Molecular characterization of the Mycobacterium tuberculosis complex (MTC) of raw milk from selected dairy farms in the Eastern Cape
- Authors: Komani, Nosiphiwo
- Date: 2013
- Subjects: Mycobacterium tuberculosis -- South Africa -- Eastern Cape , Milk -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Dairy farming -- South Africa -- Eastern Cape , Tuberculosis in cattle -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11276 , http://hdl.handle.net/10353/d1013157 , Mycobacterium tuberculosis -- South Africa -- Eastern Cape , Milk -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Dairy farming -- South Africa -- Eastern Cape , Tuberculosis in cattle -- South Africa -- Eastern Cape
- Description: Tuberculosis (TB) is an ancient infectious disease that has been infecting different populations around the globe and it has also been considered as one of the most successful human and animal disease. TB found in animals such as cattle and other known bovids is known as bovine tuberculosis. Bovine tuberculosis (BTB) is an infectious disease found in cattle mainly caused by Mycobacterium bovis. M. bovis is a member of the Mycobacterium tuberculosis complex (MTC) together with M. tuberculosis, M. africanum, and M. canetti where the natural host is humans; whereas M. caprae, M. microti and M. pinnipedii usually have animals as their natural host. In this study the molecular characterization of the MTC from cow milk in the Eastern Cape was investigated. One hundred and twenty samples (40 ml each) were collected from three dairy farms in the Eastern Cape, South Africa. These samples were processed using a modified Petroff decontamination method. Sample processing was followed by DNA isolation using a Zymo Bacterial/Fungal DNA Kit and the amplification and detection of the MTC was done using the Seeplex MTB Nested ACE assay. The drug susceptibility tests were done using GenoTypeMTBDRplus assay which detects mutations and resistance to INH (isoniazid) and RMP (rifampicin). The milk isolates were further analyzed using a spoligotyping method which is based on the PCR amplification of a highly polymorphic direct repeat locus in the M. tuberculosis genome which detects and types the MTC. A percentage of 20.8 % samples were found to be positive for MTC using the Seeplex MTB Nested ACE assay. There were 42.1 % samples that were resistant to both INH and RMP with the rest sensitive to either INH or RMP. The spoligotyping method showed that 78.3 % samples resembled Family 33 strains and the rest (21.7 %) resembled a spoligotyping signature known to be that of M.africanum. Both these strains belong to the Ancestral lineage with Indo-Oceanic and West Africa 2 lineage. The outcomes of our study showed that molecular methods for detection of MTC can be applied directly on milk samples without the need for culturing.
- Full Text:
- Date Issued: 2013
- Authors: Komani, Nosiphiwo
- Date: 2013
- Subjects: Mycobacterium tuberculosis -- South Africa -- Eastern Cape , Milk -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Dairy farming -- South Africa -- Eastern Cape , Tuberculosis in cattle -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11276 , http://hdl.handle.net/10353/d1013157 , Mycobacterium tuberculosis -- South Africa -- Eastern Cape , Milk -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Dairy farming -- South Africa -- Eastern Cape , Tuberculosis in cattle -- South Africa -- Eastern Cape
- Description: Tuberculosis (TB) is an ancient infectious disease that has been infecting different populations around the globe and it has also been considered as one of the most successful human and animal disease. TB found in animals such as cattle and other known bovids is known as bovine tuberculosis. Bovine tuberculosis (BTB) is an infectious disease found in cattle mainly caused by Mycobacterium bovis. M. bovis is a member of the Mycobacterium tuberculosis complex (MTC) together with M. tuberculosis, M. africanum, and M. canetti where the natural host is humans; whereas M. caprae, M. microti and M. pinnipedii usually have animals as their natural host. In this study the molecular characterization of the MTC from cow milk in the Eastern Cape was investigated. One hundred and twenty samples (40 ml each) were collected from three dairy farms in the Eastern Cape, South Africa. These samples were processed using a modified Petroff decontamination method. Sample processing was followed by DNA isolation using a Zymo Bacterial/Fungal DNA Kit and the amplification and detection of the MTC was done using the Seeplex MTB Nested ACE assay. The drug susceptibility tests were done using GenoTypeMTBDRplus assay which detects mutations and resistance to INH (isoniazid) and RMP (rifampicin). The milk isolates were further analyzed using a spoligotyping method which is based on the PCR amplification of a highly polymorphic direct repeat locus in the M. tuberculosis genome which detects and types the MTC. A percentage of 20.8 % samples were found to be positive for MTC using the Seeplex MTB Nested ACE assay. There were 42.1 % samples that were resistant to both INH and RMP with the rest sensitive to either INH or RMP. The spoligotyping method showed that 78.3 % samples resembled Family 33 strains and the rest (21.7 %) resembled a spoligotyping signature known to be that of M.africanum. Both these strains belong to the Ancestral lineage with Indo-Oceanic and West Africa 2 lineage. The outcomes of our study showed that molecular methods for detection of MTC can be applied directly on milk samples without the need for culturing.
- Full Text:
- Date Issued: 2013
Prevalence, seasonal trends and incidences of cattle tuberculosis and brucellosis in the Cacadu municipality, Eastern Cape, South Africa
- Authors: Nombebe, Thobeka
- Date: 2012
- Subjects: Tuberculosis in cattle -- South Africa -- Eastern Cape , Brucellosis in animals -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Mycobacterium tuberculosis -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Biostatistics and Epidemiology)
- Identifier: vital:11783 , http://hdl.handle.net/10353/d1016200 , Tuberculosis in cattle -- South Africa -- Eastern Cape , Brucellosis in animals -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Mycobacterium tuberculosis -- South Africa -- Eastern Cape
- Description: A cross-sectional study was conducted from February 2011 to August 2012 to investigate the status and identify risk factors for Bovine Tuberculosis (BTB) and cattle Brucellosis. A total of 1277 cattle from 6 selected dairy farms were tested. The farms were purposively selected on suspected risk. Samples were analysed using Rose Bengal Test (RBPT) and Complement Fixation Test (CFT) for Brucellosis and Comparative Intradermal Tuberculin Test (CITT) for BTB. The results revealed the overall prevalence of 1.6% for Brucellosis and no BTB was found. Logistic regression analysis revealed that age of cattle, herd size and number of calves a cow has had significant association with Brucellosis with p-values 0.0071, 0.0490 and 0.000 respectively. The role of location (p>0.05), gender (p>0.05) and breed (p>0.05) in the occurrence of the disease was not statistically significant at α=0.05, although higher rates were obtained in females and Jersey breed. Questionnaires were also administered to 47 individuals a total of 12 females and 35 males. Most respondents were not aware about cattle brucellosis and cattle tuberculosis. Consumption of raw or unpasteurized milk still occurs with 74.75% of respondents revealed.
- Full Text:
- Date Issued: 2012
- Authors: Nombebe, Thobeka
- Date: 2012
- Subjects: Tuberculosis in cattle -- South Africa -- Eastern Cape , Brucellosis in animals -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Mycobacterium tuberculosis -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Biostatistics and Epidemiology)
- Identifier: vital:11783 , http://hdl.handle.net/10353/d1016200 , Tuberculosis in cattle -- South Africa -- Eastern Cape , Brucellosis in animals -- South Africa -- Eastern Cape , Dairy farms -- South Africa -- Eastern Cape , Mycobacterium tuberculosis -- South Africa -- Eastern Cape
- Description: A cross-sectional study was conducted from February 2011 to August 2012 to investigate the status and identify risk factors for Bovine Tuberculosis (BTB) and cattle Brucellosis. A total of 1277 cattle from 6 selected dairy farms were tested. The farms were purposively selected on suspected risk. Samples were analysed using Rose Bengal Test (RBPT) and Complement Fixation Test (CFT) for Brucellosis and Comparative Intradermal Tuberculin Test (CITT) for BTB. The results revealed the overall prevalence of 1.6% for Brucellosis and no BTB was found. Logistic regression analysis revealed that age of cattle, herd size and number of calves a cow has had significant association with Brucellosis with p-values 0.0071, 0.0490 and 0.000 respectively. The role of location (p>0.05), gender (p>0.05) and breed (p>0.05) in the occurrence of the disease was not statistically significant at α=0.05, although higher rates were obtained in females and Jersey breed. Questionnaires were also administered to 47 individuals a total of 12 females and 35 males. Most respondents were not aware about cattle brucellosis and cattle tuberculosis. Consumption of raw or unpasteurized milk still occurs with 74.75% of respondents revealed.
- Full Text:
- Date Issued: 2012
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