Open Access Short Research Article

Heat-shock Technique for Isolation of Soil Bacillus Species with Potential of Antibiotics Secretion in Saudi Arabia

N. A. Al-Humam

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/BMRJ/2016/29126

The present research aimed to isolate soil Bacillus spp. from Al-Ahsa Province, Kingdom of Saudi Arabia, and test their potential of antibiotics production. Out of 21 soil samples, 34 bacterial isolates were obtained and following identification tests, 26 bacterial isolates in an isolation rate of 76.5% were identified as Bacillus spp by conventional methods and commercial automated identification system.  Identified species were Bacillus subtilis (7 strains), B. polymyxa (4 strains), B. licheniformis (4 strains), B. cereus (3 strains), B. mycoides (3 strains), B. pumilus (1 strains), and Bacillus spp. (4 strains). Disk diffusion test was employed to test the metabolites in sterilized cell free supernatant of isolates against some selected pathogenic bacterial species. The test indicated that B. polymyxa, B. licheniformis, B. cereus, B. mycoides and to a lesser degreeB. subtilis produced promising anti-microbial effects on tested bacterial species comparable to standard antibiotic disk.

The study suggests that some of Bacillus spp from the study area have potential to produce antibiotics that may be used to control microbial growth.

Open Access Original Research Article

Comparative Analysis of Molecular Methods for Detection of Influenza Viruses

Vikrant Sharma, Samander Kaushik

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/BMRJ/2016/28858

Aims: Influenza is a serious threat to human population worldwide therefore continuous surveillance is required to update influenza seasonal vaccines. A rapid, sensitive, specific and cost effective diagnostic method will be much helpful for patient management in the present scenario. Present study is conceptualized for detection of influenza viruses by molecular methods and compare with ‘gold standard’ virus isolation.   

Study Design: Standard strains of Influenza virus were used to standardize the molecular diagnostic assays and results were then compared with virus isolation.

Place and Duration of Study: Centre for Biotechnology, Maharshi Dayanand University, Rohtak, Haryana, India, between December 2015 and April 2016.

Methodology: Standard strains of Influenza A and B virus were used for influenza virus isolation using virus culturing in MDCK (Madin-Darby Canine Kidney) cell line by following standard tissue culture procedure. Isolated viruses were detected by Hemagglutination assay (HA) and typed by Hemagglutination inhibition assay (HI). Conventional one step RT-PCR, Taqman real time RT-PCR and RT-LAMP (Reverse transcription loop mediated isothermal amplification) were standardized on RNA extracted from standard strains. Sensitivity and specificity of these molecular methods were compared with each other as well as with virus culture (gold standard).

Results: Both influenza A and B virus strains were cultured in MDCK cells and produced cytopathic effect during virus culture. Conventional RT-PCR and real time RT-PCR detected both type of Influenza viruses. RT-LAMP also successfully detected and typed influenza viruses. RT-LAMP proved to be more rapid than other two molecular assays.  

Conclusion: Molecular diagnostic methods are useful in detection and typing of Influenza viruses and these methods provide results in short period of time when compared with traditional virus culture methods. RT-LAMP is rapid, sensitive, specific and cost effective method for influenza virus detection and subtyping.

Open Access Original Research Article

Biosorption of Heavy Metals in Dumpsite Leachate by Metal-resistant Bacteria Isolated from Abule-egba Dumpsite, Lagos State, Nigeria

Hassan Adeyemi Sanuth, Abimbola Olumide Adekanmbi

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/28011

Aim: We studied the potentials of four metal-resistant bacterial strains to reduce the concentration of selected metals in dumpsite leachate.

Methods: The bacteria which were obtained from the culture collection of the Environmental Microbiology and Biotechnology laboratory were subjected to increasing concentration of metals in a metal-supplemented medium. The bacteria were then introduced into a batch culture biosorption set-up containing the culture medium and tyndallized leachate and the residual metal concentration was determined after a 14-day incubation period using the atomic absorption spectrophotometer (AAS), after the bacterial cells had been separated from the leachate by centrifugation at 10000 rpm for 15 min.

Results: The bacterial strains demonstrated high resistance to the four selected heavy metals [lead (Pb), chromium (Cr), cadmium (Cd) and nickel (Ni)] and their combination. The Minimum Inhibitory Concentration (MIC) value for the strains on the metal-incorporated medium for all the selected metals ranged from 700-1500 µg/ml. The resistance to the metals was in the order: Pb ˃ Ni ˃ Cr ˃ Cd. Pseudomonas aeruginosa had the highest MIC to the metal combination (1300 μg/ml) while the lowest was Proteus mirabilis (800 μg/ml); Paenalcaligenes faecalis and Bordetella petrii had MIC values of 1000 μg/ml and 1200 μg/ml respectively. The biosorption set up showed that Paenalcaligenes hominis had a higher percentage reduction for Pb in the leachate with a reduction of 35.77%, while Bordetella petrii removed the highest concentration of Cd and Ni in the leachate with values of 32.81% and 34.91% respectively. However the highest percentage reduction for Cr (32.54%) was observed when the leachate was treated with a consortium of the four bacterial strains.

Conclusion: This study revealed that these metal-resistant bacteria could be very useful in the biological treatment of metal-containing wastewater.

Open Access Original Research Article

Prevalence and Antibiotic Susceptibility of Enterobacteriaceae from Fish Samples in Allahabad, India

Shreyasi Dubey, Pinki Saini, Chitralekha ., Priyanka Singh

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/28284

Aims: The present study was conducted aiming at the isolation of the pathogenic bacteria from the family enterobacteriaceae in raw, edible and apparently healthy fishes, from different fish markets of Allahabad.

Study Design: Forty fish specimens were collected from eight different locations of Allahabad region. Isolation was done using selective plating. Differentiation and characterization of different isolates was based on their growth characteristics on specific culture media, their biochemical confirmatory tests and Gram-staining reactions.

Place and Duration of Study: Fish samples were collected from eight different locations of Allahabad during January to May 2016.

Methodology: Isolation of enterobacteriaceae was done according to ISO Standard; ISO 21528-1:2004 and were confirmed by different set of biochemical tests and carbohydrate fermentation tests. Total soluble proteins of the organisms were estimated by Biuret method. Antibiotic susceptibility of the isolates was tested against amphicillin, streptomycin and ciprofloxacin at different concentrations.

Results: Strains of Klebsiella planticola, Citrobacter youngae, Citrobacter freundii, E. coli, Klebsiella ornithinolytica and Klebsiella pneumonia were identified out of the 56 isolates obtained. All the isolated strains organisms were susceptible to antibiotics such as amphicillin, streptomycin and ciprofloxacin at different concentrations of 10, 50, 80 and 100 mg/ml. The virulent proteins were highest in K. ornitholytica(103 mg) followed by Citrobacter sp (102.8 mg/ml).

Conclusion: Fishes sold in the fish markets of Allahabad showed presence of pathogenic bacteria especially from the family enterobacteriaceae; indicate poor hygienic conditions as well as improper storage environment. The results concluded that E. coli. (48.20%) represented the major part of bacterial flora, on the fish followed by Klebsiella spp. (30.2%).

Open Access Original Research Article

Ability of Trichoderma harzianum from Semi Arid Soils to Enhance Antioxidant Defense of Maize Seedlings under Water Stress

J. Chepsergon, L. A. Mwamburi, K. E. Kiprop

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/BMRJ/2016/26015

Aim: Determine the effect of different inoculum concentrations of T. harzianum from semi - arid soils on the activity of antioxidant enzymes of maize seedlings under water stress.

Methods and Results: This study employed a three-factor factorial (3×4×4) design, arranged in a completely randomized design (CRD) with three replications. Three maize varieties (H614, H629 and H6210) were treated with four concentrations of T. harzianum (0, 1x105, 1x107and 1x1010 spore/ml and thereafter grown under four osmotic potential regimes (0, -0.3, -0.6 and -0.9 MPa). Results from the study showed that T. harzianum had a significant effect on Superoxide dismutase (SOD) and catalase (CAT) activity of maize seedlings and did not enhance either maize seed germination or seedling growth. The activity of SOD and CAT was significantly enhanced by T. harzianum in all the three varieties of maize. Optimum SOD and CAT activity were recorded in seeds treated with 107 spores/ml of T. harzianum. Under normal growth conditions (0MPa), SOD and CAT activities were not enhanced by T. harzianum. However, under severe water stress (-0.9MPa), maximum activity of the enzymes was registered in all the three varieties of maize.

Conclusion: Maize seedling colonization by T. harzianum enhanced systems of antioxidative enzymes. Maximum activity of these enzymes was recorded under severe water stress (0.9MPa) mainly in seedlings treated with 107 spores/ml of T. harzianum. Consequently T. harzianum from semi-arid soils may be employed to improve maize plants’ tolerance to water stress.

Significance and Impact: With rapid increase in human population, coupled with global climate change, there is need to devise a cheap and safe option to increase the production of food crops. The ability of T. harzianum in promoting plant growth precisely maize under stress is of importance.

Open Access Original Research Article

An Exploration of Possible Consequences of Indiscriminate Consumption of Bissap Drink in Ghana

Sarah Darkwa

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/BMRJ/2016/28563

Aim: To study the chemical and microbiological profiles of dried Hibiscus sabdariffa calyces and bissap drinks prepared from these calyces and relate the findings to current prevalence of noncommunicable diseases in the Cape Coast Metropolis.

Study Design: Experimental design.

Place and Duration of Study: The chemical aspect was carried out at the School of Agriculture Laboratory of the University of Cape Coast and the microbiological aspect was at the Food Research Institute, Accra from February to April, 2016.

Methodology: Standard methods and tests were used to determine the chemical and microbiological variables. The Kjeldahl method was used to determine protein, the amount of potassium and sodium were determined using a flame photometer, the complex metric titration was used to determine the amount of calcium and magnesium. The amounts of Iron, Copper and Zinc were also determined. The pH was measured using a pH meter, titratable acidity using NaOH titration and total sugars using the colorimetric method. Total Viable Cell count and the identification of probable presence of Escherichia coli, Salmonella species and Coliforms in the bissap drinks were also carried out.

Results: Â remarkable amount of potassium was found in the range of 10874.4 –1667. 1 ug/g). Other elements in the bissap were also determined in descending order as sodium (5425.3 - 6058.4 ug/g), iron (2052.9 - 2099.6 ug/g), zinc (90.6 – 94.1 ug/g) and phosphorus (1175.5 – 1187.2 ug/g). Coliforms and Escherichia coli were identified while Salmonella species were not identified. Low pH, titratable acidity and total sugars ranged from 2.39 – 2.52; 0.27 – 0.30 and 7.2 – 14.0 respectively.

Conclusion: Hibiscus sabdariffa with its chemical constituents may promote health when used in preparing bissap drink and consumed but the acidity and high sugars present in the drink may overshadow its health benefits and cause diseases especially if taken in large quantities indiscriminately. It’s recommended that consumers drink bissap in moderation to avoid its probable health consequences.