Research Article
Volume 18 Issue 6 - 2022
Probiotic Capability of Bacillus spp. Isolated from Iru - Fermented African Locust Bean (Parkia biglobosa)
Afolake Olanbiwoninu*, Esho Deborah, Osinupebi, Ayooluwa, Theresa Awotundun and Samuel Fasiku
Department of Biological Sciences, Faculty of Natural Sciences, Ajayi Crowther University, Oyo, Oyo State, Nigeria
*Corresponding Author: Afolake Olanbiwoninu, Department of Biological Sciences, Faculty of Natural Sciences, Ajayi Crowther University, Oyo, Oyo State, Nigeria.
Received: January 25, 2022; Published: May 31, 2022


Bacillus species have diverse applications in agriculture, enzyme production and medicine to produce vaccines and probiotics. Currently, little is known about the probiotic potential of Bacillus from fermented condiments of African origin. This work aimed at isolating Bacillus spp. with probiotic potential from fermented African locust bean - iru. Iru samples were obtained from a local market in Oyo, Nigeria. The samples were pretreated by boiling to eliminate vegetative cells and inoculated using pour plate technique. Isolates were identified; their antibiotic susceptibility profile and antimicrobial activity against known pathogens (Listeria monocytogenes, Salmonella enterica and Staphylococcus aureus) were determined. Safety and technological properties (cellulase, protease, lipase and amylase activities) of the isolates were also determined. Twenty spore-forming, Gram-positive with rod shape were isolated and identified as Bacillus subtilis (8), Bacillus licheniformis (4), Bacillus pumilus (3), Bacillus polymyxa (1), Bacillus licheniformis (1), Bacillus alvei (1), Bacillus badius (1) and Bacillus sp. (1). All isolates were susceptible to levofloxacin, ciprofloxacin, ofloxacin, gentamycin and azithromycin while 16 (80%) were resistant to cefotaxime. Only 11 (55%) isolates inhibited the growth of Listeria monocytogenes, with no inhibition recorded against other pathogens. The 11 isolates synthesized at least one enzyme, with B. subtilis PA1 and B. alvei PB5 producing the four enzymes determined. γ-haemolysis was exhibited in 90% of the isolates while 10% exhibited α-haemolysis. Bacillus subtilis PA1, Bacillus subtilis PA6 and Bacillus licheniformis PA5 survived high acid, bile and simulated acid and bile. Bacillus subtilis and Bacillus licheniformis from African fermented locust beans have excellent probiotic potentials.


Keywords: Acid and Bile Tolerance; Antimicrobial Activity; Enzymes; Probiotic Potential; Safety Properties


  1. Ajayi OA., et al. “Towards the Development of Shelf Stable ‘iru’ (Parkia biglobosa) Condiment Bouillon Cubes Using Corn, Cassava and Potato Starch Extracts as Binders”. Nigerian Food Journal1 (2015): 67-72.
  2. Oloyede GK and Akintunde TI. “Fatty acids profile, physicochemical properties and antioxidant activity of unfermented and fermented Parkia biglobosa (African locust beans) seed oil”. Journal of Chemical Society of Nigeria 2 (2019): 178-186.
  3. Olanbiwoninu AA., et al. “Riboflavin Enriched iru: A fermented Vegetable Protein”. African Journal of Microbiology Research13 (2017): 546-552.
  4. Omodara TR and Aderibigbe, EY. “Microbiological, Physiochemical and Enzymatic Changes in Fermented African Locust Bean (Parkia biglobosa) Seeds Using Bacillus subtilis and Additives”. Sustainable Food Production 1 (2018): 22-29.
  5. Su Y., et al. “Bacillus subtilis: a universal cell factory for industry, agriculture, biomaterials and medicine”. Microbial Cell Factory 19 (2020): 173.
  6. Elshaghabee FMF., et al. “Bacillus as Potential Probiotics: Status, Concerns, and Future Perspectives”. Frontiers in Microbiology 8 (2017): 1490.
  7. FAO/WHO. “Guidelines for the Evaluation of Probiotics in Food”. Food and Agriculture Organization of the United Nations and World Health Organization Working Group Report. (Rome: Food and Agriculture Organization) (2002).
  8. Hong HA., et al. “The Use of Bacterial Spore Formers as Probiotics”. FEMS Microbiology Reviews 4 (2005): 813-835.
  9. Ripert G., et al. “Secreted Compounds of the Probiotic Bacillus clausii Strain O/C Inhibit the Cytotoxic Effects Induced by Clostridium difficile and Bacillus cereus Toxins”. Antimicrobial Agents and Chemotherapy 6 (2016): 3445-3454.
  10. Aspri M., et al. “Review on Non-Dairy Probiotics and Their Use in Non-Dairy Based Products”. Fermentation1 (2020): 30.
  11. CLSI Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing. (28th Edition). CLSI supplement M100. Wayne, PA (2018).
  12. Ozabor TP., et al. “Antibacterial and Antioxidant Activity of Bacillus Species Isolated from Fermented Parkia biglobosa (IRU) and Ricinuscommunis (OGIRI)- African Traditionally Fermented Food Condiments”. The Asia Journal of Applied Microbiology1 (2020): 19-29.
  13. Mangia NP., et al. “Functional and Safety Characterization of Autochthonous Lactobacillus paracasei FS103 Isolated from Sheep Cheese and its Survival in Sheep and Cow Fermented Milks During Cold Storage”. Annals of Microbiology 69 (2019): 161-170.
  14. Chaleb K., et al. “In Vitro Effect Of pH and Ethanol on Biofilm Formation by Clinical Positive Staphylococcus epidermidis Strains”. Annals of Microbiology 57 (2007): 431-437.
  15. Klingberg TD., et al. “Identification of potential probiotic starter cultures for Scandinavian-type fermented sausage”. International Journal of Food Microbiology3 (2005): 419-431.
  16. Ahmed AI., et al. “Molecular identification and in vitro evaluation of probiotic functional properties of some Egyptian lactic acid bacteria and yeasts”. Journal of Genetic Engineering and Biotechnology1 (2021): 114.
  17. Farjana I and Narayan R. “Screening, purification and characterization of cellulose from cellulase producing bacteria in molasses”. BMC Research Notes445 (2018): 36.
  18. Folashade MO and Joshua OA. “Production dynamics of extracellular protease from Bacillus species”. African Journal of Biotechnology 8 (2005): 776-779.
  19. Okorie PC and Olasupo NA. “Growth and extracellular enzyme production by microorganisms isolated from Ugba - an indigenous Nigerian fermented condiment”. African Journal of Biotechnology26 (2013): 4158-4167.
  20. Veeresh H., et al. “Isolation and characterization of lipolytic microorganisms from oil-contaminated soil”. International Journal of Advances in Engineering, Sciences and Technology (IJAEST)3 (2016): 293-297.
  21. Kalpana S., et al. “Isolation of lipolytic bacteria of waste contaminated soil: A study with regard to process optimization for lipase”. International Journal of Scientific and Technology Research 10 (2013): 214-218.
  22. Peterson E and Kaur P. “Antibiotic Resistance Mechanisms in Bacteria: Relationships Between Resistance Determinants of Antibiotic Producers, Environmental Bacteria, and Clinical Pathogens”. Frontiers in Microbiology 9 (2018): 2928.
  23. Sanz Y., et al. “Probiotics as Drugs Against Human Gastrointestinal Infections”. Recent Patents on Anti-Infective Drug Discovery2 (2007): 148-156.
  24. Iqra Y., et al. “In Vitro Probiotic Potential and Safety Evaluation (Hemolytic, Cytotoxic Activity) of Bifidobacterium strains Isolated from Raw Camel Milk”. Microorganisms 3 (2020) 354.
  25. FAO/WHO. Probiotics in Food Health and Nutritional Properties and Guidelines for Evaluation (2006).
  26. Jeon H., et al. “Probiotic Characterization of Bacillus subtilis P223 Isolated from Kimchi”. Food Science and Biotechnology 6 (2017): 1641-1648.
  27. Galie S., et al. “Biofilms in the Food Industry: Health Aspects and Control Methods”. Frontiers in Microbiology 9 (2018): 898.
  28. Lim H., et al. “Isolation and characterization of probiotic Bacillus subtilis MKHJ 1-1 possessing L-asparaginase activity”. Applied Sciences 10 (2021): 4466.
  29. Donatien K., et al. “Acid Resistance, Bile Tolerance and Antimicrobial Properties of Dominant Lactic Acid Bacteria Isolated from Traditional ‘Maari’ Baobab Seeds Fermented Condiment”. African Journal of Biotechnology5 (2012): 1197-1206.
  30. Markowiak P and Slizewska K. “Effects of Probiotics, Prebiotics, and Synbiotics on Human Health”. Nutrient9 (2017): 1021.
  31. Olanbiwoninu AA and Fasiku SA. “Production of bacterial amylases and cellulases using sweet potato (Ipomoea batatas. (L.) Lam.) peels”. African Journal of Biochemistry Research9 (2015): 104-109.
  32. Fasiku SA., et al. “Isolation of Bacteria with Potential of Producing Extracellular Enzymes (Amylase, Cellulase and Protease) from Soil Samples”. Journal of Advances in Microbiology3 (2020): 21-26.
Citation: Afolake Olanbiwoninu., et al. “Probiotic Capability of Bacillus spp. Isolated from Iru - Fermented African Locust Bean (Parkia biglobosa)”. EC Microbiology 18.6 (2021): 18-30.

PubMed Indexed Article

EC Pharmacology and Toxicology
LC-UV-MS and MS/MS Characterize Glutathione Reactivity with Different Isomers (2,2' and 2,4' vs. 4,4') of Methylene Diphenyl-Diisocyanate.

PMID: 31143884 [PubMed]

PMCID: PMC6536005

EC Pharmacology and Toxicology
Alzheimer's Pathogenesis, Metal-Mediated Redox Stress, and Potential Nanotheranostics.

PMID: 31565701 [PubMed]

PMCID: PMC6764777

EC Neurology
Differences in Rate of Cognitive Decline and Caregiver Burden between Alzheimer's Disease and Vascular Dementia: a Retrospective Study.

PMID: 27747317 [PubMed]

PMCID: PMC5065347

EC Pharmacology and Toxicology
Will Blockchain Technology Transform Healthcare and Biomedical Sciences?

PMID: 31460519 [PubMed]

PMCID: PMC6711478

EC Pharmacology and Toxicology
Is it a Prime Time for AI-powered Virtual Drug Screening?

PMID: 30215059 [PubMed]

PMCID: PMC6133253

EC Psychology and Psychiatry
Analysis of Evidence for the Combination of Pro-dopamine Regulator (KB220PAM) and Naltrexone to Prevent Opioid Use Disorder Relapse.

PMID: 30417173 [PubMed]

PMCID: PMC6226033

EC Anaesthesia
Arrest Under Anesthesia - What was the Culprit? A Case Report.

PMID: 30264037 [PubMed]

PMCID: PMC6155992

EC Orthopaedics
Distraction Implantation. A New Technique in Total Joint Arthroplasty and Direct Skeletal Attachment.

PMID: 30198026 [PubMed]

PMCID: PMC6124505

EC Pulmonology and Respiratory Medicine
Prevalence and factors associated with self-reported chronic obstructive pulmonary disease among adults aged 40-79: the National Health and Nutrition Examination Survey (NHANES) 2007-2012.

PMID: 30294723 [PubMed]

PMCID: PMC6169793

EC Dental Science
Important Dental Fiber-Reinforced Composite Molding Compound Breakthroughs

PMID: 29285526 [PubMed]

PMCID: PMC5743211

EC Microbiology
Prevalence of Intestinal Parasites Among HIV Infected and HIV Uninfected Patients Treated at the 1o De Maio Health Centre in Maputo, Mozambique

PMID: 29911204 [PubMed]

PMCID: PMC5999047

EC Microbiology
Macrophages and the Viral Dissemination Super Highway

PMID: 26949751 [PubMed]

PMCID: PMC4774560

EC Microbiology
The Microbiome, Antibiotics, and Health of the Pediatric Population.

PMID: 27390782 [PubMed]

PMCID: PMC4933318

EC Microbiology
Reactive Oxygen Species in HIV Infection

PMID: 28580453 [PubMed]

PMCID: PMC5450819

EC Microbiology
A Review of the CD4 T Cell Contribution to Lung Infection, Inflammation and Repair with a Focus on Wheeze and Asthma in the Pediatric Population

PMID: 26280024 [PubMed]

PMCID: PMC4533840

EC Neurology
Identifying Key Symptoms Differentiating Myalgic Encephalomyelitis and Chronic Fatigue Syndrome from Multiple Sclerosis

PMID: 28066845 [PubMed]

PMCID: PMC5214344

EC Pharmacology and Toxicology
Paradigm Shift is the Normal State of Pharmacology

PMID: 28936490 [PubMed]

PMCID: PMC5604476

EC Neurology
Examining those Meeting IOM Criteria Versus IOM Plus Fibromyalgia

PMID: 28713879 [PubMed]

PMCID: PMC5510658

EC Neurology
Unilateral Frontosphenoid Craniosynostosis: Case Report and a Review of the Literature

PMID: 28133641 [PubMed]

PMCID: PMC5267489

EC Ophthalmology
OCT-Angiography for Non-Invasive Monitoring of Neuronal and Vascular Structure in Mouse Retina: Implication for Characterization of Retinal Neurovascular Coupling

PMID: 29333536 [PubMed]

PMCID: PMC5766278

EC Neurology
Longer Duration of Downslope Treadmill Walking Induces Depression of H-Reflexes Measured during Standing and Walking.

PMID: 31032493 [PubMed]

PMCID: PMC6483108

EC Microbiology
Onchocerciasis in Mozambique: An Unknown Condition for Health Professionals.

PMID: 30957099 [PubMed]

PMCID: PMC6448571

EC Nutrition
Food Insecurity among Households with and without Podoconiosis in East and West Gojjam, Ethiopia.

PMID: 30101228 [PubMed]

PMCID: PMC6086333

EC Ophthalmology
REVIEW. +2 to +3 D. Reading Glasses to Prevent Myopia.

PMID: 31080964 [PubMed]

PMCID: PMC6508883

EC Gynaecology
Biomechanical Mapping of the Female Pelvic Floor: Uterine Prolapse Versus Normal Conditions.

PMID: 31093608 [PubMed]

PMCID: PMC6513001

EC Dental Science
Fiber-Reinforced Composites: A Breakthrough in Practical Clinical Applications with Advanced Wear Resistance for Dental Materials.

PMID: 31552397 [PubMed]

PMCID: PMC6758937

EC Microbiology
Neurocysticercosis in Child Bearing Women: An Overlooked Condition in Mozambique and a Potentially Missed Diagnosis in Women Presenting with Eclampsia.

PMID: 31681909 [PubMed]

PMCID: PMC6824723

EC Microbiology
Molecular Detection of Leptospira spp. in Rodents Trapped in the Mozambique Island City, Nampula Province, Mozambique.

PMID: 31681910 [PubMed]

PMCID: PMC6824726

EC Neurology
Endoplasmic Reticulum-Mitochondrial Cross-Talk in Neurodegenerative and Eye Diseases.

PMID: 31528859 [PubMed]

PMCID: PMC6746603

EC Psychology and Psychiatry
Can Chronic Consumption of Caffeine by Increasing D2/D3 Receptors Offer Benefit to Carriers of the DRD2 A1 Allele in Cocaine Abuse?

PMID: 31276119 [PubMed]

PMCID: PMC6604646

EC Anaesthesia
Real Time Locating Systems and sustainability of Perioperative Efficiency of Anesthesiologists.

PMID: 31406965 [PubMed]

PMCID: PMC6690616

EC Pharmacology and Toxicology
A Pilot STEM Curriculum Designed to Teach High School Students Concepts in Biochemical Engineering and Pharmacology.

PMID: 31517314 [PubMed]

PMCID: PMC6741290

EC Pharmacology and Toxicology
Toxic Mechanisms Underlying Motor Activity Changes Induced by a Mixture of Lead, Arsenic and Manganese.

PMID: 31633124 [PubMed]

PMCID: PMC6800226

EC Neurology
Research Volunteers' Attitudes Toward Chronic Fatigue Syndrome and Myalgic Encephalomyelitis.

PMID: 29662969 [PubMed]

PMCID: PMC5898812

EC Pharmacology and Toxicology
Hyperbaric Oxygen Therapy for Alzheimer's Disease.

PMID: 30215058 [PubMed]

PMCID: PMC6133268

News and Events

December Issue Release

We always feel pleasure to share our updates with you all. Here, notifying you that we have successfully released the November issue of respective journals and the latest articles can be viewed on the current issue pages.

Submission Deadline for Upcoming Issue

ECronicon delightfully welcomes all the authors around the globe for effective collaboration with an article submission for the upcoming issue of respective journals. Submissions are accepted on/before December 20, 2022.

Certificate of Publication

ECronicon honors with a "Publication Certificate" to the corresponding author by including the names of co-authors as a token of appreciation for publishing the work with our respective journals.

Best Article of the Issue

Editors of respective journals will always be very much interested in electing one Best Article after each issue release. The authors of the selected article will be honored with a "Best Article of the Issue" certificate.

Certifying for Review

ECronicon certifies the Editors for their first review done towards the assigned article of the respective journals.

Latest Articles

The latest articles will be updated immediately on the articles in press page of the respective journals.