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Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2019; 31(5); 688-695; doi: 10.1177/1040638719867124

Equine urinary N-acetyl-β-D-glucosaminidase assay validation and correlation with other markers of kidney injury.

Abstract: Detection of equine acute kidney injury (AKI) is hindered by limited markers of early renal damage in horses. N-acetyl-β-D-glucosaminidase (NAG), a lysosomal enzyme in renal tubular epithelium released into urine during tubular insult, has shown promise for early identification of AKI in humans and other species. We validated an assay for NAG in equine urine and measured urinary NAG in 7 azotemic and 7 non-azotemic client-owned adult horses. The enzymatic NAG assay was validated using within- and between-run coefficients of variation (CVs), recovery following standard addition, and linearity of dilution. Intra- and inter-run CVs (21% and 3.2%, respectively), average recovery following standard addition (99-109%), and linearity under serial dilution ( = 0.997) were satisfactory. Urine NAG index was significantly correlated with urinary fractional excretion of sodium (FE; ρ = 0.76, < 0.001) and plasma creatinine (ρ = 0.74, = 0.001). Median urine NAG indices were higher in azotemic horses ( = 0.006), in horses with increased urinary FE ( = 0.006), and in horses with increased urine gamma-glutamyl transferase index ( = 0.032). Urine NAG can be measured in horses and shows positive correlation with 2 current renal biomarkers. Additional work is needed to establish normal equine reference intervals and characterize the increase of urine NAG index in horses in relation to tubular injury.
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Publication Date: 2019-07-26 PubMed ID: 31347464PubMed Central: PMC6727119DOI: 10.1177/1040638719867124Google 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.

The study explores the potential of N-acetyl-β-D-glucosaminidase (NAG) as a potential marker for early detection of acute kidney injury (AKI) in horses. It presents the successful validation of a NAG assay in equine urine with significantly higher urine NAG indices observed in azotemic horses.

Objective of the Study

  • The research is primarily aimed at recognizing and investigating a potential marker for detecting early renal damages in horses, as the current procedures available have limitations.
  • The study focuses on the lysosomal enzyme N-acetyl-β-D-glucosaminidase (NAG), which is released into urine during kidney tubular damage and has shown promise as a potential early marker for acute kidney injury (AKI) in diverse species, including humans.

Methodology and Validation of the NAG Assay

  • The researchers use an enzymatic NAG assay specifically for equine urine. The assay’s validation involves evaluating its within- and between-run coefficients of variation (CVs), its recovery rate following add-on standards, and its dilution linearity.
  • Outcomes from the assay validation were determined satisfactory, with intra- and inter-run coefficients of variation at 21% and 3.2% respectively, recovery following standard addition averaging at around 99-109%, and proven linearity under serial dilution (r=0.997).

Study Findings and Correlation with Other Renal Biomarkers

  • Urinary NAG was measured in both azotemic and non-azotemic adult horses. Results have shown significant urine NAG index correlations with urinary fractional excretion of sodium (FE) and plasma creatinine.
  • It was found that the median urine NAG indices were notably higher in azotemic horses, as well as in those manifesting increased urinary FE and urine gamma-glutamyl transferase index.

Conclusions and Further Study

  • The findings posited that urine NAG can be reliably measured in horses, and has shown possible correlation with current renal biomarkers.
  • While the research results prove promising for urine NAG as a potential early renal damage marker, more research is necessary to determine the normal equine reference intervals for NAG, and to fully understand its increase in relation to tubular injury in horses.

Cite This Article

APA
Bayless RL, Moore AR, Hassel DM, Byer BJ, Landolt GA, Nout-Lomas YS. (2019). Equine urinary N-acetyl-β-D-glucosaminidase assay validation and correlation with other markers of kidney injury. J Vet Diagn Invest, 31(5), 688-695. https://doi.org/10.1177/1040638719867124

Publication

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
ISSN: 1943-4936
NlmUniqueID: 9011490
Country: United States
Language: English
Volume: 31
Issue: 5
Pages: 688-695

Researcher Affiliations

Bayless, Rosemary L
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).
Moore, A Russell
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).
Hassel, Diana M
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).
Byer, Brittney J
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).
Landolt, Gabriele A
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).
Nout-Lomas, Yvette S
  • College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Bayless).
  • College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Moore, Hassel, Byer, Landolt, Nout-Lomas).

MeSH Terms

  • Acetylglucosaminidase / urine
  • Acute Kidney Injury / diagnosis
  • Acute Kidney Injury / urine
  • Acute Kidney Injury / veterinary
  • Animals
  • Biomarkers / urine
  • Creatinine / blood
  • Female
  • Horse Diseases / diagnosis
  • Horse Diseases / urine
  • Horses
  • Humans
  • Male
  • Sodium / urine
  • gamma-Glutamyltransferase / urine

Conflict of Interest Statement

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

References

This article includes 44 references
  1. Adams R, McClure JJ, Gossett KA, Koonce KL, Ezigbo C. Evaluation of a technique for measurement of gamma-glutamyltranspeptidase in equine urine.. Am J Vet Res 1985 Jan;46(1):147-50.
    pubmed: 2857544
  2. Adelman RD, Conzelman G, Spangler W, Ishizaki G. Comparative nephrotoxicity of gentamicin and netilmicin: functional and morphological correlations with urinary enzyme activities.. Curr Probl Clin Biochem 1979;(9):166-82.
    pubmed: 446071
  3. Bohn AA. Laboratory evaluation of the equine renal system. Equine Clinical Pathology 2014:87–101.
  4. Brunker JD, Ponzio NM, Payton ME. Indices of urine N-acetyl-beta-D-glucosaminidase and gamma-glutamyl transpeptidase activities in clinically normal adult dogs.. Am J Vet Res 2009 Feb;70(2):297-301.
    pubmed: 19231965doi: 10.2460/ajvr.70.2.297google scholar: lookup
  5. Cebra C. Disorders of the urinary system. Llama and Alpaca Care: Medicine, Surgery, Reproduction, Nutrition, and Herd Health 2014:464–476.
  6. Finco DR, Duncan JR. Evaluation of blood urea nitrogen and serum creatinine concentrations as indicators of renal dysfunction: a study of 111 cases and a review of related literature.. J Am Vet Med Assoc 1976 Apr 1;168(7):593-601.
    pubmed: 1270342
  7. Garry F, Chew DJ, Hoffsis GF. Enzymuria as an index of renal damage in sheep with induced aminoglycoside nephrotoxicosis.. Am J Vet Res 1990 Mar;51(3):428-32.
    pubmed: 1969254
  8. Geor RJ. Acute renal failure in horses.. Vet Clin North Am Equine Pract 2007 Dec;23(3):577-91, v-vi.
    pubmed: 18061851doi: 10.1016/j.cveq.2007.09.007google scholar: lookup
  9. Gluhovschi G, Gadalean F, Gluhovschi C, Velciov S, Petrica L, Bob F, Bozdog G, Kaycsa A. Urinary biomarkers in assessing the nephrotoxic potential of gentamicin in solitary kidney patients after 7 days of therapy.. Ren Fail 2014 May;36(4):534-40.
    pubmed: 24456153doi: 10.3109/0886022x.2013.876349google scholar: lookup
  10. Ida S, Yokota M, Ueoka M, Kiyoi K, Takiguchi Y. Mild to severe lithium-induced nephropathy models and urine N-acetyl-beta-D-glucosaminidase in rats.. Methods Find Exp Clin Pharmacol 2001 Oct;23(8):445-8.
    pubmed: 11838319doi: 10.1358/mf.2001.23.8.662132google scholar: lookup
  11. Jansen D, Peters E, Heemskerk S, Koster-Kamphuis L, Bouw MP, Roelofs HM, van Oeveren W, van Heijst AF, Pickkers P. Tubular Injury Biomarkers to Detect Gentamicin-Induced Acute Kidney Injury in the Neonatal Intensive Care Unit.. Am J Perinatol 2016 Jan;33(2):180-7.
    pubmed: 26344007doi: 10.1055/s-0035-1563714google scholar: lookup
  12. Jung K, Pergande M, Schreiber G, Schröder K. Stability of enzymes in urine at 37 degrees C.. Clin Chim Acta 1983 Jul 15;131(3):185-91.
    pubmed: 6883714doi: 10.1016/0009-8981(83)90087-6google scholar: lookup
  13. Jung K, Schulze G, Reinholdt C. Different diuresis-dependent excretions of urinary enzymes: N-acetyl-beta-D-glucosaminidase, alanine aminopeptidase, alkaline phosphatase, and gamma-glutamyltransferase.. Clin Chem 1986 Mar;32(3):529-32.
    pubmed: 2868815
  14. Lackey MN, Belknap EB, Salman MD, Tinguely L, Johnson LW. Urinary indices in llamas fed different diets.. Am J Vet Res 1995 Jul;56(7):859-65.
    pubmed: 7574151
  15. Lapointe C, Bélanger MC, Dunn M, Moreau M, Bédard C. N-acetyl-beta-D-glucosaminidase index as an early biomarker for chronic kidney disease in cats with hyperthyroidism.. J Vet Intern Med 2008 Sep-Oct;22(5):1103-10.
    pubmed: 18700858doi: 10.1111/j.1939-1676.2008.0168.xgoogle scholar: lookup
  16. Le JM, Han YH, Choi SJ, Park JS, Jang JJ, Bae RJ, Lee MJ, Kim MJ, Lee YH, Kim D, Lee HY, Park SH, Park CB, Kang JS, Kang JK. Variation of nephrotoxicity biomarkers by urinary storage condition in rats.. Toxicol Res 2014 Dec;30(4):305-9.
    pmc: PMC4289932pubmed: 25584151doi: 10.5487/tr.2014.30.4.305google scholar: lookup
  17. Liangos O, Perianayagam MC, Vaidya VS, Han WK, Wald R, Tighiouart H, MacKinnon RW, Li L, Balakrishnan VS, Pereira BJ, Bonventre JV, Jaber BL. Urinary N-acetyl-beta-(D)-glucosaminidase activity and kidney injury molecule-1 level are associated with adverse outcomes in acute renal failure.. J Am Soc Nephrol 2007 Mar;18(3):904-12.
    pubmed: 17267747doi: 10.1681/asn.2006030221google scholar: lookup
  18. McWilliam SJ, Antoine DJ, Sabbisetti V, Turner MA, Farragher T, Bonventre JV, Park BK, Smyth RL, Pirmohamed M. Mechanism-based urinary biomarkers to identify the potential for aminoglycoside-induced nephrotoxicity in premature neonates: a proof-of-concept study.. PLoS One 2012;7(8):e43809.
    pmc: PMC3427159pubmed: 22937100doi: 10.1371/journal.pone.0043809google scholar: lookup
  19. Mishra OP, Rai AK, Srivastava P, Pandey K, Abhinay A, Prasad R, Mishra RN, Schaefer F. Predictive ability of urinary biomarkers for outcome in children with acute kidney injury.. Pediatr Nephrol 2017 Mar;32(3):521-527.
    pubmed: 27387432doi: 10.1007/s00467-016-3445-ygoogle scholar: lookup
  20. Miyai T, Ogata M. Changes in the concentrations of urinary proteins after physical exercise.. Acta Med Okayama 1990 Oct;44(5):263-6.
    pubmed: 2260497doi: 10.18926/amo/30453google scholar: lookup
  21. Morita A, Numata Y, Kosugi Y, Noto A, Takeuchi N, Uchida K. Stabilities of N-acetyl-beta-D-glucosaminidase (NAG) isoenzymes in urine: advantage of NAG isoenzyme B measurement in clinical applications.. Clin Chim Acta 1998 Nov;278(1):35-43.
    pubmed: 9877122doi: 10.1016/s0009-8981(98)00157-0google scholar: lookup
  22. Morris DD, Divers TJ, Whitlock RH. Renal clearance and fractional excretion of electrolytes over a 24-hour period in horses.. Am J Vet Res 1984 Nov;45(11):2431-5.
    pubmed: 6524740
  23. Nabity MB, Lees GE, Cianciolo R, Boggess MM, Steiner JM, Suchodolski JS. Urinary biomarkers of renal disease in dogs with X-linked hereditary nephropathy.. J Vet Intern Med 2012 Mar-Apr;26(2):282-93.
    pubmed: 22356524doi: 10.1111/j.1939-1676.2012.00891.xgoogle scholar: lookup
  24. Pennemans V, Rigo JM, Faes C, Reynders C, Penders J, Swennen Q. Establishment of reference values for novel urinary biomarkers for renal damage in the healthy population: are age and gender an issue?. Clin Chem Lab Med 2013 Sep;51(9):1795-802.
    pubmed: 23648635doi: 10.1515/cclm-2013-0157google scholar: lookup
  25. Polzin D. Chronic kidney disease. Textbook of Veterinary Internal Medicine 2005:1756–1785.
  26. Price RG. The role of NAG (N-acetyl-beta-D-glucosaminidase) in the diagnosis of kidney disease including the monitoring of nephrotoxicity.. Clin Nephrol 1992;38 Suppl 1:S14-9.
    pubmed: 1295704
  27. Raekallio MR, Saario-Paunio EM, Rajamäki MM, Sankari SM, Palviainen MJ, Siven MS, Peltoniemi SM, Leinonen ME, Honkavaara JM, Vainio OM. Early detection of ketoprofen-induced acute kidney injury in sheep as determined by evaluation of urinary enzyme activities.. Am J Vet Res 2010 Oct;71(10):1246-52.
    pubmed: 20919914doi: 10.2460/ajvr.71.10.1246google scholar: lookup
  28. Roussel AJ, Cohen ND, Ruoff WW, Brumbaugh GW, Schmitz DG, Kuesis BS. Urinary indices of horses after intravenous administration of crystalloid solutions.. J Vet Intern Med 1993 Jul-Aug;7(4):241-6.
    pubmed: 8246214doi: 10.1111/j.1939-1676.1993.tb01014.xgoogle scholar: lookup
  29. Sato R, Nakajima N, Soeta S, Sato J, Naito Y. Urine N-acetyl-beta-D-glucosaminidase activity in healthy cattle.. Am J Vet Res 1997 Nov;58(11):1197-200.
    pubmed: 9361877
  30. Sato R, Soeta S, Syuto B, Yamagishi N, Sato J, Naito Y. Urinary excretion of N-acetyl-beta-D-glucosaminidase and its isoenzymes in cats with urinary disease.. J Vet Med Sci 2002 Apr;64(4):367-71.
    pubmed: 12014584doi: 10.1292/jvms.64.367google scholar: lookup
  31. Savage CJ. Urinary clinical pathologic findings and glomerular filtration rate in the horse.. Vet Clin North Am Equine Pract 2008 Aug;24(2):387-404, vii.
    pubmed: 18652961doi: 10.1016/j.cveq.2008.05.002google scholar: lookup
  32. Sheira G, Noreldin N, Tamer A, Saad M. Urinary biomarker N-acetyl-β-D-glucosaminidase can predict severity of renal damage in diabetic nephropathy.. J Diabetes Metab Disord 2015;14:4.
    pmc: PMC4340101pubmed: 25717442doi: 10.1186/s40200-015-0133-6google scholar: lookup
  33. Smets PM, Meyer E, Maddens BE, Duchateau L, Daminet S. Urinary markers in healthy young and aged dogs and dogs with chronic kidney disease.. J Vet Intern Med 2010 Jan-Feb;24(1):65-72.
    pubmed: 20041990doi: 10.1111/j.1939-1676.2009.0426.xgoogle scholar: lookup
  34. Stockham SL, Scott MA. Urinary system. Fundamentals of Veterinary Clinical Pathology 2008:708–840.
  35. Trachtenberg F, Barregard L, McKinlay S. The influence of urinary flow rate in children on excretion of markers used for assessment of renal damage: albumin, gamma-glutamyl transpeptidase, N-acetyl-beta-D -glucosaminidase, and alpha1-microglobulin.. Pediatr Nephrol 2008 Mar;23(3):445-56.
    pubmed: 17704953doi: 10.1007/s00467-007-0568-1google scholar: lookup
  36. Trim CM, Hanson RR. Effects of xylazine on renal function and plasma glucose in ponies.. Vet Rec 1986 Jan 18;118(3):65-7.
    pubmed: 3952942doi: 10.1136/vr.118.3.65google scholar: lookup
  37. Tsuji S, Sugiura M, Tsutsumi S, Yamada H. Sex differences in the excretion levels of traditional and novel urinary biomarkers of nephrotoxicity in rats.. J Toxicol Sci 2017;42(5):615-627.
    pubmed: 28904297doi: 10.2131/jts.42.615google scholar: lookup
  38. Uechi M, Nogami Y, Terui H, Nakayama T, Ishikawa R, Wakao Y, Takahashi M. Evaluation of urinary enzymes in dogs with early renal disorder.. J Vet Med Sci 1994 Jun;56(3):555-6.
    pubmed: 7948390doi: 10.1292/jvms.56.555google scholar: lookup
  39. Uechi M, Uechi H, Nakayama T, Wakao Y, Ogasawara T, Takase K, Takahashi M. The circadian variation of urinary N-acetyl-beta-D-glucosaminidase and gamma-glutamyl transpeptidase in clinically healthy cats.. J Vet Med Sci 1998 Sep;60(9):1033-4.
    pubmed: 9795907doi: 10.1292/jvms.60.1033google scholar: lookup
  40. Wellwood JM, Ellis BG, Price RG, Hammond K, Thompson AE, Jones NF. Urinary N-acetyl- beta-D-glucosaminidase activities in patients with renal disease.. Br Med J 1975 Aug 16;3(5980):408-11.
    pmc: PMC1673872pubmed: 1156788doi: 10.1136/bmj.3.5980.408google scholar: lookup
  41. Whiting PH, Brown PA. The relationship between enzymuria and kidney enzyme activities in experimental gentamicin nephrotoxicity.. Ren Fail 1996 Nov;18(6):899-909.
    pubmed: 8948524doi: 10.3109/08860229609047716google scholar: lookup
  42. Wu X, Wei S, Wei Y, Guo B, Yang M, Zhao D, Liu X, Cai X. The reference dose for subchronic exposure of pigs to cadmium leading to early renal damage by benchmark dose method.. Toxicol Sci 2012 Aug;128(2):524-31.
    pubmed: 22610606doi: 10.1093/toxsci/kfs173google scholar: lookup
  43. Zhang X, Zhou H, Li Y. Combined Detection of Urinary Micro Albumin, α1-microglobulin and N-acetyl-β-D-glucosaminidase in the Early Diagnosis of Diabetic Nephropathy.. Pak J Med Sci 2017 Nov-Dec;33(6):1324-1327.
    pmc: PMC5768818pubmed: 29492052doi: 10.12669/pjms.336.13168google scholar: lookup
  44. Zhou X, Ma B, Lin Z, Qu Z, Huo Y, Wang J, Li B. Evaluation of the usefulness of novel biomarkers for drug-induced acute kidney injury in beagle dogs.. Toxicol Appl Pharmacol 2014 Oct 1;280(1):30-5.
    pubmed: 25034533doi: 10.1016/j.taap.2014.07.002google scholar: lookup

Citations

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  1. Venkatesan A, Roy A, Kulandaivel S, Natesan V, Kim SJ. p-Coumaric Acid Nanoparticles Ameliorate Diabetic Nephropathy via Regulating mRNA Expression of KIM-1 and GLUT-2 in Streptozotocin-Induced Diabetic Rats. Metabolites 2022 Nov 23;12(12).
    doi: 10.3390/metabo12121166pubmed: 36557204google scholar: lookup
  2. Galen GV, Olsen E, Siwinska N. Biomarkers of Kidney Disease in Horses: A Review of the Current Literature. Animals (Basel) 2022 Oct 5;12(19).
    doi: 10.3390/ani12192678pubmed: 36230418google scholar: lookup
  3. Popa R, Diaconu M, Cirlig V, Văruţ CM, Caragea DC, Codea RA, Tabaran FA, Aurori M, Gafencu M, Georgescu CC. Combination of Pentoxifylline and Ginko Biloba Nephroprotective Effect in Animal Models with Vancomycin-Induced Nephrotoxicity. Curr Health Sci J 2022 Jan-Mar;48(1):68-74.
    doi: 10.12865/CHSJ.48.01.10pubmed: 35911932google scholar: lookup
  4. Tan D, Zhao L, Peng W, Wu FH, Zhang GB, Yang B, Huo WQ. Value of urine IL-8, NGAL and KIM-1 for the early diagnosis of acute kidney injury in patients with ureteroscopic lithotripsy related urosepsis. Chin J Traumatol 2022 Jan;25(1):27-31.
    doi: 10.1016/j.cjtee.2021.10.001pubmed: 34702632google scholar: lookup
  5. van Galen G, Divers TJ, Savage V, Schott HC 2nd, Siwinska N. ECEIM consensus statement on equine kidney disease. J Vet Intern Med 2024 Jul-Aug;38(4):2008-2025.
    doi: 10.1111/jvim.17101pubmed: 38801172google scholar: lookup
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