular-diagnostic tools for strongyloid nematodes of equids: fundamental and applied implications |
Type |
Journal Article |
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Year |
2004 |
Publication |
Molecular and Cellular Probes |
Abbreviated Journal |
Mol Cell Probes |
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Volume |
18 |
Issue |
1 |
Pages |
3-16 |
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Keywords |
Animals; DNA, Helminth; DNA, Ribosomal/analysis; Equidae/*parasitology; Molecular Diagnostic Techniques/*methods; Parasitic Diseases, Animal/diagnosis; Strongylida/classification/genetics; Strongylida Infections/*diagnosis/epidemiology/etiology/veterinary |
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Abstract |
Infections of equids with parasitic nematodes of the order Strongylida (subfamilies Strongylinae and Cyathostominae) are of major veterinary importance. In last decades, the widespread use of drugs against these parasites has led to problems of resistance within the Cyathostominae, and to an increase in their prevalence and intensity of infection. Novel control strategies, based on improved knowledge of parasite biology and epidemiology, have thus become important. However, there are substantial limitations in the understanding of fundamental biological and systematic aspects of these parasites, which have been due largely to limitations in their specific identification and diagnosis using traditional, morphological approaches. Recently, there has been progress in the development of DNA-based approaches for the specific identification of strongyloids of equids for systematic studies and disease diagnosis. The present article briefly reviews information on the classification, biology, pathogenesis, epidemiology of equine strongyloids and the diagnosis of infections, highlights knowledge gaps in these areas, describes recent advances in the use of molecular techniques for the genetic characterisation, specific identification and differentiation of strongyloids of equids as a basis for fundamental investigations of the systematics, population biology and ecology. |
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Department of Veterinary Science, The University of Melbourne, 250 Princes Highway, Werribee, Victoria 3030, Australia. robinbg@unimelb.edu.au |
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0890-8508 |
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PMID:15036364 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
2636 |
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Author |
Cilnis, M.J.; Kang, W.; Weaver, S.C. |
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Title |
Genetic conservation of Highlands J viruses |
Type |
Journal Article |
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Year |
1996 |
Publication |
Virology |
Abbreviated Journal |
Virology |
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Volume |
218 |
Issue |
2 |
Pages |
343-351 |
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Keywords |
Alphavirus/*genetics; Alphavirus Infections/transmission/veterinary/virology; Amino Acid Sequence; Animals; Base Sequence; Conserved Sequence; Disease Outbreaks; Encephalitis, Viral/veterinary/virology; *Evolution, Molecular; Horses; Molecular Sequence Data; Phylogeny; RNA, Viral/genetics; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Turkeys; Variation (Genetics)/*genetics |
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Abstract |
We studied molecular evolution of the mosquito-borne alphavirus Highlands J (HJ) virus by sequencing PCR products generated from 19 strains isolated between 1952 and 1994. Sequences of 1200 nucleotides including portions of the E1 gene and the 3' untranslated region revealed a relatively slow evolutionary rate estimated at 0.9-1.6 x 10(-4) substitutions per nucleotide per year. Phylogenetic trees indicated that all HJ viruses descended from a common ancestor and suggested the presence of one dominant lineage in North America. However, two or more minor lineages probably circulated simultaneously for periods of years to a few decades. Strains isolated from a horse suffering encephalitis, and implicated in a recent turkey outbreak, were not phylogenetically distinct from strains isolated in other locations during the same time periods. Our findings are remarkably similar to those we obtained previously for another North American alphavirus, eastern equine encephalomyelitis virus, with which Highlands J shares primary mosquito and avian hosts, geographical distribution, and ecology. These results support the hypotheses that the duration of the transmission season affects arboviral evolutionary rates and vertebrate host mobility influences genetic diversity. |
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Department of Biology, University of California, San Diego, La Jolla 92093-0116, USA |
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0042-6822 |
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PMID:8610461 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
2657 |
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Author |
Chilton, N.B. |
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Title |
The use of nuclear ribosomal DNA markers for the identification of bursate nematodes (order Strongylida) and for the diagnosis of infections |
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Journal Article |
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Year |
2004 |
Publication |
Animal Health Research Reviews / Conference of Research Workers in Animal Diseases |
Abbreviated Journal |
Anim Health Res Rev |
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Volume |
5 |
Issue |
2 |
Pages |
173-187 |
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Keywords |
Animals; Birds; Cats; DNA Primers; DNA, Helminth/*analysis; DNA, Ribosomal/*analysis; Dogs; Horses; Molecular Diagnostic Techniques/veterinary; Ruminants; Strongylida/*genetics; Strongylida Infections/diagnosis/*veterinary |
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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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Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, Saskatchewan S7N 5E2, Canada. neil.chilton@usask.ca |
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English |
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1466-2523 |
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PMID:15984323 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
2628 |
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Author |
Traversa, D.; Giangaspero, A.; Iorio, R.; Otranto, D.; Paoletti, B.; Gasser, R.B. |
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Title |
Semi-nested PCR for the specific detection of Habronema microstoma or Habronema muscae DNA in horse faeces |
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Journal Article |
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Year |
2004 |
Publication |
Parasitology |
Abbreviated Journal |
Parasitology |
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Volume |
129 |
Issue |
Pt 6 |
Pages |
733-739 |
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Keywords |
Animals; DNA, Helminth/*analysis; DNA, Ribosomal Spacer/*chemistry; Feces/*chemistry; Female; Horse Diseases/*diagnosis/parasitology; Horses; Male; Polymerase Chain Reaction/*methods; Species Specificity; Spirurida Infections/diagnosis/*veterinary; Spiruroidea/*genetics |
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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. |
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Department of Biomedical Comparative Sciences, Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy. traversa@unite.it |
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0031-1820 |
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PMID:15648696 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
2631 |
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Author |
Passler, S.; Pfeffer, M. |
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Detection of antibodies to alphaviruses and discrimination between antibodies to eastern and western equine encephalitis viruses in rabbit sera using a recombinant antigen and virus-specific monoclonal antibodies |
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Journal Article |
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Year |
2003 |
Publication |
Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health |
Abbreviated Journal |
J Vet Med B Infect Dis Vet Public Health |
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Volume |
50 |
Issue |
6 |
Pages |
265-269 |
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Keywords |
Animals; Antibodies, Monoclonal/*immunology; Antibodies, Viral/*analysis/blood; DNA Primers; Encephalitis Virus, Eastern Equine/genetics/*immunology; Encephalitis Virus, Western Equine/genetics/*immunology; Encephalomyelitis, Equine/*diagnosis/*virology; Epitopes; Fluorescent Antibody Technique/*veterinary; Horses; Rabbits; Recombination, Genetic; Reverse Transcriptase Polymerase Chain Reaction/veterinary |
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Three arthropod-borne alphaviruses, western equine encephalitis viruses (WEEV), eastern equine encephalitis viruses (EEEV) and Venezuelan equine encephalitis viruses are the aetiological agents of a sometimes severe encephalomyelitis in equines and humans in the New World. With regard to the different ecology and epidemiology of these viruses, a method applied in serological screening should be able to distinguish between them as well as other related members of the genus Alphavirus in the American continent. However, this has been hampered in the past by (a) the close antigenic relationship between alphaviruses in traditional serological assays, especially in the routinely used haemagglutination-inhibition, and (b) the need of biosafety level 3 facilities to grow the viral antigens. An epitope blocking assay using an EEEV glycoprotein E1-expressing recombinant Sindbis virus and virus-specific monoclonal antibodies (mAbs) binding to the E1 of EEEV (strain NJ/60) and the E1 of Sindbis virus was established using automated flow cytometry. The test was evaluated using sera of infected and vaccinated rabbits. A cut-off value of 30% inhibition for antigenic complex-specific seroconversion was found to be sufficient for the detection of the respective infection. By using three different mAbs in parallel, we were able to detect alphavirus genus-, EEEV- and WEEV-complex-specific serum antibodies. As this test is based on the inhibition of binding of virus-specific mAbs, sera of every origin other than mouse can be tested. Thus, this assay may prove useful in the serological screening of a variety of animal species during an outbreak investigation. |
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Institute for Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, Ludwig-Maximilians-University, Munich, Germany |
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0931-1793 |
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PMID:14628996 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
2639 |
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Author |
Sebastiani, F.; Meiswinkel, R.; Gomulski, L.M.; Guglielmino, C.R.; Mellor, P.S.; Malacrida, A.R.; Gasperi, G. |
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Title |
Molecular differentiation of the Old World Culicoides imicola species complex (Diptera, Ceratopogonidae), inferred using random amplified polymorphic DNA markers |
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Journal Article |
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Year |
2001 |
Publication |
Molecular Ecology |
Abbreviated Journal |
Mol Ecol |
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Volume |
10 |
Issue |
7 |
Pages |
1773-1786 |
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Keywords |
Africa; Animals; Ceratopogonidae/*classification/*genetics; Ecology; Evolution, Molecular; Female; *Genetic Markers; Madagascar; Phylogeny; *Polymorphism, Genetic; *Random Amplified Polymorphic DNA Technique; Variation (Genetics) |
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Samples of seven of the 10 morphological species of midges of the Culicoides imicola complex were considered. The importance of this species complex is connected to its vectorial capacity for African horse sickness virus (AHSV) and bluetongue virus (BTV). Consequently, the risk of transmission may vary dramatically, depending upon the particular cryptic species present in a given area. The species complex is confined to the Old World and our samples were collected in Southern Africa, Madagascar and the Ivory Coast. Genomic DNA of 350 randomly sampled individual midges from 19 populations was amplified using four 20-mer primers by the random amplified polymorphic DNA (RAPD) technique. One hundred and ninety-six interpretable polymorphic bands were obtained. Species-specific RAPD profiles were defined and for five species diagnostic RAPD fragments were identified. A high degree of polymorphism was detected in the species complex, most of which was observed within populations (from 64 to 76%). Principal coordinate analysis (PCO) and cluster analysis provided an estimate of the degree of variation between and within populations and species. There was substantial concordance between the taxonomies derived from morphological and molecular data. The amount and the different distributions of genetic (RAPD) variation among the taxa can be associated to their life histories, i.e. the abundance and distribution of the larval breeding sites and their seasonality. |
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Department of Animal Biology, Laboratory of Zoology, University of Pavia, Piazza Botta 9, I-27100 Pavia, Italy |
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ISSN |
0962-1083 |
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PMID:11472544 |
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Equine Behaviour @ team @ |
Serial |
2647 |
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Author |
Birch, H.L.; Bailey, A.J.; Goodship, A.E. |
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Title |
Macroscopic 'degeneration' of equine superficial digital flexor tendon is accompanied by a change in extracellular matrix composition |
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Journal Article |
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Year |
1998 |
Publication |
Equine Veterinary Journal |
Abbreviated Journal |
Equine Vet J |
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Volume |
30 |
Issue |
6 |
Pages |
534-539 |
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Keywords |
Animals; Collagen/analysis; DNA/analysis; Extracellular Matrix/*chemistry; Glycosaminoglycans/analysis; Horses/injuries/*physiology; Immunohistochemistry; Rupture/veterinary; Tendon Injuries/metabolism/pathology/veterinary; Tendons/chemistry/*pathology; Water/analysis |
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Abstract |
Injuries to the superficial digital flexor tendon are common in horses required to gallop and jump at speed. Partial rupture of this tendon usually occurs in the central core of the midmetacarpal region and may be preceded by localised degenerative changes. Post mortem examination of apparently normal equine flexor tendons has revealed an abnormal macroscopic appearance in the central core, characterised by a reddish discolouration. We have previously shown that there is also physical damage to the collagen fibres. In the present study we tested the hypothesis that the abnormal appearance is accompanied by changes in the composition of the extracellular matrix of the tendon. Biochemical analysis of the extracellular matrix demonstrated an increase in total sulphated glycosaminoglycan content, increase in the proportion of type III collagen and decrease in collagen linked fluorescence in the central core of 'degenerated' tendons relative to tissue from the peripheral region of the same tendon. Dry matter content and total collagen content were not significantly different between tendon zones or normal and 'degenerated' tendons. These changes suggest a change in cell metabolism and matrix turnover in the central core of the tendon and are likely to contribute to a decrease in mechanical properties in this part of the tendon, predisposing to the characteristic partial rupture of the tendon. |
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Veterinary Basic Sciences, Royal Veterinary College, North Mymms, Hatfield, UK |
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0425-1644 |
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PMID:9844973 |
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Equine Behaviour @ team @ |
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3794 |
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