Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes
Mycoplasma mastitis raises significant concerns in the dairy industry worldwide. The study objective was to develop an accurate and rapid screening method for identification of field isolates of Mycoplasma and Acholeplasma species and investigate relative merits of conventional microbial culture ver...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Kafkas University, Faculty of Veterinary Medicine
2020-02-01
|
| Series: | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| Subjects: | |
| Online Access: | https://vetdergikafkas.org/pdf.php?id=2681 |
| _version_ | 1827922171776729088 |
|---|---|
| author | Abd Al-Bar AL-FARHA Farhid HEMMATZADEH Rick TEARLE Razi JOZANI Andrew HOARE Kiro PETROVSKI |
| author_facet | Abd Al-Bar AL-FARHA Farhid HEMMATZADEH Rick TEARLE Razi JOZANI Andrew HOARE Kiro PETROVSKI |
| author_sort | Abd Al-Bar AL-FARHA |
| collection | DOAJ |
| description | Mycoplasma mastitis raises significant concerns in the dairy industry worldwide. The study objective was to develop an accurate and rapid
screening method for identification of field isolates of Mycoplasma and Acholeplasma species and investigate relative merits of conventional
microbial culture versus PCR-based method for detecting Mycoplasma and Acholeplasma species in bovine milk. A total of 368 milk samples
collected 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-based
method. Of them, 269 (73%) tested positive. Sequencing results of 30 positive samples targeting the 16S rRNA gene, showed five different
mollicutes species involved, including Acholeplasma laidlawii, Acholeplasma axanthum, Mycoplasma arginini, Mycoplasma bovirhinis, Mycoplasma
bovis. According to these results, species-specific PCR was conducted on all samples. DNA amplifications using species-specific PCR yielded
256 (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 surveying
mollicutes using the universal PCR used in this study. The PCR system used in this study showed significant rapidity and sensitivity compared to
the conventional bacteriological culture method for screening Mycoplasma and Acholeplasma species in dairy herds. |
| first_indexed | 2024-03-13T04:38:12Z |
| format | Article |
| id | doaj.art-47dda224361c43fabe148fe46be21c41 |
| institution | Directory Open Access Journal |
| issn | 1309-2251 |
| language | English |
| last_indexed | 2024-03-13T04:38:12Z |
| publishDate | 2020-02-01 |
| publisher | Kafkas University, Faculty of Veterinary Medicine |
| record_format | Article |
| series | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| spelling | doaj.art-47dda224361c43fabe148fe46be21c412023-06-19T06:32:52ZengKafkas University, Faculty of Veterinary MedicineKafkas Universitesi Veteriner Fakültesi Dergisi1309-22512020-02-0126333734210.9775/kvfd.2019.231062681Comparison of culture and pcr for detection of field isolates of bovine milk mollicutesAbd Al-Bar AL-FARHA0Farhid HEMMATZADEH1Rick TEARLE2Razi JOZANI3Andrew HOARE4Kiro PETROVSKI5Department of Animal Production, Technical Agricultural College, Northern Technical University, Mosul, IRAQUniversity of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIADavies Centre, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIATabriz University, Department of Veterinary Clinical Sciences, Tabriz, IRANSouth East Vets, South Australia, AUSTRALIAAustralian Centre for Antimicrobial Resistance Ecology, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIAMycoplasma mastitis raises significant concerns in the dairy industry worldwide. The study objective was to develop an accurate and rapid screening method for identification of field isolates of Mycoplasma and Acholeplasma species and investigate relative merits of conventional microbial culture versus PCR-based method for detecting Mycoplasma and Acholeplasma species in bovine milk. A total of 368 milk samples collected 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-based method. Of them, 269 (73%) tested positive. Sequencing results of 30 positive samples targeting the 16S rRNA gene, showed five different mollicutes species involved, including Acholeplasma laidlawii, Acholeplasma axanthum, Mycoplasma arginini, Mycoplasma bovirhinis, Mycoplasma bovis. According to these results, species-specific PCR was conducted on all samples. DNA amplifications using species-specific PCR yielded 256 (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 surveying mollicutes using the universal PCR used in this study. The PCR system used in this study showed significant rapidity and sensitivity compared to the conventional bacteriological culture method for screening Mycoplasma and Acholeplasma species in dairy herds.https://vetdergikafkas.org/pdf.php?id=2681mycoplasmamastitisdairy cattleacholeplasma |
| spellingShingle | Abd Al-Bar AL-FARHA Farhid HEMMATZADEH Rick TEARLE Razi JOZANI Andrew HOARE Kiro PETROVSKI Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes Kafkas Universitesi Veteriner Fakültesi Dergisi mycoplasma mastitis dairy cattle acholeplasma |
| title | Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| title_full | Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| title_fullStr | Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| title_full_unstemmed | Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| title_short | Comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| title_sort | comparison of culture and pcr for detection of field isolates of bovine milk mollicutes |
| topic | mycoplasma mastitis dairy cattle acholeplasma |
| url | https://vetdergikafkas.org/pdf.php?id=2681 |
| work_keys_str_mv | AT abdalbaralfarha comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes AT farhidhemmatzadeh comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes AT ricktearle comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes AT razijozani comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes AT andrewhoare comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes AT kiropetrovski comparisonofcultureandpcrfordetectionoffieldisolatesofbovinemilkmollicutes |