Detection of Mycobacterium paratuberculosis in raw cow’s milk using polymerase chain reaction (PCR) technique
Iraqi Journal of Veterinary Sciences,
2020, Volume 34, Issue 1, Pages 83-86
AbstractParatuberculosis or Johne’s disease is a chronic debilitating disease mainly infects ruminants and caused by Mycobacterium paratuberculosis. Previous serological studies in Mosul city confirm the presence of positive reactants for paratuberculosis in cattle. However, culture methods to confirm the disease need a long incubation period and also special media. Raw cow’s milk is considered as potential source for transmission of M. paratuberculosis in cows’ herds. Accordingly, this study aimed to detect the presence of M. paratuberculosis specifically in the raw cow’s milk using polymerase chain reaction (PCR) technique as a rapid, sensitive and reliable method. A total of 50 samples of raw cow’s milk were collected from cows suffering from emaciation and unresponsive to antibiotic treatment. All the samples were subjected to DNA extraction and direct amplification PCR. The results showed that 3 (6%) out of 50 milk samples were positive for M. paratuberculosis. This is the first study in Mosul city that confirms the presence of M. paratuberculosis in raw cow’s milk using PCR technique. In conclusion, raw cow’s milk could be an important source for M. paratuberculosis infection in dairy cows, and also PCR technique could be helpful in rapid diagnosis of paratuberculosis.
- McAloon CG, Roche S, Ritter C, Barkema HW, Whyte P, More SJ, O’Grady L, Green MJ, Doherty ML. A review of paratuberculosis in dairy herds-Part 1: Epidemiology. Vet J. 2019;246:59-65. https://doi.org/10.1016/j.tvjl.2019.01.010
- Salem M, Heydel C, El-Sayed A, Ahmed SA, Zschöck M, Baljer G. Mycobacterium avium subspecies paratuberculosis: An insidious problem for the ruminant industry. Trop Anim Heal Prod. 2013;45:351-66. https://doi.org/10.1007/s11250-012-0274-2
- Kirkeby C, Græsboll K, Nielsen SS, Toft N, Halasa T. Epidemiological and economic consequences of purchasing livestock infected with Mycobacterium avium subsp. paratuberculosis. BMC Vet Res. 2017;13:202. https://doi.org/10.1186/s12917-017-1119-z
- Hasonova L, Pavlik I. Economic impact of paratuberculosis in dairy cattle herds: A review. Vet Med. 2006;51:193-211.
- Foddai A, Strain S, Whitlock RH, Elliott CT, Grant IR. Application of a peptide-mediated magnetic separation-phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis to bovine bulk tank milk and feces samples. J Clin Microbiol. 2011;49:2017-9. https://doi.org/10.1128/JCM.00429-11
- Khol JL, Braun AL, Slana I, Kralik P, Wittek T. Testing of milk replacers for Mycobacterium avium subsp. paratuberculosis by PCR and bacterial culture as a possible source for Johne's disease (paratuberculosis) in calves. Prev Vet Med. 2017;144:53-6. https://doi.org/10.1016/j.prevetmed.2017.05.013
- Laurin E, McKenna S, Chaffer M, Keefe G. Sensitivity of solid culture, broth culture, and real-time PCR assays for milk and colostrum samples from Mycobacterium avium ssp. paratuberculosis-infectious dairy cows. J Dairy Sci. 2015;98:8597-609. https://doi.org/10.3168/jds.2014-8758
- Correa-Valencia NM, Ramírez NF, Olivera M, Fernandez-Silva JA. Milk yield and lactation stage are associated with positive results to ELISA for Mycobacterium avium subsp. paratuberculosis in dairy cows from Northern Antioquia, Colombia: A preliminary study. Trop Anim Heal Prod. 2016;48:1191-200. https://doi.org/10.1007/s11250-016-1074-x
- Al-Aalim AM. Serological detection of johne's disease in cattle using ELISA techniques. Tikrit J Pure Sci. 2010;15:37-9.
- Ahmed IM. Serodiagnosis of Johne's disease by indirect ELISA in ovine. Iraqi J Vet Sci. 2010;24:41-3. https://doi.org/10.33899/ ijvs.2010.5576
- Gilardoni LR, Fernandez B, Morsella C, Mendez L, Jar AM, Paolicchi FA, Mundo SL. Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR. Brazilian J Micobiol. 2016;47:506-12. http://dx.doi.org/10.1016/j.bjm.2016.01. 013
- Dzieciol M, Volgger P, Khol J, Baumgartner W, Wagner M, Hein I. A novel real-time PCR assay for specific detection and quantification of Mycobacterium avium subsp. paratuberculosis in milk with the inherent possibility of differentiation between viable and dead cells. BMC Res Notes. 2010;3:251. https://doi.org/10.1186/1756-0500-3-251
- Al-Kass AM. Seroprevalence of Johne’s disease in goats. Iraqi J Vet Sci. 2009;23:281-3.
- Al-Farwachi M, Al-Jubory K, Flayyih A. Prevalence of paratuberculosis amange cattle with diarrhea in Ninavah, Iraq. Res Opin Anim Vet Sci. 2018;8:1-3.
- Yue R, Liu C, Barrow P, Liu F, Cui Y, Yang L, Zhao D, Zhou X. The isolation and molecular characterization of Mycobacterium avium subsp. paratuberculosis in Shandong province, China. Gut Pathol. 2016;8:9. https://doi.org/10.1186/s13099-016-0092-6
- Timms VJ, Mitchell HM, Neilan BA. Optimisation of DNA extraction and validation of PCR assays to detect Mycobacterium avium subsp. paratuberculosis. J Microbiol Meth. 2015;112:99-103. https://doi.org/10.1016/j.mimet.2015.03.016
- Al-Sanjary RA, Mohammed HA, Dahl MO. Using polymerase chain reaction technique (PCR) for detection Brucella melitensis in aborted ewes' milk in Nineveh, Iraq. Assiut Vet Med J. 2014;60:140.155-9.
- Dimareli-Malli Z. Detection of Mycobacterium avium subsp. paratuberculosis in milk from clinically affected sheep and goats. Inter J Appl Res Vet Med. 2010;8:44-50.
- Hafiz NM, Saad MF, Hanafy MH, Abdel-Latif EF. Detection of Mycobacterium avium subsp. paratuberculosis in raw buffaloe's milk. Inter J ChemTech Res. 2016;9:123-8.
- De Albuquerque PPF, de Souza Santos A, de Souza Neto OL, Kim PdCP, Cavalcanti EFTSF, de Oliveira JMB, Mota RA, Júnior JWP. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil. Brazilian J Microbiol. 2017;48:113-7. http://dx.doi.org/10.1016/j.bjm.2016.10.010
- Shrafati-chaleshtori F, Sharafati-chaleshtori R, Shakerian A, Momtaz H. Detection of Mycobacterium paratuberculosis using polymerase chain reaction (PCR) in cow raw milk samples in shahre-kord. Med Lab J. 2009;3:1-5.
- Singh AV, Singh SV, Singh PK, Sohal JS. Is Mycobacterium avium subsp. paratuberculosis, the cause of Johne’s disease in animals, a good candidate for Crohn’s disease in man? Indian J Gastroenterol. 2010;29:53-8. https://doi.org/10.1007/s12664-010-0019-y
- Zarei-Kordshouli F, Geramizadeh B, Khodakaram-Tafti A. Prevalence of Mycobacterium avium subspecies paratuberculosis IS 900 DNA in biopsy tissues from patients with Crohn’s disease: Histopathological and molecular comparison with Johne’s disease in Fars province of Iran. BMC Infect Dis. 2019;19:23. https://doi.org/10.1186/s12879-018-3619-2
- McAloon CG, Roche S, Ritter C, Barkema HW, Whyte P, More SJ, O’Grady L, Green MJ, Doherty ML. A review of paratuberculosis in dairy herds-Part 2: On-farm control. Vet J. 2019;246:54-58. https://doi.org/10.1016/j.tvjl.2019.01.009
- Slana I, Liapi M, Moravkovaa M, Kralovaa A, Pavlika I. Mycobacterium avium subsp. paratuberculosis in cow bulk tankmilk in Cyprus detected by culture and quantitative IS900 and F57 real-time PCR. Preve Vet Med. 2009;89:223-6. https://doi.org/10.1016/j.prevetmed.2009.02.020
- Mortier RA, Barkema HW, De Buck J. Susceptibility to and diagnosis of Mycobacterium avium subspecies paratuberculosis infection in dairy calves: a review. Preve Vet Med. 2015;121:189-98. https://doi.org/10.1016/j.prevetmed.2015.08.011
- Stabel J, Bradner L, Robbe-Austerman S, Beitz D. Clinical disease and stage of lactation influence shedding of Mycobacterium avium subspecies paratuberculosis into milk and colostrum of naturally infected dairy cows. J Dairy Sci. 2014;97:6296-304. https://doi.org/10.3168/jds.2014-8204
- Kawther KJ, Fawziah AA, Rasha MO. Diagnostic application of ELISA and IS900PCR using buffy coat as a source sample for the detection of Mycobacterium avium subspecies paratuberculosis in subclinical cases of bovine paratuberculosis. Basrah J Vet Res. 2018;17:186-95.
- Carvalho I, Silva Jr A, Campos V, Moreira M. Detection of Mycobacterium avium subspecies paratuberculosis by polymerase chain reaction in bovine milk in Brazil. J Dairy Sci. 2009;92:5408-10. https://doi.org/10.3168/jds.2008-1816
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