Biotechnology Journal International
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Molecular Identification of Aspergillus flavus Using Inter Transcribed Spacers (ITS)
No full textInfections caused by Aspergillus have grown in importance in the last decade. Outbreaks which are caused by Aspergillus flavus appeared to be related to the single or closely related fungal strains. The fungi produce a compound known as aflatoxin which is reputed to be the most toxic and powerful hepatocarcinogenic compound. Most of the recent studies are mainly based on, the most abundance species in the genus. However, A. flavus is the second major reason for invasive aspergillosis and direct infections in most part of the world especially tropical countries. This study is focused on the molecular identification of A. flavus. For this purpose, spores were collected from air sample. Molecular identification of the fungi was done by using universal primers ITS -1F and ITS-4R. Phylogenetic tree demonstrated that the fungal spores collected from the air lie in the clad with Aspergillus, indicating its close relationship with the group
Effects of Sun and Oven Drying Techniques on Quality of Oil Produced from Chlorella vulgaris (Microalgae) Biomass
No full textThis study involved the molecular identification of Chlorella vulgaris microalgae obtained from Kaduna State University Fish Pond for biodiesel production potential. The DNA of Chlorella microalgae was extracted and subjected to PCR. The molecular weight of the PCR product obtained was 1.8kb using 18S rDNA primer sets and BLAST analyses revealed 95% identity with Chlorella vulgaris. The Chlorella vulgaris was cultured in open aquaria tanks at the Department of Biological Sciences, Nigerian Defence Academy. The biomass harvested was subjected to varying timings of sun and oven drying techniques (25-35ºC for 72hours and 60ºC for 12hours respectively) before extraction of oil from the biomass using solvent extraction method. The values for the density (0.854 and 0.867 cm3), specific gravity (0.875 and 0.876), acid value (0.414 and 0.384 mgKOH/g), saponification value (173.3 and 170.1 mgKOH/g), kinematic viscosity (5.200 and 3.870 mm2/g at 40ºC), flash point (114 and 115ºC) and cetane number (54.00 and 47.70) for the sun and oven dried biomass oil respectively were found to be in accordance with the ASTM standard values for biodiesel and fossil diesel. GC-MS analyses of the oil extracted using the two drying methods showed that the fatty acid profiling of the oil obtained from sun dried processed biomass had C14:0, C15:0, C16:0, C18:0, C18:1 cis9 and C22:1ω9 while the oven dried biomass oil had C14:0, C16:0, C19:0, C11:1, C18:1 cis9 and C22:1ω9. Drying methods, therefore, had influenced on the composition of saturated and unsaturated fatty acids. The oven dried biomass oil possesses high monounsaturated fatty acids when compared to sun dried biomass oil though the most important fatty acids (C14:0, C16:0 and C18:1) found in standard biodiesel were present in both. The results suggested that Chlorella vulgaris microalgae can be sustainably harvested for the production of biodiesel, both drying techniques can be employed for effective extraction but oven dried biomass oil was found to be of high quality because of the balanced in saturated and unsaturated fatty acid compositions and have an easy mode of operation but it required instrumentation
AYA and HANA: New Seedless Mandarin’s Triploid Hybrids Selected in Morocco
No full textSeedlessness is a major objective in Morocco Citrus improvement programmes. Triploidy has played an important role in the development of new seedless mandarin cultivars for fresh fruit market and has received increasing attention for the consumers. For that, many series of diploid crosses were assessed. At maturity, the fruits have been harvested, and small embryos were extracted from undeveloped seed and cultured on the medium of Murashig and Skoog supplemented with 1mg/l gibberellic acid according. Triploids seedling selected by flow cytometry analysis were budded on Troyer citrange and planted in INRA domain with spacing 6 m X 3 m. Tow new seedless selections of mandarins have been evaluated in a citrus breeding program at Morocco INRA: Hana and Aya that were compared with their female parent clementine Sidi Aissa. These promising seedless mandarins were identified by easy-peeling, juicy fruits, have a pleasant taste and are sweet like the Sidi Aissa Clementine. These clones, showing commercial interest, were described and registered in Moroccan official catalogue
Genetic Diversity of Ambrosia trifida L. as Revealed by AFLP Markers
No full textAims: Invasion of alien species has become a global problem. Many invaders appear to thrive even with low levels of sequence-based genetic variation, and genetic patterns might congruent with the spatial arrangement of populations.
Study Design: The genetic diversity of giant ragweed was investigated using amplified fragment length polymorphism (AFLP). There were 5 populations(100 samples), and the distance between the two populations was more than 20 kilometers along with the mainstream of Liaohe.
Place and Duration of Study: Samples were collected from Liaohe Basin. Experiments were done in the College of Biological Science and Technology, between February 2016 and March 2017.
Methodology: The distance between the two populations was more than 20 kilometers. We sampled five populations of Ambrosia trifida L. along Liaohe river in Liaoning province (north-east of China). Leaves for AFLP experiment were stored in discoloration silica gel. Genomic DNA was extracted following the CTAB protocol.
Results: The genetic distances varied with the geographic distances. However, with the influence of many different situations, and many modes of transmission, the discrepancy of genetic distance was reflected. There was low genetic diversity during earlier period of invasion diversity. After a period of invasion, genetic diversity gradually increased. Panshanzha number of polymorphic bands and percentage of polymorphic bands was 98 and 64.47% respectively. Manduhu was 93 and 61.16%, Juliuhe was 100 and 65.79%, Shifosi was 98 and 64.47%, Shuanganqiao was 127 and 83.55%. Shuanganqiao possessed the highest polymorphism.
Conclusion: The source of genetic variation of giant ragweed in Liaohe was gene flow of population. Shuanganqiao might be the spread center
Chemical Constituents of the Essential Oil Extracted from Nicotiana tabacum Leaves
No full textGas Chromatography-Mass Spectrometric (GC-MS) was used to investigate the chemical constituents of the essential oil from fresh leaves of Nicotiana tabacum L. The result revealed thirty six (36) compounds representing 100% of the essential oil characterized from the leaves of Nicotiana tabacum L. The major constituents are 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-ol 16.37%, P-xylene 12.37%, Cyclohexane 8.43%, Farnesol isomer a 3.08%, and 1-Naphthalenepropanol 3.08%
DNA Analysis of Ralstonia solanacearum Based on 270-bp PCR-amplified Fragment from the Lowlands and Highlands of Kenya
No full textAims: Bacterial wilt, caused by Ralstonia solanacearum, is a devastating disease resulting in tremendous losses of economic crops such as plants in the Solanaceae. Studies have shown that R. solanacearum is spreading from the lowlands to the highlands. This has seen increased need for understanding the genetic diversity of R. solanacerum strains common in these areas as a basis for better strategies in their control.
Methodology: Sixty-nine bacteria isolates obtained from various wilting plant hosts (Tomato, capsicum and potato) from 11 different sites in Nyeri, Nyahururu, Kirinyaga, Kiambu, Nakuru Murang’a and Embu were subjected to molecular analysis.
Results: All the bacteria isolates were confirmed to be R. solacearum following PCR amplification of about 270-bp fragment using specific primers OLI 1-F and Y2-R. Based on the targeted 16 S rDNA sequences using primers OLI 1 and Y2, the 69 bacteria isolates had 98 to 100% identity with other DNA sequences for R. solanacearum isolates deposited in the NCBI database. Analysis of genetic differentiation showed there were total of 26 haplotypes from the 11 studied populations. The total number of segregating sites in all populations was 225.
Conclusion: Through this study, it was realized that the main causative agent of bacterial wilt in potatoes, tomatoes and capsicum grown in the lowland and highland regions in Kenya is R. solanacearum. The isolates are in two main groups (Cluster A and B) that represent mainly the phylotypes I and II respectively
Biodegradation of Indeno (1, 2, 3 - c, d) Pyrene and Dibenzo (A, H) Perylene by Aerobic Heterotrophic Bacteria and Cyanobacteria in Brackish Water
No full textBiodegradation of indeno (1, 2, 3- c, d) pyrene and dibenzo (a, h) perylene; high molecular weight PAHs by isolates of aerobic heterotrophic bacteria, cyanobacteria and its consortium from Boodo Creek characterized with brackish water was monitored for 56 days using GC- MS. The initial concentration of indeno (1, 2, 3- c, d) pyrene in treatment with aerobic heterotrophic bacteria (AHB) was 0.09 mg/l, cyanobacteria (CB) was 0.08 mg/l; AHB+CB was 0.3 mg/l and the control, C was 0.12. Dibenzo (a, h) perylene had AHB 0.10 mg/l; CB 0.07 mg/l; AHB + CB 0.21 mg/l and C 0.23 mg/l. The quantity of the PAH’s monitored reduced to 0 on day 56 in all the treatment options though fluctuation in the quantity of the HMW PAH’s was observed throughout the period monitored for biodegradation. Biodegradation of indeno (1, 2, 3- c, d) pyrene and dibenzo (a, h) perylene did not vary with time (p > 0.05)
Opuntia Extractable and Non-extractable (NEP) Residues as Co-carbon Sources for Improved Extracellular Laccase Production: Partial Purification and Characterization
No full textAim: The study of laccase production by Trametes pubescens cultured on cladode extractive residues alone, and as co-carbon source with wheat, using submerged fermentation condition.
Place and Duration of Study: Biotechnology Laboratory, Durban University of Technology, Durban, South Africa.
Methodology: Plant extraction, submerged fermentation, enzyme activity and characterization techniques were utilized.
Results: The highest laccase activity was observed at 60-80% wheat: residue ratio combinations under submerged conditions with copper and xylidine supplementation. Partially purified enzyme fractions showed similar characteristics at different temperatures and pHs. There was good retention of relative enzyme activity at temperatures near 60°C, and pH stability from pH 4.0-6.0. At optimal culture conditions laccase activity was highest at 49.01±0.21 U/ml (80% wheat: 20% residue ratio), and lower at 10.02±0.51 U/ml (80% wheat:20% NEP residue). The optimum temperature for laccase fractions was 25°C and pH optimum at 5. Highest specific activity was 3.55 U/mg of protein for the 80% wheat: 20% residue ratio laccase extract.
Conclusion: These results show the potential of cladode extractable phenol residues as potential economic growth medium for laccase production, offering a new alternative use for this agro-industrial and/or laboratory by-product
A Rapid Microwave Method for Isolation of Genomic DNA and Identification of White Rot Fungi
No full textA rapid, effective and efficient method to identify the innumerable white rot fungal strains is of utmost importance. Mycelia of the unknown as well as know isolates of WRF, after alternative washing with TE buffer and sterile water, were suspended in TE buffer. Fungi in solution were then exposed to microwave. The crude extract contained genomic DNA which was extracted and amplified using ITS primers for further identification. Based on sequencing results the identity of known cultures was confirmed, while the unknown cultures were identified as Clitopilus scyphoides (AGUM004, BankIt2098576 MH172163); Ganoderma rasinaceum (AGUM007, BankIt2098576 MH172163); Schizophyllum sp (KONA001 BankIt2098576 MH172164; AGUM011 BankIt2098576 MH172165 and AGUM021 BankIt2098576 MH172166 respectively), Coprinellus disseminatus (BANG001, BankIt2098576 MH172167) and Lentinus squarrosulus (TAMI004, BankIt2098576 MH172167). The microwave method described for isolating quality DNA of WRF without further purification steps proved a novel method requiring less than ten minutes and minimized the chances of the presence of PCR inhibitors
GIS-Based Estimate of GHG Emission from Livestock in Anambra State of Nigeria
No full textThe need for improved livestock waste management in developing countries has been emphasized by various researchers. Consideration has been given globally to advocacy for a transition from fossil energy system to Renewable Energy (RE), based on several socio-economic and environmental advantages. Green House Gas (GHG) from agricultural sector of Anambra State, with appropriate waste management strategy such as Anaerobic Digestion (AD) will definitely mitigate Methane emission hazards and will as well supply the much needed energy for domestic and industrial uses. An investigation was carried out in this study to estimate the Methane Emission Potential (MEP) of the study area as a result of poor livestock waste management system practiced in the area. This was achieved by incorporating Geographical Information System (GIS) capability to the Methane estimation. The result of the study shows that two Local Government Areas (LGA) which are Anambra West and Anambra East were the least in MEP with virtually none and 0.000646 Gg/year respectively. Idemili North LGA with MEP of 0.017962 Gg/year was observed to be the highest in Methane emission. On incorporating spatial density mapping on MEP, Onitsha North and Idemili North LGA were identified as the highest risk LGAs, with MEP of 0.00018 Gg/year and 0.000272 Gg/year respectively. The study suggests that an improved livestock waste management system using AD should be incorporated in the study area especially in the high risk zones of Onitsha and Idemili North LGA