Microbiology

ANTIMICROBIAL EVALUATION OF METABOLITES FROM CO-CULTURED NON-PATHOGENIC MICROORGANISMS AGAINST PATHOGENIC MICROORGANISMS ISOLATED FROM READY-TO-EAT FOODS

2023-07-31T11:02:16Z

ANTIMICROBIAL EVALUATION OF METABOLITES FROM CO-CULTURED NON-PATHOGENIC MICROORGANISMS AGAINST PATHOGENIC MICROORGANISMS ISOLATED FROM READY-TO-EAT FOODS BAMISI, OLUWAFERANMI ESTHER Antimicrobial activity of co-cultured Lactobacillus fermentum, Saccharomyces cerevisiae and Pleurotus ostreatus was tested against pathogenic microorganisms isolated from readyto-eat (RTE) foods. Microorganisms were isolated from RTE foods using standard microbiological techniques. The metabolites from co-cultured L. fermentum, S. cerevisiae and P. ostreatus was tested against isolated microorganisms using agar well diffusion. Bioactive compounds in metabolites was identified using Gas chromatography-Mass Spectrometry (GC-MS). Wistar albino rats were used to evaluate the in vivo antimicrobial efficacy of the metabolites. The bacteria isolated from RTE foods were Salmonella enterica, Shigella dysenteriae, Escherichia coli, Klebsiella pneumoniae (subsp ozaenae), Pseudomonas fluorescens, Bacillus cereus, Listeria monocytogens, and Staphylococcus aureus, while fungiisolated were Penicillium chrysogenum, Aspergillus flavus and Aspergillus niger. The metabolites from co-culture of P. ostreatus + S. cerevisiae and L. fermentum + P. ostreatus + S. cerevisiae showed the highest zones of inhibition of 23.00 mm and 22.00 mm against E. coli and S. aureus respectively and against Aspergillus niger and Penicillium chrysogenum at 20.00 mm and 18.70 mm respectively. Haematological parameters indicated a reduction in the WBC values (34.20×109 g/l, 35.30×109 g/l) and an increase in the PCV (35%, 36%), RBC (17.40×1012 g/l,17.40×1012 g/l) and HB (16 g/l, 16 g/l) of the infected albino rats treated with the co-cultured of P. ostreatus + S. cerevisiae and L. fermentum + P. ostreatus + S. cerevisiae respectively compared with the albino rats infected and not treated which had a WBC value (38.10×109 g/l), PCV (30%), RBC (12.90×1012 g/l) and HB (12 g/l). The histological study revealed that E. coli induced severe damage in the organs of the untreated albino rats, with observable improvements in the organs of the rats treated with the metabolites. The most abundant bioactive compounds in metabolites from co-culture of P. ostreatus + S. cerevisiae and L. fermentum + P. ostreatus + S. cerevisiae were alpha-linolenic acid (ALA), Acetic acid, 3-methylbutyl ester respectively. The pronounced antimicrobial activity against tested microorganisms could be attributed to bioactive compounds in secondary metabolites produced by the co-cultured microorganisms. The metabolites produced by microorganisms can be exploited for production of biopharmaceuticals towards better health delivery system. M.TECH THESIS

EFFECT OF ESSENTIAL OIL FROM PLANTAIN (Musa parasidiacal) AND PINEAPPLE (Ananas comosus) PEELS ON MICROORGANISMS ISOLATED FROM FUTA FISH POND

2023-07-31T10:43:56Z

EFFECT OF ESSENTIAL OIL FROM PLANTAIN (Musa parasidiacal) AND PINEAPPLE (Ananas comosus) PEELS ON MICROORGANISMS ISOLATED FROM FUTA FISH POND AROWOLO, FUNMILAYO HANNAH This study was designed to evaluate the antimicrobial activities of essential oils extracted from plantain and pineapple peels on microorganisms isolated from Federal University of Technology, Akure (FUTA) fish pond water, sediment and fish feeds. Samples were collected from fish pond water, sediment and feed at FUTA fish farm and transported to the Microbiology laboratory of FUTA for analysis. Isolation and identification of Microorganisms isolated were carried out using standard microbiological methods. The bacteria isolated were tested against commercially available antibiotics. Antibiotics residues from pond water, fish feed and pond sediment were determined using High Performance Liquid Chromatography (HPLC). Extraction of essential oil from plantain and pineapple peels was carried out using hydrodistillation method. Phytochemical screening of essential oil from plantain and pineapple peels was determined using Gas Chromatography Mass Spectrometry (GC-MS). Antimicrobial activity of essential oil against isolated microorganisms was carried out using agar well diffusion. Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were carried out. Pseudomonas aeruginosa, Esherichia coli, Staphylococcus aureus, Salmonella typhi, Bacillus sp, Shigella dysenteriae, Enterobacter aerogenes, Proteus vulgaris and Klebsiella pneumonia were isolated. Aspergillus niger (36.4%), Aspergillus flavus (42.3%) and Cladosporium sp. (23.0%) were isolated. Amoxicillin and ciprofloxacin residual content from fish pond sediment, pond water and feeds were 3.578 ppm and 2.666 ppm; 3.474 ppm and 3.143 ppm; and 3.450 ppm and 2.662 ppm respectively. The phytochemicals in the essential oils were phenols, tannins, flavonoids, saponins, alkaloids, steroids and terpenoids. Zones of inhibition by oil from pineapple and plantain peels ranged from 7.00 mm to 10.5 mm and 11.9 mm to 14.8 mm respectively. The Minimum inhibition concentration (MIC) of essential oil ranged from 6.25 mg/ml to 25.0 mg/ml and Minimum bactericidal concentration (MBC) ranged from 12.5 mg/ml to 50 mg/ml. Minimum fungicidal concentration (MFC) ranged from 25 mg/ml to 50 mg/ml. Essential oil from the peels of plantain and pineapple exhibited inhibitory activity against the test microorganisms, which can be attributed to the presence of certain secondary metabolites. Essential oil from plantain and pineapple peels possess bioactive compounds that can be exploited in formulation of fish feeds in order to suppress the growth of unwanted microorganisms. M.TECH

ANTI-INFLAMMATORY ACTIVITY OF LACTIC ACID BACTERIA ISOLATED FROM FERMENTED LOCUST BEANS ON FORMALIN INDUCED PAW OEDEMA IN WISTAR RATS.

2022-01-11T10:53:37Z

ANTI-INFLAMMATORY ACTIVITY OF LACTIC ACID BACTERIA ISOLATED FROM FERMENTED LOCUST BEANS ON FORMALIN INDUCED PAW OEDEMA IN WISTAR RATS. OLUWASOLA, HAWAH MORIYIKE Inflammation is the process by which the immune system recognizes and removes harmful stimuli and begins the healing process. Considering the potential side effects of anti- inflammatory drugs, there is a need to explore for more naturally available alternatives. Therefore, this study was designed to evaluate the anti-inflammatory activity of lactic acid bacteria (LAB) isolated from fermented locust beans (‘Iru’) using paw oedema inflammatory model induced by 1% formalin in Wistar rats. ‘Iru’ samples were purchased from Oja-Oba market, Akure, Ondo State and transported to the Microbiology laboratory of FUTA for bacteriological analyses. Isolation and identification of lactic acid bacteria (LAB) were carried out using standard microbiological techniques. The isolated LAB strains were used to treat acute inflammation in the rats. The rats were divided into six groups (A-F). Rats in group A were neither injected with formalin nor treated, while Group B rats were injected but not treated. Groups C, D and E rats received 2 × 10 7 CFU/mL of the various strains of LAB while Group F rats received diclofenac sodium (150 mg/Kg body weight) treatment following the injection of formalin. Paw thickness (mm) was checked at t = 0, 1, 4, 8, 24, 72, 168 and 336 hrs. Hematological parameters were analyzed using standard hematological techniques. Biomarkers assay for C-Reactive Protein (CRP), Interleukin-10 (IL-10) and Transforming Growth Factor-β (TGF-β) was performed on serum samples of the rats using Enzyme Linked Immunosorbent Assay (ELISA). The LAB isolated from ‘Iru’ were identified to be Weissella cibaria II-1-59, Weissella confusa JMC 1093, and Pediococcus pentosaceus DSM20336. Treatment with W. cibaria II-1-59 showed the best significant decrease in the paw oedema of the rats followed by P. pentosaceus DSM20336 and W. confusa JMC 1093 which was shown to be statistically significant at P<0.05. Oral administration of these LABs significantly decreased the WBC count in LAB treated rats. Weissella cibaria II-1-59 and Weissella confusa JMC 1093 showed a better ability to regulate leucocytes infiltration in the blood circulation of the inflamed rats than Pediococcus pentosaceus DSM20336. Also treatment with these LABs showed that they have better ability to regulate decrease in the RBC of the rats compared to diclofenac sodium. A significant decrease in the secretion of Pro-inflammatory biomarker (CRP) and increase in the serum levels of anti-inflammatory cytokines IL-10 and TGF-β were observed in all LAB treated groups. Weissella confusa JMC 1093 have a better ability to upregulate IL-10 production while Weissella cibaria II-1-59 and Pediococcus pentosaceus DSM20336 showed similar but less significant results. This suggests that these lactic acid bacteria isolated from ‘iru’ that were able to induce IL-10 production are promising anti-inflammatory agents in the treatment of inflammation, which further ascertain the health beneficial effect of the consumption of ‘iru’. M. TECH. Thesis

ANTIMICROBIAL ACTIVITIES OF STREPTOMYCES SPECIES ISOLATED FROM SOIL SAMPLES OBTAINED FROM FRUIT WASTE DUMP SITE AT SHASHA MARKET, AKURE, NIGERIA

2022-01-11T10:49:05Z

ANTIMICROBIAL ACTIVITIES OF STREPTOMYCES SPECIES ISOLATED FROM SOIL SAMPLES OBTAINED FROM FRUIT WASTE DUMP SITE AT SHASHA MARKET, AKURE, NIGERIA OLABANJI, ANUOLUWAPO OMOSEYE This research examined the antimicrobial activities of Streptomyces species isolated from soils of fruit waste dump site at Shasha Market, Akure, Ondo State. Soil samples were collected from four different fruit waste dump sites which are banana, carrot, tomato and orange dump sites from the location mentioned above with the aid of soil auger from the depth of 15 cm and Streptomyces were isolated using soil dilution technique and starch casein agar. The Streptomyces isolates were assessed for antimicrobial activity by the cross-streak technique. Production of antimicrobial metabolite was carried out in a fermentation medium. Optimization for antimicrobial metabolite production was carried out. Extraction of antimicrobial metabolite was carried out using ethyl acetate as solvent and purification of the crude extract was carried out using column and thin layer chromatography to determine the retention factor of the purified fractions. The purified extract was characterized using Fourier Transform Infrared (FTIR) analysis. Susceptibility of the test bacteria to antibiotics was carried out. The antimicrobial activities of the crude and purified extracts of the Streptomyces species was evaluated by agar well diffusion assay on plant and human bacteria and fungi. Ten Streptomyces species were isolated from fruit waste dump sites. Streptomyces rimosus TOMT and Streptomyces antibioticus TOMT were isolated from tomato dump site. Streptomyces citreus ORNG, Streptomyces albus ORNG, Streptomyces famosus ORNG, Streptomyces viridoflavus ORNG, Streptomyces flavovirens ORNG and Streptomyces rochei ORNG were isolated from orange dump site. Also, Streptomyces spiralis CART and Streptomyces griseoflavus BANA were isolated from carrot and banana dump sites respectively. In the preliminary screening of the antimicrobial activities of the Streptomyces species, five out of the ten isolates possessed antimicrobial activities against the test organisms. They were Streptomyces albus ORNG, Streptomyces viridoflavus ORNG, Streptomyces spiralis CART, Streptomyces flavovirens ORNG and Streptomyces rochei ORNG. Out of the five isolates, three were Streptomyces albus ORNG, Streptomyces flavovirens ORNG and Streptomyces rochei ORNG. Optimum production of antimicrobial metabolite by Streptomyces albus ORNG, Streptomyces flavovirens ORNG and Streptomyces rochei ORNG occurred at pH of 8. Culture medium amended with glucose served as good carbon source for maximum production of antimicrobial metabolites. Effect of ultraviolet radiation on the Streptomyces isolates resulted to decrease in inhibition zones against the plant and human bacteria and fungi. The crude extract showed antimicrobial activity against all the test bacteria and fungi. The purified extracts of Streptomyces albus ORNG, Streptomyces flavovirens ORNG and Streptomyces rochei ORNG gave retention factors of 0.47 cm, 0.58 cm and 0.55 cm respectively. The maximum antibacterial activity of Streptomyces albus ORNG was observed against Streptococcus spp. The maximum antifungal effect of Streptomyces flavovirens ORNG was observed against Sclerotium russia, while maximum antifungal activity of Streptomyces rochei ORNG was observed on Aspergillus fumigatus. Ciprofloxacin was the most active antibiotic with broad spectrum of activity. Nystatin showed maximum activity against Candida albicans. Mancozeb showed maximum activity against Colletotrichum spp. The minimum inhibitory concentration of the purified extracts ranged between 3.1 mg/ml and 50 mg/ml. The infrared analysis revealed the possibility of the presence of phenolic compounds in the purified extract. The antimicrobial metabolites obtained from Streptomyces species in this study showed significant inhibition on the selected bacteria and fungi. This emphasizes the role of soil Streptomyces in modern drug discovery, production of novel antimicrobial compounds to target resistant bacteria and biocontrol of plant pathogens. M. TECH. Thesis