Open Access Original Research Article

Genetic Profiling of Cynodon dactylon Species Using ISSR Markers and Its Pharmacological Activities

Chandrika S. Tanthry, Saswati Mandal, S. D. Shruthi

Asian Journal of Biotechnology and Genetic Engineering, Page 1-10

Aim: Cynodon dactylon is a grass variety of the tropical and sub-tropical region belonging to the family Poaceae. In this study genetic profiling of 3 different varieties of Cynodon dactylon were done using ISSR primers.

Methodology: Polymorphism patterns were observed by amplifying DNA with 8 sets of primers such as OPA16, OPB11, OPC06 etc. PyElph tool was used to understand the genetic similarity of the samples and accordingly the relatedness was shown by dendrogram construction by DendroUPGMA. The grass samples were also used to check their antibacterial activity against potential pathogenic bacteria like Pseudomonas aeruginosa, Salmonella typhi, Chrysobacterium gleum etc. The plant species had showed a very satisfactory inhibition against the bacteria. In vitro anti-diabetic activity of Cynodon dactylon was checked by calculating its ability to inhibit the enzyme alpha-amylase and the anti-inflammatory activity was carried out by calculating the inhibition percentage of albumin degradation.

Outcome: All these experiments were done to study the pharmacological property as well as to gather knowledge on the genetic make-up of Cynodon dactylon. Overall, the final outcome supports the ethanobotanical claim of this plant.

Open Access Original Research Article

Bar Coding and Genetic Diversity Analysis of Millet Crop Species of Elusine Genus Using RAPD Markers

Aarti R. Desai, B. Ramya, S. D. Shruthi

Asian Journal of Biotechnology and Genetic Engineering, Page 11-18

Aim: Millets are a group of highly variable small seeded grasses, grown around the world as cereal crops or grains for fodder and human food. In this study, screening of crop primers with 8 genotypes of millet crop species belonging to genus Panicum, Eleusine, Echinochloa and Brachiaria was done. Main objective was to understand the genetic diversity among 7 millet crops varieties and a wild grass using RAPD markers by targeting nuclear gene. To find out the relationship between species of the same genus based on RAPD markers and to construct phylogenetic tree using PyElph 1.4 and Past326b softwares.

Methodology: PCR analysis was done using the selected RAPD to identify the polymorphic loci between the 8 genotypes. Primers OPC06, OPD13, OPW04, OPC18 were selected for analysis based on the band patterns, for further RAPD analysis. Bar Coding and Sequencing was done for validation of species with the help of softwares BLAST and Clustal OMEGA. Phylogenetic tree was constructed using all 4 primers with the help of software tools PyElph 1.4 and Past326b. The point parameter, which guided the joining linkage rule, is unweighted pair group average (UPGMA) and the genetic distance was computed from raw data.

Outcome: Among the 4 primers used, OPW04 and OPD13 showed maximum scoring of bands based on the phylogeny were established between the crop species of Eleusine species and the Indian Goose grass. The dendrogram showed that the varieties could be grouped into clusters. The sample C (Elusine indica), D (Elusine coracana) is diverged from the same root and C, D and G (Indian goose grass) are from the same root. Sample G is found to be very closely related to the C and D.

Open Access Original Research Article

Curative, Suppressive and Prophylactic Phyto-Medicinal Therapy/Synergistic Efficacy of Alstonia boonei/Capsicum frutescens Ethanolic Extracts against Plasmodium berghei (NK 65)/Salmonella typhi (ATCC 35723) Infected Swiss Albino Mice

Oludare Temitope Osuntokun, Isaac Tolulope Faniyi

Asian Journal of Biotechnology and Genetic Engineering, Page 19-34

The purpose of this research work is to evaluates the medicinal activity of Alstonia boonei and Capsicum frutescens extracts for Prophylactic, Curative & Suppressive phytomedicinal therapy against  Plasmodium berghei (NK 65)/Salmonella typhi (ATCC 35723) Infected Swiss Albino Mice. Alstonia boonei belongs to the Apocynaceae family and has several medicinal properties, for the treatment of various diseases while Capsicum frutescens is a tropical plant, an important agricultural crop and a popular African vegetables. The stem bark of Alstonia boonei and fruit of Capsicum frutescens were collected from Adekunle Ajasin University’s reserve forest. Both plant samples were authenticated in the herbarium unit of Forest Research Institute of Nigeria (FRIN) with identification number FHI 109806,109872 respectively. Plasmodium berghei (NK 65) and Salmonella Typhi (ATCC 35723) from the Institute of Advanced Medical Research and Training (IAMRAT), College of Medicine, University of Ibadan, Nigeria. The animals were acclimatized for two weeks and fed equally on rat chow and water throughout the period of the research work. A standard inoculum of 1×107 of parasitized erythrocytes from a donor mouse and 1x10-6 innoculum size of Salmonella typhi in volumes of 0.2 ml were used to infect the experimental animals intra-peritoneally. The suppressive antimalarial test also called“Test on Early Malaria Infection” was used in this study. Twenty five Male Swiss albino mice weighing between 18-20g were obtained from the animal house, the experimental  animal were divided into five groups of five mice each, which were inoculated with the parasite and the organism (Plasmodium berghei/Salmonella typhi) on the first day of the experiment (day 1). 72 hours. Group’s 1-3 mice received 100, 200 and 400mg/kg body weight of the extract per day orally. While the 4th group served as the positive control received 10mg/kg Chloroquine/Ciprofloxacin orally, 5th group received 0.2ml distilled water and served as negative control for the same period. Result obtained on observed the establishment of malaria and typhoid infection by administration of treatments dosage with different concentration of Alstonia boonei and Capsicum frutescens. The synergism of Alstonia boonei stem bark and Capsicum frutescens extract with significant (P< 0.05) dose dependent reduction in percentage parasitemia level at the three (100, 200, and 400) mg/kg doses, While P. berghei parasitaemia was the highest, Chloroquine /Ciprofloxacin, a significant weight loss compared to the other mice groups, suggesting that weight loss is a key criterion in this model. The synergism of Alstonia boonei of stem bark and Capsicum frutescens friut  extracts exhibits a significant curative, suppressive and prophylactic potency against Plasmodium berghei (NK 65)/Salmonella typhi (ATCC 35723) in infected mice as demonstrated by the reduction in the level of parasitaemia dose dependently. It is evident based on these findings that Alstonia boonei / Capsicum frutescens possess promising and potent antimalarial and anti-typhoid potential which justifies its usage in folk medicine for the management of malaria and typhoid infection.

Open Access Original Research Article

Spermatozoa Motility of Boer Buck Semen with Addition of Sweet Orange Essential Oil to Tris Yolk Extender

Sukma Aditya Sitepu, Zaituni Udin, . Jaswandi, . Hendri

Asian Journal of Biotechnology and Genetic Engineering, Page 35-38

Aims: The sweet orange essential oil contains antioxidants and antibacterials that can inhibit bacterial growth and prevent damage due to cold shock so that spermatozoa motility increases. Determine the effect of the addition sweet orange essential oil to tris yolk extender to spermatozoa motilty Boer Buck frozen semen.

Study Design: Randomized Block Design.

Place and Duration of Study: Laboratory Reproduction of Loka Penelitian Kambing Potong Sei Putih Indonesia, between January and March 2019.

Methodology: The research procedure starts with the preparation of semen extender, collection of fresh semen, dilution of semen, equilibration, freezing of semen, and thawing. Semen collected from 3 boer bucks were analyzed for the study using randomized block design as experimental design. Sweet orange essential oils as treatment used at 4 treatment level at (P0) control group (P1) 0,25%, (P2) 0,5%, (P3) 0,75% and (P4) 1% on the tris yolk extender. The observed variables was motility spermatozoa evaluated before freezing and after freezing (Post-Thawing).

Results: The results showed that the addition of sweet orange essential oil had a very significant effect (P <0.01). The results of adding sweet orange essential oil to the extender (Post-Thawing) were 36% (P0), 40% (P1), 40% (P2), 44% (P3) and 50% (P4).

Conclusion: The best values ​​the motility 50% (P4). It can conclude that the best result of adding sweet orange essential oil to tris yolk extender is (P4) 1%.

Open Access Original Research Article

Relationship of Testicular Biometrics with Body Weight in Rams: A Slaughter House Study in Yobe State, Africa

Musa Isa Mabu, Jibrin Musa Mabu, Bello Abubakar Anka, Yunusa Saheed

Asian Journal of Biotechnology and Genetic Engineering, Page 39-46

The research was a slaughter house survey of a testicular biometrics in relation to body weight in Ram. A total of 100 samples (Testes) were collected from rams of age ranges of 1 – 3 years. The data collected on each animal include; breed, color, sex, age, scrotal circumference, scrotal length, testes weight and body weight. The study was aimed to determine the relationship between testicular weight and body weight in sheep, and to predict the body weight in sheep from these testicular measurements. The data were subjected to the analysis of variance (ANOVA) using a general linear model SPSS (2001), significant means were separated using a Duncan multiple range test, a regression model was used to predict the body weight of rams with these testicular measurements as the independent variables. Descriptive method was used for the interpretation of the result. The relationship between body weight and testicular measurements were highly positive and significant at (P<0.01) except the relationship between body weight and testicular weights in Balami was significant at (P<0.05) level while the relationship between body weight and scrotal length was not significant. From the result of the study obtained farmers especially in Nigeria and other developing countries of the world can select superior breeds of rams in the field and predict the probable weights using testicular measurements especially; scrotal length scrotal circumference and testicular weight in Balami and Yankasa in the absence of weighing scale, and at the end to proffer suggestion for selection based on the testicular size as an indirect criterion for selection for genetic improvement.