Open Access Original Research Article

Characterization and Screening of Lactic Acid Bacteria (LAB) and Yeast from Fermented Sorghum and Maize Products for use as Starter Culture

Oluwatosin Charles Ayodeji, Afolabi Folake Titilayo, Abdulkadir Musliu

Journal of Advances in Microbiology, Page 1-11
DOI: 10.9734/jamb/2021/v21i930379

This study focused on the isolation, Characterization and screening of lactic acid bacteria and yeast for use as starter culture in order to eradicate the problem of excessive microbial contamination and the presence of mycotoxins.

Lactobacillus plantarum and Saccharomyces cerevisiae were the predominant microorganisms isolated from the samples collected. The isolates were screened for their ability to produce enzymes and metabolites. Lactic acid bacteria produce a variety of antimicrobial compounds such as lactic acid, hydrogen peroxide, diacetyl on them as a natural competitive means to overcome other microorganism sharing the same niche. They were also screened for their ability to withstand some physiological stress like acid tolerance, temperature, salt concentration and antibacterial activity; the isolates produced significant values of enzymes and, antimicrobial metabolites. The antimicrobial activity of Lactobacillus plantarum is mainly attributed to the low pH due to Lactic acid production. The antimicrobial activity of the Saccharomyces cerevisiae appears to be mainly due to the competition with the other microorganisms. In addition, depletion of Oxygen and production of CO2, competition for nutrients and the production of antimicrobial substances could have been responsible for the overall antimicrobial activity of both cultures. According to the results obtained, both lactic acid bacteria and yeast proved to be a good source of starter culture.

Open Access Original Research Article

Characterization and Plant Growth Promoting Properties of a Bacillus sp. Isolated from Maize Roots

Lynda Kelvin-Asogwa, Frank C. Ogbo

Journal of Advances in Microbiology, Page 12-24
DOI: 10.9734/jamb/2021/v21i930380

Aim: To isolate plant growth promoting Bacillus strains from maize roots and to characterize using molecular methods, the strain with greatest potential for plant growth promotion.

Place and Duration of Study: Department of Applied Microbiology and Brewing, Nnamdi Azikiwe University, Awka, between February 2019 and March 2020.

Methodology: The isolation of plant growth promoting rhizobacteria (PGPR) from maize roots was made using Nitrogen Free Bromothymol Blue (NFB) broth. They were screened for Phosphate solubilizing activities on Pikovskaya (PVK) agar. Quantitative determination and solubilization of different types of Phosphates was carried out using Pikovskaya broth. Optimization of factors affecting phosphate such as NaCl concentration, initial pH of the medium, size of inoculum, was done using pvk broth. Evaluation of other plant growth promoting properties were carried out such as IAA, Ammonia, cellulase and HCN production. 

Results: Eleven Nitrogen fixing bacteria were isolated using NFB broth based on colour change of the medium from green to blue.  Test for phosphate solubilization abilities of the organisms revealed that nine of the isolates solubilized phosphate on PVK agar. Organism coded with IS52 gave the least solubilization index of 1.14 while isolate IS19 gave the highest index of 3.4. Isolate IS19 yielded the highest amount of 73.5µg/ml P, while isolate IS30 was the weakest solubilizer in PVK broth, yielding 19.4µg/ml P. The best isolate IS19, produced the plant growth hormone Indole Acetic acid at a concentration of 105.4μg/ml. The organism also gave a positive result for ammonia and cellulase production but did not produce Hydrogen cyanide. It was identified as Bacillus subtilis using the 16S rRNA gene sequencing.

Conclusion: Bacillus subtilis fixed Nitrogen qualitatively and solubilized insoluble phosphates in addition to other plant growth promoting properties, thus Bacillus subtilis has potential for plant growth promotion, making it an efficient strain for biofertilizer production

Open Access Original Research Article

Appraisal on Microbiologal Qualities of Hospital Wastewaters from off a Local Government Area of Kwara State, Nigeria

Usman Kolawole Muftau, Arotupin Daniel Juwon, Ekundayo, Fatuyi Olanipekun

Journal of Advances in Microbiology, Page 25-38
DOI: 10.9734/jamb/2021/v21i930381

Aims: This study investigates the microorganisms associated with hospital wastewaters collected from the Offa Local Government Area of Kwara State, Nigeria, during the wet and dry seasons.

Study Design: This project was a cross sectional descriptive study in which subjects were hospital wastewater samples collected from the study site.

Place and Duration of Study: the samples were analyzed in the department of microbiology, Federal University of Technology, Akure, Ondo State, Nigeria.

Methodology: Microbiological study of one hundred and twenty-six samples of hospital wastewater collected both during wet season and dry season periods in the years 2018 – 2019 from Offa Local Government Area of Kwara State, Nigeria, was carried out using conventional and molecular techniques respectively and the Global Positioning System (GPS) of each collection site was accurately recorded.

Results: The microbial load of wet season samples collected from Offa Local Government Area ranged between 7±4,00 and 150±43.59 while that of dry season samples ranged between 10±2.00 x 105 and 225±67.27 (x 105cfu/ml). The bacteria isolated from wet season samples included; Alcaligenes faecalis, A. aquatilis and Staphylococcus saprophyticus with percentage occurrences of 65.4, 19.2 and 15.4 respectively, while bacteria isolated from dry season sample were A. faecalis and S. saprophyticus with percentages occurrence of 79.17 and 20.89, respectively.

Conclusion: The findings from this study showed that hospital wastewater collected during dry season period had more bacterial load than that of wet season period. The findings also confirmed A. faecalis as the most predominant and prevailing bacteria inhabiting hospital wastewater. Thus, care must be taken by avoiding hospital wastewater from getting into the municipal water supply to prevent infections associated with A. faecalis, A. aquatilis and S. saprophyticus.

Open Access Original Research Article

Phytochemical Analysis and In vivo Evaluation of Individual Activity of Ethanolic Leaf Extracts of Azadirachta indica, Senna occidentalis and Momordica balsamina against Plasmodium berghei Infected Mice

Adam Musa Bature, Karderam Bukar Dikwa, Abdullah Isyaku Alhaji, Deboral Madi Dibal

Journal of Advances in Microbiology, Page 39-49
DOI: 10.9734/jamb/2021/v21i930382

Malaria is a major cause of morbidity and mortality in tropics and subtropics region with Nigeria accounting for the highest proportions in Africa. This is accompanied with emerging resistance to available drugs, posing it a public health concern. This study is aimed at determining the invivo activity of the individual ethanolic leaf extracts of the Azadirachta indica, Senna occidentalis and Momordica balsamina. The leaves of A. indica, S. occidentalis and M. balsamina were subjected to preliminary phytochemical screening. Ethanolic extraction of leaves of plants was carried out and invivo evaluation of the individual activity of extracts determined using standard procedures. 55 mice were randomly divided into 11 groups lettered A – K; positive group, negative group and 9 extract groups. Results showed that M. balsamina had the highest yield of 7.6%, followed by A. indica with 6.5% and S. occidentalis with 5.7%. The preliminary phytochemical screening revealed the presences of alkaloids, flavonoids, saponins, steroids, phenolics and tannin in all plants. The comparison of the individual study groups showed that Senna occidentalis is more effective at 600mg/kg dosage and prolonged survival of the mice in its group in the study period. This plant possessed significant (P-value <0.05) antiplasmodial activity, thus lowered parasitaemia in infected mice.

Open Access Original Research Article

Prevalence of Methicillin Resistant Staphylococcus aureus Bacteriuria among Pregnant Women Attending Secondary Health Hospitals in Ilorin, Nigeria

Muritala I. Bale, Shola K. Babatunde, S. Awe

Journal of Advances in Microbiology, Page 50-57
DOI: 10.9734/jamb/2021/v21i930383

Background: Urinary tract infection is one of the most frequently acquired infections in both community and hospitals and is common among the adolescents and the old genders.

Aim: To determine the prevalence of methicillin resistant Staphylococcus aureus bacteriuria among pregnant women attending secondary health hospitals in Ilorin, Nigeria

Study Design: An experimental study which involve a random selection of consented pregnant women.

Place and Duration of Study: Department of Biosciences and Biotechnology Kwara State University Malete between January 2018 and June 2019.

Methodology: In this study, a total of 856 pregnant women mid stream clean catch early morning voided urine samples for two consecutive days (383 of the samples were collected from Sobi Specialist Hospital, 278 from Adewole Cottage Hospital and 195 collected from Ajikobi Cottage Hospital) were randomly screened for the presence of Staphylococcus aureus bacteriuria using standard microbiological procedures such as growth on mannitol salt agar, Gram reaction, catalase and coagulase tests. The Kirby–Bauer disk diffusion method was used to determine the antibiotic sensitivity profile of S. aureus isolated using oxoid antibiotic discs.

Results: Out of 856 samples screened 56 samples (6.5%) showed significant Staphylococcus aureus bacteriuria, 16- 25 years  has a prevalence rate of 5.6%, 26- 35 years (5.8%) while 35-45 years have the  prevalence rate of 10.9%. A total of 7 (12.5%) methicillin resistant Staphylococcus aureus were isolated. Antibiotic sensitivity profile shows that 26.8% were resistance to gentamicin, 44.6% to tetracycline, 19.6% to chloramphenicol, 33.9 % to erythromycin, 67.9% to amoxicillin, 32% to augmentin, 12.6% to ceftriaxone, 5.3% to ciprofloxacin and 100% susceptibility to both nitrofurantoin and vancomycin.

Conclusion: The study shows the high prevalence of MRSA and high susceptibility of nitrofurantoin and vancomycin to all the MRSA isolated.

Open Access Original Research Article

Bioethanol Production from Cassava Peels Inoculated with Saccharomyces cerevisiae and Zymomonas mobilis

Kehinde Tope Adegbehingbe, Foluso Faparusi, Bartholomew Saanu Adeleke

Journal of Advances in Microbiology, Page 58-67
DOI: 10.9734/jamb/2021/v21i930384

Aim: This study was designed to determine the bioethanol production from fermented cassava peel using Saccharomyces cerevisiae and Zymomonas mobilis.

Methodology: Cassava peels were collected from cassava processing sites, washed, surface sterilized, dried, milled into flour, pretreated, and fermented. Saccharomyces cerevisiae and Z. mobilis suspensions were aseptically inoculated into the fermenting medium and allowed to stand for seven days. The pH, total reducing sugar, chemical composition, and bioethanol composition of the fermenting substrates were determined.

Results: A pH decrease from 5.2 to 4.1 was recorded in the sample fermented with S. cerevisiae while the least pH value of 3.8 was obtained from the sample fermented with Z. mobilis, respectively. The total reducing sugar (glucose) of fermented samples decreased from 3.4% to 1.5% (Z. mobilis) and 3.4% to 1.88% (S. cerevisiae) compared with the control sample. The chemical composition showed high protein and fat contents in the fermented samples. High percentage yield of 30% with ethanol volume of 45 mL was recovered from cassava peel inoculated with S. cerevisiae while flash point, i.e. the lowest temperature at which fuel produces enough vapor to cause ignition leading to flame generation of 24oC, was recorded for both fermented samples inoculated with S. cerevisiae and Z. mobilis.

Conclusion: The ability of the bacterium and the yeast isolates exhibiting high potential for bioethanol production could be promising in various industrial processes as an alternative to fossil transportation fuel.

Open Access Original Research Article

Bioremediation of Soil Contaminated with Spent Engine Oil using Pig Dung

A. A. Ibiene, O. Aleruchi, L.C. Nnodim, R. U. Ihunwo

Journal of Advances in Microbiology, Page 68-78
DOI: 10.9734/jamb/2021/v21i930385

Studies were carried out to investigate the bioremediation potential of pig dung in a soil contaminated with spent engine oil. Soil samples were obtained from the Ofrima complex, University of Port Harcourt. The soil samples were contaminated with various concentrations (50 ml and 100 ml) of spent engine oil and allowed for 21 days for proper exposure, mimicking natural spill. This was followed by the addition of the pig dung. The experimental setup was labeled sample A (1 kg soil + 100 g pig dung + 50 ml spent engine oil) and sample B (1 kg soil + 100 g pig dung + 100 ml spent engine oil). The physicochemical parameters and the microbiological analysis were done using standard methods. The total petroleum hydrocarbon was analyzed using gas chromatographic methods. Analyses were carried out at 14 days intervals for 28 days. The physicochemical parameter results showed a reduction in pH values in the contaminated soil samples, ranging from 6.21 - 6.65 in sample A and 6.57 - 6.87 in sample B. Temperature values were constant at 230C from day 1 to day 14 in sample A and increased at day 28 to 24 0C, also for sample B, the temperature was constant at 230C from day 1 to day14 and increased at day 28 to 26 0C. The amount of heavy metal (Lead) content decreased from 4.3645 - 1.93676 (mg/kg) and 6.18361 - 3.89654 (mg/kg) for samples A and B, respectively. There was also a significant reduction in the amount of Total Petroleum Hydrocarbon, from 16631.86 - 3280.83 mg/kg for sample A and 18464.73 - 6784.60 mg/kg for sample B. The THB counts for samples A and B ranged from 7.73 - 7.91 and 7.05-8.20 (Log cfu/g), respectively. The fungal counts ranged from 3.99–4.58 and 5.12 - 7.93 (Log cfu/g) for samples A and B respectively. HUB counts ranged from 4.52–5.09 and 4.93- 5.55 (Log cfu/g) for samples A and B, respectively. The HUF counts ranged from 4.12 - 5.49 and 4.13 - 4.70 (Log cfu/g) for samples A and B, respectively. The results clearly showed that microorganisms capable of utilizing total petroleum hydrocarbon were present, also the pig dung showed both bio-stimulation and bio-augmentation tendency to attract high microbial load which supported the bioremediation of the spent engine oil contaminated soil.

Open Access Original Research Article

Public Health Implications of Coliform Contaminants in Non-packaged, Commercially Hawked Herbal Remedies Sold in Port Harcourt

C. L. C. Ndukwu, N. P. Akani, S. A. Wemedo, T. Sampson

Journal of Advances in Microbiology, Page 79-87
DOI: 10.9734/jamb/2021/v21i930386

The long history of use of coliforms as indicators of microbiological quality of water and food materials constitutes a veritable rationale for the ascertainment of the microbiological status of locally brewed, liquid herbal remedies, commercially hawked and widely consumed within Port Harcourt metropolis as envisaged in this study. Seventy two samples of twelve different non-packaged, oral herbal remedies commonly known as “Agbo” – the Yoruba word for medicine - were purchased from six different localities within Port Harcourt City and Obio/Akpor local governments of Rivers state. They were analyzed within two hours for total heterotrophic bacterial counts and total coliform counts. The bacterial isolates were characterized and identified by Gram staining and Biochemical tests using standard methods. The total coliform count is 4.53±1.11 Log10cfu/ml ranging from 3.69±0.85 for Sexually Transmitted Diseases (STD) remedies to 5.46±1.15 Log10cfu/ml for the cure-all (Gbogbonise) remedies. The mean total coliform counts (TCC) for the remedies are 4.53±1.00 Log10CFU/ml. Gbogbonise had the highest TCC of 5.46±1.15 Log10 cfu/ml, while STD remedies had the least TCC of 3.69±0.85 Log10cfu/ml. Out of a total of one hundred and fifty six isolates there were seventy three coliforms constituting 46.8% of all      isolates. They include Enterobacter aerogenes (19; 12.2%), Enterobacter cloacae (18; 11.5%), Klebsiella pneumoniae (10; 6.4%) Citrobacter rodentium  (7;4.5) Enterobacter pyrinus:  (5;3.2%) Enterobacter hormaechei (3; 1.9%)  Klebsiella oxytoca (3; 1.9%) Serratia marcescens  (3;1.9%) Serratia rubidaea  (3; 1.9%) Hafnia alvei (1; 0.6%) and Pantoea dispersa (1; 0.6%)  The high prevalence of coliform contamination is an indication of poor microbial quality, implying that consumption of the products may pose potential health hazards. Education and enlightenment of handlers on the principles of basic hygiene and good manufacturing principles, is advised to ensure that products have tolerable levels of bacterial contaminants.