Open Access Original Research Article

Partial Purification and Characterization of Angiotensin Converting Enzyme Inhibitory Alkaloids and Flavonoids from the Leaves and Seeds of Moringa oleifera

A. M. Abdulazeez, O. S. Ajiboye, A. M. Wudil, H. Abubakar

Journal of Advances in Biology & Biotechnology, Page 1-11
DOI: 10.9734/JABB/2016/21974

Aim: In the present study, the alkaloid, flavonoid and saponin-rich extracts of the leaves and seeds of Moringa oleifera were partially-purified and the ACE inhibitory activities and patterns of inhibition determined.

Methodology: Alkaloids, flavonoids and saponins were extracted from the leaves and seeds of Moringa oleifera. They were then partially-purified via thin layer chromatography and column chromatography. The Angiotensin Converting Enzyme (ACE) inhibitory activities of each fraction was determined using Cushman and Cheung method with some modifications on the assay conditions, while the mechanisms of inhibition investigated using Lineweaver-Burk plots.

Results: The alkaloid-rich extracts of the leaves (6.27±0.12 mg/ml) and seeds (2.04±0.95 mg/ml) as well as flavonoid-rich extracts of the leaves (1.20±0.31 mg/ml) and seeds (2.16±0.56 mg/ml) of M. oleifera inhibited ACE at concentrations significantly (P <.05) lower than the saponin-rich extracts (26.01±1.02 and 24.32±2.31 mg/ml for the leaves and seeds, respectively). Thus, both alkaloid and flavonoid-rich extracts were further purified via thin layer chromatography and column chromatography. Of the four fractions obtained from the alkaloid-rich leaf extract, the ACE inhibitory activity of fraction 2 (0.46±0.01 mg/ml) was significantly higher (P <.05), while fraction 1 (0.07±0.01 mg/ml) of the alkaloid-rich seed extract significantly (P <.05) inhibited ACE compared to other fractions. For the flavonoid-rich extracts, fractions 3 (0.03±0.01 mg/ml) and 4 (0.42±0.03 mg/ml) obtained from the leaves and seeds, respectively, significantly (P <.05) inhibited ACE than other fractions. From the kinetic studies, fractions 1 and 2 exhibited uncompetitive types of inhibition with a decrease in Vmax and corresponding decrease in Km at 0.5 mg and 1.0 mg of inhibitor, respectively. Fractions 3 and 4 were competitive inhibitors of ACE, as the Km increased with increasing concentration of inhibitor, while Vmax remain unchanged. In conclusion, this study has demonstrated the ACE inhibitory activity of alkaloids and flavonoids from the seeds and leaves of Moringa oleifera and their potential as source of ACE inhibitors that may be beneficial for the management of hypertension.


Open Access Original Research Article

The Effect of Endosulfan on Ovaries of Mus musculus: A Histological Study

Arti Kumari, Preety Sinha

Journal of Advances in Biology & Biotechnology, Page 1-7
DOI: 10.9734/JABB/2016/17787

Background: Today pesticidal poisoning is an important health issue. The term “pesticides” covers a wide range of substances of differing chemical composition. Although we know these are useful in agricultural sector. However, it’s increased and unprotected use in agricultural practices has greatly enhanced the chances of its exposure to animals and man. This increases the risk of health disorders.

Objective: The aim of present study was designed to examine the effect of pesticide an Endosulfan on the ovary of female Mus musculus.

Materials and Methods: They were administered orally according to their body weight with Endosulfan @ 3 mg/kg body weight for 2 weeks, 4 weeks, 6 weeks and 12 weeks. The animals were sacrificed on day next to the last day of treatment and ovaries were extirpated. Ovaries were studied histologically after haematoxylin - eosin staining.

Results and Discussion: Observations showed structural defects during histological examinations of Graafian follicles after exposure of Endosulfan. Degeneration in follicular cells was seen and ova were often detached. Degeneration of ova occured in some follicles, irrespective of the stage of their development. The organization of granulosa cells was also disturbed in longer duration of treatment. The abnormalities gradually increased with dose duration time. Such types of abnormalities have not been seen in the control ovary.

Conclusion: It may be concluded from this study, Endosulfan has adverse effects on ovary at cellular level. Damaged cells of ovarian follicles, destroys steroid hormone production, which can result into improper ovarian function.


Open Access Original Research Article

Effect of Earthworm (Perionyx excavatus) Powder on Selected Bacteria and Fungi

Govindra P. Punu, Abdullah Ansari, Sirpaul Jaikishun, Diana Seecharran

Journal of Advances in Biology & Biotechnology, Page 1-15
DOI: 10.9734/JABB/2016/22572

Aims: Investigations were carried out at University of Guyana Turkeyen Campus during the year 2014-15 and focused on the chemical composition of earthworm (Perionyx excavatus) powder and its effect on microbes.

Study Design and Methodology: Dried earthworm powder was prepared from adult Perionyx excavatus and tested against bacteria (Pseudomonas aeruginosa ATCC27583, Escherichia coli ATCC25922 and Staphylococcus aureus ATCC25923) as well as fungi (Candida albicans ATCC24055; Aspergillus flavus and Aspergillus niger). Petri plates inoculated with known amount of bacterial and fungal colonies on Mueller Hinton agar media were treated with disks impregnated with different concentrations of aqueous solvent extract and ethyl acetate solvent extract of earthworm powder by following the disk diffusion suspecting tests technique. Earthworm powder was subjected to analysis of Magnesium (Mg), Calcium (Ca), Iron (Fe), Manganese (Mn), Zinc (Zn), Nitrogen (N) and Copper (Cu) using standard procedures.

Results: Ethyl acetate extract of earthworm powder at concentration 1:5 was more effective against Gram negative aerobic bacteria Pseudomonas aeruginosa ATCC27583 while aqueous extract at concentration 1:5 was more effective on Gram negative anaerobic bacteria Escherichia coli ATCC25922. Aqueous extract of earthworm powder proved to be more effective on bacteria Staphylococcus aureus ATCC25923 in comparison to ethyl acetate extract of earthworm powder.  Aqueous extract of earthworm powder had greater antifungal activity on Candida albicans ATCC24055 and Aspergillus flavus, whereas ethyl acetate extract at higher concentration have shown higher antifungal activity on Aspergillus niger.

Conclusion: This study conclusively proved that the earthworm powder prepared from Perionyx excavatus has antifungal and antibacterial properties.


Open Access Original Research Article

Bio-ethanol Production as Bio-solvent (Antifermenter), Antiseptic and Bio-fuel from Date Fruit Waste and its Suitable Properties Identification

A. B. M. Sharif Hossain, Mohammed Saad Aleissa

Journal of Advances in Biology & Biotechnology, Page 1-13
DOI: 10.9734/JABB/2016/22332

Significance of the Study: Biomass derived bio-ethanol is biodegradable, nontoxic and suitable substitute for fossil fuels. It can be used as bio-solvent and antiseptic in the pharmaceutical industry and can also be used to reduce greenhouse gas emission like CO2, CO, HC, SOex and NOX.  Dates fruit biomass is an important subsistence crop in arid and semiarid regions of the world and a useful feedstock for bioethanol production. It is rich in monosaccharide and can be fermented easily.

Aims: The present study was conducted to investigate the effects of pH, temperatures, durations, yeast and enzyme concentration, water percent and parts of date fruit on bioethanol yield and its different properties.

Methodology: Fermentation bioprocess was used to produce bioethanol from date fruit waste.

Results: The best yield (22.5%) of bioethanol in different parameters such as pH, temperatures, fermentation period and different yeast (S. cerevisiae) concentrations was found 5.8, 28°C, 5 days and 4 g/l, respectively. Moreover, viscosity and acid value were found within the limits prescribed by the latest American Standards for Testing Material (ASTM). There was no toxic elements found in the produced bioethanol and maintained the quality of ASTM standard. The elements found in bioethanol were Fe, Cu, Sn, Mn, Ag, Mo, Zn, P, Ca, Mg, Si and Na. The green house gas emissions such as hydrocarbons, sulfur dioxide, carbon dioxide, and nitrogen oxides were reduced while using the bioethanol, blended with pure fuel.

Conclusion: From the results it could be concluded that waste date fruit could be used as a potential feedstock for bioethanol production and suitable for 10% bioethanol as fuel for engine use without any modification of the engine.


Open Access Original Research Article

Isolation and Identification of Keratinolytic Bacteria that Exhibit Feather-degrading Potentials

F. O. Iruolaje, J. Ogbeba, M. Y. Tula, J. A. Ijebor, B. A. Dogo

Journal of Advances in Biology & Biotechnology, Page 1-9
DOI: 10.9734/JABB/2016/22082

Aims: To isolate and identify feather degrading bacteria from soils collected from the feather dumping site.

Study Design: Isolation and preliminary identification of bacterial isolates with keratinolytic potentials.

Place and Duration of Study: Mudalawal poultry processing site, Bauchi state, Nigeria, between January 2014 to October, 2014.

Methods: Soil samples from feather dumping sites were screened for bacterial growth. The isolated bacteria were identified based on their morphological and biochemical characteristics and were tested for their ability to degrade whole intact feather and powdered feather samples.

Results: The results showed that three (3) of the isolates belonged to the genus Bacillus which include; Bacillus licheniformis, Bacillus subtilis and Bacillus cereus, while one isolate is a Gram-negative rod, Serratia marcencens. Although B. licheniformis demonstrated higher feather degrading ability but with no statistical difference from other isolates in degrading both powdered and intact whole feathers samples. Synergy among the isolates to degrade powdered feather showed no statistical difference (p=0.317). Also, synergy among all the isolates resulted in degradation of intact feather significantly higher than that of S. marcescens, but not significantly different from those of B. licheniformis (p=0.389), B. subtilis (p=0.096) and B. cereus (p=0.096). Moreover, the keratinolytic ability of the isolates on intact whole feather was time dependent. It was also observed that the rate of degradation of powder feather samples was significantly higher than the rate of degradation of intact whole feather by all the isolates (p=0.026).

Conclusion: These isolates are therefore promising organisms for the management of chicken feather waste through biotechnological processes.