Showing papers in "Microbiology Research Journal International in 2021"

Microbiology and Heavy Metal Content of Wetlands Impacted by Crude Oil Pollution in Rivers State, Southern Nigeria

Abstract: Wetland soils constitute vast, under-exploited and sometimes undiscovered ecologies in many countries of the World, including Nigeria. A total of 54 wetland soil samples including surface and subsurface soil at depths of 0-15 cm and 15-30 cm were collected using a sterile hand auger for a period of three months between August and October and subjected to standard and analytical microbiological procedures. The wetland soil samples were further subjected to atomic absorption spectroscopy (AAS) to check for presence and concentration of heavy metals. Results obtained showed that apart from heterotrophic bacterial and fungal counts, hydrocarbon utilizing bacteria (HUB) counts were higher in the surface soil ranging from 12.06±3.43X10 cfu/g at Iwofe to 6.19±2.67 a X10 7 cfu/g at Chokocho while subsurface soil had HUB ranging from 8.91±6.67 a X10 3 cfu/g at Eagle Island to 4.93±3.95X10cfu/g at Chokocho. Heavy metals such as Fe, Pb, Cd and Ni were recorded in concentrations above FEPA permissible limit in the surface and subsurface soil across the three wetlands. The heavy metal concentration in each wetland however, decreased with an increase in soil depth. According to literatures, elevated levels of heavy metals in soils decrease Original Research Article Chibuike et al.; MRJI, 31(2): 53-63, 2021; Article no.MRJI.66612 54 microbial population, diversity and activities. However, the microbial population in this study increased with increasing heavy metal concentration which indicates that the microbes can tolerate or utilize heavy metals in their systems; as such can be used for bioremediation of heavy metal polluted soils.

Evaluation of Antimicrobial activity and Genotoxic Potential of Capparis spinosa (L.) Plant Extracts

Abstract: Objective: The aims of this study were to evaluate the antimicrobial activity and the genotoxic effect of both ethanolic and aqueous extracts of stem and leaf of Capparis spinosa (C. spinosa) plant on Escherichia coli (E. coli) ATCC 25922, Staphylococcus aureus (S. aureus) ATCC 6538P, clinical isolate of Methicillin-resistant S. aureus (MRSA) and Klebsiella pneumoniae (K. pneumoniae) and Candida albicans (C. albicans) ATCC 90028. Materials and Methods: The antimicrobial activity was determined using microbroth dilution method, while the genotoxic effect was investigated using randomly amplified polymorphic DNA (RAPD)-PCR and enterobacterial repetitive intergenic consensus (ERIC)-PCR. Results: The MIC values of both ethanolic and aqueous leaf and stem extracts of C. spinosa plant had a range 6.25 mg/ml to 100 mg/ml. In addition, it was found that ethanolic extract more effective than aqueous extract. The genotoxic activity of aqueous leaf extract, showed changes in both Random Amplified Polymorphic DNA (RAPD)-PCR and Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR profiles of E. coli strain treated with extract compared to untreated (negative) control. These changes included an alteration in the intensity, absence or appearance of new amplified fragments. Original Research Article Adwan and Omar; MRJI, 31(1): 48-57, 2021; Article no.MRJI.67085 49 Conclusions: Results of this study strongly show the genotoxic effect of aqueous leaf extract from C. spinosa plant on E. coli. The findings draw awareness to the possible toxic effect use of C. spinosa plant in traditional medicine and point out the capability of using C. spinosa to treat bacterial or fungal infections. More studies are needed to detect the exact ingredients of this plant as well as the mechanisms responsible for genotoxicity. Further in vivo genotoxicity studies are recommended to ensure and to evaluate the safety of using plants for therapeutic purposes. In addition, results of this study showed that molecular fingerprinting based on ERIC-PCR can be used to evaluate the genotoxic effect in the model bacterial species E. coli.

Influence of Cropping Practices on the Persistence and Vertical Migration of Escherichia coli from Wastewater in Hydromorph Soil in Wet Tropical Zone

Abstract: Little attention is paid to the influence of cultivation practices on the persistence and vertical migration of undesirable bacteria in hydromorphic soils as they have increased the risk of crop recontamination. Therefore the objective of this study was to determine the implication of some cultural practices on the persistence and vertical migration of E. coli in the soil. In this study, raw Original Research Article Ntangmo Tsafack et al.; MRJI, 31(2): 86-97, 2021; Article no.MRJI.66676 87 sewage (single application) and stream water (multiple application) were applied on lettuce, carrot and aubergine plots. The results revealed that overall, E. coli persisted longer on plots with crops and were more persisted in the rainy season on all cultivated plots that had received wastewater from the sewage treatment plant until harvest. While in the dry season, it was only detected at harvest on lettuce plots. The E. coli rate increased gradually overtime on the plots that had received water from the watercourse. Aubergine was the only plant that significantly facilitates the vertical migration of E. coli to the water table. On the whole, crops favored the persistence of E. coli on the soil surface and therefore increase the health risk related to the use of wastewater in agriculture.

Entamoeba gingivalis and Trichomonas tenax in Periodontal Disease

Abstract: This review is aimed at elucidating the role of Entamoeba gingivalis and Trichomonas tenax in the aetiology of periodontal disease. Periodontal disease results from localised inflammation of the periodontium due to plaque accumulation and if left untreated can lead to loss of teeth. Although dental plaque is composed mainly of bacteria, Entamoeba gingivalis and Trichomonas tenax, both of which are protozoan parasites have been found in plaque and implicated in periodontal disease. E. gingivalis is an amoeba associated with poor oral hygiene while T. tenax is a pyriform flagellate that lives in the tartar around the teeth, cavities of carious teeth, necrotic mucosal cells in the gingival margins of gums and pus pockets in tonsillar follicles. These parasites are transmitted by close contact, saliva, droplet spray and kissing or use of contaminated dishes, cups, spoons and Review Article Oladokun et al.; MRJI, 31(3): 61-72, 2021; Article no.MRJI.68707 62 forks as well as drinking water. Age, gender, socio-economic status, dental condition and gingival pathology have been reported to influence the presence of the parasite. Genetic variability and stress are also some of the factors that determine the transition of the periodontium at some gingival sites from healthy to inflame. Researchers have observed that the prevalence and severity of periodontitis is higher in developing countries than developed countries. But with good oral hygiene, regular scaling and polishing and use of antiparasitic drugs, periodontal disease caused by these parasites can be prevented and periodontal health restored.

Strain-level Identification of Beneficial Lactobacilli of Dairy Origin using 16S rRNA Sequencing: A Biotechnology Approach

Abstract: Aim: This study investigated the cultural method and 16S rRNA gene analysis to reveal the composition and diversity of lactic acid bacteria (LAB) from dairy origin (spontaneous fermented cow milk) in Nigeria. Methods: Six dairy samples which includes two raw cow milk, two raw goat milk and two fermented cow milk (nono) were collected and subjected to standard microbiological investigation using both cultural and molecular methods. The dairy samples were cultured on MRS media, and the isolates were identified using physiological and biochemical parameters. DNA of four selected probiotic LAB isolates from nono were amplified using PCR while the amplicons were electrophoresed in agarose gel, pre-stained with ethidium bromide and characterized by 16S rRNA gene analysis. The result of the DNA sequencing were analyzed using NCBI BLAST. Results: A total of 55 presumptive LAB were isolated. Twenty nine (29) Lactobacillus plantarum representing 52.7%, Pediococcus acidilactici 15(27.2%), Lactobacillus casei and Lactobacillus Original Research Article Aforijiku et al.; MRJI, 31(2): 13-21, 2021; Article no.MRJI.58975 14 brevis 4(7.3%) while Lactobacillus fermentum 3(5.5%) were recorded respectively. Result of the gel electrophoresis revealed DNA size of approximately 1500bp. The selected probiotic LAB from nono used in this study were confirmed as Lactobacillus plantarum N17, Lactobacillus plantarum N24, Lactobacillus brevis N10 and Lactobacillus casei N1 based on 16S rRNA gene sequence analysis while the phylogenetic analysis revealed a 98-100% similarity with a high homology level which affirms the strain of the organism. Conclusion: This study has demonstrated the diversity of LAB existing in dairy samples known as fermented cow milk (nono) which could be harnessed as valuable sources for LAB isolation and potential probiotic organisms.

Assessment of Physicochemical and Bacteriological Quality of Well Water Samples in Ido Community, Nigeria

Abstract: This study was carried out to investigate the physicochemical and bacteriological quality of well water samples in Ido community. Thirty water samples were collected from five different wells ( six samples from each well) and subjected to standard microbiological and physicochemical analysis. All physicochemical properties showed difference (p<0.05) in all the wells analyzed except temperature and turbidity. Temperature ranged between 25.3±3.5 0 C and 26±2.35 0 C; pH, 6.3±0.7 and 6.9±0.3; electrical conductivity, 127.6±1.9 and 157.8±7.7μs/cm; total suspended solids, 25.2±8.9 and 53.2±1.8mg/l; turbidity, 0.15±0.7 and 1.20±1.4NTU; dissolved oxygen, 4.40±2.8 and 5.35±2.1 mg/l; Biological Oxygen Demand, 9.40±2.8 and15.4±2.8mg/l; Chemical Oxygen Demand, 177.2±1.6 and 260.3±1.6mg/l; chloride, 59.8±8.5 and 101.2±2.6mg/l and total hardness, 246.6±1.9 and 395.6±2.7mg/l. All values fell within WHO standards except Chemical Oxygen Demand (WHO Standard 40mg/l) and total hardness (WHO Standard 100mg/l). Results of microbial population did not show any difference (p>0.05) across the wells. However, Total Heterotrophic Bacteria ranged from 2.15±0.91×10 4 to 5.3±0.86×10 4 cfu/ml; total coliform, 3.00±0.77×10 4 to 6.18±0.73×10 4 cfu/ml; Total feacal coliform, 2.61±0.71×10 to 4.39±0.76×10 cfu/ml; Total vibrio count, 2.68±0.81× 10 to 4.4±0.86×10 3 cfu/ml; Total salmonella shigella count. 2.02±0.84×10 3 to 4.8±0.95×10 3 cfu/ml. Total Original Research Article Akani et al.; MRJI, 31(1): 1-10, 2021; Article no.MRJI.63945 2 coliform bacteria ranged from 220 to > 1600 coliform MPN index /100ml, while thermo tolerant coliform bacteria ranged from 220 to 1600 coliform MPN index /100ml -1 . A total of forty bacterial isolates belonging to ten genera were identified. They include species of Bacillus 22 (26.4%), Staphylococcus 14(16.8%), Vibrio 13(15.7%), Serratia 6(7.3%), Enterobacter 6(7.3%), Chromobacterium 4(4.8%), Salmonella 4 (4.9%), Shigella 4(4.8%) and E. coli 4(4.8%). This result highlights the fact that well water in Ido community are not safe microbiologically for drinking without additional treatment such as boiling or disinfection and this could lead to outbreak of water borne diseases. Good and proper environmental and personal hygiene is advocate especially by the users of those wells to prevent their contamination with bacterial pathogens.

Susceptibility Pattern of Pathogens Causing Blood Stream Infections in a Tertiary Care Hospital: A Two-year Retrospective Study from Southern Nigeria

Abstract: Aim: Bloodstream infections are a major cause of morbidity and mortality worldwide. The prevalence of causative microorganisms varies from one geographical region to another. This study was aimed at determining the etiological agents prevalent in our environment and their susceptibility profile. Study design: This is a retrospective study carried out at the University of Calabar Teaching Hospital, Calabar, Nigeria. Original Research Article Ekeng et al.; MRJI, 31(3): 53-60, 2021; Article no.MRJI.68874 54 Methodology: Blood culture results of patients documented over a two-year period were retrieved and analyzed. Blood culture positive isolates were detected using conventional method and Oxoid signal blood culture systems. Antimicrobial sensitivity tests were carried out by Kirby-Bauer disc diffusion method. Methicillin resistance in Staphylococcus aureus and coagulase negative Staphylococcus species (CoNS) was detected by disk diffusion method using 30 μg cefoxitin disk. ESBL production was detected by phenotypic confirmatory disc diffusion test (PCDDT) and the double disc synergy test (DDST). Results: A total of 413 blood culture antimicrobial susceptibility test results were analyzed, of which 116 (28.09%) were identified as culture positive. Sixty-nine (59%) of the positive isolates were from female patients. Out of 116 positive cultures, 58.62% (68/116) were Gram positive organisms, 40.52% (47/116) were Gram negative organisms, non albicans Candida accounted for 0.86% (1/116). Staphylococcus aureus (n=41, 35.3%) was the predominant isolate and showed high sensitivity to levofloxacin (100%), Linezolid (100%) and Amikacin (100%). Twelve isolates of S. aureus were methicillin resistant, while 1 isolate was inducible clindamycin resistant. Of the 116 isolates identified in this study, forty-three (43) were multidrug resistant with highest number of multidrug resistant isolates from Staphylococcus aureus (n=20). 21.28% (n=10) of the Gramnegative isolates were positive for extended spectrum beta lactamases. Conclusion: A high rate of antimicrobial resistance is observed among microorganisms causing blood stream infections. This emphasizes the need for antimicrobial sensitivity testing in the management of blood stream infections.

A Review of the Innate Immune Evasion Mechanisms and Status of Vaccine Development of Klebsiella pneumonia

Abstract: Klebsiella pneumoniae (KP) is a human pathogen causing a broad spectrum of diseases such as urinary tract infections (UTI), pneumonia, pyogenic liver abscess, bloodstream infections, and sepsis. Neonate, geriatric and immunocompromised individuals are the most vulnerable to KP infections. The success of KP as an infectious agent is due to the evolution of various mechanisms to evade the host's immune system. These diverse mechanisms have led to the dominance of KP infections in community settings where hypervirulent strains predominate and in hospital-acquired infections where multidrug-resistant strains predominate. KP infections in the past decades have been increasingly associated with high morbidity and mortality due to the emergence of multidrugresistant and hypervirulent strains capable of evading both the internal immune defense mechanisms and external antimicrobial agents. The pharmaceutical industries have very few and often expensive new antibiotics in the pipeline, offering little hope for antibiotic therapy. The development of new therapeutic strategies such as polyvalent, biconjugate vaccines that can provide protective immunity, especially against vulnerable populations, can mitigate the effects of Review Article Tiria and Musila; MRJI, 31(1): 33-47, 2021; Article no.MRJI.66311 34 KP infections. In this review, we discuss the virulence mechanisms of KP and how it evades the innate host immunity, and the interplay between the virulence and immune evasion strategies. The progress in the search for a vaccine to protect against KP infections will also be highlighted.

murA, uhpT, glpT and fosA Genes of Fosfomycin Resistance in Multi-drug Resistant E. coli Isolated from Hematological Malignancies' Patients with Blood Stream Infection

Abstract: Aim: To estimate the prevalence of murA, uhpT, glpT and fosA resistance genes of fosfomycin in multidrug resistant Escherichia coli isolated from patients suffering from hematological malignancies with on top blood stream infection together with correlating this distribution to the rate of expression of AmpC, ESBLs and MBLs in such isolates. Methods: 205 blood samples collected from patients with underlying hematological malignancies were cultured to isolate E. coli strains. Multidrug resistance was detected. PCR was done to determine fosfomycin resistance genes; murA, uhpT, glpT and fosA. Results: A total of 83 (40.5%) E. coli strains were isolated from blood samples. 58 (69.9%) were found to be multidrug resistant. AmpC beta-lactamase production was deduced in 15 (25.8%) isolates. 43 (74.1%) isolates were ESBLs producers whereas 9 (15.5%) were MBL class A carbapene-mases producers. Depending on PCR results, murA gene was detected in one isolate (1.7%), uhpT gene in 3 (5.2%), glpT gene in 4 (6.9%) whereas fosA gene was found in 7 E. coli strains (12.1%). Conclusion: Fosfomycin is a promising antibiotic with limited degree of prevalence of its resistant genes; however disseminating resistance might increase in multidrug resistant E. coli

Exploitaton of the Functional Properties of Leuconostoc mesenteroides and Lactobacillus plantarum as Probiotic using Soyabeans Flour as Vehicle

Abstract: Lactic acid bacteria produce a variety of antimicrobial compounds such as acetic acid, hydrogen peroxide, diacetyl on them as a natural competitive means to overcome other microorganism sharing the same niche. Seven different strains of Leuconostoc mesenteroides and Lactobacillus plantarum were screened for their ability to produce enzymes and metabolites. They were also screened for their ability to withstand some physiological stress like acid tolerance, temperature, salt concentration and antibacterial activity. Leuconostoc mesenteroides S3 and Lactobacillus plantarum Yh1produced values significantly different to other five isolates and they were selected. These functional properties were exploited in the fermentation of soyabean in order to obtain a probiotic vehicle. The selected isolates were used as starter both singly and in consortium. This research work also finds out the suitability of soya beans flour as a vehicle for probiotic microorganisms (Lactobacillus plantarum and Leuconostoc mesenteroides). The cleaned, dried and roasted soya beans were grinded to flour. The soya beans flour was fermented with Lactobacillus plantarum and Leuconostoc mesenteroides and there was mass increase in lactic acid counts in the fermented soya beans flour store at room temperature. All lactic acid fermented soya beans flour recorded increase at room temperature but slight reduction in number at refrigerator temperature from day 0 to day 14 of storage. Lactobacillus plantarum fermented soya beans flour (LPFSB) recorded increase from 5.9x105 (day 0) to 10.60x105cfu/g (day 14), while the L. plantarum stored at refrigeration temperature had 5.9x105cfu/g on (day 0) to 5.5x105cfu/g on day 14. The L. plantarum and consortium of both starters used were observed to have considerable increase in cell growth after storage, therefore this satisfies the criteria for good probiotic bacteria. Leuconostoc mesenteroides fermented soya beans slightly reduced in cell number at pH of 2 but others survived well at acidic pH and 10% bile. The cell number reduced from 4.8x106 (initial) to 4.2x106 (final). The pH of the intestine ranges from 2.5 to 3.5 and the starter culture used survive this pH and the bile product which shows greatly that they can serve as probiotic in the treatment of infection in gastrointestinal tract .

Role of PCR in Rapid Detection Group B Streptococcus in Pregnant Females in Al-Quwayiyah General Hospital, Riyadh, Saudi Arabia

Abstract: Background: Group B streptococcus is one of the most common causes of severe neonatal infections. Aim: To detect the prevalence of group B. Streptococcus and their antimicrobial susceptibility and to assess the role of PCR as a rapid method of its detection. Place and Duration of the Study: A cross sectional and prospective cohort study was carried out from September 2019 to February 2020 in Gynaecology and Obstetrics OPD and inpatient units in AlQuwayiyah General hospital, Riyadh, Saudi Arabia Methodology: Paired rectal/vaginal specimens were collected from 540 pregnant females with gestational age 35 or more, Each swab was inoculated into selective medium, Todd Hewitt, One swab is streaked onto blood agar plates incubated in 5% CO2 for 24h at 36oC. β-hemolytic colonies growth is identified by Gram’s stain, colony morphology and CAMP test. The confirmation and antimicrobial susceptibility were done by Vitek II machine, The second swab was used for PCR to identify atr gene. Original Research Article Khater and Abdel-Motaal; MRJI, 31(3): 45-52, 2021; Article no.MRJI.68764 46 Results: Out of 540 pregnant women 87 (16.1%) were colonized with GBS isolates. The positive GBS women aged 25 (22-34) and negative GBS women aged 23 (24-35) with no statistical difference. Patients aged more than 35 years old has the higher rate of positive GBS, 46.2%. No significant association detected between GBS and gestational age at delivery, antenatal visits, BMI and gravidity. The GBS strains isolated from pregnant women was 100% susceptible to linezolid and vancomycin followed by ampicillin (93.1%) and tobramicin (83.9%) then gentamicin (81.6%) and levofloxacin(78.2%) and showed least antibiotic susceptibility to erythromycin (26.4%). The PCR was positive in 145 (26.9%). Using culture as gold method, PCR sensitivity was 100% (95% CI: 91.62-100), while specificity was 83.2% (95% CI: 82.61-91.02). Negative and positive predictive values were 100% and 61% respectively. Kappa between the two methods was 0.71, which indicate major agreement. Conclusion: The GBS prevalence among the pregnant females in Al Quwayiyah General Hospital was 16.1%. Detection of GBS using new PCR technique was found to have high sensitivity and faster results, allowing efficient management of GBS and reduction in newborn morbidity and mortality however the cost is high for some laboratories. Further studies should be assessed to be both low cost and accurate rapid screening.

Scardovia wiggsiae and Streptococcus sobrinus Prevalence among Orthodontic and Non-Orthodontic Patients

Abstract: Background: Dental cavities or caries have been identified as among the most prevalent of preventable oral conditions. However, studies are discovering new information regarding the incidence and prevalence of several cariogenic organisms, including Streptococcus mutans (SM), the recently discovered Scardovia wiggsiae (SW), as well as Streptococcus sobrinus (SS). These studies have revealed varying prevalence among different populations, such as those undergoing orthodontic treatment. Based upon this information, the main goal of the current study was to assess the prevalence of specific cariogenic organisms (SS and SW) within saliva samples originally obtained from a dental school-based clinic. Original Research Article Trumbo et al.; MRJI, 31(2): 1-12, 2021; Article no.MRJI.66401 2 Methods: The protocol for this retrospective study of DNA isolated from previously collected saliva samples was reviewed and approved by the Institutional Review Board (IRB) as exempt research. In brief, clinical DNA samples were screened for SS and SW using quantitative polymerase chain reaction (qPCR). Demographic and subgroup (Orthodontic, non-Orthodontic) analysis was also performed. Results: This study found that pediatric (12-17 year old patient) samples were much more likely to harbor either SW or SS compared with adult (>18 year old patient) samples. In addition, this study found many more SW-positive samples among pediatric orthodontic patients compared with either adult or pediatric non-Orthodontic patients, which may suggest this population may be at higher risk for SW-related caries or other negative oral health outcomes. Finally, this study found these microbial populations to be strongly linked within the same patient samples. Conclusions: This study has demonstrated that prevalence of SW and SS may be more highly associated with specific population subgroups, including SS observed in non-orthodontic patients and SW found among pediatric orthodontic patients. These results also differ from previous evidence, which found only minor and partially overlapping prevalence of these and other oral microbes. The results of this current study may suggest that SS and SW may be more strongly correlated within similar oral microbial communities and their presence may be directly or indirectly linked through one or more behavioral, microbial or other factors – although more research will be needed to determine these mechanisms.

Presence and Virulence Potential of Aeromonas hydrophila in Selected Water Sources for Household Consumption in Makurdi, Benue State

Abstract: This study was conducted to investigate the prevalence of Aeromonas hydrophila in selected drinking water sources in Makurdi. A total of 100 water samples (Tap, river, stream, well, pond and borehole water) were collected from different locations in Makurdi. Isolation and identification of the organism was performed using standard microbiological techniques. Further confirmation of the isolates as Aeromonas hydrophila was carried out using the Microbact 24E detection kit and polymerase chain reaction (PCR). A. hydrophila was detected in 12 (12%) out of the 100 samples; 6.67%, 8.82%, 7.14%, 25%, 30% and 20% of tap, well, borehole, river, pond and stream water samples respectively. The highest isolation rate of A. hydrophila (30%) was from pond water. All A. hydrophila isolated exhibited heamolysin, protease and lipase activity. The findings of this study revealed that treated and untreated drinking water sources in Makurdi are contaminated with potentially virulent A. hydrophila strains which may pose a health risk to consumers. Therefore, basic water treatment should be applied to drinking water to reduce public health threat posed by this finding. Original Research Article Tersoo-Abiem et al.; MRJI, 31(3): 31-39, 2021; Article no.MRJI.67457 32

Screening of Antioxidant Activity of 20 Kinds of Mushrooms from Southwest of China

Abstract: The ethanol and water extracts of 20 kinds of mushrooms from Sichuan Province, Southwest of China, were investigated for their antioxidant activity using DPPH assay. The results showed the ethanol extract of Serpula lacrymans, Daldinia concentrica, and Scleroderma verrucosum had well DPPH free radical scavenging activities 91.33%, 79.76% and 62.82%, respectively. Besides, the water extracts of Serpula lacrymans, Armillaria luteo-virens, and Lentinus edodes also possessed pretty high DPPH free radical scavenging activity were 95.59%, 88.76% and 86.93%, respectively. Based on the above comparison, the EC50 and total phenolic content of the ethanol extracts of Serpula lacrymans, Daldinia concentrica, and Scleroderma verrucosum were also measured. Their EC50 and total phenolic content valued of 17.87mg·mL -1 , 11.19mg·mL -1 and 35.01mg·mL -1 , as well as 0.0769μg·L, 0.0673μg·L and 0.0545μg·L, respectively. The results showed there was a correlation between antioxidant activity and total phenolic content. Besides, the reaction time of the DPPH test affected the free radical scavenging, which reflected the difference of the extract component would impact the test method. Original Research Article He et al.; MRJI, 31(1): 26-32, 2021; Article no.MRJI.65905 27

Microbiological Assessment on Stalk, Endocarp, Epicarp, Tip and Vascular Tissue of Different Varieties of Banana Fruit (Musa spp.)

Abstract: Aim: The aim of this study was to carry out microbiological assessment on Stalk, Endocarp, Epicarp, Tip and Vascular Tissue of Different Varieties of Banana fruit (Musa spp.) (Cavendish, Dwarf Cavendish, Red, Lady Finger and Grand Nain Banana). Study Design: The study employs statistical analysis of the data and interpretation. Place and Duration of Study: Three major markets Oil Mill (OM), Fruit Garden (FG), Mile Three (MT), all located in the city of Port-Harcourt, Nigeria. Sample collection lasted for a week and the analysis was carried out every day and it lasted for six months. Methodology: A total of forty-five (45) banana (Cavendish, Dwarf Cavendish, Red, Lady Finger and Grand Nain Banana) fruit samples were collected for a period of three months from three different markets (Oil Mill, Fruit Garden and Mile Three Markets) in Port Harcourt, Nigeria. The collected samples were grouped into three (Unripe, Healthy-Looking and Spoiled) and were subjected to standard microbiological procedures which includes standard plate counts, biochemical and molecular identification of the isolates. Original Research Article Nrior et al.; MRJI, 31(3): 1-23, 2021; Article no.MRJI.67241 2 Results: A total of 124 isolates were isolated from the different sampled parts (Cut stalk, Tip, Endocarp, Vascular Tissue and Epicarp) of the banana fruit. Staphylococcus aureus count in the unripe sampled Banana fruits from Oil Mill market showed high prevalent rate in the Cut Stalk of the Cavendish Banana at 4.17±0.31x102 CFU/g followed by the Tips of the Grand Nain and Cavendish Banana at 4.03±0.21x102 CFU/g and 4.00±0.20x102 CFU/g, respectively. While the TCC in the Healthy-Looking sampled fruits from Fruit Garden showed more microbial load in the Epicarp of the Dwarf Cavendish Banana at 2.93±1.15x103 CFU/g followed by the Tip and Epicarp of the Grand Nain Banana at 2.70±0.82x103 CFU/g and 2.60±0.46x103 CFU/g. The total fungal count (TFC) in Healthy-Looking sampled Banana fruits showed high prevalent rate in Oil Mill market at 2.79x103 CFU/g > Mile Three market, 2.31x103 CFU/g > 1.35x103 CFU/g at Fruit Garden market. While the result of the total Staphylococcal count in the unripe sampled Banana fruits showed that Staphylococcus aureus was more prevalent in Creek Road market at 1.65x102 CFU/g > Mile One at 1.64x102 CFU/g and the least seen in Mile Three market at 1.50x102 CFU/g. The result of the Coliform count in the spoiled sampled Banana fruits showed that coliform was more prevalent in Mile One market at 2.14x103 CFU/g > 1.98x103 CFU/g in Fruit Garden market and less prevalent in Creek Road market at 1.75x103 CFU/g. The Grand Nain Banana had the most microbial load at 33.9% > Dwarf Cavendish Banana, 25% > Cavendish Banana, 16.9% > Lady Finger Banana, 14.5% > Red Banana at 9.7%. The study location with the most microbial load is the Oil Mill market at 21.23% > Mile One, 20.64% > Creek Road, 20.01% > Mile Three, 19.23% > Fruit Garden, 18.88%. Conclusion: Grand Nain banana variety has the highest microbial load thus consumption of it should be washed or cleansed thoroughly. Secondly, of the different parts sampled, Cut stalk of banana is associated with highest microbial load, therefore it should checked properly or cut off when peeling/during consumption, The high load of Staphylococcus aureus, Escherichia coli and Bacillus is of great concern. These organisms associated with different parts and varieties of banana fruits (Musa spp.) in Port Harcourt Nigeria, poses serious threat to consumers. Prohibition of anthropogenic activities within the markets and farms should be encouraged in order to reduce the level of contamination of these fruits. Also, public awareness on safe and hygienic practices in the handling and distribution of Banana fruits from the farms to the markets should be encouraged.

Bacteriological and Parasitological Assessment of Water use in Selected Abattoirs in Port Harcourt Metropolis, Rivers State, Nigeria

Abstract: In the abattoir, large amount of water is being used in processing operations; and this produces huge amount of wastewater which empty into the surrounding water bodies. The cross-sectional study followed the conventional microbiological techniques of macroscopy, microscopy, isolation, and identification to assess the bacteriological and parasitological examination of water sources in selected abattoirs in Port Harcourt Metropolis, Rivers State, Nigeria. The result of microbial population showed Total Coliform ranging from 5 x 10 1 cfu/ml – 2.80 x 10 2 cfu/ml and Faecal Coliform bacteria of 2.80 x 10 cfu/ml -1.85 x 10 cfu/ml. Total Heterotrophic bacteria ranged from 7.8 x10 1 cfu/ml to 4.2 x10 2 cfu/ml. The mean microbial counts; 4.86 x10 1 cfu/ml Fecal Coliform, 9.65 x 10cfu/ml Total Coliform and 1.82 x 10cfu/ml Heterotrophic Count. The isolates identified were Klebsiella spp, Shigella spp., Salmonella typhymurium, E. coli, Proteus spp., Salmonella paratyphi, Vibrio cholera. Nkpor village abattoir recorded highest occurrence of bacteria isolate with a total of 50(22.03%) followed by Iwofe 48(21.15%). Water sample from ponds had a higher parasitic contamination while samples from the borehole were free from parasitic infestation. Water samples Original Research Article Nwadiuto et al.; MRJI, 31(1): 19-25, 2021; Article no.MRJI.65446 20 for parasitological assessment were filtered and examined for parasitic organism using standard methods. Four different parasites were identified in the study, they include Ascaris lumbricoides (32.5%), Entamoeba histolytica (22.5%), Giardia lamblia (17.5%) and the least Taenia spp (7.5%). Nkpor-village abattoir had the highest level of contamination with (30.0%) parasite occurrence, followed by Mgboshimini abattoir (15.0%). The use of unsafe water for meat processing by butchers, portends a serious public health risk to consumers who purchase the meat, from these abattoirs. Therefore, government should endeavour to make borehole water available to all the abattoirs in Port Harcourt.

Susceptibility to Antibiotics and Reactive Oxygen Species in Escherichia coli: A Survey of Clinical and Environmental Isolates

Abstract: Aims: Some bacterial responses to oxidative stress also diminish antibiotic susceptibility; also, some antibiotics do increase oxidative stress within bacterial cells. Linkage or cross-resistance to prooxidants and antibiotics could facilitate the selection of antibiotic resistance and/or virulence. We made this survey in order to detect this possible linkage in Escherichia coli isolates. Methodology: The susceptibility of 102 E. coli clinical (causative of urinary or gastrointestinal infections) and environmental (rural or urban dust) isolates towards paraquat, H2O2, and antibiotics was measured using disc assays. Catalase and superoxide-dismutase (SOD) activities were measured. Results: Susceptibility to prooxidants was similar across isolates of all four sources, but urinary and urban dust isolates were more resistant to antibiotics. H2O2 \"resistant\" organisms had more antibiotic resistance phenotypes, particularly towards sulfadiazine and tetracycline. Paraquat \"resistance\" seems associated to beta-lactam resistance; but paraquat \"susceptibility\" seems associated to resistance towards chloramphenicol, gentamicin, ciprofloxacin and nitrofurantoin. Prooxidant disc assays correlate to catalase and superoxide-dismutase activities. A weak relationship H2O2/antibiotic-resistance, but not superoxide/antibiotic-resistance, is suggested. Conclusion: Overall, antibiotics exerting their action through oxidative stress, do not seem to have resulted in the co-selection of oxidative stress resistance, or vice versa. However, a possible link between resistance to some antibiotics and to H2O2 might contribute to co-selection between these two chemical insults.

Serosurvey of Crimean-Congo Haemorrhagic Fever Virus (CCHFV) in Cattle in Livestock Areas of Côte d'Ivoire, West Africa

Abstract: Background: Crimean-Congo haemorrhagic fever virus (CCHFV) is an arbovirus transmitted by Ixodid ticks and causes a highly pathogenic disease called Crimean-Congo haemorrhagic fever with a mortality rate of up to 50% in humans. Original Research Article Valery et al.; MRJI, 31(1): 11-18, 2021; Article no.MRJI.66906 12 Methods: From January to December 2019, tick and blood samples were taken from seven cattle farms located in the towns of Korhogo, Bondoukou, Man and Bouaflé. In each of the farms, samples were taken from twenty cattle. These tick and blood samples were sent to the national reference laboratory of the Department of Epidemic Viruses of the Pasteur Institute of Côte d'Ivoire for diagnosis of CCHFV as part of the surveillance of haemorrhagic fever viruses in Côte d'Ivoire. Molecular detection of the Crimean-Congo haemorrhagic fevervirus in ticks was performed by realtime PCR. For the serological analysis, the ELISA was used for the detection of antibodies to CCHFV (IgM and IgG). Results: The study allowed the identification of 411 ticks divided into three genera Amblyomma, Hyalomma and Rhipicephalus. The predominant species were Rhipicephalus (Boophilus) microplus with (47.4%) and Amblyomma variegatum with (42.8%). No ticks gave positive results for the research of the CCHFV.Serological analysis showed that all 140 blood samples were IgM negative and 53.57% were IgG positive. Cattle from Bouaflé (90%) and Korhogo (70%) showed a higher seroprevalence than cattle from other regions. There was no significant difference in seropositivity between female bovines (54.1%) and male bovines (52.5%) (P>0.05). There is an association between age and anti-CCHF IgG positivity (P≤0.05), as cattle less than 3 years old (59.3%) showed a higher seroprevalence. Conclusion: CCHFV serological tests have been detected in cattle, one of the livestock known to play a role in the amplification of CCHFV. Our results demonstrate that the prevalence of CCHFV is high in livestock areas of Côte d'Ivoire and suggest that surveillance for CCHFV should be

Comparative Analyses of ACE2 Receptor Binding Corona Viruses that Cause Mild versus Severe Acute Respiratory Syndrome in Humans

Abstract: Aim: To analyze the spike proteins and Replication-Transcription Complexes (RTCs) of the Mild and Severe Acute Respiratory Syndrome (SARS) and SARS-related coronaviruses (CoVs) to find out the similarities and differences between them, as both of groups bind to angiotensin-converting enzyme 2 (ACE2) receptor for human cell entry. Study Design: Bioinformatics, Biochemical, Site-Directed Mutagenesis (SDM), X-ray crystallographic, cryo-Electron microscopic (cryo-EM) and Mass Spectrometric (MS) data were analyzed. Methodology: The protein sequence data for spike proteins and the proteins of the RTCs, viz. the RNAdependent RNA polymerases (RdRps), primases and the nonstructural protein 7 (NSP7) were obtained from PUBMED and SWISS-PROT databases. The advanced version of Clustal Omega was used for protein sequence analysis. Along with the conserved motifs identified by the bioinformatics analysis, the data already available by biochemical and SDM experiments and X-ray Original Research Article Palanivelu; MRJI, 31(2): 64-85, 2021; Article no.MRJI.66560 65 crystallographic and cryo-EM studies on these proteins were used to confirm the possible amino acids involved in ACE2 receptor binding and active sites of the RTCs. For identification of probable N-linked and O-linked glycosylation sites, NetNGlyc 1.0 and NetOGlyc 4.0 tools of Technical University of Denmark were used. ExPASy tool was used for pI analysis. Results: The spike protein of human CoV (HCoV)-NL63 is ~90 amino acids longer than the spike proteins of SARS and SARS-related CoVs. The additions are mostly found in the N-terminal regions and few insertions are also found in the crucial receptor binding domain (RBD). The SARS and SARS-related CoVs and HCoV-NL63 showed several conserved residues, motifs and large peptide regions. The most important aspect between the recent pandemic causing SARS-CoV-2 and HCoVNL63 is a unique but different tetrapeptide insertions very close to the S1/S2 cleavage region, i.e., PRRAand -IPVR-, respectively. The next cleavage point S2’ and the transmembrane domains are conserved between the two groups. The RdRps are highly conserved between the two groups. The catalytic regions, catalytic amino acids and the NTP selection tripeptide regions are completely conserved between SARS-CoVs and HCoV-NL63. However, one of the metal binding sites, viz. the universal –GDDreported in all RdRps is aligning with– KDGin the RdRp of HCoV-NL63. The other metal binding site, viz. –SDDis completely conserved in both the groups. The NiRAN domains of the RdRps differed from the possible catalytic amino acid and NTP selection tripeptide regions. The primases (NSP8) and the NSP7 subunits of the RTC are highly conserved in both the groups. The NSP8 and NSP7 subunits exhibit closer similarities between the MERS-CoV and HCoV-NL63. Unlike other SARS and SARS-related CoVs, the HCoV-NL63 possesses only a single accessory protein. Interestingly, a large number of amino acids are replaced with Ns in the spike proteins (which is also reflected in the number of N-linked glycosylation sites in it) as well as in the RTC. Conclusions: Detailed analysis revealed several unique features in the HCoV-NL63 pathogen. As all the pandemic strains like SARS-CoV-1, SARS-CoV-2 and the milder HCoV-NL63 strain, use the same ACE2 receptor for entry into human cells, the frequent infection of humans by HCoV-NL63, especially in children, suggests that there is an ample opportunity for highly pathogenic variants to evolve in the future.

Evaluation of Antibacterial Properties of Different Leaf Extracts of Hyptis suaveolens (L) Poit

Abstract: Aim: Hyptis suaveolens is well known for its immense medicinal properties and are beneficial as folk medicine. The objective of the study is to investigate the efficacy of the leaves for it antimicrobial activity. Material and Methods: The leaves of Hyptis suaveolens was subjected to extraction process using n-hexane, ethanol and distilled water as solvents and the antimicrobial activity was analyzed against different bacterial strains viz.; Escherichia coli, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella aerogenes, Pseudomonas aeruginosa, Salmonella typhi, Enterococcus faecalis by agar well diffusion method. Results: Aqueous extracts showed inhibitory effect against the different tested bacteria organisms with variable zone of inhibitory range 6 -21 mm. Conclusion: The aqueous extracts of Hyptis suaveolens exhibited the presence of highly effective bio-active compounds in these extracts. These can further be evaluated and characterize to improve upon what has been done to create a novel compound that can be useful for various medicinal purposes. Short Research Article Erinle et al.; MRJI, 31(3): 40-44, 2021; Article no.MRJI.68177 41

Characterisation of Some Selected Bacterial Isolates from Vegetable Oil Contaminated Soil

Abstract: Microbial lipases occupy a place of prominence among biocatalysts and are often used for various biotechnological applications. Because of huge variation in applications, the availability of lipases with specific characteristics is still a limiting factor. There is therefore need for extensive characterisation of lipase for various applications. This work was carried out to characterise lipases from some selected bacterial isolates. Isolates identified as Bacillus subtilis, Bacillus licheniformis, Pseudomonas cepacia Pseudomonas fluorescens, Alcaligenes sp. and Flavobacterium sp. from a vegetable oil contaminated soil were characterized. Temperature, pH and ion concentration, (NaNO3 and MgSO4), incubation time, agitation speed, carbon sources and nitrogen sources were optimised for growth and lipase activity. Increase in microbial growth does not necessarily suggest increase in lipolytic activity as generally observed from this study. Temperature, pH, incubation time and agitation speed which had optimum enzyme activities for crude enzyme of Pseudomonas fluorescens (0.8 U/mL), were 27 o C, 7.0, 24 h, and 0 rpm respectively. Growth was not generally supported by AgN03 in all the organisms selected but supported by KNO3. However MgSO4 generally supported lipase Original Research Article Popoola and Olanbiwoninu; MRJI, 31(2): 22-37, 2021; Article no.MRJI.66637 23 production. Olive oil and peptone as sources of carbon and nitrogen respectively supported both growth and lipase production in the selected organisms. These bacterial isolates characterized had lipolytic activities, hence they have high potential for various biotechnological applications.

Community-Acquired Methicillin-Resistant Staphylococcus Aureus an Important Enemy against us; Why Investigation of Their Main Toxins Reveals Contradictory Data

Abstract: Community-Acquired Methicillin-resistant Staphylococcus aureus (CA-MRSA) strains are serious human pathogens because of their micro floral abilities; resistance to clinically important antibiotics and ability to evade the host immune defences. Panton–Valentine leucocidin (PVL) and Phenol-soluble modulins (PSMs) are the main important virulence factors of CA-MRSA. The aim of this work was to provide an explanation on why there are contradictory findings in studies of PVL and PSMs. Several factors such as differences in growth media or in injection mode, species-specific interaction, contamination in culture supernatants, the concentration of toxin, and exposure time that have an effect on conducting of these studies were discussed in this paper.

Antibiogram of Biofilm Producing Bacteria Isolated from Urine of Patients in Three Hospitals in Port Harcourt, Rivers State

Abstract: Aim: The aim of this study was to determine the antibiogram of biofilm producing bacteria isolated from urine of patients in three hospitals in Port Harcourt, Rivers State. Study Design: The study employs statistical analysis of the data and interpretation Place and Duration of Study: The study was conducted at three (3) hospitals; University of Port Harcourt Teaching Hospital (UPTH), Meridian Hospital D / line branch (MRD1) and Meridian Hospital Ikoku branch, all located in Port Harcourt, Rivers State. Sample collection was for three (3) months, analysis was carried out daily and it lasted for six (6) months. Methodology: A total of Forty-five (45) urine samples were collected for a period of three (3) months from the three (3) hospitals. The samples were labelled properly, according to date and time of collection. The collected samples were subjected to standard microbiological procedures which includes standard plate counts, identification, biofilm screening, sensitivity testing using Kirby-Bauer disk diffusion method, Phenotypic screening of extended spectrum beta lactamase and molecular characterization of the isolates Results: The results of the bacterial population of urine samples from the hospitals showed that the total heterotrophic bacterial counts for Meridian Hospital D/line (MRD1), Meridian Hospital Ikoku (MRD2) and University of Port Harcourt Teaching Hospital (UPTH) ranged from 4.93 - 6.30 x107cfu/ml. The Total coliform count ranged from 1.89-3.04 x106cfu/ml for Meridian Hospital D/line (MRD1), Meridian Hospital Ikoku (MRD2) and University of Port Harcourt Teaching Hospital (UPTH). Total faecal coliform counts ranged from 0.78-1.11 x105CFU/ml for Meridian Hospital D/line (MRD1), Meridian Hospital Ikoku (MRD2) and University of Port Harcourt Teaching Hospital (UPTH). A total of fifty-eight (58) bacterial isolates were isolated from urine of patients and 36(62.1%) isolates were identified as biofilm producers. The biofilm bacteria identified were 17.2% Staphylococcus,6.9% E. coli, 10.3% Pseudomonas, 6.9% Proteus ,10.3% Bacillus and 10.3% Enterococcus species. Biofilm forming ability of bacteria is considered a virulent factor and it is implicated to being a possible cause of increased resistance to most antibiotics. Varying susceptibility pattern was observed among biofilm isolates. Biofilm bacteria were resistant to several groups of antibiotics. Ofloxacin, Gentamycin, Imipenem and Nitrofurantoin can be used as drug of interest for most bacterial biofilm urinary tract infections. CTX-M and TET A gene were identified in the biofilm bacteria in this study to be possible factors that confer resistance to antibiotics. The presence of icaD and papC gene in the isolates whose genome were studied have been found to be possible factors that confers biofilm producing ability. This study indicates the emergence and rapid spread of biofilm producing bacteria and their resistance to antibiotics. Therefore, strict infection control practices as well as therapeutic guidance for confirmed infections should be rapidly initiated.

Optimal Conditions for Expression of Enzymatic Activities in Bacillus Strains Essential for Production of Soumbara, a Functional Fermented Food Involved in Blood Pressure Homeostasis

Abstract: Background: Soumbara is a traditional fermented food involved in the blood pressure homeostasis. The functional properties of this food strongly depend on the fermentative microbiota. In this study, we screened and identified at molecular level, some potential starters strain among the main microbiota associated with soumbara in Cote d’Ivoire, and investigated the conditions for optimal expression of their functional performance. Methods: We screened and identified by ribosomal gene sequencing, interesting microbial strains and conditions for optimal expression of their functional performance notably proteolytic, lipolytic, pectinolytic, amylolytic and cellulolytic activities was investigated using semi quantitative method. Results: The isolated microbiota was composed of 90.41 % of Bacillus strains, the rest being yeast Original Research Article Kouakou et al.; MRJI, 31(2): 38-52, 2021; Article no.MRJI.66454 39 and lactic acid bacteria. A total of 4 performant strains, specifically Bacillus subtilis (BS14P), Bacillus subtilis (KS16P), Bacillus velezensis (HS27M) and Bacillus pumilis (PS10P) were extracellular enzymes producing strains with halo diameter ranging between 1.9 and 2.8 cm. These strains grew optimally in the temperature range of 3035C at pH 7-8. The largest enzyme producer Bacillus velezensis strain (HS27M) was remarkably able to grow well at relatively high temperature (40C) and in a larger pH range (6-9). Proteolysis enzymes were produced optimally at 40-45C, pH 7 whereas lipolysis occurred maximally at 40C, pH 7. Likewise maximum pectinolytis, cellulolysis and amylolysis occurred at 45C, pH range 7-8. Conclusion: This study suggests Bacillus velezensis HS27M as valuable starter cultures for the production of soumbara with bioactive and organoleptic quality.

Characterization of Microbes in Protein Supplements Available Commercially

Abstract: Supplements have been used widely since several years and their production has greatly increased with many different types, e.g. (Vitamins, Collagen, Proteins. etc...), and for multiple uses. Supplements were made to complete the nutrients deficiency that people have, or to prevent the deficiency condition, and some are using it to boost their body with nutrients like the athletes. Protein is one of the most selling supplements in the market used as powder or drink, and also been added to many different foods and snacks to make it healthier. This paper involves a food safety and quality study to identify and characterize the microbial pathogens in protein powder supplements available commercially.

Bacteriological Quality of Potable Water Consumed in Cape Coast and Takoradi Metropolis, Ghana

Abstract: Background: Bacterial contamination of potable water remains a global canker and has been reported to result in deaths from gastrointestinal infections. Treatment of gastrointestinal infections is becoming difficult due to antimicrobial resistance. This study sought to assess the bacteriological quality of potable water consumed in Cape Coast and Takoradi Metropolis of Ghana. Methodology: Eighty-seven (87) samples of potable water were collected from various vicinities of Cape Coast (43) and Takoradi (44). The samples were analyzed for their bacterial loads using various laboratory bacteriological procedures and the resulting colonies were subjected to standard identification techniques. Antimicrobial Susceptibility Testing (AST) was carried out to determine the susceptibility patterns of the various isolates. Results: A total of 220 bacterial isolates were identified comprising 18 species, with Bacillus cereus. (13.6%), Staphylococcus aureus (8.5%), and Klebsiella sp. (10%), being predominant and Pseudomonas sp. (2.3%), Streptococcus sp. (1.8%), and Serratia sp. (0.5%) being less frequent. Antimicrobial Sensitivity Testing (AST) revealed multiple antimicrobial-resistant bacteria including, Escherichia coli, Pseudomonas sp., and Klebsiella sp. The average HPC and TCC of the various samples ranged from 0.20 - 1.94 x 108 CFU/ml and 0.00 - 2.39 x 108 CFU/ml respectively. Conclusion: Some potable water in Cape Coast and Takoradi metropolis including most sachet water sold on the streets were found to be highly contaminated with bacteria.