Original Scientific Article
Prevalence of subclinical mastitis pathogens in small dairy farms in Republic of North Macedonia
Aleksandar Janevsкi*,
Iskra Cvetkovikj,
Sanja Kiprijanovska,
Aleksandar Dimovski,
Katarina Davalieva,
Ljupco Angelovski,
Miroslav Kjosevski,
Dine Mitrov

Mac Vet Rev 2020; 43 (1): 23 - 30


Received: 20 October 2019

Received in revised form: 10 January 2020

Accepted: 13 January 2020

Available Online First: 24 January 2020

Published on: 15 March 2020

Correspondence: Aleksandar Janevsкi, aleksandar_janevski@yahoo.com


Subclinical mastitis is an asymptomatic udder infection distributed worldwide with enormous losses in the dairy industry. The study’s objective was to determine the presence of this pathological condition in small dairy farms in the R. of N. Macedonia and to identify the most common associated bacteria. Milk samples were obtained from 96 dairy cows (378 udder quarters) in seven dairy farms, in 3 consecutive samplings 24-72 hours apart. The samples were cultured on routine bacteriological growth media and incubated for 24-48 hours. The isolates were identified by AximaiD Plus MALDITOF MS Platform. Subclinical mastitis was found in 49 animals (51%) and 104 infected quarters (27%). The most frequent isolated bacteria on cow level were Streptococcus uberis (19.4%), Staphylococcus haemolyticus (13.4%), Staphylococcus aureus (7.4%) and Staphylococcu ssimulans (7.4%). On quarter level, the most isolated pathogen was Streptococcus uberis (35.6%) followed by Staphylococcu shaemolyticus and Staphylococcus aureus (10.3% and 9.2% respectively). Subclinical mastitis was found to be highly present in the selected small dairy farms. The most prevalent bacteria identified in the dairy farms (Streptococcus uberis, Staphylococcus aureus and coagulase–negative staphylococci) indicate that poor management and udder health practices, inadequate milking procedures and lack of mastitis control strategies greatly contribute to occurrence and persistence of subclinical mastitis.

Keywords: subclinical mastitis, MALDI-TOF, bacteria, milk


  1. Gianneechini, R., Concha, C., Rivero, R., Delucci, I., Moreno López J. (2002). Occurrence of clinical and sub-clinical mastitis in dairy herds in the west littoral region in Uruguay. Acta Vet Scand. 43, 221-230. https://doi.org/10.1186/1751-0147-43-221 PMid:12831175 PMCid:PMC1764198
  2. Gonçalvesa, J.L., Kamphuisb, C., Martinsa, C.M.M.R., Barreiroa, J.R., Tomazia, T., Gameiroa, A.H., Hogeveenb, H., dos Santos, M.V. (2018). Bovine subclinical mastitis reduces milk yield and economic return. Livest Sci. 210, 25-32. https://doi.org/10.1016/j.livsci.2018.01.016
  3. Forsbаck, L., Lindmark-Mansson, H., Andren, A., Kerstedt, M.A., Svennersten-Sjaunja, K. (2009). Udder quarter milk composition at different levels of somatic cell count in cow composite milk. Animal 3(5): 710-717. https://doi.org/10.1017/S1751731109004042 PMid:22444450
  4. Sharif, A., Muhammad, G. (2008). Somatic cell count as an indicator of udder health status under modern dairy production: A review. Pakistan Vet J. 28(4): 194-200.
  5. Persson, Y., Nyman, A.K.J., Grönlund-Andersson, U. (2011). Etiology and antimicrobial susceptibility of udder pathogens from cases of subclinical mastitis in dairy cows in Sweden. Acta Vet Scand. 53(1): 36. https://doi.org/10.1186/1751-0147-53-36 PMid:21649936 PMCid:PMC3118135
  6. Tongel, P., Broucek, J. (2010). Influence of hygienic condition on prevalence of mastitis and lameness in dairy cows. Slovak J Anim Sci. 43(2): 95-99.
  7. Abdel-Rady, A., Sayed, M. (2009). Epidemiological studies on subclinical mastitis in dairy cows in Assiut Governorate. Vet World. 2(10): 373-380. https://doi.org/10.5455/vetworld.2009.373-380
  8. Sanotharan, N., Pagthinathan, M., Nafees, M.S.M. (2016). Prevalence of bovine subclinical mastitis and its association with bacteria and risk factors in milking of Batticaloa district in Sri Lanka. IJSRIT 3 (6): 137-150.
  9. Botrel, M.A., Haenni, M., Morignat, E., Sulpice, P., Madec, J.Y., Calavas, D. (2010). Distribution and antimicrobial resistance of clinical and subclinical mastitis pathogens in dairy cows in Rhone-Alpes, France. Foodborne Pathog Dis. 7(5): 479-487. https://doi.org/10.1089/fpd.2009.0425 PMid:19919286
  10. Tenhagen, B.A., Koster, G., Wallmann, J., Heuwieser, W. (2006). Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg, Germany. J Dairy Sci. 89, 2542-2551. https://doi.org/10.3168/jds.S0022-0302(06)72330-X
  11. Sharma, N., Singh, N. K., Bhadwal, M. S. (2011). Relationship of somatic cell count and mastitis: an overview. Asian-Aust J Anim Sci. 24(3): 429 - 438. https://doi.org/10.5713/ajas.2011.10233
  12. Hristov, S., Relich, R., Stancovich, B., Vukovich, D. (2006). Influence of individual factors on somatic cell count in cow's milk. Proceedings of Scientific Papers, 12 (3-4): 47-57. [in Serbian]
  13. Bojkovic Kovacevic, S., Lazarevic, N. (2009). Correlation between CMT test, subclinical mastitis and somatic cell count. Proceedings of Scientific Papers, 15 (3-4) 29-37. [in Serbian]
  14. Manzi, M. de P., Nуbrega, D.B., Faccioli, P.Y., Troncarelli, M.Z., Menozzi, B.D, Langoni, H. (2012). Relationship between teat-end condition, udder cleanliness and bovine subclinical mastitis. Res Vet Sci. 93, 430-434. https://doi.org/10.1016/j.rvsc.2011.05.010 PMid:21669449
  15. National Mastitis Council (2012). Interpreting bacteriological culture results to diagnose bovine intramammary infections. Research Committee Report. [Internet], Verona, Winskonsin, USA. [Accessed November 2018] https://www.nmconline.org/wp-content/uploads/2016/08/Interpreting-Bacteriological-Culture-Results.pdf
  16. Katic, V., El Tayeb, H., Babic, L., Popovic J. (1994). Effect of mastitis on milk quality. Vet Glas. 48(3-4): 171-176. [in Serbian]
  17. Knight, CH., Peaker, M., (1991). Mammary gland physiology. Flemishe Vet J. 62, (Suppl. 1): 33-41.
  18. Berglund, I., Pettersson, G., Ostensson, K., Svennersten-Sjaunja, K. (2007). Quarter milking for improved detection of increased SCC. Reprod Dom Anim. 42, 427-432. https://doi.org/10.1111/j.1439-0531.2006.00803.x PMid:17635782
  19. Food and Veterinary Agency of Republic of Macedonia (2012). Rulebook for specific requirements for safety and hygiene and the manner of conducting official controls of milk and milk products. Official Gazette of Republic of Macedonia No. 26/2012. [in Macedonian]. [Accessed November 2019] http://fva.gov.mk/images/stories/zakoni/pdf/2012_026%20pravilnik_posebni_baranja_sluzbeni_kontroli_mleko.pdf
  20. Commission of European Communities: Council regulation 853/2004/EC on laying down specific hygiene rules for on the hygiene of foodstuffs (2004). Official Journal L139/55 [Accessed November 2019] https://eurlex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2004:139:0055:0205:en:PDF
  21. National Mastitis Council (2004). Microbiological procedures for the diagnosis of bovine udder infection and determination of milk quality. microbiological procedures for the diagnosis of bovine udder infection and determination of milk quality. [Internet]. Verona, Winskonsin, USA. [Accessed November 2018] https://extension.umaine.edu/veterinarylab/wp-content/uploads/sites/23/2014/07/Procedures-for-Collecting-Milk-Samples-NMC.pdf
  22. Pitkala, A., Haveri, M., Pyorala, S., Myllys, V., Honkanen-Buzalski, T. (2004). Bovine mastitis in Finland 2001 - prevalence, distribution of bacteria and antimicrobial resistance. Dairy Sci. 87, 2433-2441. https://doi.org/10.3168/jds.S0022-0302(04)73366-4
  23. Wilkins, C. L., O Lay, J. (2006) Identification of microorganisms by mass spectrometry. Hoboken, New Jersey: Wiley-Interscience. https://doi.org/10.1002/0471748641
  24. AnagnosTec [Internet]. Potsdam, Germany: Spectral archive and microbial identification system user manual (2009). Availlable from: http://www.anagnostec.eu/products-services/reference- databases.html
  25. ISO 13366-2:2006 Milk-enumeration of somatic cells, Part 2-guidance on the operation of fluoro-opto-electronic counters. 2nd ed. Geneva: ISO; 2006. 15p.
  26. Sylejmani, D., Ramadani, N., Robaj, A., Hamidi, A. (2015). Prevalence and antimicrobial susceptibility of bacterial isolates from subclinical mastitis in dairy farms in Kosovo. BJVM 9(4): 299-307. https://doi.org/10.15547/bjvm.938
  27. Dohoo, R., Meek, A.H. (1982). Somatic cell in bovine milk. Can Vet J. 23, 119-125.
  28. Tomazi, T., Gonchalves, J. L., Barreiro, J. R, Arcari, M. A., dos Santos, M. V. (2015). Bovine subclinical intramammary infection caused by coagulase-negative staphylococci increases somatic cell count but has no effect on milk yield or composition. Dairy Sci. 98, 3071-3078. https://doi.org/10.3168/jds.2014-8466 PMid:25726098
  29. Djabri, B., Bareille, N., Beaudeau, F., Seegers, H. (2002). Quarter milk somatic cell count in infected dairy cows: a meta-analysis. Vet Res. 33, 335-357. https://doi.org/10.1051/vetres:2002021 PMid:12199362
  30. Kochyigit, R., Yilmaz, O., Ozench, E., Uchar, M. (2016). Effect of some risk factors on subclinical mastitis in dairy cows. Vet J. 9(3): 185-193. https://doi.org/10.5578/kvj.27875
  31. Slyzius, E., Juozaitiene, V., Juozaitis, A., Zilaitis, V., Slyziene, B., Cereskiene, E. (2014). Udder quarters morphological and milking traits risk factors influencing productivity and subclinical mastitis in dairy cows. Bulgarian Journal of Agricultural Science 20(6): 1502-1507.
  32. Мalik, T.A., Мohini, M., Mir, S.H., Ganaie, B.A, Singh, D., Varun, T.K., Howal, S., Thakur, S. (2018). Somatic cells in relation to udder health and milk quality-a review. J Anim Health Prod. 6(1): 18-26. https://doi.org/10.17582/journal.jahp/2018/
  33. Čačić, Z., Kalit, S., Antunac, N., Čačić, M. (2003). Somatic cells and factors affecting their number in milk. Mljekarstvo 53(1): 23-36. [in Serbian]


© 2020 Janevski A. This is an open-access article published under the terms of the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Conflict of Interest Statement

The authors have declared that no competing interests exist.

Citation Information

Macedonian Veterinary Review. Volume 43, Issue 1, Pages 23-30, e-ISSN 1857-7415, p-ISSN 1409-7621, DOI: 10.2478/macvetrev-2020-0010, 2020