Abstract
Aims: To develop a biological assay utilising proteolytic properties to determine pepsinogen concentrations in equine serum and assess the feasibility of the assay for determining pepsinogen concentrations in horses with and without gastric ulceration.
Methods: A micronised method of pepsinogen activity was assessed involving the incubation of duplicate equine serum samples at 37°C in a 0.1MHCl / glycine and Bovine Serum Albumin buffer (pH 1.6). Total protein concentrations following 0.5hr and 3.5hrs of incubation were determined by linear regression analysis using a commercial protein assay kit. Proteolytic activity per hour was compared to that of porcine pepsinogen standards and estimation of pepsinogen concentration was obtained by quadratic regression analysis. Pepsinogen concentrations were measured in serum samples collected from apparently healthy horses (n=9) and horses with gastroscopic evidence of gastric ulceration (n=5). Results: Pepsinogen concentrations measured were variable in both healthy horses (range; 0-99.5 iu/l; mean 40.0; SD ± 36.1) and horses with gastric ulceration (range; 0-99.2iu/l; mean 67.6; SD ± 41.2). No statistical difference was found between healthy and diseased horses using a one-way ANOVA analysis. Conclusions and Practical Significance: The assay utilised has the advantages of using small volumes of equine serum (100μl) and a fairly short total running time (6 hours) and may easily be adapted to many laboratories. Pepsinogen concentrations in healthy horses and in horses with low-grade gastric ulceration appear to be at the low end of detection for this assay. The assay may have an application as a methodology for the detection of gastric ulceration, however further validation of the test is required using increased numbers of horses. Acknowledgements: The Horse Trust for funding this project and Prof. Michael Stear and Dr. Cathy Wyse for their laboratory assistance.
Methods: A micronised method of pepsinogen activity was assessed involving the incubation of duplicate equine serum samples at 37°C in a 0.1MHCl / glycine and Bovine Serum Albumin buffer (pH 1.6). Total protein concentrations following 0.5hr and 3.5hrs of incubation were determined by linear regression analysis using a commercial protein assay kit. Proteolytic activity per hour was compared to that of porcine pepsinogen standards and estimation of pepsinogen concentration was obtained by quadratic regression analysis. Pepsinogen concentrations were measured in serum samples collected from apparently healthy horses (n=9) and horses with gastroscopic evidence of gastric ulceration (n=5). Results: Pepsinogen concentrations measured were variable in both healthy horses (range; 0-99.5 iu/l; mean 40.0; SD ± 36.1) and horses with gastric ulceration (range; 0-99.2iu/l; mean 67.6; SD ± 41.2). No statistical difference was found between healthy and diseased horses using a one-way ANOVA analysis. Conclusions and Practical Significance: The assay utilised has the advantages of using small volumes of equine serum (100μl) and a fairly short total running time (6 hours) and may easily be adapted to many laboratories. Pepsinogen concentrations in healthy horses and in horses with low-grade gastric ulceration appear to be at the low end of detection for this assay. The assay may have an application as a methodology for the detection of gastric ulceration, however further validation of the test is required using increased numbers of horses. Acknowledgements: The Horse Trust for funding this project and Prof. Michael Stear and Dr. Cathy Wyse for their laboratory assistance.
| Original language | English |
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| Pages | 413-414 |
| Number of pages | 2 |
| Publication status | Published - 2007 |
| Event | 46th British Equine Veterinary Association Congress - Edinburgh, United Kingdom Duration: 12 Sept 2007 → 15 Sept 2007 |
Conference
| Conference | 46th British Equine Veterinary Association Congress |
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| Country/Territory | United Kingdom |
| City | Edinburgh |
| Period | 12/09/07 → 15/09/07 |