Open Access Original Research Article

Statistical Optimization of Fermentation Parameters for Cellulase Production Utilizing Banana Peel

Modhuleena Mandal, Uma Ghosh

Journal of Advances in Biology & Biotechnology, Page 1-9
DOI: 10.9734/JABB/2016/26982

Aim: To optimize the physical parameters of solid state fermentation (SSF) by implementing statistical tool for production of cellulase, one of the potential biocatalyst in industries.

Study Design: Fermentation was carried out by the employment of response surface methodology (RSM) based on the Box-Behnken design (BBD) available in software Design-Expert (Version 7) Stat-Ease, Inc.

Place and Duration of Study: Department of Food Technology and Biochemical Engineering, Jadavpur University, Kolkata, India, between June 2014 and December 2014.

Methodology: The optimization of environmental factors for production of cellulase (FPase) was carried out by response surface methodology (RSM) based on the Box-Behnken design (BBD). The design included a total of 29 experimental trials that comprised time and temperature of fermentation, amount of substrate and hydration ratio as model factors for three levels.

Results: The mutual interaction between the independent variables under optimized conditions yielded FPase at the level of 8.05 U/ gds and total dissolved protein at the level of 1.4 mg/ml which were close to the predicted values from the model (7.95 U/gds and 1.39 mg/ml respectively for FPase and protein). The ideal parameters for cellulase production by Aspergillus sp. were 25% of hydration, 33ºC, 3.1 g of substrate and 7 days of fermentation.

Conclusion: A good correspondence between the predicted values from the model and the experimental values were seen. Consequently, we can use this methodology to adequately describe the interaction effect between the important independent variables influencing the fermentation process and the responses.


Open Access Original Research Article

Effect of Phosphate Solubilizing Bacteria on Growth and Development of Pearl Millet and Ragi

B. Harinathan, S. Sankaralingam, S. Palpperumal, D. Kathiresan, T. Shankar, D. Prabhu

Journal of Advances in Biology & Biotechnology, Page 1-7
DOI: 10.9734/JABB/2016/26290

In the present investigation, the effect of Bacillus sp. (C2) and Pseudomonas sp. (C7) isolated from Calotropis gigantea and Cyanodon dactylon from Shenbagathoppu hills, Srivilliputhur, Tamil Nadu, India on growth and development of pearl millet and ragi were studied. The effect of these isolates was recorded for morphological characters like plant height, fresh and dry weight. In the present study, maximum fresh weight and dry weight of pearl millet and ragi were recorded when treated with isolate C2 compared to C7 on all schedules of 30 and 45 days. Plant height recorded maximum in pearl millet on 45 days when treated with isolate C7 while ragi recorded maximum growth when treated with C2 isolate. Physiological characters like total chlorophyll, IAA, starch, fresh and dry weight were recorded on 45 day after sowing both in pearl millet and ragi treated with C2 and C7 isolates. It was found that, the isolate C2 treated plants recorded maximum total chlorophyll, IAA, starch, fresh and dry weight compared to C7 isolate treated plants both in pearl millet and ragi. Among the parameters tested, ragi plants recorded better total chlorophyll, IAA, starch, fresh and dry weight compared to pearl millet plants.


Open Access Original Research Article

Molecular Based Detection, Validation of a LAMP Assay and Phylogenetic Analysis of Capripoxvirus in Kenya

D. C. A. Omoga, M. Macharia, E. Magiri, J. Kinyua, J. Kasiiti, T. Holton

Journal of Advances in Biology & Biotechnology, Page 1-12
DOI: 10.9734/JABB/2016/27178

Aims: To optimize the different available molecular based techniques, validate the Loop-mediated isothermal amplification (LAMP) assay for detection of Capripoxviruses (CaPVs) DNA and deep sequencing to confirm the results of the LAMP assay and ensure that an effective and reliable detection method is recommended and adopted for use in Capripox diagnosis. The phylogenetic analysis was also to determine CaPVs genome variability, evolution, and diversity.

Study Design:  It included sample collection, DNA extraction, analysis and sequencing.

Place and Duration of Study: Biosciences East and Central Africa-International Livestock Research Institute (BecA-ILRI) Hub, between May 2014 and March 2015.

Methodology: We analyzed 130 samples including blood, skin nodules and cell cultures from symptomatic and suspected sheep, goats and cattle. The samples were tested by, conventional PCR, LAMP assay and real time PCR as the gold standard method, the results obtained were compared and used to determine sensitivity and specificity. The Limit of detection (LOD) of real time PCR and LAMP assay was determined using a serially diluted positive sample. Deep sequencing of 18 LAMP positive samples from different parts of the country was done and the obtained sequenced data analyzed using bioinformatics platform.

Results: The disease prevalence in the population was 61%, The validated LAMP assay had a 97% sensitivity and a specificity of 100%. Using the Fisher's Exact Tests, a P-value of (P = .8) at 95% interval was obtained. This shows that there is no significant difference between the validated LAMP Assay and the gold standard method. The Phylogenetic analysis of the 3 important CaPVs genes revealed the distribution of CaPVs into the 3 major specific lineages.

Conclusion: This study supports the adoption of LAMP assays for CaPVs diagnosis as a rapid and robust method of detection, disease monitoring and controlling outbreaks. The sequence data confirms the possibility of cross infection, nonhost specificity of CaPVs and also provides a reference in genomic research and a basis for molecular epidemiological studies in Kenya. 


Open Access Original Research Article

Screening for Mycotoxins and Major Bioactive Molecules in Mould Infested Brown Variety (BV) and Speckled Flecked Spotted Variety (SFSV) Vigna subterranea (Bambara Nut) in Nigeria

A. A. Brooks, P. T. Nnaji

Journal of Advances in Biology & Biotechnology, Page 1-9
DOI: 10.9734/JABB/2016/24948

Aims: This research was carried out with the sole aim of screening for mycotoxins and identifying the major bioactive molecules in two varieties of mould infested Vigna subterranea.

Place and Duration of Study: Department of microbiology, University of Calabar, Cross River State Nigeria was used for this research within the space of four months.

Methodology: Two varieties of Vigna subterranea were collected into a clean sealed container from major markets in South Eastern Nigeria. Samples were properly labeled and conveyed to the laboratory. After four months, samples were blended and dissolved in methanol and ethanol at a ratio of 80:20 volumes / volume (v/v). Bioactive molecules and mycotoxins were screened for using Gas chromatography/ Mass spectrometry.

Result: There was high level of similarities in major bioactive molecules of fatty acids, sugars, amino acids, phenols, alcohol and antioxidants in the two varieties. The pH and moisture content were within unfavourable range for mycotoxin production. Moisture content of the varieties was 2.46% and 8.03% while the pH was 7.01 and 6.9 respectively. Some biomolecules were peculiar to only one variety. Brown Variety (BV) and Specked Flecked Spotted Variety (SFSV) had Oleic acid with peak percentage area of 37.91% and 34.40% as the highest bimolecules respectively. Palmitic acid had percentage area above 15% in the two varieties.

Conclusion: This study juxtaposes the fact that mycotoxin production in V. subterranea is a synergistic effect of mould contamination and the necessary favourable environment.


Open Access Original Research Article

In vivo Evidence of Mice Hippocampal Alterations Resulting from High Dose Caffeine Exposure during Intrauterine Life

J. O. Owolabi, A. J. Olanrewaju, S. Y. Olatunji

Journal of Advances in Biology & Biotechnology, Page 1-7
DOI: 10.9734/JABB/2016/26543

Caffeine ingestion during pregnancy requires attention and investigation towards assessing the level of safety and possible effects on the brain and mental health. Caffeine ingestion during pregnancy has been discouraged despite concerns about lack of adequate facts to support this presumption and that such position is being taken based on speculations or inadequate facts. To this end, this research investigated the effects of caffeine exposure on hippocampus at various dosages during intrauterine life. Eighteen pregnant mice were divided into three groups A-C. Group A were fed ad libitum on mice feed pellets throughout the experimental duration to serve as control. Group B were administered the lower dosage [50 mg/kg body weight] of caffeine while Group C were administered high dosage [120 mg/kg body weights] of caffeine during intrauterine life by oral gavage. Brain tissues of the animals were excised after being sacrificed by cervical dislocation at Day 12 of postnatal life. The tissues were processed using the Haematoxylin and Eosin staining technique and the results were subject to histomorphological analysis. Caffeine at the high dosage substantially compromised hippocampal formation and dentate gyrus structural integrity; particularly by limiting cellular differentiation and elaboration of the cells.  The differentiation of neurons into the typical pyramidal cells of the hippocampus was largely limited.