- Title
- Molecular characterization of Mycobacterium bovis isolated from cattle and sputum specimens from humans in the Eastern Cape province, South Africa : public health and economic implication
- Creator
- Bhembe, Nolwazi Londiwe
- Subject
- Mycobacterium tuberculosis -- South Africa -- Eastern Cape Public health -- South Africa -- Eastern Cape
- Date Issued
- 2016
- Date
- 2016
- Type
- Thesis
- Type
- Doctoral
- Type
- PhD
- Identifier
- http://hdl.handle.net/10353/13238
- Identifier
- vital:39625
- Description
- Tuberculosis (TB) is a disease caused by the members of the Mycobacterium tuberculosis complex (MTBC), which infects both animals and humans. The primary host of Mycobacterium bovis is cattle. It remains a major global health problem. About 2 billion people are thought to be infected with TB and about 1.3 million decease each year from the disease. This study is aimed at determining the prevalence of the disease, characterizing the MTBC isolates and increasing the understanding of the genetic diversity MTBC in the Eastern Cape Province of South Africa, from studying both samples from both humans and cattle. A total of 376 cattle lymph nodes were collected from two commercial abattoirs for investigations. Suspected 6 000 sputum specimens were also collected from humans (cattle farmers). All collected samples were cultured, and isolates were confirmed with a multiplex polymerase chain reaction (PCR) targeting the mpb64 and IS6110 genes. Positive isolates were tested for resistance against anti-TB drugs using the standard Lowenstein Jensen proportion method. Isolates from cattle lymph nodes were further characterized to species level, using the region of deletion 1 (RD1) and oxyR restriction fragment length polymorphism analyses. MIRU-VNTR typing and spoligotyping assays were used to further examine the genetic diversity of all strains. Obtained spoligotype patterns were matched with isolates published in the universal spoligotyping database (SITVIT2). MTBC was detected in 162 (43.09 percent) cattle lymph nodes and 156 sputum specimens. Some isolates 25.9 percent (42/162) from the cattle lymph nodes samples were resistant to rifampicin (RIF) and 98.7 percent (154/156) were resistant to isoniazid (INH) and rifampicin (RIF) (of the human sputum specimens). All cattle lymph nodes isolates exhibited the RD1 gene, and the oxyR analyses classified 96.9 percent of the isolates as Mycobacterium tuberculosis complex and 3.1 percent as Mycobacterium bovis. Twenty seven spoligotype patterns were isolated from cattle lymph nodes, with 10 shared types (SIT) consigned to 5 lineages, including Bov_4-Caprea (1.9 percent), Microti (0.6 percent) and Beijing (17.9 percent) constituting East Asian, Latin-American-Mediterranean (LAM) (3.7 percent), X (4.3 percent), MANU (3.1percent), S (0.6 percent) and 67.9 percent were orphans that were further categorized to their designated lineages (Indo-Oceanic, Euro-American and West-African 2). Ninety one percent of the isolates from humans (farmers) were grouped into 7 clusters by spoligotyping, while the remaining 9 percent was unclustered. SITs were observed in 89.6 percent of the isolates from the Beijing, LAM, X, T and S families. The Beijing family was the most prevalent strain from both human isolates (77.2 percent) and cattle lymph nodes isolates (17.9 percent), and the T (0.6 percent) and Microti (0.6 percent) families were the least detected genotypes in both humans and cattle lymph nodes. In the cattle lymph nodes, spoligotyping showed a higher clustering rate of 82.1 percent, with the lowest (HGDI) = 0.485. The 12 MIRU-VNTR clustering rate was 64.8 percent, showing a higher HGDI of 0.671. However, a combination of both methods showed a decreased clustering rate of 53.7 percent; however these methods resulted in a higher HGDI (0.676). Isolates from human sputum specimens spoligotyping showed a higher clustering rate of 89 percent with an HGDI = 0.822. The 12 MIRU-VNTR showed the lowest clustering rate of 69 percent and an HGDI = 0.940 and the combination of the methods showed a clustering rate of 78 percent and an HGDI = 0.951. The results of this study show the diversity of MTBC strains in the Eastern Cape Province and the low clustering rate indicates continuing transmission in the province. The detection of MTBC strains from slaughtered cattle lymph nodes in abattoirs confirms the risk of providing TB infected meat in markets which has serious implications for the control of TB in South Africa. The continuing spread of MTBC in South Africa threatens both the public and economic health.
- Format
- 246 leaves
- Format
- Publisher
- University of Fort Hare
- Publisher
- Faculty of Science and Agriculture
- Language
- English
- Rights
- University of Fort Hare
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UFH Department of Biochemistry & Microbiology
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