Crude extracts activity of three selected medicinal plants from the Venda region against some pathogenic organisms.

Background: The emerging of antimicrobial resistance has become a problem as it is threatening public health worldwide. Objectives: To extract crude extracts from three different medicinal plants, test activity against Mycobacterium smegmatis, Staphylococcus aureus and Escherichia coli and screen for phytochemicals of those that showed activity against the targeted bacteria. Methods: KirkiaacuminataOliv., Dichrostachyscinerea (L.) Wight &Arn. and MimusopszeyheriSond. plants were collected at Thengwe area, Mafukani village, Limpopo Province, South Africa. The plant materials collected were extracted using four solvents. Antimicrobial screening was accomplished using the agar well diffusion method and the crude extracts that showed activity against the targeted organisms were screened for phytochemicals using different tests. Results: With all solvents used for extraction, methanol had a greater yield of 14.1% from Dichrostachyscinerea crude extracts. Kirkiaacuminata and Dichrostachyscinerea were medicinal plants that inhibited Mycobacterium smegmatis and Staphylococcus aureus at the lowest concentration of 2.5 mg/ml and 1.25 mg/ml. Conclusions: The results from this study show that the selected medicinal plants are active against Mycobacterium smegmatis and Staphylococcus aureus and their pharmacological properties can be further analyzed for the development of new drugs.

Characterization of mutations in the rpoB gene conferring rifampicin resistance in Mycobacterium tuberculosis complex isolated from lymph nodes of slaughtered cattle from South Africa.

Tuberculosis (TB) is an ongoing public health care, with the state of affairs exacerbated by the growth of anti-TB drug-resistant forms in South Africa. Not much attention is given to zoonotic TB. Thus, this study aimed to determine the presence of rpoB mutations among Mycobacterium tuberculosis complex (MTBC) isolates of lymph nodes from slaughtered cattle. A count of 14,950 carcasses from selected abattoirs were examined for nodular lesions and enlarged lymph nodes; 376 lymph nodes were cultured for MTBC. Positive isolates were tested for drug sensitivity against three anti-TB drugs, rifampicin, isoniazid, and ethambutol, using the Lowenstein-Jensen proportion method. Rifampicin-resistant isolates were sequenced, and spoligotyping was performed for lineage classification. A total of 162 isolates were confirmed as MTBC and 42 isolates were resistant to rifampicin. All rifampicin-resistant isolates carried the H526D rpoB mutation, and almost all of them carried an additional nonsynonymous nucleotide substitution in the hot spot region, in three other codons (510, 516 and 522). In total, 5 different mutations at four codons are reported, including one isolate showing 3 of them which has never been reported in South Africa. In addition, we report 4 different spoligo patterns, with 34 isolates known and 8 unknown spoligotype international types. From the known clades, 5 (11.9%) isolates were identified as Bov_4 caprae lineage, 29 (69%) Beijing, and 8 (19.1%) remaining unknown clades. The detection of MTBC-resistant patterns from cattle lymph nodes (Eastern Cape, South Africa) necessitates the investigation of other possible routes of MTBC transmission.

Clonality and genetic profiles of drug-resistant Mycobacterium tuberculosis in the Eastern Cape Province, South Africa.

In this study, we investigated the diversity of drug-resistant Mycobacterium tuberculosis isolates from families who own cattle in the Eastern Cape Province of South Africa using spoligotyping and mycobacterial interspersed repetitive-unit-variable number tandem repeat (MIRU-VNTR) typing. The Mycobacterium tuberculosis was investigated using MIRU-VNTR and the Mycobacterium tuberculosis families were evaluated using spoligotyping. Spoligotyping grouped 91% of the isolates into seven clusters, while 9% of the deoxyribonucleic acid (DNA) from TB isolates were unclustered from a total of 154 DNA used. Previously described shared types were observed in 89.6% of the isolates, with the Beijing family, SIT1, the principal genotype in the province, while the families T, SIT53 and X1, SIT1329 were the least detected genotypes. MIRU-VNTR grouped 81% of the isolates in 23 clusters while 19% were unclustered. A combination of the VNTR and spoligotyping grouped 79% of the isolates into 23 clusters with 21% unclustered. The low level of diversity and the clonal spread of drug-resistant Mycobacterium tuberculosis isolates advocate that the spread of TB in this study may be instigated by the clonal spread of Beijing genotype. The results from this study provide vital information about the lack of TB control and distribution of Mycobacterium tuberculosis complex strain types in the Eastern Cape Province of South Africa.

Prevalence of tuberculous lymphadenitis in slaughtered cattle in Eastern Cape, South Africa.

OBJECTIVE: To detect the prevalence of Mycobacterium tuberculosis complex (MTBC) in the lymph nodes of slaughtered cattle collected from selected abattoirs in Eastern Cape Province of South Africa. METHODS: A total of 376 lymph nodes were collected from slaughtered cattle over a period of 12 months. Certain characteristics (sex, age, body condition score, and breed) were observed to be associated with MTBC among slaughtered cattle. Collected samples were cultured and tested for acid-fast bacilli (AFB). DNA was isolated, purified, and quantified using a spectrophotometer. Quantified DNA was confirmed to be MTBC by multiplex PCR targeting two genes (IS6110 and mpb64). RESULTS: Of the 376 collected lymph nodes, 182 were positive when tested by Ziehl-Neelsen stain and 162 were confirmed positive for MTBC by PCR. MTBC was isolated from lymph nodes with nodular lesions (72.8%, 118/162) and inflamed lymph nodes (27.1%, 44/162). All detected MTBC isolates were positive for region of deletion 1 (RD1). No isolate was detected to have Mycobacterium bovis bacille Calmette-Guérin (BCG). However, 3.1% had M. bovis and 96.9% had M. tuberculosis. CONCLUSIONS: The presence of live Mycobacterium strains in slaughtered cattle poses a health risk to beef consumers and abattoir workers.

Molecular detection and characterization of resistant genes in Mycobacterium tuberculosis complex from DNA isolated from tuberculosis patients in the Eastern Cape province South Africa.

BACKGROUND: Tuberculosis (TB) in both animals and humans is caused by Mycobacterium tuberculosis complex (MTBC) primarily transmitted by inhalation of aerosolized droplets containing the organism. Multi-drug resistance (MDR) and extensive drug resistance (XDR) are evolutionary features of Mycobacterium tuberculosis to subvert the antibiotic regimes in place. The heavy burden of TB worsened by HIV endemic in South Africa motivated for the investigation of MTBC prevalence among TB patients in Port Elizabeth and the amplification and sequencing of the DNA amplicons known to confer resistance to TB drugs. METHODS: Three thousand eight hundred and ten (3810) sputum specimens were processed and DNA was isolated from sputum specimens collected from different hospitals and health care places in the Eastern Cape Province, South Africa. DNA was amplified using the Seeplex® MTB Nested ACE detection assay. The agar-dilution proportion method was used to perform drug-sensitivity testing using 7H10 Middlebrook medium. Target genes known to confer resistance to first and second-line drugs were amplified and the amplicons sequenced. RESULTS: One hundred and ninety (5%) DNA samples tested positive for MTBC and from the resistant profiles of the 190 positive samples, we noted that multidrug-resistant TB was identified in 189 (99.5%) with 190 (100%) patients infected with MTB resistant to isoniazid and 189 (99.5%) having MTB resistant to rifampicin. Other percentages of drug resistance observed including 40% pre-XDR and 60% of XDR. CONCLUSION: This study provides valuable data on the different kinds of mutations occurring at various target loci in resistant MTBC strains isolated from samples obtained from the Eastern Cape Province. The results obtained reveal a high incidence of MDR amongst the positive samples from Eastern Cape Province, South Africa.