Open Access Original Research Article

Compositional and Amino Acid Profile of Nicker Bean (Entada gigas) Seeds

H. N. Ogungbenle, O. T. Oyadipe

Biotechnology Journal International, Page 43-50
DOI: 10.9734/BBJ/2015/14681

The proximate, anti-nutritional factors, functional properties, minerals and amino acid composition of nicker bean (Entada gigas) were determined. The sample contained crude protein and carbohydrate of 24.8±0.02% and 47.2±0.10% respectively. The sample exhibited good emulsion capacity and emulsion stability of 62.3±0.03% and 36.2±0.01% respectively. The foaming capacity was 35.0±0.02% while the least gelation concentration was 4.00±0.01%. It contained nutritionally valuable minerals. The values of the anti-nutritional factors were: phytate (6.59±0.50 mg/g), oxalate (3.50±0.10 mg/g) and tannin (0.26±0.02%). Glutamic acid was the most abundant amino acid with the value of 51.1 g/100 g crude protein while methionine (4.10 g/100 g crude protein) was the least abundant amino acid in the sample. Entada gigas is a potential source of nutrients that should be cultivated and its consumption encouraged.  

Open Access Original Research Article

Response Surface Methodology Based Optimization of a New Isolate Bacillus pumilus ZR LS S2 with Fibrinolytic Activity

C. V. Reshma, K. Fathimathu Zuhara

Biotechnology Journal International, Page 51-61
DOI: 10.9734/BBJ/2015/15263

Aims: The work aims to isolate potential fibrinolytic enzyme producing isolates from various samples with the aim of developing a suitable optimization strategy using response surface methodology.

Study Design:  Plackett- Burman and Face centered central composite design was used.

Place and Duration of Study: Department of Life Sciences, University of Calicut.

Methodology: Various samples were screened using fibrin agar plates for the isolation of fibrinolytic enzyme producing bacteria and was identified by 16 s rRNA sequencing. Further the physical parameters and media components were optimized using Plackett- Burman design and face centered central composite design.

Results: A novel isolate with fibrinolytic potential was isolated and was identified as Bacillus pumilus ZR LS S2. A novel media was formulated using this data, for the production of fibrinolytic enzyme, which contains peptone, casein, MgSO4 and NaCl. The isolate produced 6738.384 U/ml of enzyme in optimized conditions. 

Conclusion: Initial studies were performed for the production of the fibrinolytic enzyme by the novel isolate, further studies are required to effectively validate the potential of the fibrinolytic enzyme and to develop effective application for the same.     

Open Access Original Research Article

Bioremediation of Hydrocarbon Polluted Mangrove Swamp Soil from the Niger Delta using Organic and Inorganic Nutrients

Charles Chibueze Ezekoye, Ebiokpo Rebecca Amakoromo, Abiye Anthony Ibiene

Biotechnology Journal International, Page 62-78
DOI: 10.9734/BBJ/2015/15083

Aim: To achieve bioremediation of hydrocarbon impacted mangrove soil using organic (sterile and non-sterile poultry wastes) and inorganic nutrient (NPK).

Study Design: Three treatments and the control designs were setup in three replicates and kept in the laboratory at room temperature (28±2°C) throughout the investigation period (6 weeks). Three of the set-ups designated Treatments (SPW, NSPW and NPK) were treated with sterile poultry wastes, non-sterile poultry wastes, nitrogen – phosphorus- potassium, respectively, while the fourth set – up with no treatment were designated control (CTRL). Set ups SPW, NSPW and NPK were designed to determine the effects of sterile poultry wastes, non-sterile poultry wastes and NPK in bioremediation of hydrocarbon polluted soil, respectively. However, the control (CTRL) was designed to determine the contribution made by indigenous (autochthonous) microorganisms and natural attenuation to the soil.

Place and Duration of Study: Department of Microbiology, Faculty of Biological Sciences, University of Port Harcourt, Rivers State, Nigeria, between August, 2012 and June, 2013.

Methodology: A laboratory-scale study was carried out using organic (sterile and non-sterile poultry wastes) and inorganic nutrient (NPK) to achieve bioremediation of hydrocarbon impacted mangrove soil.

Results: In a 42 day study, the sterile poultry wastes (SPW) treated option had an increase in total logarithmic heterotrophic bacterial count from 5.19±0.01 to 7.64±0.08 while non-sterile poultry wastes (NSPW) increased from 5.22±0.11 to 7.65±0.06. The logarithmic heterotrophic bacterial population of the NPK set-up increased from 5.23±0.11 to 7.90±0.10. The untreated set up had its total logarithmic heterotrophic bacterial count increased from 5.14±0.07 to 6.63±0.08. The total logarithmic hydrocarbon utilizing bacteria in SPW, NSPW and NPK treated options increased from 3.70±0.10 to 7.11±0.10, 3.82±0.10 to 7.20±0.10 and 3.93±0.08 to 7.73±0.07, respectively, at which time the corresponding value obtained for untreated increased from 3.63±0.06 to 5.56±0.06. Statistical analyses showed significant difference at p<0.05 level for the four conditions. Several genera of bacteria were isolated as hydrocarbon utilizing bacteria. They include Bacillus sp., Citrobacter sp., Corynebacterium, Escherichia sp., Flavobacterium, Micrococcus, Pseudomonas sp., Salmonella sp. and Vibrio sp. The conductivity values nitrate and phosphate concentrations in SPW, NSPW, and NPK options decreased progressively with slight decrease in the untreated. At day 42, the changes in total petroleum hydrocarbon (TPH) decreased to 498.14±0.01 ppm, 389.42±0.01 ppm, 285.38±0.01 ppm and 1087.00±0.01 ppm in SPW, NSPW, NPK and untreated, respectively.

Conclusion: The use of organic sources such as poultry wastes (sterile and non-sterile) and inorganic nutrient (NPK) has shown good promises in the bioremediation of hydrocarbon impacted mangrove soil.

Open Access Original Research Article

Investigation of Genetic Diversity among Medicago species Using RAPD Markers

Komal Murtaza, Khushi Muhammad, Mukhtar Alam, Ayaz Khan, Zainul Wahab, Muhammad Shahid Nadeem, Nazia Akbar, Habib Ahmad, Waqar Ahmad

Biotechnology Journal International, Page 79-86
DOI: 10.9734/BBJ/2015/14338

Aims: Medicago is known as the Queen of forage with potential economic importance to our society. The present study aimed at the use of RAPD-PCR DNA marker to identify the genetic fingerprints affinities of six species of Alfalfa.

Place and Duration of Study: The study was conducted at the Department of Genetics, Garden Campus, Hazara University, Mansehra Pakistan during February, 2011 to August, 2013.

Methodology: In this study, six species of Medicago namely TWAL (Tetraploid Wisconsin Alfalfa Line), Medicago arborea, Medicago falcata, Medicago sativa, Medicago lupulina and Medicago polymorpha were used to explore the diversity of alfalfa. Seven out of 120 decamers produced 34 polymorphic loci with 100% polymorphism to identify the different species of Medicago crop. The range of polymorphic loci was observed from 300 to 700 bp. Eleven species specific loci were generated by seven decamers. Primer B-18 generated single specific locus 700 bp against genomic DNA of M. lupulina and it is important to identify particular species of Alfalfa. The bivariate data were recorded as the presence of locus 1 and absence 0 and then this data was transferred into A and C respectively to make it suitable for DNAMAN software (version; Applied Biostatistics Inc). Moreover, cluster analysis was performed using sequence alignment and divergence function of the DNAMAN against the bivariate data collected from the products of decamers. All members clustered in a unique pattern except M. falcata and M. lupulina those shared 86% homology. Three distinct groups were observed during UPGMA (Unweighted pair Group Method with Arithmetic Mean). During the phylogenetic study, TWAL was observed to have genetic diversity from other five species of Alfalfa.

Conclusion: So, the present study is enabling us to discriminate different species of Alfalfa and it could be useful to identify and authenticate different species of the same genus of medicinal important plant from the Flora of Pakistan.

Open Access Original Research Article

Genetic and Phenotypic Identification of Vancomycin-Resistant Staphylococcus aureus Isolates from Retail Poultry Carcasses in Omu-Aran, North-Central Nigeria

C. E. Okolie, U. C. Essien, J. Idoko

Biotechnology Journal International, Page 87-92
DOI: 10.9734/BBJ/2015/13145

Staphylococcus aureus is a well-known agent of zoonotic infections. Livestock-associated methicillin-resistant S. aureus (LA-MRSA) had been receiving public health attention for over a decade. Recently, the genomes of some MRSA strains evolved further by enabling acquisition of vanA gene from enterococcus which drives the emergence of vancomycin-resistant S. aureus (VRSA), thus signaling a higher threat to antimicrobial chemotherapy and diagnostic microbiology. This study was designed to examine slaughtered chicken carcasses in Omu-Aran, North-Central Nigeria for the presence of VRSA using vancomycin agar screen (VAS) as recommended by the Clinical and Laboratories Standards Institute (CLSI). To provide independent witness to further support the evidences from VAS, a 235 bp marker for vanA gene was simultaneously detected by PCR. From April 2013 through May 2014, chicken carcasses (n=784) were collected and studied. Among 155 (19.8%) samples which yielded S. aureus, VAS and vanA PCR methods unequivocally identified VRSA in 22 (14.2%). Compared with 46.2% VRSA report from Zaria, North-Western Nigeria, the incidence of VRSA is much less in Omu-Aran chicken carcasses than those of Zaria. Further investigation in other parts of Nigeria is recommended in order to generate nation-wide data on VRSA in this country.