Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes

Petrovski, Kiro; Hoare, Andrew; Hemmatzadeh, Farhid; Jafari Jozani, Raziallah; Al-Farha, Abd Al-Bar; Tearle, Rick

doi:10.9775/kvfd.2019.23106

Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes

Abd AL FARHA, (Department of Animal Production, Technical Agricultural College, Northern Technical University, Mosul, Iraq)

Farhid HEMMATZADEH, (University of Adelaide, School of Animal and Veterinary Sciences, South Australia, Australia)

Rick TEARLE, (Davies Centre, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, Australia)

Razi JOZANI, (Tabriz University, Department of Veterinary Clinical Sciences, Tabriz, Iran)

Andrew HOARE, (South East Vets, South Australia, Australia)

Kiro PETROVSKI (Australian Centre for Antimicrobial Resistance Ecology, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, Australia)

Kafkas Üniversitesi Veteriner Fakültesi Dergisi

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Yıl: 2020 Cilt: 26 Sayı: 3 Sayfa Aralığı: 337 - 342 Metin Dili: İngilizce DOI: 10.9775/kvfd.2019.23106 İndeks Tarihi: 01-12-2020

Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes

Öz:

Mycoplasma mastitis raises significant concerns in the dairy industry worldwide. The study objective was to develop an accurate and rapidscreening method for identification of field isolates of Mycoplasma and Acholeplasma species and investigate relative merits of conventionalmicrobial culture versus PCR-based method for detecting Mycoplasma and Acholeplasma species in bovine milk. A total of 368 milk samplescollected at individual cow level from a single dairy farm in South Australia, 192 (52%) tested positive for mollicutes using a conventional culturebased method. DNA extracted directly from milk and used for amplification through specifically designed universal mollicutes PCR-basedmethod. Of them, 269 (73%) tested positive. Sequencing results of 30 positive samples targeting the 16S rRNA gene, showed five differentmollicutes species involved, including Acholeplasma laidlawii, Acholeplasma axanthum, Mycoplasma arginini, Mycoplasma bovirhinis, Mycoplasmabovis. According to these results, species-specific PCR was conducted on all samples. DNA amplifications using species-specific PCR yielded256 (70%) positive mollicutes samples. The developed universal PCR demonstrated best concordance with species-specific PCR (Cohen’s Kappa= 0.747±0.031). Co-infection by two or more of the above-mentioned mollicutes showed highest prevalence. It is recommended surveyingmollicutes using the universal PCR used in this study. The PCR system used in this study showed significant rapidity and sensitivity compared tothe conventional bacteriological culture method for screening Mycoplasma and Acholeplasma species in dairy herds.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Pfutzner H, Sachse K: Mycoplasma bovis as an agent of mastitis, pneumonia, arthritis and genital disorders in cattle. Rev Sci Tech, 15 (4): 1477-1494, 1996. DOI: 10.20506/rst.15.4.987
2. Sulyok K, Kreizinger Z, Wehmann E, Lysnyansky I, Bányai K, Marton S, Jerzsele Á, Rónai Z, Turcsányi I, Makrai L, Jánosi S, Nagy S, Gyuranecz M: Mutations associated with decreased susceptibility to seven antimicrobial families in field and laboratory-derived Mycoplasma bovis strains. Antimicrob Agents Chemother, 61 (2): e01983-16. 2017. DOI: 10.1128/aac.01983-16:AAC.01983-16
3. Lysnyansky I, Ayling RD: Mycoplasma bovis: Mechanisms of resistance and trends in antimicrobial susceptibility. Front Microbiol, 7:595, 2016. DOI: 10.3389/fmicb.2016.00595
4. González RN, Wilson DJ: Mycoplasmal mastitis in dairy herds. Vet Clin North Am Food Anim Pract, 19 (1): 199-221, 2003. DOI: 10.1016/s0749- 0720(02)00076-2
5. Fox LK, Kirk JH, Britten A: Mycoplasma mastitis: A review of transmission and control. J Vet Med B Infect Dis Vet Public Health, 52 (4): 153-160, 2005.
6. George L, Divers T, Ducharme N, Welcome F: Diseases of the Teats and Udder. In, Rebhun’s Diseases of Dairy Cattle. 2nd ed., Elsevier Health Sciences, 2007.
7. Boonyayatra S, Fox Lk, Gay J, Sawant A, Besser T: Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction. J Vet Diagn Invest, 24 (1): 7-13, 2011. DOI: 10.1177/1040638711425936
8. Jasper D, Dellinger J, Rollins M, Hakanson H: Prevalence of mycoplasmal bovine mastitis in California. Am J Vet Res, 40, 1043-1047, 1979.
9. Al-Farha A, Hemmatzadeh F, Khazandi M, Hoare A, Petrovski K: Evaluation of effects of Mycoplasma mastitis on milk composition in dairy cattle from South Australia. BMC Vet Res, 13:351, 2017. DOI: 10.1186/ s12917-017-1274-2
10. Windsor HM, Windsor GD, Noordergraaf JH: The growth and long term survival of Acholeplasma laidlawii in media products used in biopharmaceutical manufacturing. Biologicals, 38 (2): 204-210, 2010. DOI: 10.1016/j.biologicals.2009.11.009
11. Kirk J, Glenn K, Ruiz L, Smith E: Epidemiologic analysis of Mycoplasma spp. isolated from bulk-tank milk samples obtained from dairy herds that were members of a milk cooperative. J Am Vet Med Assoc, 211, 1036-1038, 1997.
12. Hotzel H, Frey J, Bashiruddin J, Sachse K: Detection and differentiation of ruminant mycoplasmas. In, Sachse K, Frey J (Eds): PCR Detection of Microbial Pathogens. Methods in Molecular Biology™, 231- 246, Humana Press, 2003. DOI: 10.1385/1-59259-344-5:231
13. Pinnow CC, Butler JA, Sachse K, Hotzel H, Timms LL, Rosenbusch RF: Detection of Mycoplasma bovis in preservative-treated field milk samples. J Dairy Sci, 84, 1640-1645, 2001. DOI: 10.3168/jds.s0022- 0302(01)74599-7
14. Schnee C, Schulsse S, Hotzel H, Ayling RD, Nicholas RAJ, Schubert E, Heller M, Ehricht R, Sachse K: A novel rapid DNA microarray assay enables identification of 37 Mycoplasma species and highlights multiple Mycoplasma infections. PLoS One. 7 (3):e33237, 2012. DOI: 10.1371/ journal.pone.0033237
15. Parker AM, House JK, Hazelton MS, Bosward KL, Sheehy PA: Comparison of culture and a multiplex probe PCR for identifying Mycoplasma species in bovine milk, semen and swab samples. PLoS One, 12 (3):e0173422, 2017. DOI: 10.1371/journal.pone.0173422
16. Gioia G, Werner B, Nydam DV, Moroni P: Validation of a Mycoplasma molecular diagnostic test and distribution of mycoplasma species in bovine milk among New York State dairy farms. J Dairy Sci, 99 (6): 4668- 4677, 2016. DOI: 10.3168/jds.2015-10724
17. Bashiruddin JB, Frey J, Königsson MH, Johansson KE, Hotzel H, Diller R, de Santis P, Botelho A, Ayling RD, Nicholas RAJ, Thiaucourt F, Sachse K: Evaluation of PCR systems for the identification and differentiation of Mycoplasma agalactiae and Mycoplasma bovis: A collaborative trial. Vet J, 169 (2): 268-275, 2005. DOI: 10.1016/j.tvjl.2004.01.018
18. Kobayashi H, Hirose K, Worarach A, Paugtes P, Ito N, Morozumi T, Yamamoto K: In vitro amplificatin of the 16S rRNA genes from Mycoplasma bovirhinis, Mycoplasma alkalescens and Mycoplasma bovigenitalium by PCR. J Vet Med Sci, 60 (12): 1299-1303, 1998. DOI: 10.1292/jvms.60.1299
19. Al-Farha A, Petrovski K, Jozani R, Hoare A, Hemmatzadeh F: Discrimination between some Mycoplasma spp. and Acholeplasma laidlawii in bovine milk using high resolution melting curve analysis. BMC Res Notes, 11:107, 2018. DOI: 10.1186/s13104-018-3223-y
20. Srinivasan R, Karaoz U, Volegova M, MacKichan J, Kato-Maeda M, Miller S, Nadarajan R, Brodie EL, Lynch SV: Use of 16S rRNA gene for identification of a broad range of clinically relevant bacterial pathogens. PLoS One, 10 (2):e0117617, 2015. DOI: 10.1371/journal.pone.0117617
21. Szacawa E, Niemczuk K, Dudek K, Bednarek D, Rosales R, Ayling R: Mycoplasma bovis infections and co-infections with other Mycoplasma spp. with different clinical manifestations in affected cattle herds in eastern region of Poland. Bull Vet Inst Pulawy, 59 (3): 331-338, 2015. DOI: 10.1515/bvip-2015-0049
22. Nicholas R, Ayling R, McAuliffe L: Mycoplasma Diseases of Ruminants. Wallingford, CABI; 2008.
23. Thurmond MC, Tyler JW, Luiz DM, Holmberg CA, Picanso JP: The effect of pre-enrichment on recovery of Streptococcus agalactiae, Staphylococcus aureus and Mycoplasma from bovine milk. Epidemiol Infect, 103 (3): 465-474, 1989. DOI: 10.1017/s0950268800030879
24. Al-Farha A, Khazandi M, Hemmatzadeh F, Jozani R, Tearle R, Hoare A, Petrovski K: Evaluation of three cryoprotectants used with bovine milk affected with Mycoplasma bovis in different freezing conditions. BMC Res Notes, 11:2016, 2018. DOI: 10.1186/s13104-018-3325-6
25. Markey B, Leonard, F, Archambault, M, Cullinane A, Maguire D:Clinical Veterinary Microbiology, Elsevier Health Sciences, 2013.
26. Sievers F, Higgins DG: Clustal omega. Curr Protoc Bioinformatics, 48, 3.13.1-16, 2014. DOI: 10.1002/0471250953.bi0313s48
27. Lorenz T: Polymerase chain reaction: basic protocol plus roubleshooting and optimization strategies. J Vis Exp, (63):e3998, 2012. DOI: 10.3791/3998
28. Radaelli E, Castiglioni V, Losa M, Benedetti V, Piccinini R, Nicholas RAJ, Scanziani E, Luini M: Outbreak of bovine clinical mastitis caused by Mycoplasma bovis in a North Italian herd. Res Vet Sci, 91 (2): 251-253, 2011. DOI: 10.1016/j.rvsc.2011.01.006
29. Justice-Allen A, Trujillo J, Corbett R, Harding R, Goodell G, Wilson D: Survival and replication of Mycoplasma species in recycled bedding sand and association with mastitis on dairy farms in Utah. J Dairy Sci, 93 (1): 192-202, 2010. DOI: 10.3168/jds.2009-2474
30. Maunsell FP, Woolums AR, Francoz D, Rosenbusch RF, Step DL, Wilson DJ, Janzen ED: Mycoplasma bovis infections in cattle. J Vet Intern Med, 25, 772-783, 2011. DOI: 10.1111/j.1939-1676.2011.0750
31. Jasper, D: The role of Mycoplasma in bovine mastitis. J Am Vet Med Assoc, 181, 158-162, 1982.
32. Peredeltchouk M, Wilson David SA, Bhattacharya B, Volokhov DV, Chizhikov V: Detection of Mycoplasma contamination in cell substrates using reverse transcription-PCR assays. J Appl Microbiol, 110 (1): 54-60, 2011.
33. Waters AP, McCuthan TF: Ribosomal RNA: Nature’s own polymeraseamplified target for diagnosis. Parasitol Today, 6 (2): 56-59, 1990. DOI: 10.1016/0169-4758(90)90071-b
34. D’Inzeo T, De Angelis G, Fiori B, Menchinelli G, Liotti FM, Morandotti GA, De Maio F, Nagel D, Antonaci M, Sanguinetti M, Spanu T: Comparison of mycoplasma IES, mycofast revolution and mycoplasma IST2 to detect genital mycoplasmas in clinical samples. J Infect Dev Ctries, 11 (01): 98- 101, 2017. DOI: 10.3855/jidc.8039

APA	Al-Farha A, Hemmatzadeh F, Tearle R, Jafari Jozani R, Hoare A, Petrovski K (2020). Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. , 337 - 342. 10.9775/kvfd.2019.23106
Chicago	Al-Farha Abd Al-Bar,Hemmatzadeh Farhid,Tearle Rick,Jafari Jozani Raziallah,Hoare Andrew,Petrovski Kiro Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. (2020): 337 - 342. 10.9775/kvfd.2019.23106
MLA	Al-Farha Abd Al-Bar,Hemmatzadeh Farhid,Tearle Rick,Jafari Jozani Raziallah,Hoare Andrew,Petrovski Kiro Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. , 2020, ss.337 - 342. 10.9775/kvfd.2019.23106
AMA	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. . 2020; 337 - 342. 10.9775/kvfd.2019.23106
Vancouver	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. . 2020; 337 - 342. 10.9775/kvfd.2019.23106
IEEE	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K "Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes." , ss.337 - 342, 2020. 10.9775/kvfd.2019.23106
ISNAD	Al-Farha, Abd Al-Bar vd. "Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes". (2020), 337-342. https://doi.org/10.9775/kvfd.2019.23106

APA	Al-Farha A, Hemmatzadeh F, Tearle R, Jafari Jozani R, Hoare A, Petrovski K (2020). Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 26(3), 337 - 342. 10.9775/kvfd.2019.23106
Chicago	Al-Farha Abd Al-Bar,Hemmatzadeh Farhid,Tearle Rick,Jafari Jozani Raziallah,Hoare Andrew,Petrovski Kiro Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 26, no.3 (2020): 337 - 342. 10.9775/kvfd.2019.23106
MLA	Al-Farha Abd Al-Bar,Hemmatzadeh Farhid,Tearle Rick,Jafari Jozani Raziallah,Hoare Andrew,Petrovski Kiro Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, vol.26, no.3, 2020, ss.337 - 342. 10.9775/kvfd.2019.23106
AMA	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2020; 26(3): 337 - 342. 10.9775/kvfd.2019.23106
Vancouver	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2020; 26(3): 337 - 342. 10.9775/kvfd.2019.23106
IEEE	Al-Farha A,Hemmatzadeh F,Tearle R,Jafari Jozani R,Hoare A,Petrovski K "Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes." Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 26, ss.337 - 342, 2020. 10.9775/kvfd.2019.23106
ISNAD	Al-Farha, Abd Al-Bar vd. "Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes". Kafkas Üniversitesi Veteriner Fakültesi Dergisi 26/3 (2020), 337-342. https://doi.org/10.9775/kvfd.2019.23106