Open Access Short Research Article

Effects of Temperature on Seed Germination of Invasive Plant Rorippa amphbia (L.) Besser

Yufeng Xu, Miao Liu, Yuanhao Lv, Qiuli Wang, Bo Qu, Meini Shao

Biotechnology Journal International, Page 1-7
DOI: 10.9734/bji/2018/v22i330058

Aims: Global warming and biological invasion are major environmental issues faced in the world. In the study, Rorippa amphibia, a perennial invasive clone plant in northern China, was used as a material to study the germination characteristics of the seeds at different temperatures.

Study Design: Germination test of R. amphibia seeds at different temperature was studied by means of laboratory culture. The germination percentage, germination index, germination potential, bud height and root length of the seeds were determined.

Place and Duration of Study: Samples were collected from the west side of the swimming pool of Shenyang Agricultural University of Liaoning Province in August 2017. Experiments were done in the College of Biological Science and Technology, between October 2017 and June 2018.

Methodology: The petri dish method was used in the experiment. Fifty seeds were randomly selected and soaked in distilled water for 12h. The seeds were placed in a petri dish covered with double filter paper, cultured at 10℃, 15℃, 20℃, 25℃, 30℃, 35℃ and 40℃ in light incubators for 12h darkness and 12h light (4000lux), with 3 repeats per processing. Seed germination was based on embryo root breakthrough seed coat ≥ 1 mm. During the experiment, the numbers of seed germination were recorded every day, and the filter paper was kept moist until there was no new seed germination for 2 consecutive days, which was regarded as the end of germination. The numbers of seed germination should be counted regularly every day, and the beginning and duration of germination should be recorded. The germination rate, daily germination rate, germinating potential, germinating index and vigor index of R. amphibia seeds at different temperature were calculated by measuring bud height and radicle length on the 10th day after germination.

Results: The temperature range of seed germination of R. amphibia was wide, which could germinate at 15 - 40℃. Lower temperature delayed the peak period of seed germination at some extent and the germination rates of R. amphibia peak at 30 - 35℃, which were 44.67% and 50% respectively. At 35℃, germination potential and germination index were 25.33% and 29.46, reaching the maximum value.

Conclusion: The reason for the wide temperature range of seed germination and the low germination rate might be the candidate method for clonal plant population establishment in temperate zone. The higher germination rate of high temperature condition suggested that clone invasive plants in temperate regions were more invasive during global warming.

Open Access Original Research Article

Molecular Variability of Ocimum gratissimum L. Accessions Using RAPD Marker

O. J. Olawuyi, O. F. Kariunwi, A. O. Akanmu

Biotechnology Journal International, Page 1-11
DOI: 10.9734/BJI/2018/45381

Aim: This study investigated the molecular variability among accessions of Ocimum gratissimum from selected states in Nigeria and Mali using RAPD marker.

Study Design: The experimental design was complete randomized design (CRD) with three replicates.

Materials and Methods: Twenty accessions of Ocimum gratissimum were collected from nineteen selected Local Governments in four South-western States of Nigeria (Ogun, Oyo, Osun and Lagos) and Mali, to assess their genetic diversity and phylogenetic relationship. Molecular statistics of binary data generated from Random Amplified Polymorphic DNA (RAPD) marker was conducted using numerical taxonomic and multivariate analysis (NTSYS-PC) package, while dendrogram was constructed by Jaccard’s similarity coefficient using unweighted paired group method of arithmetic mean (UPGMA).

Results: Accession Y3 from Ona-Ara yielded the highest total volume of DNA concentration (736.9 µ/l), while the highest genomic DNA concentration of 2.44 ng/ was recorded in accession L-04 from Agege.

Out of total number of 52 bands from three primers of RAPD, 48 produced polymorphic amplified products. OPO-08 primer was highly polymorphic with 94.73%, and had the highest allele numbers, gene diversity and polymorphic information contents of 16.0, 0.914 and 0.909 respectively, while OPO-06 produced the highest number of 20 polymorphic bands. Cluster II was the highest group in the dendogram, and comprised of two states (Oyo and Lagos) and Mali which constituted seven accessions; Y-03 (Ona-Ara), Y-04 (Egbeda), Y-05 (Ido), L-01 (Surulere), L-03 (Ifako-Ijaye), L-04 (Agege) and M (Mali).

Accession S-03 from Ife-North was the most distant with highest similarity index of 1.188.

Conclusion: The RAPD is highly polymorphic, and could be useful in characterizing and revealing wide range of genomic variation and phylogenetic relationship among different accessions of O. gratissimum with broad genetic base.

Open Access Original Research Article

Feasibilitiy Study of Bioethanol Production: A Case of Spent Mushroom and Waste Paper as Potential Substrate

H. O. Stanley, A. O. Erewa, C. N. Ariole

Biotechnology Journal International, Page 1-8
DOI: 10.9734/bji/2018/v22i330057

Aims: The aim of the present study is to produce ethanol from waste paper and spent mushroom using Saccharomyces cerevisiae as enzymes for the fermentation process.

Study Design: Waste paper and spent mushroom samples were subjected to fermentation and hydrolysis by Aspergilus niger and Saccharomyces cerevisiae to produce bioethanol.

Place and Duration of Study: This study was carried out in the Environmental Microbiology Laboratory, University of Port Harcourt, Nigeria, between May and October 2017.

Methodology: Waste paper and spent mushrooms samples were hydrolyzed by Aspergillus niger, and the hydrolysate from each set up subjected to fermentation by Saccharomyces cerevisiae. Ethanol was extracted by fractional distillation, and qualitatively determined by Gas Chromatography with Mass Spectrometry.

Results: After 8 days of fermentation, there was decrease in glucose content in waste paper hydrolysate ranging from (0.51-0.1 mg/l), and spent mushroom substrate (0.3-0.07 mg/l). Upon extraction of the bioethanol, the highest yield was recorded for waste paper hydrolysate which after characterization with GC-MS ethanol concentration was 28.01 mg/l, followed by spent mushroom hydrolysate 26.8 mg/l.

Conclusion: This study revealed that bioethanol can be obtained from fermentation of waste paper using Saccharomyces cerevisiae and ethanol can be obtained after the paper has been used in growing edible mushroom; if adopted, this could be a way to achieving environmental sustainability.

Open Access Original Research Article

Reproductive Toxicity and Biomarker Response to a Daily Dose of Tramadol in Male Albino Rats (Rattus norvegicus)

E. Oriakpono, Obemeata, M. Ajah, Ogechi

Biotechnology Journal International, Page 1-7
DOI: 10.9734/bji/2018/v22i330059

This study was designed to evaluate the effect of a daily dose of tramadol on selected biomarkers viz: haematological parameters, sperm count, kidney and liver damage in male albino rats. Twenty four wistar rats were divided randomly into two groups: control group and treated groups, the treated group were further divided into four groups and housed in cages. Clean drinking water was served to control (group 1), and 1.6 mg/kg bodyweight of tramadol was administered to group 2 (7 days treatment), group 3 (14 days treatment), group 4 (21 days treatment) and group 5 (21 days treatment +7 days withdrawal) in addition to a daily standard diet for all groups. Treatment of rats with tramadol caused significant decrease (P<0.05) in WBC, platelet and lymph. in group 2, on bicarbonate, AST and protein, it showed significant decrease (P<0.05) in group 3, and it showed significant decrease (P<0.05) in group 5 on Cl-, AST, ALT, bicarbonate, AST, PCV, Hb, RBC, WBC, platelet, lymphocytes and sperm count. The results indicates that tramadol has negative effects on the liver which may induce severe liver damage when used for a prolonged period, the results also shows that tramadol can cause anaemia as seen by the observed negative changes in the blood parameters evaluated. Therefore, administration should be with great caution and from a licensed pharmacist or doctor while self prescription or over the counter administration should be avoided considering the associated adverse effects.

Open Access Original Research Article

Preliminary Studies on Isolation, Bile Tolerance and Antibiogram of Potential Probiotics (Probionts) from Locally Fermented Food Products at Beach Market, Nsukka Metropolis, Enugu State, Nigeria

D. P. Berebon, K. C. Ofokansi, A. A. Attama, C. O. Eze, R. C. Onwusoba, I. C. Ugwoke

Biotechnology Journal International, Page 1-10
DOI: 10.9734/bji/2018/v22i330060

Aims: The study aims to isolate, evaluate bile tolerance and antibiogram studies of potential probiotics (Lactobacillus spp) from locally fermented Food Products (Akamu, Aqua Rafa® Yoghurt, Ogiri, Okpeye) and Kunu at Beach Market, Nsukka.

Study Design: A ten - fold serial dilution and spread plate method using De Man, Rogosa and Sharpe (MRS) medium was adopted for isolation of potential Probionts.

Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, University of Nigeria Nsukka, between March - September, 2018.

Methodology: Only catalase negative and Gram positive isolates characteristic of lactobacilli were used. Bile tolerance test was performed by monitoring the bacterial growth at different Bile salt concentrations (0.2%, 0.3% and 2%). The antibiogram of the isolates was assessed using the Kirby-Bauer disc diffusion method against commercial antibiotic discs of ampicillin, vancomycin, gentamycin, ciprofloxacin, methicillin and erythromycin.

Results: All the 18 screened isolates were tolerant to bile salt at 0.2 % and 0.3 % with inhibition of growth at 2 % bile concentration. All isolates were observed to be resistant to methicillin (100 %) but very sensitive to gentamycin (11%) and ciprofloxacin (22%) respectively. The isolates showed intermediate resistance to other antibiotics: vancomycin (33%), erythromycin (33%) and ampicillin (44%). The decreasing pattern of resistance was thus: methicillin > ampicillin > vancomycin and erythromycin > ciprofloxacin > gentamycin. Isolates from Yoghurt (66.67%) and Ogiri (53.33%) provided most of the resistant isolates. Methicillin would provide best antagonist potential as all the isolates exhibited very high level of resistance (100 %).

Conclusion: These results suggest that all the eighteen potential Lactobacillus spp strain show potential for probiotic applications and the locally fermented food products are rich sources of probionts.