Open Access Short Research Article

Plasmid and Restriction Endonuclease Analyses of Bacteria Associated with Pneumonia in HIV/AIDS Patients in Ekiti State, Nigeria

A. O. Oluyege, O. Ojo-Bola

Biotechnology Journal International, Page 94-101
DOI: 10.9734/BBJ/2015/16656

Bacterial infections are an important cause of morbidity and mortality in individuals with the human immunodeficiency virus. The emergence of multiple-drug resistant bacteria has been documented by many researches. This study was therefore carried out to determine whether the resistances of bacterial isolates from HIV positive and HIV negative patients are plasmid mediated or chromosomal mediated. The Plasmid, Post Plasmid-curing Sensitivity and Restriction enzymes endonuclease were done using standard methods. The result of plasmid analysis showed that Plasmid-mediated resistance was observed in both populations and the molecular weight of the plasmid DNA was 1000 base pairs. Plasmid mediated resistance was common, and this was observed in all isolates from HIV/AIDS patients with exceptions of P. aeruginosa in which the resistance was chromosomally mediated. Restriction endonuclease analysis from E. coli revealed 3 distinct clusters. The result of restriction enzymes analysis indicate that the pneumonia infection in HIV/AIDS patients is likely to be hospital acquired in the study location. The study also suggests a common source of infection of the patients.

Open Access Original Research Article

Protective Effects of CYP2E1 Inhibitors on Metabolic Syndrome-induced Liver Injury in Guinea Pigs

Volodymyr V. Rushchak, Ganna M. Shayakhmetova, Anatoliy V. Matvienko, Mykola O. Chashchyn

Biotechnology Journal International, Page 57-67
DOI: 10.9734/BBJ/2015/17756

The present work reports the effects of CYP2E1-inhibitors (quercetin, 4- methylpyrazole and disulfiram) on the indices characterizing state of the liver in guinea pigs with metabolic syndrome (MS) induced by protamine sulfate repeated administrations.

The investigation of quercetin, 4-methylpyrazole and disulfiram effects on hepatic cytochrome P450 2E1 (CYP2E1) protein and activity changes was conducted.  Simultaneously, the content of reactive oxygen species (ROS) and markers of liver damage were determined in experimental animals’ blood. The link between increased hepatic expression of CYP2E1, prominent ROS generation and liver damage in animals with MS has been discovered. It has been demonstrated that CYP2E1 protein content and activity in guinea pigs with MS rose almost 3 times compared to intact animals. These events were accompanied by increase in ROS generation and metabolism and liver disturbances symptoms: increase in serum glucose and cholesterol contents (2 and 2.6 times respectively), alanine aminotransferase (2.8 times), aspartate aminotransferase (6.4 times), and alkaline phosphatase (1.8 times) elevations. Our investigation suggests that administration of quercetin, 4-methylpyrazole, and disulfiram in guinea pigs with MS caused decrease in this isoenzyme protein expression (2.5, 1.7 and 2.4 respectively) as well as its enzymatic activity in liver (6.6, 1.1, and 1.7 respectively). The content of blood ROS was partially restored or normalized by all three CYP2E1 inhibitors. In turn, suppression of CYP2E1 activity and ROS generation led to decrease in hepatic MS manifestation. It is apparent from the present observation that quercetin has the highest efficiency among the investigated substances. Further studies on various quercetin doses and administration regimens could provide relevant information for the development of MS-related nonalcoholic fatty liver disease treatment.

Open Access Original Research Article

Optimization of Operating Conditions Affecting Microbiologically Influenced Corrosion of Mild Steel Exposed to Crude Oil Environments Using Response Surface Methodology

K. K. Salam, S. E. Agarry, A. O. Arinkoola, I. O. Shoremekun

Biotechnology Journal International, Page 68-78
DOI: 10.9734/BBJ/2015/16810

In this study, the influence of four operating parameters (pH, salinity, nitrate concentration and immersion time) and their interactions on the microbiologically influenced corrosion (MIC) rate of mild steel in simulated crude oil environments were investigated by response surface methodology (RSM). 4-level historical data design: pH (A) at 4, 6, 8, 10, salinity (B) at 25, 50, 75 and 100 g/l, nitrate (C) at 25, 50, 75 and 100 g/l and immersion time (D) at 168, 336, 504 and 672 h, was employed to correlate the factors with the corrosion rate as response. A polynomial regression model was developed and validated prior to optimization studies. The result showed that pH has the most influential effect on the response and that the predicted data had a reasonable agreement with the experimental data with the values of R2 = 0.9660 and Adj-R2 = 0.9516. The optimum conditions of the crude oil environments were observed at: pH (9.37), salinity (94.73 g/l), nitrate concentration (37.97 g/l) and immersion time of mild steel (168 h) in order to achieve minimum corrosion rate of 0.155196 mpy. The study has revealed that the historical data RSM design is an efficient statistical technique for predicting the optimum operating conditions of crude oil environments required to minimize mild steel corrosion in oil pipelines by incorporating all factors under consideration.

Open Access Original Research Article

Comparing the Effect of Two Promoters on Cassava Somatic Embryo at Transient GUS Assay Level

Olufemi O. Oyelakin, Jelili T. Opabode, Emmanuel O. Idehen

Biotechnology Journal International, Page 79-84
DOI: 10.9734/BBJ/2015/17092

35S promoter from the Cauliflower Mosaic Virus (pCaMV) is a constitutive promoter commonly used in plant genetic transformation while Cassava Mosaic Virus (pCsVMV) is another promoter which is underutilized. The combination of the two promoters was used to form (pOYE153). The method adopted includes the insertion of a β–glucuronidase reporter gene (UidA) into a promoter cassette comprising the CsVMV promoter. The second construct (pCAMBIA2310) had (pCaMV) used for the selectable marker and gene of interest. This construct was mobilized into Agrobacterium tumefaciens strain LBA4404 and then tested for expression of the UidA gene in transient assays in cassava somatic embryos. After co-cultivation of these Agrobacterium with the plant tissues, histochemical β–glucuronidase (GUS) assays were performed to determine the level of UidA gene expression in transient assays. The results showed that the pCsVMV was able to drive high gene expression of β–glucuronidase reporter gene (UidA) in the transient assays in cassava somatic embryo. Expression of the gene also increases with the increase in the day of co-cultivation and likewise expression of the gene was higher for the sample in the light than the dark.

Open Access Original Research Article

Antibacterial Activity of Phytoalexins from Infected Theobroma cacao L

Emmanuel Dayo Fagbohun, Babatunde Idowu Aderiye, Saburi Adejimi Adesanya

Biotechnology Journal International, Page 85-93
DOI: 10.9734/BBJ/2015/17373

Aims: To study the antimicrobial activities of the compounds produced due to reactions of cocoa (Theobroma cacao) with Phytophthora palmivora during infection.

Place and Duration of Study: Department of Microbiology, Ekiti State University, Ado Ekiti, Nigeria and the Department of Pharmacognosy, Obafemi Awolowo University, Ile-Ife, Nigeria between September, 2009 and July, 2011.

Methodology: Phytophthora palmivora was used to infect healthy cocoa pods. The phytoalexins were extracted using solvent extraction and purified using standard methods. Agar diffusion and paper disc methods were used to study the antibacterial activities of the compounds extracted.

Results: Two major compounds were subsequently isolated, purified and characterized as FC-3-B21 and FC-4-B22 using the 1HNMR and 13CNMR as well as the 2D cosy data. Compound FC-3-B21 was characterized as 7,8,9,-trihydroxy-2,8-dihydroxy naphtha-10-one while FC-4-B22 was characterized as ester of glycerol: Bacillus subtilis, Escherichia coli, Micrococcus luteus, Pseudomonas aeruginosa and Staphylococcus aureus, were all sensitive to 7,8,9,-trihydroxy-2,8-dihydroxy naphtha-10-one even at 20 mg/ml. Pseudomonas aeruginosa showed the least sensitivity with 2.0 mm zone of inhibition at 20 mg/ml and 12 mm at 100 mg/ml while B. subtilis was the most sensitive with zone of inhibition of 4.0 mm at 20 mg/ml and 21.0 mm at 100 mg/ml. With FC-4-B22 (ester of glycerol) using paper disc method, P. aeruginosa was the most resistant with no zone of inhibition at 20 mg/ml. Meanwhile, S. aureus was the most sensitive with zone of inhibition of 3.0 mm at 20 mg/ml. However, B. subtilis exhibited a zone of 15.0 mm at 100 mg/ml. Using the agar diffusion method, 7,8,9,-trihydroxy-2,8-dihydroxy naphtha-10-one showed an appreciable effect on the tested pathogens. Pseudomonas aeruginosa was the least sensitive with the zone of inhibition of 3.0 mm at 20 mg/ml and 8.0 mm at 100 mg/ml while S. aureus was the most sensitive to the extract at 20 mg/ml with the zone of inhibition of 6.0 mm at 100 mg/ml. Bacillus subtilis was most sensitive to the extract at 100 mg/ml with the zone of inhibition of 19.0 mm. Testing the efficacy of ester of glycerol using agar diffusion method, P. aeruginosa showed the least resistance while B. subtilis was the most sensitive.

Conclusion: From this study, it can be concluded that the two novel compounds exhibited differential antibacterial activities towards the test bacteria.