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BMC veterinary research2020; 16(1); 384; doi: 10.1186/s12917-020-02574-2

Characterization of total adenosine deaminase activity (ADA) and its isoenzymes in saliva and serum in health and inflammatory conditions in four different species: an analytical and clinical validation pilot study.

Abstract: Measurement of adenosine deaminase (ADA) can provide information about cell-mediated immunity. This report's objective was to study the enzymatic activity of total ADA (tADA) and its isoenzymes ADA1 and ADA2 in canine, equine, porcine, and bovine serum and saliva and their changes in different inflammatory situations in each species. Besides, an automated method for ADA2 measurement was developed and validated. Results: tADA was present in serum and saliva of healthy animals of the four species. Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) concentration of 0.47 mM was needed for ADA1 inhibition in canine and porcine samples (serum and saliva) and bovine saliva, whereas for equine saliva 0.94 mM was needed. ADA2 activity was not detected in bovine serum and was very low or absent in equine serum and bovine saliva. An automated procedure to measure ADA2 consisting of adding EHNA to a commercial reagent for tADA measurement provided repetitive (coefficients of variation < 8.8% in serum and  0.90) results, being equivalent to a manual incubation of the sample with EHNA at a similar concentration. Salivary tADA, as well as ADA1 and ADA2, were higher in dogs with leishmaniosis, horses with acute abdominal disease and pigs with lameness than in healthy animals. tADA and isoenzymes in saliva showed a positive significant correlation with serum ferritin in dogs (r = 0.602, P < 0.01; r = 0.555, P < 0.05; and r = 0.632, P < 0.01; respectively for tADA, ADA1 and ADA2) and serum C-reactive protein in pigs (r = 0.700, P < 0.01, for both tADA and ADA1; r = 0.770, P < 0.001, for ADA2), whereas salivary ADA2 significantly correlated with serum amyloid A in horses (r = 0.649, P < 0.01). In cows, salivary tADA and ADA1 significantly increased after calving, correlating with total white blood cell count (r = 0.487, P < 0.05, for both tADA and ADA1). Conclusions: The activity of total ADA and its different isoenzymes, can be measured in serum and saliva of dogs, horses, pigs and cows by a simple and fast procedure described in this report. When measured in saliva, these analytes correlated with other biomarkers of inflammation and it could potentially be used as a biomarkers of inflammation and immune activation in the species of this study.
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Publication Date: 2020-10-12 PubMed ID: 33046093PubMed Central: PMC7549231DOI: 10.1186/s12917-020-02574-2Google Scholar: Lookup
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  • Journal Article
  • Validation Study

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.

This research pilot study investigates the enzymatic activity of total adenosine deaminase (tADA) and its isoenzymes ADA1 and ADA2 in dogs, horses, pigs, and cows. The researchers also develop and validate an automated method for ADA2 measurement, and observe the changes in ADA levels in different inflammatory situations within each species.

ADA Activity and its Isoenzymes in Different Species

  • An enzyme called adenosine deaminase (ADA) can provide information about the immune response within an organism. This study focuses on the measurement of this enzyme and its different types (ADA1 and ADA2) within the blood and saliva of four different species: dogs, horses, pigs, and cows.
  • The total ADA (tADA) enzyme and its forms were found to be present in the serum and saliva of healthy animals across all four species.
  • The researchers note that certain concentrations of a substance called Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) were necessary to inhibit ADA1 activity in specific species and their serum or saliva samples. However, ADA2 activity could not be detected in the cow’s blood and was very low or absent in horse blood and cow saliva.

Automated Procedure for ADA2 Measurement

  • The researchers also developed an automated method to measure ADA2. This method included adding EHNA to a commercial reagent for tADA measurement, which successfully provided consistent and accurate results.
  • This automated procedure was shown to be equivalent to a manual process of sample incubation with EHNA at a similar concentration.

ADA Levels in Health and Disease

  • Observations revealed that the levels of tADA, as well as ADA1 and ADA2, were higher in dogs with leishmaniosis, horses with acute abdominal disease, and pigs with lameness, compared to their healthy counterparts.
  • Notably, the ADA levels in saliva were significantly correlated with other disease markers such as ferritin in dogs, C-reactive protein in pigs, and serum amyloid A in horses. Meanwhile, in cows, increased levels of ADA in saliva were observed after calving, which showed a correlation with their total white blood cell count.

Overall Conclusion

  • In conclusion, this study successfully demonstrated that the activity of ADA and its isoenzymes can be measured in both serum and saliva samples of dogs, horses, pigs, and cows. Moreover, they showed this measurement could be obtained through a simple and fast procedure.
  • When measured in saliva, the ADA levels correlated with other known biomarkers for inflammation, suggesting that they could potentially serve as additional indicators of inflammation and immune activation in these species.

Cite This Article

APA
Contreras-Aguilar MD, Tvarijonaviciute A, Monkeviciene I, Martín-Cuervo M, González-Arostegui LG, Franco-Martínez L, Cerón JJ, Tecles F, Escribano D. (2020). Characterization of total adenosine deaminase activity (ADA) and its isoenzymes in saliva and serum in health and inflammatory conditions in four different species: an analytical and clinical validation pilot study. BMC Vet Res, 16(1), 384. https://doi.org/10.1186/s12917-020-02574-2

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 16
Issue: 1
Pages: 384
PII: 384

Researcher Affiliations

Contreras-Aguilar, María Dolores
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Tvarijonaviciute, Asta
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Monkeviciene, Ingrida
  • Department of Anatomy and Physiology, Research Center of Digestive Physiology and Pathology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes str. 18, LT-47181, Kaunas, Lithuania.
Martín-Cuervo, María
  • Animal Medicine, Faculty of Veterinary Medicine of Cáceres, University of Extremadura, Av. De la Universidad s/n, 10005, Cáceres, Spain.
González-Arostegui, Luis Guillermo
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Franco-Martínez, Lorena
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Cerón, José Joaquín
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Tecles, Fernando
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain.
Escribano, Damián
  • Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Campus of Excellence Mare Nostrum, University of Murcia. Campus de Espinardo s/n, 30100 Espinardo, Murcia, Spain. det20165@um.es.

MeSH Terms

  • Adenine / analogs & derivatives
  • Adenosine Deaminase / blood
  • Adenosine Deaminase / metabolism
  • Adenosine Deaminase Inhibitors
  • Animals
  • Automation
  • Biomarkers / blood
  • Biomarkers / metabolism
  • Cattle / blood
  • Cattle / metabolism
  • Clinical Enzyme Tests / methods
  • Clinical Enzyme Tests / veterinary
  • Dogs / blood
  • Dogs / metabolism
  • Horses / blood
  • Horses / metabolism
  • Inflammation / blood
  • Inflammation / enzymology
  • Inflammation / veterinary
  • Isoenzymes / blood
  • Isoenzymes / metabolism
  • Saliva / metabolism
  • Swine / blood
  • Swine / metabolism

Grant Funding

  • 19849/GERM/15 / Fundaciu00f3n Su00e9neca
  • R-605/2016 / Universidad de Murcia
  • R-605/2016 / Universidad de Murcia
  • IJC2018-035105-I / Ministerio de Economu00eda, Industria y Competitividad, Gobierno de Espau00f1a

Conflict of Interest Statement

The authors declare that they have no competing interests.

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Citations

This article has been cited 26 times.
  1. Rodrigues M, López-Martinez MJ, Ortin-Bustillo A, Cerón JJ, Martinez-Subiela S, Muñoz-Prieto A, Lamy E. Changes in the Saliva Proteome of Pigs with Diarrhoea Caused by Escherichia coli. Proteomes 2023 Apr 3;11(2).
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