Comparative Clinical Pathology

, Volume 28, Issue 1, pp 151–156 | Cite as

Evaluation of leukocyte esterase test strips for rapid diagnosis of subclinical mastitis in dairy cows

  • Abdolah Mirzaei
  • Amirarsalan Khorsand
  • Abolfazl Hajibemani
  • Hassan SharifiyazdiEmail author
Original Article


Subclinical mastitis is difficult to detect because it has no visible indications and requires special tests to be detected. Therefore, an untimely and precise diagnosis of subclinical mastitis has been suggested. The purpose of this paper was to estimate the performance of the Analyticon reagent strip test application for the diagnosis of subclinical mastitis as a cow-side test. Milk samples from 111 dairy cows were collected and the leukocyte esterase (LE) test was performed with the reagent strip test. Leukocyte esterase activity was estimated following 10 (LE10) and 15 (LE15) min. Also, milk somatic cell count (SCC) obtained by an automated Fossomatic machine was considered as the gold standard for diagnosing subclinical mastitis. The specificity (Sp), sensitivity (Se), positive likelihood ratio (+LR), negative likelihood ratio (−LR), area under the receiver operating curve (AUC), and kappa coefficient (κ) were determined. The 75.7% (84/111) milk samples with SCC ≥ 200,000/mL were considered as the subclinical mastitis milk. A strong statistically significant correlation was found between SCC, LE test marker, and SCC scoring (0–3) in all milk samples. Based on the cutoff point, the LE15 test had Se = 80%, Sp = 100%, +LR = ∞, and −LR = 0.20. There was a very close relation between LE15 and SCC test with a kappa coefficient of 0.70 (P < 0.0001). Also, according to the cutoff point of 0.5, the LE10 test had Se = 64%, Sp = 100%, +LR = ∞, and −LR = 0.36. In addition, there was a very close relation between LE10 and SCC tests with a kappa coefficient of 0.50 (P < 0.0001). The LE activity was significantly higher (P < 0.05) in subclinical mastitis samples compared to that of healthy milk samples at the time of 10 and 15 min. In summary, these results showed a strong correlation among the LE test markers with the reagent strip test of Analyticon and SCC using the Fossomatic machine. Based on such findings, it is suggested that the LE activity could be used as a clinically useful cow-side test and a valid method for screening the subclinical mastitis.


Subclinical mastitis Somatic cell count Leukocyte esterase activity Cow-side test 



This study was funded by the Research Council of Shiraz University and School of Veterinary Medicine, Shiraz University (Grant No. 87-GR-VT-47).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This experiment was performed under the approval of the state committee on animal ethics, Shiraz University, Shiraz, Iran. Also, the recommendations of European Council Directive (86/609/EC) of November 24, 1986, regarding the protection of animals used for experimental purposes, were considered.


  1. Blowey RW, Edmondson P (2010) Mastitis control in dairy herdsCabi: an illustrated and practical guide. Farming Press Books, IpswichCrossRefGoogle Scholar
  2. Booth R, Morse G, Treece J, Sammelwitz P (1965) Relationship of a-esterase activity blood-serum albumin and leukocyte count during experimental exposure to streptococcus agalactiae (cornell strain 48). J Dairy Sci 822Google Scholar
  3. Cheong S, Nydam D, Galvao K, Crosier B, Ricci A, Caixeta L, Sper R, Fraga M, Gilbert R (2012) Use of reagent test strips for diagnosis of endometritis in dairy cows. Theriogenology 77:858–864CrossRefGoogle Scholar
  4. Couto G, Vaillancourt D, Lefebvre R (2013) Comparison of a leukocyte esterase test with endometrial cytology for diagnosis of subclinical endometritis in postpartum dairy cows. Theriogenology 79:103–107CrossRefGoogle Scholar
  5. Djabri B, Bareille N, Beaudeau F, Seegers H (2002) Quarter milk somatic cell count in infected dairy cows: a meta-analysis. Vet Res 33:335–357CrossRefGoogle Scholar
  6. Forster T, Montgomery M, Montoure J (1961) Some factors influencing the activity of the A-, B-, and C-esterases of bovine milk1. J Dairy Sci 44:1420–1430CrossRefGoogle Scholar
  7. Hajibemani A, Mirzaei A, Sharifiyazdi H, Ghasrodashti AR (2016) Vaginal fornix discharge cellularity and its leukocyte esterase activity for diagnosis of endometritis in dairy cows. J Fac Vet Med Istunbul Univ i 42:57–64Google Scholar
  8. Hiss S, Mueller U, Neu-Zahren A ,Sauerwein H (2007) Haptoglobin and lactate dehydrogenase measurements in milk for the identification of subclinically diseased udder quarters. Vet Med -praha-;52:245Google Scholar
  9. Iraguha B, Hamudikuwanda H, Mushonga B, Kandiwa E, Mpatswenumugabo JP (2017) Comparison of cow-side diagnostic tests for subclinical mastitis of dairy cows in Musanze district, Rwanda. J S Afr Vet Assoc 88:1–6CrossRefGoogle Scholar
  10. Jones GM ,Bailey TL (2009) Understanding the basics of mastitisGoogle Scholar
  11. Kandeel SA, Ebied MH, Arnaout FK, Galila EM, Megahed AA, Constable PD (2017) Clinical utility of two leukocyte esterase reagent strips for the cow-side diagnosis of subclinical mastitis in lactating dairy cattle. Assiut Vet Med J:63Google Scholar
  12. KrrcHEN B (1981) Review of the progress of dairy science: bovine mastitis: milk compositional changes and related diagnostic test. J Dairy Res 48:167–188CrossRefGoogle Scholar
  13. Lam T, Riekerink RO, Sampimon O, Smith H (2009) Mastitis diagnostics and performance monitoring: a practical approach. Ir Vet J 62:S34CrossRefGoogle Scholar
  14. Luedecke L, Forster T, Ashworth U (1967) Relationship between California mastitis test reaction and leucocyte count, catalase activity, and A-esterase activity of milk from opposite Quarters1. J Dairy Sci 50:1592–1596CrossRefGoogle Scholar
  15. Marquardt R, Forster T, Spencer G, Stabenfeldt G (1966) A-esterase activities of milk from cows with experimentally induced mastitis1, 2. J Dairy Sci 49:631–635CrossRefGoogle Scholar
  16. Philpot WN, Nickerson SC (2000) Winning the fight against mastitis. Westfalia Surge, Inc., Napervile, ILGoogle Scholar
  17. Pillai S, Kunze E, Sordillo L, Jayarao B (2001) Application of differential inflammatory cell count as a tool to monitor udder health. J Dairy Sci 84:1413–1420CrossRefGoogle Scholar
  18. PortaCheck (2010) PortaSCC® cow test - research." Welcome to PortaCheck Porta Check, INC Web 28 Nov. <
  19. PortaCheck (2011) The PortaSCC® milk test is a somatic cell count test for early on-farm mastitis detection in cows. Moorestown, NJ.
  20. Pyörälä S (2003) Indicators of inflammation in the diagnosis of mastitis. Vet Res 34:565–578CrossRefGoogle Scholar
  21. Rahman M, Islam M, Uddin M, Aktaruzzaman M (2010) Prevalence of subclinical mastitis in dairy cows reared in Sylhet district of Bangladesh. Int J Bio Res 1:23–28Google Scholar
  22. Rodrigues A, Cassoli L, Machado P, Ruegg P (2009) Evaluation of an on-farm test to estimate somatic cell count. J Dairy Sci 92:990–995CrossRefGoogle Scholar
  23. Ruegg P, Reinemann D (2002) Milk quality and mastitis tests. Bovine Pract 36:41–54Google Scholar
  24. Salvador R, Soliven R, Balagan E, Abes N, Gutierrez C, Mingala C (2014) Evaluation of a portable somatic cell counter in the diagnosis of bubaline subclinical mastitis. Thai J Agric Sci 47:205–209Google Scholar
  25. Schukken YH, Wilson DJ, Welcome F, Garrison-Tikofsky L, Gonzalez RN (2003) Monitoring udder health and milk quality using somatic cell counts. Vet Res 34:579–596CrossRefGoogle Scholar
  26. Schultz L, Broom R, Jasper D, Berger RM, Natwke R, Philpot W, Smith J, Thompson P (1978) Current concepts of bovine mastitis. In: The National Mastitis Council;1–16Google Scholar
  27. Smolenski G, Haines S, Kwan FY-S, Bond J, Farr V, Davis SR, Stelwagen K, Wheeler TT (2007) Characterisation of host defence proteins in milk using a proteomic approach. J Proteome Res 6:207–215CrossRefGoogle Scholar
  28. St John A, Boyd JC, Lowes AJ, Price CP (2006) The use of urinary dipstick tests to exclude urinary tract infection: a systematic review of the literature. Am J Clin Pathol 126:428–436Google Scholar
  29. Viguier C, Arora S, Gilmartin N, Welbeck K, O’Kennedy R (2009) Mastitis detection: current trends and future perspectives. Trends Biotechnol 27:486–493CrossRefGoogle Scholar
  30. YalÇin C (2000) Cost of mastitis in Scottish dairy herds with low and high subclinical mastitis problems. Turk J Vet Anim Sci 24:465–472Google Scholar
  31. Zadoks RN, Schukken YH (2006) Use of molecular epidemiology in veterinary practice. Vet Clin: Food Anim Pract 22:229–261CrossRefGoogle Scholar
  32. Zhao X, Lacasse P (2008) Mammary tissue damage during bovine mastitis: causes and control. J Anim Sci 86:57–65CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Clinical Sciences, School of Veterinary MedicineShiraz UniversityShirazIran
  2. 2.Department of Clinical Sciences, Faculty of Veterinary MedicineTabriz UniversityTabrizIran

Personalised recommendations