Browsing by Author "Green, Ezekiel"

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    Antibacterial activities of crude secondary metabolite extracts from pantoea species obtained from the stem of solanum mauritianum and their effects on two cancer cell lines
    (MDPI, 2019) Uche-Okereafor, Nkemdinma; Sebola, Tendani Edith; Tapfuma, Kudzanai; Mekuto, Lukhanyo; Green, Ezekiel; Mavumengwana, Vuyo
    ENGLISH ABSTRACT: Endophytes are microorganisms that are perceived as non-pathogenic symbionts found inside plants since they cause no symptoms of disease on the host plant. Soil conditions and geography among other factors contribute to the type(s) of endophytes isolated from plants. Our research interest is the antibacterial activity of secondary metabolite crude extracts from the medicinal plant Solanum mauritianum and its bacterial endophytes. Fresh, healthy stems of S. mauritianum were collected, washed, surface sterilized, macerated in PBS, inoculated in the nutrient agar plates, and incubated for 5 days at 30 °C. Amplification and sequencing of the 16S rRNA gene was applied to identify the isolated bacterial endophytes. These endophytes were then grown in nutrient broth for 7–14 days, after which sterilized Amberlite® XAD7HP 20–60 mesh (Merck KGaA, Darmstadt, Germany) resin was added to each culture to adsorb the secondary metabolites, which were later extracted using ethyl acetate. Concentrated crude extracts from each bacterial endophyte were tested for antibacterial activity against 11 pathogenic bacteria and two human cancer cell lines. In this study, a total of three bacterial endophytes of the Pantoea genus were identified from the stem of S. mauritianum. The antibacterial test showed that crude secondary metabolites of the endophytes and stem of S. mauritianum possessed antibacterial properties against pathogenic microbes such as Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa, with concentrations showing inhibition ranging from 0.0625 to 8.0000 mg/mL. The anticancer analysis showed an increase in cell proliferation when A549 lung carcinoma and UMG87 glioblastoma cell lines were treated with both the plant and endophytes’ crude extracts. As far as we know, this is the first study of its kind on Solanum mauritianum in South Africa showing S. mauritianum endophytes having activity against some of the common human pathogenic organisms.
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    Antimicrobial resistance phenotype of staphylococcus aureus and escherichia coli isolates obtained from meat in the formal and informal sectors in South Africa
    (ps202304, 2020-09) Jaja, Ishmael Festus; Jaja, Chinwe-Juliana Iwu; Chigor, Nnamdi Vincent; Anyanwu, Madubuike Umunna; Maduabuchi, Ezealisiji Kenneth; Oguttu, James Wabwire; Green, Ezekiel
    Background: Foodborne diseases (FBD) caused by resistant pathogens are a global public health problem. One main driver of the increasing FBD incidence is the transfer of pathogenic organisms from animal guts to carcasses during processing and subsequent transfer from meat products to consumers. Methods: In this study, meat samples from abattoirs in the formal meat sector (FMS) (n = 140) and slaughter points in the informal meat sector (IMS) (n = 104) were collected for microbial detection and phenotypic AMR determination using polymerase chain reaction. Results. The antibiogram of Staphylococcus aureus isolates revealed that resistance to clindamycin (74.3%) and ampicillin (59.5%) was highest in the FMS, while resistance to penicillin (83.8%) and tetracycline (82.1%) was highest in the IMS. Escherichia coli isolates show significant resistance to chloramphenicol (90.7%) and tetracycline (82.3%) in the FMS. Likewise, resistance to tetracycline (92.3%) and sulfamethoxazole/trimethoprim (87.5%) was highest in the IMS. The multiple antibiotic resistance index (MARI) for S. aureus and E. coli ranged from 0.3 to 0.8 and 0.2 to 0.5, respectively. Conclusion: This study suggests high-level contamination of meat with resistant pathogens and highlights the public health consequences associated with consuming such unhygienic products
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    Cytotoxic activity of crude extracts from Datura stramonium’s fungal endophytes against A549 lung carcinoma and UMG87 glioblastoma cell lines and LC-QTOF-MS/MS based metabolite profiling
    (BMC (part of Springer Nature), 2019-11-21) Tapfuma, Kudzanai Ian; Uche-Okereafor, Nkemdinma; Sebola, Tendani Edith; Hussan, Raeesa; Mekuto, Lukhanyo; Makatini, Maya Mellisa; Green, Ezekiel; Mavumengwana, Vuyo
    Background: Endophytic fungi are a proven source of bioactive secondary metabolites that may provide lead compounds for novel drug discovery. In this study, crude extracts from fungal endophytes isolated from Datura stramonium were evaluated for cytotoxic activity on two human cancer cell lines. Methods: Fungal endophytes were isolated from surface sterilized aerial parts of D. stramonium and identified using molecular, morphological and phylogenetic methods. Ethyl acetate crude extracts from these isolates were evaluated for cytotoxic activity on A549 lung carcinoma and UMG87 glioblastoma cell lines. Metabolite profiling was then performed by liquid chromatography coupled to quadrupole time-of-flight with tandem mass spectrometry (LC-QTOF-MS/MS) for the cytotoxic crude extract. Results: Eleven fungal endophytes were identified from D. stramonium. Significant cytotoxicity was only observed from the crude extract of Alternaria sp. KTDL7 on UMG87 glioblastoma cells (IC50 = 21.49 μg/ml). Metabolite profiling of this crude extract tentatively revealed the presence of the following secondary metabolites: 1,8-dihydroxynaphthalene (1), anserinone B (2), phelligridin B (3), metacytofilin (4), phomopsidin (5) and vermixocin A (6). Compounds 2 and 3 have been shown to be cytotoxic in literature. Conclusion: The findings in this study suggest that the crude extract of Alternaria sp. KTDL7 possesses compound(s) cytotoxic to glioblastoma multiforme cells. Future studies to isolate and characterize the cytotoxic compound(s) from this fungus could result in lead development of a fungal-based drug for glioblastoma multiforme treatment.
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    Diversity and antimicrobial activity of culturable fungal endophytes in Solanum mauritianum
    (MDPI, 2020-01) Pelo, Sharon; Mavumengwana, Vuyo; Green, Ezekiel
    Plant endophytes are microbial sources of bioactive secondary metabolites, which mimic the natural compounds chemistry of their respective host plants in a similar manner. This study explored the isolation and identification of fungal endophytes, and investigated the antibacterial and antimycobacterial activity of their crude extracts. Fungal endophytes were isolated from Solanum mauritianum, identified using morphological traits and internal transcribed spacer ribosomal-deoxyribonucleic acid (ITS-rDNA) sequence analysis. Eight fungal endophytes were identified as Aureobasidium pullulans, Paracamarosporium leucadendri, Cladosporium sp., Collectotrichum boninense, Fusarium sp., Hyalodendriella sp., and Talaromyces sp., while Penicillium chrysogenum was isolated from the leaves and unripe fruits. Good activity was observed for the crude extracts of Paracamarosporium leucadendri inhibiting Mycobacterium bovis, Klebsiella pneumoniae, and Pseudomonas aeruginosa at 6 µg/mL. Crude extracts of Fusarium sp., showed activity at 9 μg/mL against M. bovis, M. smegmatis and K. pneumonia. In general, the crude extracts showed great activity against Gram-negative and Gram-positive bacteria and novel results for two mycobacteria species M. bovis and M. smegmatis. The results provide evidence of diverse fungal endophytes isolated from Solanum mauritianum, and evidence that fungal endophytes are a good source of bioactive compounds with pharmaceutical potential, particularly against Mycobacterium tuberculosis.
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    Evaluating antibacterial and anticancer activity of crude extracts of bacterial endophytes from Crinum macowanii Baker bulbs
    (Wiley Open Access, 2019) Sebola, Tendani Edith; Uche‐Okereafor, Nkemdinma C.; Tapfuma, Kudzanai I.; Mekuto, Lukhanyo; Green, Ezekiel; Mavumengwana, Vuyo
    The results from this study revealed that crude extracts isolated from bacterial endophytes obtained from Crinum macowanii bulbs showed activity against both Gram-positive and Gram-negative pathogenic bacteria, while Acinetobacter guillouiae crude extracts displayed anticancer activity. This study aimed to isolate and characterize bacterial endophytes and their crude extracts from C. macowanii bulbs. Endophytes were isolated using validated surface sterilization techniques, followed by phenotypic and genotypic profiles of the isolates. Crude extracts were extracted from the endophytes using ethyl acetate, while methanol:dichloromethane (1:1) was used to obtain crude extracts from the bulbs. Antibacterial activity of crude extract from each endophyte was investigated against selected pathogenic strains using the broth microdilution method, and anticancer activity against U87MG glioblastoma and A549 lung carcinoma cells was determined by the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. Acinetobacter guillouiae, Pseudomonas moraviensis, Pseudomonas sp., Rahnella aquatilis, Bacillus cereus, Novosphingobium sp., Raoultella ornithinolytica, and Burkholderia tropica were successfully isolated. The crude extracts from the majority of endophytes showed antibacterial activity, ranging from 0.125 to >16.00 mg/ml against Gram-negative and Gram-positive pathogenic bacteria. Acinetobacter guillouiae extracts showed a high bioactive potential against U87MG glioblastoma cell lines by reducing their growth by 50% at concentrations of 12.5, 6.25, and 3.13 µg/ml. Crude extracts isolated from C. macowanii bulbs showed potential for possible drug lead against common pathogenic bacteria.