Open Access Original Research Article

Utilization of Genetic Algorithm to Optimize Biogas Production from Livestock Waste to Use in a CHP Plant in Agricultural Farms

Mostafa Kamalinasab, Alireza Vakili

Biotechnology Journal International, Page 1149-1164
DOI: 10.9734/BBJ/2014/12755

Utilization of Genetic Algorithm to Optimize Biogas Production from Livestock Waste to Use in a CHP Plant in Agricultural Farms

The optimization of biogas production with respect to external influences and various process disturbances is essential for efficient plant operation. However, the optimization of such plants is a challenging issue due the underlying nonlinear and complex digestion processes. One approach to solving this problem is to use the flexibility and power of computational intelligence methods such as Genetic Algorithms (GAs). The present study utilizes GA as tools for simulating and optimizing of biogas production process. Considering the effect of digester operational parameters, such as temperature (T), total solids (TS), volatile fatty acid (VFA), pH and A/TIC-ratio (amount of Acids (A) compared to Total Inorganic Carbon (TIC)), the optimal amount of biogas was converged to be 53910 cubic meters per month.

In order to reach the main goals on the energy problems, it is important to study and analyze the distributed CHP plant for agricultural companies and farms. This paper also describes a feasibility study of a biogas CHP (Cogeneration of Heat and Power )plant in a cow farm in Iran. With the developed model, it is specified that using 53,910 cubic meters of biogas, an internal combustion engine with electrical power of 375 kW can be operated continuously. Obtained results illustrated how the utilization of gaseous product from cow farm effluent (biogas) as fuel for heat and power generation can reduce primary energy consumption and its associated costs.Genetic algorithm

Open Access Original Research Article

Study on Flowering Time SSR Marker RM 248 in Traditional Rice Accessions in Sri Lanka

A. L. Ranawake, N. Mori

Biotechnology Journal International, Page 1165-1174
DOI: 10.9734/BBJ/2014/12569

Study on Flowering Time SSR Marker RM 248 in Traditional Rice Accessions in Sri Lanka

Aims: To study the diversity of flowering time locus RM248 in some traditional rice cultivars in Sri Lanka and compare it with days to flowering of studied rice cultivars.

Study Design: A field experiment was carried out to evaluate the days to flowering of the studied traditional rice cultivars. Rice plants were transplanted according to randomized complete block design with four replications and 20 plants per plot with 15 cm X 20 cm plant spacing. Diversity of flowering time locus RM 248 was analyzed by performing PCR using SSR marker RM 248 with genomic DNA of different eighty-nine rice genotypes. Gel electrophoresis was carried out in vertical gel electrophoresis unit for three hours. Gel plates were Silver stained and banding pattern was scored.

Place and Duration of Study: Field data were collected in Sri Lanka. The experiment was carried out during 2011/2012 Maha season and 2012 Yala season at the Faculty of Agriculture, University of Ruhuna, Sri Lanka. In Sri Lanka Yala is referred as dry season while Maha is referred as wet season. Laboratory data were collected at the Laboratory of Genetics, Graduate school of Science and Technology, Kobe University, Japan from October 2012 – December 2012.

Methodology: Days-to-flowering of traditional rice cultivars was studied in a field experiment and diversity of flowering time locus, RM 248 was studied by PCR amplification of SSR marker RM 248. SSR marker was amplified using the Gene Amp PCR System 9700 thermocycler. Amplified products were fractionated by electrophoresis through 6% denaturing Polyacrylamide gels and stained using Silver sequence DNA sequencing system staining regents (Promega, USA).

Results: Silver stained Polyacrylamide gel showed six different allele types in traditional rice accessions in RM 248 locus.

Conclusion: There is a great diversity in the RM248 locus among the traditional rice cultivars in Sri Lanka.

Open Access Original Research Article

In vitro Antimicrobial Activity of Crude Tannins Isolated from the Stem Bark of Annona senegalensis

Wurochekke Abdullahi Usman, Mahmoud Suleiman Jada, Romanus Moses Jideofor

Biotechnology Journal International, Page 1175-1181
DOI: 10.9734/BBJ/2014/11885

In vitro Antimicrobial Activity of Crude Tannins Isolated from the Stem Bark of

Annona senegalensis

The aim of this study is to determine the antimicrobial activity of crude tannins from the stem bark of Annona senegalensis on some diarrhoea causing bacteria strains. The study was carried out in the laboratory of Biochemistry department, Modibbo Adama University of Technology, Yola, Adamawa State, Nigeria, between July and October, 2013. The antimicrobial activity was carried out using agar disc diffusion method. The phytochemical analysis revealed the presence of tannins, alkaloids, saponins, cardiac glycosides, steroids, tarpenoids and phenols. Quantitatively, crude tannins of 170.16mg were found to be present in 500mg of the extract which makes up a percentage composition of 34.03%. The extracted crude tannins were used to test for antimicrobial effect against Escherichia coli, Salmonella typhi, and Staphylococcus aureus and were only effective on Staphylococcus aureus with 19mm as the inhibition zone using the concentration of 100mg/ml. The Minimum Inhibitory Concentration (MIC) of the extracted crude tannins was observed at 12.5mg/ml on Staphylococcus aureus. This suggests that the isolated crude tannins have an antistaphylococci activity and can be used to treat infections caused by the organism.

Open Access Original Research Article

In vivo and In vitro spasmolytic Effect of Ficus sur Forssk Ethanol Leaf Extract on the Gastrointestinal Tract

S. C. Akomas, S. N. Ijioma, C. U. Emelike

Biotechnology Journal International, Page 1182-1190
DOI: 10.9734/BBJ/2014/11919

In vivo and In vitro spasmolytic Effect of

Ficus sur Forssk Ethanol Leaf Extract on the Gastrointestinal Tract

Aim: Sequel to the report that Ficus sur ethanol leaves extract (FELE) has anti-diarrheal property and has been used in some parts of Nigeria for the treatment of the disease, this work examined the effect of Ficus sur ethanol leaves extract (FELE) on gastrointestinal motility.

Experimental Design: In vivo and in vitro Animal experiments were employed on whole rats and isolated intestinal tissues.

Methods: In the in vivo work, 25 adult rats were divided into 5 groups of 5 rats each. Group 1 was given 0.2ml normal saline and served as control. Group 2 received Atropine (0.1mg/kg), while groups 3, 4 and 5 were treated with 150, 300 and 500mg/kg of FELE respectively. Thirty minutes later, 0.2ml of charcoal meal was administered to all the rats used. The rats were sacrificed in another 30 minutes time. Each animal was opened to measure the distance travelled by the charcoal meal against the whole length of the small intestine. For the in vitro study, isolated guinea pig ileum and rabbit jejunum were used to study the effects of FELE on the gastrointestinal tract.

Results: In the in vivo studies, FELE exhibited a significant (P<0.05) dose dependent reduction in gastrointestinal tract motility as 150, 300 and 500mg/kg inhibited intestinal motility in treated rats by 58.36, 62.06 and 63.35% respectively and compared favorably with Atropine which inhibited same by 53 30%. On the isolated tissues, FELE exhibited relaxation effects and also significantly (P<0.05) blocked Acetylcholine induced intestinal contractions.

Conclusion: The relaxation effect of FELE on the gastrointestinal tract suggests that the extract contain principles with anticholinergic property and could be valuable in the management of diarrhea and gastrointestinal problems associated with hyper parasympathetic innervation. The results therefore agree with traditional claim that Ficus sur leaves extract has antidiarrheal property.

Open Access Original Research Article

Lipase Production by Fungal Isolates Grown in Palm Oil Mill Effluent

P. N. Ibegbulam-Njoku, O. K. Achi

Biotechnology Journal International, Page 1191-1200
DOI: 10.9734/BBJ/2014/12854

Lipase Production by Fungal Isolates Grown in Palm Oil Mill Effluent

Aims: The present work is aimed at determining the optimum conditions of pH, temperature and nitrogen concentration for lipase production by Candida rugosa and Geotrichum candidum in POME. 

Methodology: Five milliliter of sterile water was added to respective agar slant containing the two different fungal isolates while developed growth was scrapped with sterile needle and subsequently transferred into nutrient broth contained in 250 mL Erlenmeyer flasks. These flasks were further incubated for 48 h at 28ºC on rotary shaker at 180rmp. At this stage 0.1 mL of inoculum was transferred to the production medium (POME) and incubated for 144 h.

Results: At initial pH6.0, C. rugosa produce maximum lipase activity of 26.37 UmL-1 while G. candidum showed highest activity of 29.4 UmL-1 at pH 7. C. rugosa alo showed maximum lipase activity of 27.8 UmL-1 at 30ºC while G. candidum produced highest lipase activity (24.9 UmL-1) at 35ºC. The use of soybean meal in the optimization of production lipase revealed that at concentration of 3.5%w/v, the respective fungi isolates were best supported for lipase yield with maximum activity of 25.97 UmL-1 by C. rugosa and 28.32 UmL-1 by G. candidum.

Conclusion: This work evaluated the effect of three culture conditions (pH, temperature and nitrogen conc.) on lipase production by G. candidum and C. rugosa cultivated in POME. Results reveal that the factors were critical to growth and lipase production by the organisms and may be useful indices in the production of lipase even from other oil processing effluents. 

Open Access Original Research Article

Biotechnological Potential of Alpha Amylase Production by Bacillus subtilis Using Cassava Peel Powder as a Substrate

Ebiamadon Andi Brisibe, Helen Bankong

Biotechnology Journal International, Page 1201-1211
DOI: 10.9734/BBJ/2014/10856

Cassava peels, a major agricultural waste associated with the processing of cassava tubers into value-added products such as industrial starch and derived food items including garri and foofoo, were used as raw material for the production of α-amylase. The major step of the process is solid state fermentation of the mash prepared from this by-product by Bacillus subtilis that was isolated from a solid municipal waste disposal site. The effects of varying durations of incubation, temperature, medium pH and substrate levels were characterized. The maximum α–amylase activity of 7.12 lU/ml/min was recorded in a medium containing 50g of dried cassava peel powder as substrate after 24 hours at a pH of 7.0 and temperature of 35°C. The crude α–amylase produced was confirmed by using it to hydrolyze industrial starch which yielded maltose, a demonstration that the enzyme produced can be used in different biotechnological processes. It can be concluded that the use of cassava peels as substrates for the production of α–amylase does not only add value and decrease the amount of this agro-industrial waste from the environment but also reduces the general cost of amylase production that is desired for various biotechnologically-based industrial applications.

Open Access Original Research Article

Production of Some Polysaccharides from Kojic Acid Producing Aspergillus oryzae var. effusus NRC14 Biomass

Abd El-Naby M. Saad, Helmy M. Hassan, Mahmoud M. Hazzaa, Eman I. Ibrahim

Biotechnology Journal International, Page 1212-1222
DOI: 10.9734/BBJ/2014/13740

Production of Some Polysaccharides from Kojic Acid Producing Aspergillus oryzae var. effusus NRC14 Biomass

Aims: The present investigation describes the utilization of Aspergillus oryzae var. effusus NRC14 biomass producing kojic acid in the production of fine chemicals that have valuable applications such as chitin, chitosan, β-1, 3 glucan and /or their derivatives.

Methodology: Experiments of cell growth and substrate utilization were conducted in static cultures with identified medium composition. The technology involved in the extraction of their biopolymers is quite simple and the size scale is compatible with those currently used. This work was performed in Department of Microbial Chemistry, NRC.

Results and Conclusion: The utilization of A. oryzae var. effusus NRC14 biomass waste actually will cover a part of the cost production of kojic acid; therefore, clean cell walls preparations were isolated from the fungus biomass at different stages during kojic acid fermentation in a glucose salts medium. The major chitin constituent was extracted from the prepared cell walls from six days cultured A. oryzae var. effusus NRC14 biomass by cold conc. HCl or by 5% of LiCl/ DMAA solvent in yields 4.78% and 2.44%, respectively. Among the chitin family, chitosan has received special attention since it has several applications. Chitosan was extracted from alkali insoluble material (cell wall) obtained from 12 days cultured biomass by using different concentrations of acetic acid (0.5-2.5%) the results showed that the highest yield of chitosan was 4.65% at a concentration of 2% acetic acid. The isolated product was characterized by infrared spectrum. Chitosan- glucan complex was isolated in 17.94% yield by treating the A. oryzae var. effusus NRC14 biomass produced from kojic acid production for 12 days with 40% aqueous NaOH for 6h. at 95°C. Chitosan-glucan complex was fractionated into its components. The amount of extracted chitosan was 3.5%; the residue obtained after solubilization of chitosan by acid treatment was characterized as being mostly insoluble glucan (94%). Chitin-glucan complex was isolated from 6 days cultured A. oryzae var. effusus NRC14 biomass in yield 17.034%. The lyophilized product was fractionated into its components (chitin and glucan) in yields 16.36% chitin, 82.72% β, 1-3 glucan. The infrared spectra of the isolated products as well as of standard samples proved the identity of the isolated materials as chitosan, chitin and glucan.

Open Access Original Research Article

Enhancement of Bone Formation by Bone Morphogenetic Protein-2 Released from Poly (L-lactic-co-glycolic acid) Microsphere

Wen Zhao, Yuying Liu, Xudong Huang

Biotechnology Journal International, Page 1223-1237
DOI: 10.9734/BBJ/2014/12949

Enhancement of Bone Formation by Bone Morphogenetic Protein-2 Released from Poly

(L-lactic-co-glycolic acid) Microsphere 

An appropriate carrier acting as a sustained delivery vehicle for bone morphogenetic proteins (BMPs) is required for the maximal clinical effectiveness of these osteogenic proteins to enhance bone formation. The purpose of this study was to evaluate a low-molecular-weight poly(L-lactic-co-glycolic acid) (PLGA) copolymer as a synthetic, biodegradable carrier for the sustained delivery of bone morphogenetic protein-2 (BMP-2), and then to address the hypothesis that BMP-2 delivery from this vehicle could promote cell proliferation in vitro and ectopic bone formation in vivo. The BMP-2 was entrapped in microspheres of PLGA by using an improved water-in-oil-in water double-emulsion-solvent-extraction technique. The in vitro release kinetics of rhBMP-2 was determined by ELISA. Then we verified the effect of the sustained delivery vehicle on MSC cell proliferation. The ectopic bone induction in intramuscular implants of mice was evaluated at 2 and 4 weeks post-implantation. The results showed the PLGA microsphere released a total of 14.2%±0.71% rhBMP-2 at the initial phase followed by a prolonged release for 28 days. The rhBMP-2 released from the PLGA microsphere stimulated an increase in alkaline phosphatase (ALP) activity of MSC cells for 5 days in vitro, suggesting that the delivery vehicle releases BMP-2 for a prolonged period in an active form. Moreover, the released rhBMP-2 from the PLGA microsphere significantly promoted MSC cells proliferation after days 5 in culture. In vivo bone formation studies showed the rhBMP-2-loaded PLGA microsphere induced ectopic bone formation to a much greater extent than did rhBMP-2 treated mice. These results demonstrated that the PLGA copolymer material is capable of potentiating the osteogenic efficacy of BMP-2 and, as such, represents a promising delivery vehicle for BMP-2 for orthopedic and dental repair.