Open Access Original Research Article

Genotyping and Nucleotide Sequences of Growth Hormone Releasing Hormone and Its Receptor Genes in Egyptian Buffalo

Othman E. Othman, Mohamed F. Abdel-Samad, Nadia A. Abo El-Maaty, Karima M. Sewify

Biotechnology Journal International, Page 62-71
DOI: 10.9734/BBJ/2015/11619

Genotyping and Nucleotide Sequences of Growth Hormone Releasing Hormone and Its Receptor Genes in Egyptian Buffalo

Aim: The hypothalamic hormone, growth hormone-releasing hormone, is the principal stimulator of pituitary growth hormone (GH) synthesis and secretion. GHRH and its receptor (GHRHR) provide important functions in the regulation of the GH axis and in the development and proliferation of pituitary somatotropic axis. This study aimed to identify the genotypes and nucleotide sequences of two multifunctional genes; growth hormone-releasing hormone (GHRH) and its receptor (GHRHR) in Egyptian buffalo.

Methodology: Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and El-Gmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 296-bp fragment from GHRH gene and a 425-bp fragment from GHRHR gene of Egyptian buffalo. The PCR-amplified fragments were digested with HaeIII (GHRH) and Eco57I (GHRHR), electrophoresed and analyzed on agarose gels stained with ethidium bromide. The two amplified fragments were also sequenced and aligned with published sequences.

Results: Depending on the presence of the restriction site at 241^242 position (GG^CC) in 296-bp amplified fragments of GHRH, we genotyped all tested buffalo animals as AA. Due to the absence of the restriction site at position 300^301 ([CTGAAG(N)16^] in the amplified fragment of GHRHR (425-bp), we genotyped the tested animals as AA. The Egyptian buffalo GHRH and GHRHR nucleotide sequences were submitted to NCBI/Bankit/GenBank and have the accession numbers JN967799 and KC295414, respectively.

Conclusion: The Egyptian buffaloes are characterized by best production traits like high milk fat content as well as higher average daily gain and body weight where they are possess with fixed GHRHAA and GHRHRAA genotypes which were reported as desired genotypes for milk and growth production traits in different cattle breeds and the cattle are genetically homologous with buffaloes. To the best of our knowledge, these polymorphic sites are not identified in other buffalo populations. The identification of genotypes and nucleotide sequences of these two multifunctional genes may be useful in future marker-assisted selection (MAS) for more efficient breeding and genetic conservation programs of Egyptian buffalo.

Open Access Original Research Article

Comparative Studies on Some Edible Oils for Biodiesel Production in Nigeria

Humphrey Ibifubara, Nsikan I. Obot, Michael A. C. Chendo

Biotechnology Journal International, Page 72-83
DOI: 10.9734/BBJ/2015/14333

With the growing population and subsequent increasing demand for energy in Nigeria, coupled with the under-developed electrical energy platforms and the vast problems of the convectional energy sources such as continuous gas flaring, energy crisis is still lingering and enormous. The aim of this work is to compare the quality of different blends of biodiesel produced from several vegetable oils. The biodiesel was obtained from oil of animal fat and vegetable oils of bleached palm, corn, cottonseed, groundnut and soya through transterification reaction. The physiochemical properties of four blends were measured to ascertain their adherence with the ASTM standard for biodiesel. The results show a high percentage yield from most of the feedstock with the highest yield of 95% from bleached palm oil while the lowest value of 61.5% is from animal fat. Viscosity obtained at temperatures of 10ºC to 60ºC, with the highest value of 7.717mm2/sec is from B100 of animal fat at 10ºC and lowest value of 1.840mm2/sec is from B20 of corn oil at 60ºC. Also other values for density, flash point, cold point and sulfur content all conform to the ASTM standard range.

Open Access Original Research Article

In vitro Antitheilerial Activity of Paluther (Artemether 80) Against Theileria lestoquardi

Hayat M. Farah, Tigani H. Elamin, Hassan E. Khalid, Abdel Rahim M. El. Hussein, Zakia A. Mohammed, Husna M. El Basheir

Biotechnology Journal International, Page 84-91
DOI: 10.9734/BBJ/2015/10605

In vitro Antitheilerial Activity of Paluther

(Artemether 80) Against Theileria lestoquardi

Aim: The aim of this study was to screen Artemether 80 for activity against Theileria lestoquardi (Apicomplexa: Theileridae) using buparvaquone as a standard drug.

Study Design: In vitro study under laboratories conditions.

Place and Duration of Study: Veterinary Research Institute, between 2006 and 2008.

Methodology: Artemether 80 was screened for the first time to investigate activity against T. lestoquardi at different concentrations. Blood was collected separately from normal sheep and sheep infected naturally with Theileria. Normal lymphocyte cells and lymphocyte cells infected with Theileria were isolated from heparinized blood with Ficoll-paque. Isolated cells were grown in Minimum Essential Medium (MEM), supplemented with 20% calf serum and sub cultured. The parasite was identified with indirect fluorescent antibody test (IFA). A volume of 2.7 ml of lymphoblast cell suspension at concentration of 5x104 cell/ ml was distributed in tissue culture plates, and then 0.3 ml of drug at concentrations of 0.1, 1.0, 10 and 100 mg/L was added separately. A volume of 0.3 ml MEM was added to infected untreated control.

Results: The in vitro antitheilerial activity of Artemether 80 against T. lestoquardi 48 h after exposure was 0%, 14%, 30% and 45% at concentrations of 0.01, 0.1, 1.0 and 10 mg/L, respectively as compared with activity of buparvaquone at the same concentrations being 74%, 83%, 92% and 100%, respectively. Both Artemether 80 and buparvaquone caused in vitro partial cytotoxic effect at the highest concentrations. Activity and/ or partial cytotoxic effect of both drugs caused changes in the morphology of macroscizonts and host lymphoblast cells, decreased the number of macroschizonts/cell, mean number of dividing cells, increased the number of cells with extra cellular macroschizonts.

Conclusion: It was concluded that Artemether 80 is slightly effective in vitro against T. lestoquardi.

Open Access Original Research Article

Identification of Suitable Condition for Mannanase Production by Bacillus sp. GA2(1)

Sudathip Chantorn, Suda Natrchalayuth, Kulwadee Phannachet, Jirawan Apiraksakorn

Biotechnology Journal International, Page 92-97
DOI: 10.9734/BBJ/2015/15121

Identification of Suitable Condition for Mannanase Production by Bacillus sp. GA2(1)

Aims: The effects of 1% (w/v) supplementation of additional 5 agricultural wastes, corn cob, bagasses, coffee residues, soybean meal, and copra meal for mannanase production by Bacillus sp.GA2(1) were studied. Hence, partial characterization of mannanase was determined.

Methodology: The 1%(v/v) overnight cultured of Bacillus sp. GA2(1) was transferred into the basal medium and shaken at 150 rpm for 18 h at 37ºC. The additional of 5 AWs, corn cob, bagasses, coffee residues, soybean meal, and copra meal for the mannanase production were investigated. The cell suspension was centrifuged, and the crude mannanases were collected and stored at –20ºC for enzyme assay. The mannanase activities were measured by the dinitrosalicylic acid method. The optimal pH of mannanase were studied by measuring enzyme activity at pH 3-10 using 50 mM of following buffers; citrate (pH 3.0-6.0), phosphate (pH 6.0-8.0), and glycine-NaOH (pH 8.0-10.0). The optimal temperature was measured at 30-80ºC. Under standard assay conditions, locust bean gum was used as substrate to determine the optimal pH and temperature of the reaction. Thermostability was determined by preincubating the enzyme at different temperatures (30-80ºC) for 1 h. The residual mannanase activities were measured under standard condition.

Results: Among bagasses, coffee residues, soybean meal, corn cob and copra meal, the coffee residues was the most effective carbon source, the maximum yield of mannanase activity was 0.26 U/ml. The optimal temperature and pH for mannanase activity was pH 6.0 and 50ºC of 0.44 and 0.35 U/ml, respectively. The stability of enzyme was determined at 30-80ºC for 60 min. The results revealed that mannanase retained more than 96% of remaining activity after incubation of 60 min at 50ºC.

Conclusion: The maximum mannanase production was found when the medium was supplemented with coffee residues. Crude mannanase showed the highest activities of 0.44 U/ml at pH 6.0 and of 0.35 U/ml at 50ºC. The mannanase from Bacillus sp. GA2(1) retained more than 90% of theirs activities at 30-60ºC after preincubated for 60 min and then rapidly decreased.

Open Access Original Research Article

Evaluation of Hypoglycemic Efficacy of Methanolic Extracts of Moringa Oleifera and Phyllanthus amarus in Diabetic Rats

I. Oluwole Oyewole, A. Olufunke Taiwo, O. Nurudeen Quadri

Biotechnology Journal International, Page 98-102
DOI: 10.9734/BBJ/2015/13164

Evaluation of Hypoglycemic Efficacy of Methanolic Extracts of Moringa Oleifera and Phyllanthus amarus in Diabetic Rats

Aims: This study was undertaken to determine the hypoglycemic efficacy of methanolic extracts of Moringa oleifera and Phyllanthus amarus in alloxan-induced diabetic rats.

Study Design: Experimental Animal Study.

Place and Duration of Study: Department of Biochemistry, Osun State University, Osogbo Nigeria between September and March, 2013.

Methodology: Twenty four rats sorted into 4 groups were used for the study. Rats in control group (group 1) received distilled water while diabetes was induced in groups 2-4 rats by intraperitoneal administration of alloxan. Animals in groups 3 and 4 were treated with 500 mg/kg bw of methanolic leaf extract of Moringa oleifera and whole plant extract of Phyllanthus amarus respectively for 14 days while group 2 rats were left untreated. Serum glucose and total protein concentrations were measured in the rats after treatment.

Results: The two extracts reversed the alloxan-induced hyperglycaemic condition in rats as there was a significant reduction in blood glucose levels with Moringa oleifera having a more pronounced effect. Level of serum total protein was also significantly reduced in rats treated with the two extracts.

Conclusion: This study is a further scientific validation of the widely claimed use of Moringa oleifera and Phyllanthus amarus as useful ethnomedical treatment for diabetes mellitus.