Open Access Original Research Article

In vitro Antioxidant Activities and Effect of Hydroethanolic and Aqueous Extracts of Terminalia avicennioides (Combretaceae) on Salmonella

Louis-Claire Ndel Famen, Benjamin Tangue Talom, Richard Simo Tagne, Gabriel Tchuente Kamsu, Norbert Kodjio, Stephen Tamekou Lacmata, Donatien Gatsing

Microbiology Research Journal International, Page 1-14
DOI: 10.9734/mrji/2020/v30i130185

Today, Typhoid fever remains a public health problem in developing countries due to the poor quality of lifestyle associated with abusive and inappropriate use of antibiotics.

Aims: Considering the ethnopharmacological relevance of Terminalia avicennioides                              (T. avicennioides) (Combretaceae), this study was designed to investigate the in vitro antisalmonella and antioxidant activities of various extracts of this plant.

Methodology: The microdilution method was used to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of T. avicennioides extract. These extracts were also subjected to in vitro antioxidant tests such as diphényl-2-picrylhydrazyle (DPPH) radical scavenging test, ferric reducing-antioxidant power (FRAP), hydroxyl radical (OH) nitric oxide (NO) and Hydrogen Peroxide Scavenging Capacity.

Results: In vitro antisalmonella activity reveals that T. avicennioides stem bark extracts presented MIC values ranging from 64 to 512 μg / mL on tested microorganisms. This extract exhibited a good ability to trap DPPH with an IC50 of 8.30 μg / mL. The iron reducing power obtained with this extract had ODs ranging from 0.96 to 1.63. Phytochemical screening showed the presence of alkaloids, flavonoids, saponins, phenols anthocyanin and anthraquinone in all the extracts.

Conclusion: The results suggest that stem extract of T. avicennioides contains antisalmonella and antioxidant substances, which could be used for the treatment of typhoid fever and another salmonellosis.

Open Access Original Research Article

Using of Some Agro-industrial Wastes for Improving Carotenoids Production from Yeast Rhodotorula glutinis 32 and Bacteria Erwinia uredovora DSMZ 30080

Gehan F. Galal, Rania F. Ahmed

Microbiology Research Journal International, Page 15-25
DOI: 10.9734/mrji/2020/v30i130186

Some agro-industrial wastes such as clarified cane molasses, high test molasses, sweet whey, potato starch and corn steep liquor were tested as carbon sources or nitrogen source for growth and carotenoid accumulation using bacteria Erwinia uredovora DSMZ 30080 and yeast Rhodotorula glutinis number 32. Erlenmeyer flasks containing 100 ml of production media, the flasks were inoculated with 1 ml of standard inoculum and incubated at 150 rpm for 4 days at 30°C. Samples were collected periodical every 24h, cell dry weight and carotenoids concentration were determined. Sweet whey and highest molasses gave the highest growth being 2.85 and 7.34 gl-1, respectively and scored the same layout on carotenoids conc. which reach the peak during stationary phase (72 h of fermentation). Using of high test molasses and sweet whey as carotenoid production media were incremented carotenoid conc. about 1.7 and 2 fold (with respect to reference media). Increasing high test molasses conc. to give 5% initial sugar led to up great growth, carotenoids conc., productivity, yield and Yc/x from Rhodo. glutinis 32 to be 7.31 gl-1, 2.67 mgl-1, 0.037 mgl-1h-1, 0.067% and 0.365, respectively. Furthermore, using corn steep liquor (30%) as nitrogen sources augmented carotenoids concentration about 3.8 and 4fold for incomplete and complete production media using Rhodo. glutinis32. Also, a negligible effect on growth was observed with dark incubation with both strains which dropped about 75 and 48% with regard to control for E. uredovora DSMZ 30080 and Rhod. glutinis 32, respectively, whereas, carotenoids conc. was increased about 21% for E. uredovora DSMZ 30080 in dark condition.

Open Access Original Research Article

Assessment of Biofilm Formation among the Clinical Isolates of Escherichia coli in a Tertiary Care Hospital

Bedobroto Biswas, Naik Shalini Ashok, Deepesh Nagarajan, Md Zaffar Iqubal

Microbiology Research Journal International, Page 26-32
DOI: 10.9734/mrji/2020/v30i130187

Aims: Identification and grading of the Escherichia coli according to their biofilm production capability.

Study Design:  Cross-sectional study.

Place and Duration of Study: This was conducted in Department Microbiology at M.S. Ramaiah Medical college and Hospital, Bengaluru from March 2017 to August 2017.

Methodology: A total of 55 non repetitive Escherichia coli isolates were identified from various clinical samples like urine, pus ,tissue and peritoneal fluids .All the organisms were isolated in pure culture and biofilm formation was detected in vitro by Gold standard TCP (Tissue culture plate) method. Organisms were incubated for an extended period of 48 hours and the biofilms were detected by acetone alcohol elution method. Organisms were categorized as strong, moderate, weak and no biofilm producers based on the obtained OD value of the elute.

Results: Majority of the isolates of Escherichia coli were obtained from catheterized urine culture (67.03%) followed by pus (25.50%).Most of the isolates were capable of forming biofilm in vitro by tissue culture plate method except a few (9.1%). 40% of the isolates were strong biofilm formers which had >4 ODC. 25.5% showed medium biofilm-forming capability and rest 25.5% showed weak biofilm formations in vitro.

Conclusion: The ability to form biofilm from a species can give us a better understanding of the biofilm-related infections pertaining to the particular group. Detection of biofilms remains a most important determinant to approximate the incidence of such infections. Categorization of organisms according to their biofilm formation may help us understand the frequency of biofilm-associated infections, and thus take necessary precautions to avoid the problem. Further studies involving the detection of biofilm may be conducted and the tests can be implemented in routine diagnostic microbiology to assess the usefulness of the methods in detection of biofilm-related infections.

Open Access Original Research Article

Bio-stimulation Approach in Bioremediation of Crude Oil Contaminated Soil Using Fish Waste and Goat Manure

V. G. Awari, D. N. Ogbonna, R. R. Nrior

Microbiology Research Journal International, Page 33-46
DOI: 10.9734/mrji/2020/v30i130188

Aim: This study aimed to evaluate the ability of Fish waste and Goat manure to bio-stimulate the degradation process during bioremediation of crude oil-contaminated soil.

Study Design: Research was designed to evaluate and compare the strength of the organic nutrients (Goat manure and fish waste)   to stimulate the biodegradation of crude oil contaminated soil within 56 days.

Place and Duration of Study: Study was carried out in Rivers State University Farm, Rivers state, Nigeria for 56 days from July to September 2018.  Analyses were carried out weekly (per 7 days interval).

Methodology:  Eight (8) experimental set-up were employed, each having 5kg farm soil, all were left fallow for 6 days before contamination with crude oil on the 7th day in the respective percentages. Four of the set-ups were contaminated with 5% Crude oil while the other four were contaminated with 10% Crude oil. The contaminated plots were further allowed for 21 days for proper contamination and exposure to natural environmental factors to mimic a crude oil spill site before the application of bio stimulating agents (fish waste and goat manure). The set-ups of 5% Crude Oil Contaminated Soil (5% COCS) and 10% Crude Oil Contaminated Soil (10% COCS) were then stimulated with nutrient organics; Goat Manure (GM) and Fish Waste (FW) except two setups (one 5% COCS and the other 10% COCS) which were used as controls. The treatments (setups) were as follows: 5% COCS    (control 1), 5% COCS + GM, 5% COCS + FW, 5% COCS + GM + FW and 10% COCS (Control 2), 10% COCS +GM, 10% COCS + FW, 10% COCS + GM + FW. Physiochemical and microbiological status of the soil before and after contamination was evaluated while parameters including Nitrate, Sulphate, Phosphate and Total Petroleum Hydrocarbon (TPH), as well as Microbial analyses, were monitored throughout the experimental period. Bioremediation efficiency was estimated from percentage (%) reduction of Total Petroleum Hydrocarbon (TPH) from day 1 to the residual concentration at day 56 of bio-stimulation setups with the control. The bio-stimulating potentials of goat manure and fish waste were compared using statistical tools.

Results: The results revealed decrease in TPH with increasing time. The Amount (mg/kg) and Percentage (%) of Total  Petroleum  Hydrocarbon (TPH) remediated within the period of this study for 5% Crude Oil Contaminated Soil were as follows: 5% COCS-Ctrl 1 (563.52 mg/kg; 8.60%) < 5% COCS + GM (3608.84 mg/kg; 55.11%) < 5% COCS + FW (4156.49 mg/kg; 63.47%)  < 5% COCS + GM + FW (4350.69 mg/kg; 66.44%) while 10% crude oil contaminated soil were: 10% COCS-Ctrl 2 (125.71 mg/kg; 1.21%) < 10% COCS + GM (4422.75 mg/kg; 42.82%) < 10%COCS + FW (5542.16 mg/kg; 53.66%) < 10% COCS + GM + FW (6168.66 mg/kg; 59.72%). This result shows that combination treatment with goat manure and fish wasteis more effective and has more bio-stimulating potentials than the single treatments. With respect to individual bio-stimulating agent, fish waste proves more effective and had a higher bioremediation efficiency than goat manure. The results of colonial counts obtained revealed that the total heterotrophic bacterial and total fungal counts generally increased during the study across the trend. The counts obtained from day 7 to 56 in the respective experimental set ups were as follows: Total Heterotrophic Bacteria  counts increased from 6.32 to 8.20 Log10CFU/g (Control) < 6.32 to 9.05 Log10CFU/g  (COCS+FW) < 6.41 to 9.13 Log10CFU/g (COCS+GM) < 6.32 to 9.58 Log10CFU/g (COCS+FW+GM). Similar progression was observed for total fungi, hydrocarbon utilizing bacteria and hydrocarbon utilizing fungi in all the experimental set ups although irregular differences were observed in the control set ups.

Conclusion: The combination of organic nutrient such as goat manure and fish waste as bio-stimulating agents have shown to have higher percentage (%) bioremediation efficiency than when applied singly. It was also observed that the microbial biomass increased with time; moreover the nutrient monitoring analysis revealed a continuous gradual increase of the soil nutrient as bioremediation increases with time. The nutrient inherent in the bio-stimulating agents’ fish waste and goat manure resulted in increased soil nutrient (from day 7 to 56) as bioremediation period increase thereby enhancing soil nutrients at end of experiment. It is therefore recommended that bio-stimulating agents such as fish waste and goat manure should be employed in bioremediation of crude oil-contaminated soil especially due to its soil nutrient enhancement after bioremediation exercise. It’s a very good nutrient amendment option.

Open Access Original Research Article

Oral Site Specific Sampling Reveals Differential Location for Scardovia wiggsiae

Graydon Carr, Arvin Alexander, Linh Nguyen, Karl Kingsley

Microbiology Research Journal International, Page 47-55
DOI: 10.9734/mrji/2020/v30i130189

Introduction: The newly discovered cariogenic pathogen Scardovia wiggsiae has been the subject of intense scientific interest due to the role it may play in the development or progression of caries and oral disease. The primary objective of this study was to perform DNA microbial screening from five specific oral sites, including the gingival crevice between the upper central incisors, biofilm of the upper first molar and lingual incisor, as well as the dorsum of the tongue – for comparison with unstimulated saliva. These data may provide significant insights into site-specific oral locations that harbor S. wiggsiae.

Methods: More than one hundred previously collected clinical samples (n=105) were identified for inclusion in this study. DNA isolates were screened using a NanoDrop spectrophotometer to determine overall DNA quantity and quality. Samples with sufficient quality and quantity were screened for the presence of S. wiggsiae using validated PCR primers.

Results: More than one hundred patient samples (n=105) were identified, which were comprised of mostly female (57%) versus male (43%) and minority (71%) versus White (29%). The average DNA concentrations ranged between 13.74 and 14.69 mg/mL, with A260:A280 ratios ranging between 1.62 – 1.70. Results of molecular screening using S. wiggsiae specific primers demonstrated only a small percentage of pooled samples (7.6%) harbored this DNA, which was highly concentrated among the samples from tooth surfaces (Upper First Molar, Lingual Incisor) and saliva compared with the gingival crevice and dorsum of the tongue.

Discussion: These data provide novel information regarding specific oral locations, including tooth surfaces that harbor S. wiggsiae. In addition, these sites also provide new information regarding oral sites that do not appear to harbor this organism, including the gingival crevice and dorsum of the tongue. This information may be particularly useful to oral health researchers as they strive to limit and reduce the cariogenic microbiome among high-risk populations.

Open Access Original Research Article

Prevalence of Hepatitis B Surface Antigen and Hepatitis C Antibodies in Sickle Cell Disease Children under Sixteen in Two University Hospitals of Lome

S. Dossim, Koffi Mawusse Guedenon, A. Chick, K. Tanga, M. Kolou, V. Kponvi, T. Amavi, A. D. Gbadoe, A. Y. Segbena, P. Bonnabry, A. Y. Dagnra

Microbiology Research Journal International, Page 56-61
DOI: 10.9734/mrji/2020/v30i130190

Introduction: Sickle cell disease causes chronic anemia with the need for transfusions. The risk in children to get transfusions transmitted infections is high.

Aims: Determine the prevalence of HBsAg and HCV antibodies in sickle cell disease children under sixteen in Lome (Togo).

Study Design: It is a cross-sectional study.

Place and Duration of Study: Sample: Haematology Unit of Campus University Hospital and Paediatric Unit of Sylvanus Olympio University Hospital of Lomé collected between February to May 2016. Sample processing: Campus Hospital Laboratory.

Methodology: We collected blood in sickle cell disease patients in Campus and Sylvanus Olympio university hospitals at Lomé and informations about sickle cell type, transfusion, and hepatitis B vaccination. Sera were tested with Cobas e411 Roche® in the determining of hepatitis B surface antigen (HbSAg) and hepatitis C antibodies (HCVAb). Epi Info was used for statiscal analysis ®. Significant associations were found when P<0.05.

Results: Total of 172 patients from Campus Hospital and 79 from Sylvanus Olympio were included. Sex ratio and SS phenotype were 0.93, 69.8% and 1.32, 64.5% respectively. HBsAg was detected in 1.7% from Campus and 7.6% in Sylvanus Olympio.  One patient from Campus carried HCVAb. Significant association between hepatitis B and sex (P=0.02) and hepatitis B and vaccination were found (P= 0.0003). Males were more infected and patients who were unvaccinated carried HBsAg.

Conclusion: Vaccination against viral hepatitis and best blood donation screening are necessary to avoid these viral diseases in sickle cell disease children.

Open Access Original Research Article

Isolation and Characterization of Carbonoclastic Bacteria Diversity in Oil-contaminated Soil in Cape Coast Metropolis, Ghana

H. D. Nyarko, G. C. Okpokwasili, O. F. Joel, I. A. K. Galyuon

Microbiology Research Journal International, Page 62-74
DOI: 10.9734/mrji/2020/v30i130191

Aims: The study aimed at the quantification, isolation and characterization of hydrocarbon degrading bacteria in oil-contaminated and pristine soils.

Methodology: Soil samples from petroleum hydrocarbon polluted sites at auto-mechanic workshops, a mechanic village, as well as pristine (control) soils, comprising of 14 sampling locations within Cape Coast Metropolis in the Central Region of Ghana were collected using standard sampling techniques. Collected soil samples were treated and cultured while enumerations, isolations and characterization of carbonoclastic bacteria associated were evaluated.

Results: Bacterial populations isolated from hydrocarbon-polluted sites had higher aerobic counts ranging from 7.24-8.02 log10 cfu/g of soil when compared with the pristine sites (from 6.79-7.61 log10 cfu/g of soil). Also, soil samples from the mechanic village (8.76 to 7.48 log10 cfu/g of soil) recorded more bacterial counts than those from the mechanic garages (8.02 to 7.24 log10 cfu/g of soil). The calculated percentage profiles of all the hydrocarbon utilizing bacteria in the total culturable heterotrophic bacteria were low throughout the study, even though the percentage scores were all above 50%. A total of 19 hydrocarbon degraders were isolated. The isolates identified belong to the genera Pseudomonas, Proteus, Bacillus and Enterobacter.

Conclusion: The outcome of the study based on the bacteria populations, identification profiles, coupled with their survival and multiplications in designated medium amended with crude oil as the carbon and energy sources, suggest their petroleum hydrocarbon degrading capabilities, hence may be used in bioremediation applications.