Scholarly works in Pharmaceutical Microbiology
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Item Extracts of Trichilia heudelotii (Meliaceae) planch, a Nigerian medicinal plant have antibacterial and antifungal activity(Sciencedomain International, 2024) Adeniyi, B. A.; Adagbasa, O. O.; Idowu, P. A.; Igbokwe, C. O.; Moody, J. O.; Aiyelaagbe, O. O.There is no overemphasizing the need for novel phytotherapeutic agents to combat the menace of drug resistance in microbial pathogens. Many plant species so far, have yielded some bioactive compounds with great promise for such drugs. Trichilia heudelotii (Meliaceae) is commonly used in traditional medicine in Nigeria for the treatment of many microbial infections ranging from gastrointestinal infections to gonorrhea. This study is thus designed to determine the phytochemicals present in T. heudeloti and evaluate the plant’s extracts' antimicrobial activity on some bacterial and fungal isolates. The phytochemical screening was done using standard procedures. Soxhlet extracts using n-hexane and methanol were drying in vacuo. The methanol extract was partitioned into petroleum ether, chloroform, and aqueous fractions. The antibacterial and antifungal activity of the extracts determined using the agar-well diffusion method. The MICs were determined for the extracts using the agar dilution method. The qualitative phytochemical screening revealed the presence of tannins, saponins, alkaloids, cardenolides, and anthraquinones in the leaf, stem bark, and root bark of T. heudelotii. The extracts showed considerable activity against Gram-positive and Gram-negative organisms, and fungi with dermatophytes including Klebsiella spp, Escherichia coli, Proteus spp, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Neisseria gonorrhoeae, Mycobacterium smegmatis, Candida albicans, Aspergillus niger, Rhizopus stolonifer, Penicillium spp, Microsporium canis, and Trichophyton mentagrophytes. The mean diameter of zones of inhibition exhibited by the extracts ranged between 10mm+ 0.5 and 32+ 0.33mm. The methanol extracts compared favourably with the gentamycin (standard control). The minimum inhibitory concentration (MIC) ranged between 0.157mg/mL and 20mg/mL. The crude methanol extracts and methanol residue showed the highest activity of all the extracts while the hexane extract showed the lowest activity and the Petroleum ether fraction was inactive. These results showed the potential of T. heudelotii as a possible candidate for bioactive compounds for the discovery and development of new drugs for the treatment of diseases caused by test pathogens.Item Antimicrobial, phytochemical and antioxidant screening of acalypha fimbriata Leaf Extract for alternative antimicrobial therapy(Faculty of Physical Sciences and Faculty of LifeSciences, Univ. of Ilorin, Nigeria, 2023) Okunye, O. L.; Idowu, P. A.; Okanlawon, B. M.; Adejumo, O. E.; Saka, A. S.; Oyinloye, O. E.; Ayedun, J. S.; Adeyemo, O. M.; Kaseem, L. S.; Idowu, A. O.In folklore, Acalypha fimbriata have been associated with ‘cure all’ properties, but without sufficient empirical ethnopharmacological scientific backups. This research work is therefore necessitated to determine the antimicrobial, phytochemicals and antioxidant activity of Acalypha fimbriata. The leaf of Acalypha fimbriata were Soxhlet extracted, reconstituted, and screened for phytochemical constituent of antimicrobial importance. Conventional biochemical characterization was carried out on the isolates of Staphylococcus aureus, Escherichia coli Pseudomonas aeruginosa and Klebsiella pneumoniae obtained for the research work and the antimicrobial activity of the plant sample was determined using the agar well diffusion technique. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) of the extract were determined by broth dilution method on the isolates. The antioxidant activity of the extract was determined using 2,2-diphenyl-1 picrylhydrazyl (DPPH) method and Graphpad prism were used to interprets the data. Alkaloids, flavonoid, anthraquinones, tannins and saponins was found in the leaf extract. Staphylococcus aureus exhibited highest zone of growth inhibition (28mm) at 100mg/ml while Pseudomonas aeruginosa had the lowest (14mm) at 100mg/ml from the antimicrobial assay. In the determination of the minimum inhibitory concentration and minimum bactericidal concentration, Staphylococcus aureus exhibited MIC and MBC at 0.625μg/ml and 12.5 μg/ml, while Pseudomonas aeruginosa elicited MIC and MBC at 10 μg/ml and >10 μg/ml of MIC and MBC respectively.Themethanol extract of the plant acted as hydrogen/electrons donor or scavenger of radicals with fifty percent inhibitory concentration (IC50) of 59.83 μg/ml while that of Ascorbic acid (standard) was found to be 92.70 μg/ml using.The varied MIC’s and MBC’s obtained coupled with the values recorded for the antioxidant radicals validate the antimicrobial and antioxidant potential of Acalypha fimbriata that can be explored for therapeutic option, if further purified and optimally processed.Item Antibacterial effects of the leaves and twigs of Turraea vogelii on some enteric pathogens(Faculty of Science, Bauchi State University, Gadau (BASUG), Nigeria, 2023) Olufadi-Ahmed, H. Y.; Idowu, P. A.; Audu, H. J.Qualitative and quantitative phytochemical analysis as well as antibacterial analysis of extracts of Turraea vogelii (Hook F.) leaves and twigs on some enteric pathogens were reported in the current study. The qualitative phytochemical screening of the leaf extracts showed the presence of saponins, flavonoids, cardiac glycosides, anthraquinones, terpenoids, steroids and alkaloids. In addition, the quantitative phytochemical screening showed that the greatest percentage yield was from the methanol extract with alkaloids (8.8%) and terpenoids (8.7%) showing the highest concentrations. The antibacterial effects of hexane, ethyl acetate and methanol extracts of Turraea vogelii at concentrations 80 mg/mL, 40 mg/mL and 20 mg/mL each were studied by the pour plate method. Clinical isolates of Escherichia coli, Salmonella typhi and Proteus mirabilis (five each) obtained from University College of Health (UCH), Ibadan, Nigeria were employed as test organisms. Gentamicin was used as control at concentration of 10 μg/mL. Preliminary antimicrobial assay using only methanol showed antimicrobial activity in both leaves and twigs. Results showed that Escherichia coli was most susceptible to all extracts of the leaves while Proteus mirabilis was the least susceptible. For the twigs, Salmonella typhi was most susceptible to the hexane extract, E. coli was most susceptible to ethyl acetate extract and Proteus mirabilis was most susceptible to the methanol extract. Minimum Inhibitory Concentration (MIC) of leaves showed that its hexane extract is bacteriostatic at 40 mg/mL but at 20 mg/mL, it showed bacteriostatic activity against Proteus mirabilis. The methanol extract of the twigs had an MIC of 20 mg/mL for most of the isolates collected except the Salmonella typhi that had an MIC of 40 mg/mL. Minimum Bactericidal Concentration (MBC) of the ethyl acetate extract of leaves and methanol extract of twigs was 40 mg/mL. This study has demonstrated the antibacterial effect of leaves and twigs extracts of T. vogelii on some enteric pathogens.Item Plasmid DNA mediated vancomycin-resistant Staphylococcus aureus (VRSA) from cases of urinary tract infection(Faculty of Pharmaceutical sciences, University of Jos, 2022) Okunye, O. L.; Idowu, P. A.; Okanlawon, B. M.; Durowaiye, M. T.; Adejumo, O. E.; Babalola, C. O.; Ayedun, J. S.; Oyinloye, O. E.; Osungunna, M. O.Vancomycin resistance commonly occurs with Staphylococcus aureus, a pathogen that exhibits intrinsic virulence. Sixty isolates of Staphylococcus aureus tested positive as vancomycin possessive isolates on Brain Heart Infusion Agar medium fortified with 6 μg/mL vancomycin from 100 clinical samples of urine from patients with cases of UTIs via agar diffusion method. Standard 30 μg vancomycin disc served as control. Increase in zones of growth inhibition in relation to vancomycin concentrations was noticed in some of the isolates while it was reversed in others, despite the increase in concentration. Typed strain was susceptible to six concentrations of vancomycin exposed and to the control. Thirty one of the sixty isolates were resistant to vancomycin control. Resistant isolates from standard vancomycin discs were then subjected to molecular investigation. Of the 31 isolates exposed to gel electrophoresis, 14 (47%)elicited plasmids of varied molecular weights ranging from 0.79-23.13 kb. The magnitudes of vancomycinresistant isolates from the clinical samples studied, coupled with some incoherent zones of inhibition and the plasmid DNA obtained from the resistant isolates, suggest the need for infection control practitioners and epidemiologist to devise strategies to curtail the spread of this pathogen both in hospital and community settings.Item A study of skin sepsis amongst abbatoir workers in Moniya, Ibadan, Oyo State, Nigeria(Association of Resident Doctors (ARD), University College Hospital (UCH), Ibadan, 2022) Okunye, O. L.; Babalola, C. O.; Adeleke, O. E.; Idowu, P. A.; Coker, E. M.; Ayedun, J. S.; Durowaye, M. T.Background: Skin sepsis is a pyodermal infection caused by Lancefield’s group streptococci and Staphylococcus aureus . It is characterized by discolored and mottled skin, cellulitis, impetigo and multi-systemic collagen muscularitis and can be transmitted from person to person. Objective: This study sampled the skin of consented abattoir workers in Moniya Ibadan, for clinical sepsis, with a view to establishing a causal relationship between the infection obtained and the abattoir workers examined. Methodology: A total of 100 meat handler’s hands and forearms were examined. Swabs were taken from lesions which appeared clinically to be infected and then propagated on selective culture media designed for staphylococci and streptococci. Conventional biochemical tests and Lancefield determination were carried out as considered appropriate. Results: Of the 100 swabs from the categories of abattoir workers examined, 43 streptococci (35 from lesion 8 from wound) and 36 Staphylococcus aureus (20 from lesion and16 from wound) were obtained. The regression analysis from the grouping of gender, causative agents and specific infection as a predictor of infection were recorded to be significant ( = 0.18; t = 1.74; p < 0.05) for the nature of but non-significant ( =-0.067; t =-0.649; p> 0.05) for the gender. In-vitro antigen antibody reaction on StreptexM kit elicited varied reactions to Lancefield’s serological grouping (A (56%), B (9%) C (7%) G (22%) and L (7%). Resistance of Staphylococcus aureus in varying percentages (Tetracycline and gentamicin 62%, meropenem and ceftriaxone 100%, amikacin, 10%, and vancomycin 80%) to conventional antibiotics were observed. Conclusion: From this study point of view, the causal relationship between theinfection and the infected has been establish from the pathogens of pyodermal origin contacted from cows, that causes sepsis across all the category of abattoir worker studied. There is a need to provide an ideal functioning abattoir fully equipped with required facilities for safety and ease of execution of duties.Item Molecular Investigation of Clinical Isolates of Staphylococcus aureus from Cases of Boil Infection(Lagos University Medical Society (LUMS), 2022) Okunye, O. L.; Lawal, S. A.; Idowu, P. A.; Okanlawon, B. M.; Adejumo, O. E.; Fagboun, A. B.; Babalola, C. O.; Saka, A. S.; Oyinloye, O. E.Background: Boil, a pyodermal infection caused by Staphylococcus aureus is a very common skin disease. It is characterized by pus filled lump at specific anatomical loci. Objectives: This study evaluated the antimicrobial a c t i v i t y o f v a n c o m y c i n p o w d e r i n v a r i e d concentrations alongside with 30μg vancomycin standard discs, determined the minimum inhibitory concentration and plasmid profiles of the resistant isolates of Staphylococcus aureus from boil infection. Methods: A total of one hundred (100) exudates from boils were collected with a sterile swab for bacteriological examination; the samples were culture on mannitol salt agar, followed by Gram staining and other conventional biochemical characterization. The isolates were tested by agar diffusion method against varied concentrations of vancomycin and a standard 30 μg vancomycin disc as a control. The resistant isolates from standard vancomycin disc were subjected to molecular investigation to determine the source of their vancomycin resistance. Results: Remarkable zones of growth inhibitions to varied concentrations of vancomycin powder that ranged from 32μg/mL-1.0 μg/mL were recorded, a l t h o u g h , v a n c o m y c i n a n t i b i o t i c s a r e n o t concentration dependent. The minimum inhibitory concentration for the 60 isolates examined elicited varied values. Of the 60 isolates exposed to plasmid investigation, fourteen(14) elicited resistance that were plasmid mediated which molecular weight ranged from 0.82 kb - 27.22kb. Conclusion: The resistance of the isolates observed from plasmid patterns with varied molecular weights could aid the transferability of that factor to other related bacteria which could be a threat to therapeutic management of boil infection.Item Bioassay-Guided Isolation of an Antibacterial Flavone and Cytotoxicity of the Leaf Extracts of Vitex rivularis Gurke (Lamiaceae)(Nigerian Association of Pharmacists in Academia (NAPA), 2022) Idowu, P. A.; Samuel, B. B.; Mbachu, C. C.Background: The global challenge of resistance to several antibacterial agents has necessitated the continuous search for new antibacterial molecules. Natural products have proven to be a major source of new therapies for infections. Objectives: This study aimed at investigating the antibacterial and cytotoxicity activities of Vitex rivularis leaf extract and isolate its bioactive principle based on its local application in the treatment of bacterial infections. Materials and Methods: Methanol extract was tested against Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6638, Staphylococcus aureus ATCC 59232, Proteus mirabilis ATCC 15290 and Pseudomonas aeruginosa ATCC 15442. Cytotoxicity activity of the crude extract was done using Brine Shrimp Lethality Assay (BSLA). Crude methanolic extract was subjected to column chromatography on silica gel and that afforded eight fractions (F1 – F8). The pooled fractions were monitored for antimicrobial activity against Escherichia coli and Bacillus subtilis. Minimum inhibitory concentration (MIC) of the crude and the most active fraction were determined by agar-dilution method using Gentamicin as the positive control. Fraction 7 was purified by recrystallisation in ethyl acetate to obtain Compound F7. The isolated bioactive compound was characterized by spectroscopic data (IR, 1H and 13C NMR, COSY, HMQC). Results: The MIC of the extract and isolated compound, F7, were between 0.25-0.50 mg/ml on E. coli and B. subtilis. Cytotoxicity activity showed LC50 of leaf extract to be 168.66μg/ml. The structure of the isolated bioactive compound was confirmed to be 5-hydroxy-3',4',6,7-tetramethoxyflavone Conclusion: The observed antibacterial activity of Vitex rivularis leaf extract justified the traditional use of the plant’s leaf in the treatment of bacterial infections by local users. The isolated compound can serve as a lead compound in the development of new antimicrobials agents.Item Detection of extended spectrum beta-lactamase escherichia coli and histamine contents in raw Mackerel (Scomber japonicus) sold in open markets in Sagamu, Nigeria(Faculty of Basic Medical Sciences, College of Medicine, University of Lagos (UNILAG), 2022) Okunye, O. L.; Idowu, P. A.; Okanlawon, B. M.; Adejumo, O. E.; Lawal, A. S.; Ademola, S. R.Background: A food borne sickness called histamine fish poisoning is frequently brought on by eating some fish species that have high levels of histamine and other biogenic amines in their tissues. When fish is improperly stored and preserved, its natural makeup makes it polluted. Objectives: This study was carried out to examine the histamine contents, determine the extended spectrum beta-lactamase producing isolates of Escherichia coli from the gut of raw mackerel fish obtained in Sagamu markets and relate the plasmid isolated, if present, with Extended Spectrum Beta Lactamase (ESBL). Materials and Methods: Fifty (50) mackerel fish were dissected and their guts were removed for the isolation of E. coli on eosin methylene blue agar medium. The isolates obtained were Gram stained, biochemically characterized, and thereafter plated on Mueller Hinton agar impregnated with ESBL discs by double disc synergy methods. Ten gram (10 g) of each part of fish weighing 100 g was blended for histamine determination by spectrophotometry and plasmid analysis of some selected ESBL resistance amongst the isolates were determined. Results: Escherichia coli were found in all fifty mackerel fish and 31 (62%) of those were ESBL producers. All of these were plasmid-free. In 14 (28%) of the 50 fish analysed, histamine concentrations of more than 100 mg/100 g exceeded the tolerance threshold of 10 mg/100 g, Conclusion: The results from the study showed that some of the fish sold in the markets of Sagamu contain ESBL producing Escherichia coli which may be considered as reservoirs for resistant bacteria. Significant level of histamine recorded surpassed histamine tolerance level in fish for human consumption. There is a need to provide storage facilities and raise hygiene awareness in markets where fish is sold.Item Evaluation of some selected non sterile pharmaceutical products for bacterial and fungi of clinical importance(Faculty of Pharmaceutical Sciences, University of Nigeria, Nsukka, 2022) Okunye, O. L.; Idowu, P. A.; Okanlawon, B. M.; Adejumo, O. E.; Coker, E. M.; Ayedun, J. S.; Osungunna, M. O.; Adeyemo, O. M.; Oyinloye, O. E.; Adeleke, O. E.This study investigated, identified, and quantified microbial contamination of 12 non-sterile pharmaceuticals products frequently made available to Akala Primary Health Care Centre Ibadan, for the possibility of detecting harmful and non-pathogenic microorganisms. Though sterility is not a requirement in official compendia for non-sterile pharmaceuticals, their bioburdens should not exceed the acceptable limit. The representative, syrups, tablets, capsules, and disinfectants from the dispensing unit were selected as guided by the conventional protocol for the study type. Bacteria and fungi of clinical potential were isolated and enumerated using standard microbiology procedures. Ten (10) of the twelve (12) non-sterile pharmaceutical products examined elicited microbial contamination beyond USP acceptable bio-burden standard. The isolates of bacteria identified comprised Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli elicited varied resistance to gentamicin, ciprofloxacin, tetracycline, and erythromycin but susceptible to augmentin, amoxicillin, cloxacillin, and chloramphenicol, while the Candida albicans fungi isolated, were susceptible to ketoconazole and fluconazole at every concentration exposed. The differences in means for CFU/mL and zones of inhibition among the microorganisms isolated were considered, data collected were analyzed using SPSS 15 and the graph was plotted using Graph Pad prism 8.1 Version 5 for Windows. Ten of the twelve (83%) of non-sterile pharmaceuticals products examined were presumably contaminated which could be an indication of improper handling, poor dispensing, poor repackaging, and or non-adherence to Good Manufacturing Practices. Therefore, training and educating the dispensers, as well as patients, on the proper handling and use of medicines to reduce or prevent microbial contamination are hereby advocated.Item A Study of microbial infestation of currency notes in circulation from selected handlers with specific professions in Sagamu community of Ogun State(Centre National de la Recherche Scientifique et Technologique (CNRST), 2022) Okunye, O. L.; Idowu, P. A.; Okanlawon, B. M.; Adejumo, O. E.; Alao, I. O.; Adeyemo, O. M.; Ayedun, J. S.Bacteria are ubiquitous and the ease of survival, adaptation and transmission of some nutritionally non-exacting species amongst the microbes had made it a force to be reckoned with. This study evaluated bacterial contaminants on selected naira denomination, officially recognized currency of transaction in circulations and determined the resistance of the isolates to conventional antibiotic in use. A total of 160 samples of currency notes 20 each of 8 existing denomination were collected from selected subjects of various professions for bacteriological examination and antibiogram determination with a view to identifying possible risk factors associated with these contaminated notes. Equal percentage of polymers to paper currency was collected, the microbial load was found to be higher in lower denominations irrespective of their polymer status. The total bacterial count per milliliter variedbetween 2.28x104 and 4.20x107. The ratios of percentage distributions of isolates;Staphylococcus aureus (36.8%), Escherichia coli (31.5%), Bacillus spp (3.7%) andPseudomonas aeruginosa (27.5%) and varied resistance to antibiotics used were recorded. The alarming resistance of bacterial of clinical status obtained from thisstudy to conventional antibiotics, serves an indication of potential threat of contaminated currency notes to the public health.