Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms
Mastitis is a cow disease usually signalized by irritation, swelling, and soreness of the udder. It is characterized by physical, chemical, and biological changes in the udder and milk. The aim of this study was to detect and characterize pathogens causing subclinical mastitis (SCM) from the milk of...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
De Gruyter
2021-08-01
|
| Series: | Open Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1515/biol-2021-0080 |
| _version_ | 1797998065401987072 |
|---|---|
| author | Khasapane Ntelekwane G. Nkhebenyane Jane S. Kwenda Stanford Khumalo Zamantungwa T. H. Mtshali Phillip S. Taioe Moeti O. Thekisoe Oriel M. M. |
| author_facet | Khasapane Ntelekwane G. Nkhebenyane Jane S. Kwenda Stanford Khumalo Zamantungwa T. H. Mtshali Phillip S. Taioe Moeti O. Thekisoe Oriel M. M. |
| author_sort | Khasapane Ntelekwane G. |
| collection | DOAJ |
| description | Mastitis is a cow disease usually signalized by irritation, swelling, and soreness of the udder. It is characterized by physical, chemical, and biological changes in the udder and milk. The aim of this study was to detect and characterize pathogens causing subclinical mastitis (SCM) from the milk of dairy cows of small-scale farmers through culture and molecular techniques. Milk was collected from 32 cows belonging to 8 small-scale farmers around Harrismith District, South Africa. The results showed that screening of SCM by California mastitis test and somatic cell counts (SCC) was 21.87 and 25%, respectively. Culture methods revealed the presence of Staphylococcus aureus at 93% followed by Streptococci spp. and Escherichia coli at 36.4 and 13.3%, respectively. The PCR could only detect E. coli, while single-molecule real-time sequencing showed a total of 2 phyla, 5 families, 7 genera, and 131 species. Clostridiaceae was the most abundant family, while Romboutsia was the most abundant genus followed by Turicibacter spp. The present study has documented the occurrence of SCM causing pathogens in milk collected from cows of small-scale farmers in Harrismith, indicating that SCM may be present at higher levels than expected. |
| first_indexed | 2024-04-11T10:42:33Z |
| format | Article |
| id | doaj.art-ce62630c607247ddad7863fe44cd73e2 |
| institution | Directory Open Access Journal |
| issn | 2391-5412 |
| language | English |
| last_indexed | 2024-04-11T10:42:33Z |
| publishDate | 2021-08-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Open Life Sciences |
| spelling | doaj.art-ce62630c607247ddad7863fe44cd73e22022-12-22T04:29:08ZengDe GruyterOpen Life Sciences2391-54122021-08-0116180080810.1515/biol-2021-0080Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farmsKhasapane Ntelekwane G.0Nkhebenyane Jane S.1Kwenda Stanford2Khumalo Zamantungwa T. H.3Mtshali Phillip S.4Taioe Moeti O.5Thekisoe Oriel M. M.6Department of Life Sciences, Centre for Applied Food Sustainability and Biotechnology, Central University of Technology, Bloemfontein, 9300, South AfricaDepartment of Life Sciences, Centre for Applied Food Sustainability and Biotechnology, Central University of Technology, Bloemfontein, 9300, South AfricaSequencing Core Facility, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South AfricaSequencing Core Facility, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South AfricaSequencing Core Facility, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South AfricaEpidemiology, Parasites and Vectors, Agricultural Research Council – Onderstepoort Veterinary Research (ARC-OVR), Onderstepoort 0110, South AfricaUnit for Environmental Sciences and Management, North-West University, Potchefstroom, 2531, South AfricaMastitis is a cow disease usually signalized by irritation, swelling, and soreness of the udder. It is characterized by physical, chemical, and biological changes in the udder and milk. The aim of this study was to detect and characterize pathogens causing subclinical mastitis (SCM) from the milk of dairy cows of small-scale farmers through culture and molecular techniques. Milk was collected from 32 cows belonging to 8 small-scale farmers around Harrismith District, South Africa. The results showed that screening of SCM by California mastitis test and somatic cell counts (SCC) was 21.87 and 25%, respectively. Culture methods revealed the presence of Staphylococcus aureus at 93% followed by Streptococci spp. and Escherichia coli at 36.4 and 13.3%, respectively. The PCR could only detect E. coli, while single-molecule real-time sequencing showed a total of 2 phyla, 5 families, 7 genera, and 131 species. Clostridiaceae was the most abundant family, while Romboutsia was the most abundant genus followed by Turicibacter spp. The present study has documented the occurrence of SCM causing pathogens in milk collected from cows of small-scale farmers in Harrismith, indicating that SCM may be present at higher levels than expected.https://doi.org/10.1515/biol-2021-0080microbiotamastitis16s ribosomal rnadna sequencingmicrobial milk composition |
| spellingShingle | Khasapane Ntelekwane G. Nkhebenyane Jane S. Kwenda Stanford Khumalo Zamantungwa T. H. Mtshali Phillip S. Taioe Moeti O. Thekisoe Oriel M. M. Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms Open Life Sciences microbiota mastitis 16s ribosomal rna dna sequencing microbial milk composition |
| title | Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| title_full | Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| title_fullStr | Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| title_full_unstemmed | Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| title_short | Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| title_sort | application of culture pcr and pacbio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms |
| topic | microbiota mastitis 16s ribosomal rna dna sequencing microbial milk composition |
| url | https://doi.org/10.1515/biol-2021-0080 |
| work_keys_str_mv | AT khasapanentelekwaneg applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT nkhebenyanejanes applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT kwendastanford applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT khumalozamantungwath applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT mtshaliphillips applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT taioemoetio applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms AT thekisoeorielmm applicationofculturepcrandpacbiosequencingfordeterminationofmicrobialcompositionofmilkfromsubclinicalmastitisdairycowsofsmallholderfarms |