FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Metabolites / Automated Spectrophotometric Assays for the Measurement of A...
Metabolites2024; 14(3); doi: 10.3390/metabo14030147

Automated Spectrophotometric Assays for the Measurement of Ammonia and Bicarbonate in Saliva of Horses: Analytical Validation and Changes in Equine Gastric Ulcer Syndrome (EGUS).

Abstract: Ammonia (NH3) and bicarbonate (HCO3) have been related to gastric ulcers in humans. Ammonia is considered a possible cause of gastric ulcers, whereas bicarbonate has a protective function. The presence of ulcers in the stomach of horses is defined as Equine Gastric Ulcer Syndrome (EGUS), which is a frequent disease in this species, and it has been associated with changes in saliva composition, such as in analytes related to inflammation, immune system and oxidative stress. The objectives of this study were (1) to perform an analytical validation of two automated spectrophotometric assays, one for ammonia and one for bicarbonate, in the horses' saliva and (2) to evaluate their possible variations with EGUS. Analytical validation of the automated assays for ammonia and bicarbonate in the saliva of horses showed that both assays were precise and accurate. In addition, significantly higher values of ammonia and lower values of bicarbonate were found in the saliva of horses with EGUS compared to healthy horses. It can be concluded that ammonia and bicarbonate can be measured in the saliva of horses and that ammonia increases and bicarbonate decreases in this sample type could be related to the presence of EGUS in this species.
Get Full Text
Publication Date: 2024-02-28 PubMed ID: 38535307PubMed Central: PMC10972414DOI: 10.3390/metabo14030147Google 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.
LinkedInCopy
  • Journal Article

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 paper presents an analysis and validation of two automated spectrophotometric assays for measuring ammonia and bicarbonate in horse saliva, as well as evaluates their fluctuations in relation to Equine Gastric Ulcer Syndrome (EGUS).

Objective and Process

The key goals of this study, explained in the abstract were:

  • To perform an analytical validation of two automated spectrophotometric assays, specifically for ammonia and bicarbonate, in horse saliva. Spectrophotometric assays are procedures used to measure the concentration of particular chemicals (in this case, ammonia and bicarbonate) based on light absorption properties.
  • To evaluate the changes in the presence and concentration of these chemicals in relation to Equine Gastric Ulcer Syndrome (EGUS), a common ailment in horses associated with alterations in saliva composition.

Analytical Validation Results

The results of the validation of the automated assays for ammonia and bicarbonate in the saliva showcase that these assays are precise and accurate. Such precision and accuracy are crucial to reliable measurements in any kind of chemical analysis and scientific experiments, making these assays effective methods of measuring ammonia and bicarbonate levels.

Correlation with Equine Gastric Ulcer Syndrome (EGUS)

The researchers discovered that horses with EGUS had notably higher levels of ammonia and lower levels of bicarbonate in their saliva compared to healthy horses. This suggests a relationship between:

  • The increase in ammonia levels, which has been considered a possible cause of gastric ulcers in humans
  • The decrease in bicarbonate levels, which is known to have a protective function against gastric ulcers by neutralizing acid in the stomach.

This also indicates that these changes in the levels of these two chemicals can potentially be linked to the presence of EGUS.

Conclusion

The study concludes that ammonia and bicarbonate levels can be measured in horse saliva using the validated assays, and that changes in these levels could be connected to the presence of Equine Gastric Ulcer Syndrome in horses. Thus, these measurements might explore new avenues for understanding, diagnosing, and treating EGUS.

Cite This Article

APA
Muñoz-Prieto A, Llamas-Amor E, Contreras-Aguilar MD, Ayala I, Martín Cuervo M, Cerón JJ, Hansen S. (2024). Automated Spectrophotometric Assays for the Measurement of Ammonia and Bicarbonate in Saliva of Horses: Analytical Validation and Changes in Equine Gastric Ulcer Syndrome (EGUS). Metabolites, 14(3). https://doi.org/10.3390/metabo14030147

Publication

Metabolites
ISSN: 2218-1989
NlmUniqueID: 101578790
Country: Switzerland
Language: English
Volume: 14
Issue: 3

Researcher Affiliations

Muñoz-Prieto, Alberto
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo s/n, 30100 Murcia, Spain.
Llamas-Amor, Eva
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo s/n, 30100 Murcia, Spain.
Contreras-Aguilar, María Dolores
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo s/n, 30100 Murcia, Spain.
Ayala, Ignacio
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo s/n, 30100 Murcia, Spain.
Martín Cuervo, María
  • Department of Animal Medicine, Faculty of Veterinary, University of Extremadura, Avda de la Universidad s/n, 10003 Cáceres, Spain.
Cerón, José Joaquín
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo s/n, 30100 Murcia, Spain.
Hansen, Sanni
  • Section Medicine and Surgery, Department of Veterinary Clinical Sciences, University of Copenhagen, Agrovej 8, 2630 Taastrup, Denmark.

Grant Funding

  • 19894/GERM/15 / Fundación Séneca - Agencia de Ciencia y Tecnología de la Región de Murcia
  • RYC2021-033660-I / Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (AEI)

Conflict of Interest Statement

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

References

This article includes 36 references
  1. Nascimento M.M., Gordan V.V., Garvan C.W., Browngardt C.M., Burne R.A.. Correlations of Oral Bacterial Arginine and Urea Catabolism with Caries Experience. Oral. Microbiol. Immunol. 2009;24:89–95.
    doi: 10.1111/j.1399-302X.2008.00477.xpmc: PMC2742966pubmed: 19239634google scholar: lookup
  2. Bhogadia M., Edgar M., Hunwin K., Page G., Grootveld M.. Detection and Quantification of Ammonia as the Ammonium Cation in Human Saliva by 1H NMR: A Promising Probe for Health Status Monitoring, with Special Reference to Cancer. Metabolites 2023;13:792.
    doi: 10.3390/metabo13070792pmc: PMC10383521pubmed: 37512499google scholar: lookup
  3. Kopstein J., Wrong O.M.. The Origin and Fate of Salivary Urea and Ammonia in Man. Clin. Sci. 1977;52:9–17.
    doi: 10.1042/cs0520009pubmed: 23916google scholar: lookup
  4. Boivin R., Bost J.. Saliva Ammonia and Urea in the Sheep. Effects of Alimentary Nitrogen Additives (Author’s Transl). Ann. Rech. Vet. 1977;8:143–152.
    pubmed: 596793
  5. Huizenga J.R., Gips C.H.. Determination of Ammonia in Saliva Using Indophenol, an Ammonium Electrode and an Enzymatic Method: A Comparative Investigation. Clin. Chem. Lab. Med. 1982;20:571–574.
    doi: 10.1515/cclm.1982.20.8.571pubmed: 7142917google scholar: lookup
  6. Bardow A., Madsen J., Nauntofte B.. The Bicarbonate Concentration in Human Saliva Does Not Exceed the Plasma Level under Normal Physiological Conditions. Clin. Oral Investig. 2000;4:245–253.
    doi: 10.1007/s007840000077pubmed: 11218496google scholar: lookup
  7. Sand H.F.. Source of the Bicarbonate of Saliva. J. Appl. Physiol. 1951;4:66–76.
    doi: 10.1152/jappl.1951.4.2.66pubmed: 14888619google scholar: lookup
  8. Lundström T., Lingström P., Wattle O., Carlén A., Birkhed D.. Equine Saliva Components during Mastication, and in Vivo PH Changes in the Oral Biofilm of Sound and Carious Tooth Surfaces after Sucrose Exposure. Acta Vet. Scand. 2020;62:21.
    doi: 10.1186/s13028-020-00518-2pmc: PMC7245034pubmed: 32446309google scholar: lookup
  9. Kearney D.J., Ritchie K., Peacock J.S.. Gastric-Juice Ammonia Assay for Diagnosis of Helicobacter Pylori Infection and The Relationship of Ammonia Concentration to Gastritis Severity. Am. J. Gastroenterol. 2000;95:3399–3403.
    doi: 10.1111/j.1572-0241.2000.03351.xpubmed: 11151868google scholar: lookup
  10. Tsujii M., Kawano S., Tsuji S., Fusamoto H., Kamada T., Sato N.. Mechanism of Gastric Mucosal Damage Induced by Ammonia. Gastroenterology 1992;102:1881–1888.
    doi: 10.1016/0016-5085(92)90309-Mpubmed: 1587407google scholar: lookup
  11. Hata M., Yamazaki Y., Ueda T., Kato T., Kohli Y., Fujiki N.. Influence of Ammonia Solution on Gastric Mucosa and Acetic Acid Induced Ulcer in Rats. Eur. J. Histochem. 1994;38:41–52.
    pubmed: 7517730
  12. Vadgama P., Alberti K.G.M.M.. The Possible Role of Bicarbonate in Mucosal Protection and Peptic Ulceration. Digestion 1983;27:203–213.
    doi: 10.1159/000198954pubmed: 6653920google scholar: lookup
  13. Højgaard L., Nielsen A.M., Rune S.J.. Peptic Ulcer Pathophysiology: Acid, Bicarbonate, and Mucosal Function. Scand. J. Gastroenterol. 1996;31:10–15.
    doi: 10.3109/00365529609094555pubmed: 8726273google scholar: lookup
  14. Su T., Song Y., Pan X., Zhang Y., Shen Z., Lu J., Du Y., Li Z.. Efficacy of Omeprazole and Sodium Bicarbonate Suspension in the Treatment of Peptic Ulcer: A Multicenter Clinical Trial. Chin. J. Dig. 2022;12:6–13.
  15. Skoczylas T., Yandrapu H., Poplawski C., Asadi M., Wallner G., Sarosiek J.. Salivary Bicarbonate as a Major Factor in the Prevention of Upper Esophageal Mucosal Injury in Gastroesophageal Reflux Disease. Dig. Dis. Sci. 2014;59:2411–2416.
    doi: 10.1007/s10620-014-3099-1pubmed: 24610537google scholar: lookup
  16. Vokes J., Lovett A., Sykes B.. Equine Gastric Ulcer Syndrome: An Update on Current Knowledge. Animals 2023;13:1261.
    doi: 10.3390/ani13071261pmc: PMC10093336pubmed: 37048517google scholar: lookup
  17. Merritt A.M.. Appeal for Proper Usage of the Term ‘EGUS’: Equine Gastric Ulcer Syndrome. Equine Vet. J. 2009;41:616.
    doi: 10.2746/042516409X454574pubmed: 19927576google scholar: lookup
  18. Muñoz-Prieto A., Cerón J.J., Rubio C.P., Contreras-Aguilar M.D., Pardo-Marín L., la Peña I.A., Martín-Cuervo M., Henriksen I.-M.H., Arense-Gonzalo J.J., Tecles F.. Evaluation of a Comprehensive Profile of Salivary Analytes for the Diagnosis of the Equine Gastric Ulcer Syndrome. Animals 2022;12:3261.
    doi: 10.3390/ani12233261pmc: PMC9740180pubmed: 36496782google scholar: lookup
  19. Henneke D.R., Potter G.D., Kreider J.L., Yeates B.F.. Relationship between Condition Score, Physical Measurements and Body Fat Percentage in Mares. Equine Vet. J. 1983;15:371–372.
    doi: 10.1111/j.2042-3306.1983.tb01826.xpubmed: 6641685google scholar: lookup
  20. Shawaf T., El-Deeb W.M., Elgioushy M.. The Contribution of Specific and Nonspecific Biomarkers in Diagnosis of Equine Gastric Ulcer Syndrome (EGUS) Under Field Condition. J. Equine Vet. Sci. 2020;84:102853.
    doi: 10.1016/j.jevs.2019.102853pubmed: 31864460google scholar: lookup
  21. Sykes B.W., Hewetson M., Hepburn R.J., Luthersson N., Tamzali Y.. European College of Equine Internal Medicine Consensus Statement-Equine Gastric Ulcer Syndrome in Adult Horses. J. Vet. Intern. Med. 2015;29:1288–1299.
    doi: 10.1111/jvim.13578pmc: PMC4858038pubmed: 26340142google scholar: lookup
  22. Wise J.C., Wilkes E.J.A., Raidal S.L., Xie G., Crosby D.E., Hale J.N., Hughes K.J.. Interobserver and Intraobserver Reliability for 2 Grading Systems for Gastric Ulcer Syndrome in Horses. J. Vet. Intern. Med. 2021;35:571–579.
    doi: 10.1111/jvim.15987pmc: PMC7848314pubmed: 33284465google scholar: lookup
  23. Muñoz-Prieto A., Contreras-Aguilar M.D., Cerón J.J., de la Peña I.A., Martín-Cuervo M., Eckersall P.D., Henriksen I.-M.H., Tecles F., Hansen S.. Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome. Animals 2023;13:1367.
    doi: 10.3390/ani13081367pmc: PMC10135270pubmed: 37106929google scholar: lookup
  24. Huizenga J.R., Vissink A., Kuipers E.J., Gips C.H.. Helicobacter Pylori and Ammonia Concentrations of Whole, Parotid and Submandibular/Sublingual Saliva. Clin. Oral. Investig. 1999;3:84–87.
    doi: 10.1007/s007840050083pubmed: 10803116google scholar: lookup
  25. Gröschl M.. Stability of Salivary Steroids: The Influences of Storage, Food and Dental Care. Steroids 2001;66:737–741.
    doi: 10.1016/S0039-128X(01)00111-8pubmed: 11522335google scholar: lookup
  26. . Guidance for Industry: Bioanalytical Method Validation 2001. .
  27. Huizenga J.R., Teelken A.W., Tangerman A., de Jager A.E.J., Gips C.H., Jansen P.L.M.. Determination of Ammonia in Cerebrospinal Fluid Using the Indophenol Direct Method. Mol. Chem. Neuropathol. 1998;34:169–177.
    doi: 10.1007/BF02815078pubmed: 10327416google scholar: lookup
  28. Murakami M., Saita H., Teramura S., Dekigai H., Asagoe K., Kusaka S., Kita T.. Gastric Ammonia Has a Potent Ulcerogenic Action on the Rat Stomach. Gastroenterology 1993;105:1710–1715.
    doi: 10.1016/0016-5085(93)91067-Rpubmed: 8253347google scholar: lookup
  29. Miller K.E., Balbás J.C., Benton R.L., Lam T.S., Edwards K.M., Kriebel R.M., Schechter R.. Glutaminase Immunoreactivity and Enzyme Activity Is Increased in the Rat Dorsal Root Ganglion Following Peripheral Inflammation. Pain. Res. Treat. 2012;2012:414697.
    doi: 10.1155/2012/414697pmc: PMC3250962pubmed: 22229088google scholar: lookup
  30. Foster A., Bhattacharjee P., Tresoldi E., Pakusch M., Cameron F.J., Mannering S.I.. Glutamine Deamidation Does Not Increase the Immunogenicity of C-Peptide in People with Type 1 Diabetes. J. Transl. Autoimmun. 2023;6:100180.
    doi: 10.1016/j.jtauto.2022.100180pmc: PMC9811213pubmed: 36619657google scholar: lookup
  31. Rumiantsev V.A., Esaian L.K., Tolstova O.O., Khokhlova A.S.. Oral Acid-Basic Balance Peculiarities in Patients with Gastrointestinal Tract Pathology. Stomatologiia 2009;88:27–30.
    pubmed: 20037535
  32. Namiot Z., Stasiewicz J., Markowski A.R., Namiot D.B., Jaroszewicz W., Kemona A., Górski J.. Gastric Acid and Salivary Bicarbonate. Is There a Relationship in Duodenal Ulcer Patients?. Rocz. Akad. Med. Bialymst. 2004;49:75–79.
    pubmed: 15631318
  33. Guslandi M., Ballarin E.. Assessment of the ‘Mucus-Bicarbonate’ Barrier in the Stomach of Patients with Chronic Gastric Disorders. Clin. Chim. Acta. 1984;144:133–136.
    doi: 10.1016/0009-8981(84)90046-9pubmed: 6529851google scholar: lookup
  34. Malhotra S.L.. Protective Action of Saliva in Peptic Ulceration. Scand. J. Gastroenterol. 1967;2:95–104.
    doi: 10.3109/00365526709180053pubmed: 20184475google scholar: lookup
  35. van den Boom R.. Equine Gastric Ulcer Syndrome in Adult Horses. Vet. J. 2022;283–284:105830.
    doi: 10.1016/j.tvjl.2022.105830pubmed: 35472513google scholar: lookup
  36. Bochnia M., Goetz F., Wensch-Dorendorf M., Koelln M., Zeyner A.. Chewing Patterns in Horses during the Intake of Variable Quantities of Two Pelleted Compound Feeds Differing in Their Physical Characteristics Only. Res. Vet. Sci. 2019;125:189–194.
    doi: 10.1016/j.rvsc.2019.06.014pubmed: 31254716google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.