PhD (Microbiology) / Department of Microbiology / See the attached abstract below

MSc (Microbiology) / Department of Microbiology / Background: The medicinal value of plants lies in some chemical substances that produce a definite physiological action in the human body. The secondary metabolites help the plants to survive hash conditions and could be used by humans as supplements of their health, as foods additives or for medicinal purposes. This bioactive compounds are not always beneficial to human beings, and some of this plants bioactive compounds can be toxic or genotoxic to human cells. This study used several methods to evaluate of phytochemical constituents and mutagenic properties of Coccinia rehmanni and Jatropha zeyheri plant extracts. Methodology: Methanol was used for extraction of the bioactive compounds from the two selected plants, filtered with Whatman filter paper and evaporated with rotary evaporator. The extracts were fractionated using open column chromatography. Chemical and TLC methods were used to determine phytochemicals of the study plants extracts and fractions. The plants extracts and fractions were tested against Vero cell lines in order to evaluate cytotoxicity and genotoxicity of the plants. NucRed and LTR Hoechst 33342 dyes were used for cytotoxicity and genotoxicity respectively. For the evaluation of cytotoxicity and genotoxicity Quantification of live and dead cells for the screening assay was performed using the ImageXpress Micro XLS Widefield Microscope and acquired images analyses using the MetaXpress software and Multi-Wavelength Cell Scoring Application Module. Antimutagenicity of plants extracts was observed using PARP universal colorimetric assay kit. Acquired data was transferred to an EXCEL spreadsheet and data was analyzed. Results and discussion: C. rehmanni (12.03%) yielded more extract than J. Zeyheri (8.20%). the two plants had different compound composition and were in different stages of maturity. The study revealed the domination of Terpenoids, Cardiac glycosides, Phenolic and tannis. With an exception of two fraction fractions all the fractions was found to be toxic to an extent were genotoxicity of such fraction could not be concluded. The reason for such extreme toxicity could be due to the influence of the retained alcohol during rotary evaporation. xvi | P a g e Conclusion: this study provides and add to existing knowledge on the phytochemicals mutagenicity and anti-mutagenicity of C. rehmanni and J. Zeyheri medicinal plants. The study serves as scientific proof that extensive use of this plant in traditional medicine for treatment of various ailments may lead to some irreversible damages. / NRF

PhD (Microbiology) / Department of Microbiology / Background: Currently, Human Immunodeficiency Virus/ Acquired Immune Deficiency Syndrome (HIV/AIDS) is one of the deadliest diseases in the world. In Gabon, the prevalence is estimated at 4.1%. Because of the several side effects of highly active antiretroviral therapy on the health of people living with HIV, an emphasis on the utilization of based plant treatments have been noticed highlighted with testimonies of patient health improvement. That led researchers to conduct ethnobotanical surveys aiming at reporting medicinal plants used to treat HIV related opportunistic infections. From the outcomes, 20 plants have been selected based on the frequency of their use but also on the absence of papers in the literature related to HIV research conducted in Gabon. This study aims at evaluating the cytotoxicity, antioxidant, antimicrobial, anti-HIV and immunomodulatory activities of the 20 selected plants and to establish the phytochemical profile of these plants. Methodology: The plants were extracted using absolute methanol and distilled water. Qualitative (phytochemical screening) and quantitative (TPC and TFC) analysis were performed on the 40 extracts obtained. The extracts were assessed for cytotoxicity on Vero and HeLa cells using MTT and dual staining/fluorescence microscopy. The antimicrobial activity of the extracts was evaluated using well diffusion assay and micro-broth dilution assay. DPPH free radical scavenging assay and ferric reducing power were used to determine the antioxidant potential of the extracts. The anti-HIV effect was evaluated on HIV infected blood using reverse transcriptase inhibitory assay and p24 antigen inhibitory assay. The immunomodulatory effects of the extracts were evaluated on HIV infected blood using IL-2, IL-6 and TNF-α ELISA. Coula edulis and T. iboga methanol extracts were selected for fractionation using column chromatography. LC-MS was used to establish the phytochemical profile of the two selected extracts. Results: The results revealed that saponins, alkaloids, terpenoids, phenols, tannins, steroids, flavonoids and cardiac glycosides were detected in almost all the plant extracts. The extracts were found to contain more phenolics than flavonoids especially the methanolic extracts. Both alcoholic and aqueous extracts of C. edulis, M. cecropioides, S. ochocoa, S. kamerounensis, U. guineensis and C. religiosa, the methanolic extract of A. hirtella and R. vomitoria and the aqueous extracts of U. klainei presented IC50 significantly lower (p<0.05) than ascorbic acid (38.87±1.54 μg/ml). Both Coula edulis methanolic and aqueous extracts, methanolic extracts of S. kamerounensis, S. gabonensis, M. monandra, U.guineensis and A. klaineana showed antioxidant activity significantly xii | P a g e (p<0.01) higher than Ascorbic acid whose EC50 was 152.16±1.09 μg/ml. For the fluorescence microscopy the extracts of interest for Vero cells inhibition were A. klaineana, T. iboga, U. klainei and C. edulis while for Hela cells the cytotoxic activity was significant (p<0.05) for the following extracts: U. klainei, U. guineensis, P. soyauxii, A. klaineana, V. conferta, M. monandra, R. vomitoria, R. africanum, C. edulis and S. ochocoa. The extracts of C. edulis, M. cecropioides, S. ochocoa, S. gabonensis, A. hirtella, R. vomitoria, S. kamerunensis, P. soyauxii, U. guineensis and T. iboga displayed the highest antimicrobial activity against all the selected bacteria. For the anti-HIV the extracts of interest were V. conferta (47%), C. lucanusianus (47%), C. religiosa (44%), A. hirtella (42%), S. kamerunensis (41%), M. puberula (41%) as well as both aqueous and methanolic extracts of T. iboga (48% and 45% respectively), U. guineensis (46% and 41% respectively) and A. klaineana (41% and 44% respectively). The results for RT inhibition assay revealed that out of 24 selected plant extracts selected only C. edulis, C. religiosa, C. lucanusianus, R. africanum, P. soyauxii and V. conferta were able to decrease RT by more than 50%. The modulatory effect of the extracts on the secretion of IL-2, IL-6 and TNF-α by HIV infected cells was found to be insignificant (p>0.05). The fractionation of T. iboga and C. edulis methanol extracts eluted 4 fractions each. The LC-MS analysis revealed at negative ionization the presence of phenolic compounds (Ellagic acid, Gallic acid, and quercetin) in C. edulis samples while alkaloids (ibogaine, ibogaline, voacangine) were mainly found in T. iboga samples at positive ionization. The two crude extracts and the fractions showed some levels of antimicrobial, antioxidant, anti-HIV and anti-inflammatory activities. Conclusion: The present study has validated the use of the selected plants in the management of AIDS in Gabon and provided an explanation of the improvement in HIV individual’s life reported. Some of these plants could constitute good candidate for promising anti-HIV molecules. / NRF