Open Access Short Research Article

A Simple and Effective Phenol-Chloroform Method of DNA Extraction from Mammalian Feces

Abinaya Nadarajan, Karthy Sivapushanam, Kanchana Ramasamy, Debasis Jana, Pradeep Anbazhagan

Asian Journal of Biotechnology and Genetic Engineering, Page 1-8

Aims: To non-invasively collect the fecal samples of 12 different mammals and devise a simple method of fecal DNA extraction using a modified phenol-chloroform procedure of DNA isolation for species identification.

Study Design:  The experiment was laid out to check the applicability of the devised protocol. Morphological identification was done in the field to collect samples from the wild. Molecular characterization was carried out in the Molecular laboratory for species identification.

Place and Duration of Study: Advanced Institute for Wildlife Conservation (AIWC), Tamil Nadu Forest Department, Vandalur, Chennai, Tamil Nadu. The samples were collected between the time period of September 2019 and March 2020. The molecular analysis was performed between July 2020 and July 2021.

Methodology: We devised a scat/ fecal DNA isolation protocol and tested its applicability on 81 samples (30 herbivores, 15 omnivores, 36 Carnivores). Fresh and old samples were collected from wild (n=41) and captive (n=40) areas and used for the study. Independent isolation for each species was carried out with extraction control. The DNA isolated samples were quantified for concentration using Nanodrop Spectrophotometer to calculate the optimum DNA concentration for amplification. Independent PCR amplification of mitochondrial regions of cytochrome b and 12S rRNA were performed and gel electrophoresis was carried out for each sample for positive amplicons. The PCR products were sequenced using genetic analyzer.

Results: The protocol was validated by checking the strength of the devised method to work on species belonging to different ecological types. The sample size is n=81, positive amplification in cytochrome b region is 71 and 12S rRNA region is 79. Success of devised DNA extraction protocol on different population kinds, such as wild (n=41) and captive (n=40) were evaluated with a ratio of ‘Positive PCRs of samples’ against ‘Total samples for PCR’. It is 97.56 % (cytochrome b) and 100 % (12S rRNA) of Wild population and 60.0 % (cytochrome b) and 95% (12S rRNA) of captive population.

Conclusion: The devised protocol successfully worked on both wild and captive populations of herbivores, omnivores and carnivores. The success rate is better in 12S rRNA region on comparison with Cytochrome b. The applicability and reliability of the protocol has been tested and validated by checking the obtained sequences in the NCBI database and submitting the same to the database.  

Open Access Original Research Article

Identification of Novel Bioactive Compounds from Banana Fruit (Musa sapientum) as Antidepressant in Pregnant Women: Molecular Docking, Physiochemical and ADMET Evaluation

K. R. Sharmili Banu, I. Mohana Priya, A. S. Azar Zochedh

Asian Journal of Biotechnology and Genetic Engineering, Page 9-24

Mood disorder in women during pregnancy is a critical public issue. Generally, pregnancy and childbirth has major impact in the development of mental depression in women. There are many factors in which depression occur, such as post pregnancy depression after past deliveries, event of depression in the family, limited or lack of support by the partner and environment, unplanned pregnancy, pregnancy during young age, previous miscarriage, low level or lack of education and unemployment. Depression may occur due to the influence of estrogen and progesterone in the neurotransmitter system of serotonin, dopamine and norepinephrine. Banana plant (Musa sapientum) is known to be utilized in Indian folklore medication and Ayurveda for the treatment of various diseases. Various parts of banana plant consist of many bioactive compounds. In this current investigation, eighteen bioactive compounds were retrieved from banana fruit and docked against ER and PR using PyRx tool. From the eighteen bioactive compounds five bioactive compounds were chosen for further study based on the binding affinity. The predicted the physiochemical and ADMET properties were used to assess drug-likeness. According to the results campesterol, folic acid, quercetin, rutin and stigmasterol shown good molecular docking scores (> -8 Kcal/mol) and these compounds may be used a lead compounds to regulate ER and PR in the neurotransmitter system against depression in pregnant women.

Open Access Original Research Article

Preliminary Investigation of Haemoglobin Polymorphism among Indigenous Cattle in North Western Nigeria

Bunjah Umar Danladi, Adejoh-Ubani Elizabeth Ojoma, Rufina Obioma Okeke, Edward Christopher Dung, Japheth Joel Kalang, Danjuma Reuben Tom, Stanley David Oziegbe

Asian Journal of Biotechnology and Genetic Engineering, Page 25-29

The goal of this research was to identify the genetic pool of the Bunaji and Sokoto-Gudali cows
using haemoglobin (Hb) polymorphism. Blood samples were collected from a total of 150 mature Bunaji, Sokoto-Gudali and HolsteinXBunaji cows in north western Nigeria. To reveal the band patterns of haemoglobin, the red cell lysates were treated to cellulose acetate electrophoresis and a particular staining process. Individual cattle were found to have three co-dominant alleles, resulting in three genotypes (AA, AB, and AC). The A and B alleles had gene frequencies of 0.62 and 0.38, respectively. In the cattle population, the corresponding genotype frequencies for AA, AB, and AC were 0.34, 0.57, and 0.10, respectively. The variation between the observed and the expected genotype number was not significant (P>0.05) thereby conforming with Hardy-Weinberg frequencies. There is a need for further study to determine the association between morphological traits with haemoglobin polymorphism using high resolution DNA markers for detailed understanding of health status and breed characteristics of indigenous cattle in north western Nigeria.

Open Access Original Research Article

Assessment of Heavy Metal Contamination and Physicochemical Parameters of Sachet Water Consumed in Bukuru Metropolis of Jos South Local Government Area of Plateau State, Nigeria

Y. Denkok, V. G. Linus, U. S. Abara, K. F. Oyebade

Asian Journal of Biotechnology and Genetic Engineering, Page 52-60

Aim: The study investigated various concentrations of heavy metals present in some sachet water consumed in Bukuru metropolis of Jos South Local Government Area of Plateau State, Nigeria. Materials and Methods: Twenty five (25) sachet water were obtained from different selling spots and were grouped into five according to their commercially christened names as: SW1, SW2, SW3, SW4 and SW5 with each of these groups having five samples. Metallic concentrations were analyzed using AAS and standard analytical procedure were used to assay for temperature, conductivity, dissolved oxygen, total dissolved solid and PH.

Result: The results obtained from this study reveals that lead has a concentration of (0.73± 0.01, 0.75± 0.01, 0.68± 0.01, 0.75± 0.01 and 0.78± 0.01)ppm) for samples of SW1,SW2, SW3, SW4 and SW5 respectively. All of these concentration were relatively higher when compared to the WHO control. Zinc was detected at (0.003±0.2, 0.01±0.4, 008±0.5, 0.003±0.6 and0.005±0.2) ppm for SW1,SW2,SW3,SW4 and SW5 respectively. Iron was detected at (0.002±0.00, 0.03±0.00, 0.05±0.01, 0.03±0.00 and 0.03±0.00) ppm for samples of SW1,SW2,SW3, SW4 and SW5 respectively Iron concentrations were relatively lower in all the samples when compared to the control. Chromium was detected at (0.195 ±0.01, 0.104±0.00, 0.149±0.00, 0.204±0.03 and 0.138±0.00) ppm for samples of SW1, SW2, SW3, SW4 and SW5 respectively. Chromium concentrations were significantly elevated in all the samples relative control. Different concentrations of cadmium were assayed at (0.670±0.01, 0.695±0.01, 0.670±0.00, 0.735±0.01 and 0.628±0.01) ppm for samples of SW1, SW2, SW3, SW4 and SW5 respectively. Cadmium levels were significantly higher in all the groups when compared to the control. Arsenic was detected at (0.160±0.00, 0.120±0.00, 0.320±0.00, 0.060±0.01 and 0.180±0.01) ppm for samples of SW1, SW2, SW3, SW4 and SW5 respectively. They were all significantly higher in concentration relative control. The conductivity, temperature, total dissolved solid and PH values were lower when compared to the control.The dissolved oxygen concentrations were higher in all the water samples relative control.

Conclusion: The outcome of this investigation shows that the metallic characterization of the various branded sachet water do not meet the recommended standard due to elevated concentration of heavy metals beyond the permissible level. The physicochemical parameters were however, very low in concentration except for dissolved oxygen which was relatively high.

Open Access Review Article

Plant Bioactives and Biotechnological Approaches to Enhance Their Production and Bioactivity

G. Janarny, H. A. C. O. Hettiarachchi, N. N. G. Chiranthika, K. D. P. P. Gunathilake

Asian Journal of Biotechnology and Genetic Engineering, Page 30-51

Plants considered as an excellent source for discovering new compounds with medicinal significance in drugs/ functional foods/ nutraceutical formulations. Bioactive compounds of found in plants are those secondary metabolites exhibiting health benefit in human. It is obvious that studies related to plant bioactives engaging to develop in many aspects involving researchers from different scientific disciplines. In the explore for alternatives for producing and enhancing the desirable bioactives from plants, biotechnological approaches such as fermentation, plant tissue culture, metabolic engineering, genetic transformation, plant cell elicitation, microencapsulation (as delivery mode) are found to have potential in industrial production of bioactive plant metabolites. The present review discusses the different categories of plant bioactives along with an overview of their medicinal applications as antioxidants, anti-inflammatory agents, anti-diabetic and anti-cancer agents. The current trend on applications of recent biotechnological tools in the production of plant bioactives has also been discussed.