Biotechnology Journal International
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In vitro Pepper (Capsicum annuum L.) Anther Culture: Can be Affected Via Vitamins B?
No full textPepper (Capsicum annuum L.) is a vegetable in family Solanaceae and has a very important place and development potential in the field of plant breeding and biotechnology. It is at the forefront of haploid plant acquisitions in plant biotechnology, especially in the sense that it helps traditional wisdom. In vitro haploid studies have shown that the genetic structure is \u27recalcitrant\u27 and the rate of embryo formation is low during the vegetative stages of embryo formation and applications are being made to increase it. In this study, four different pepper cultivars and vitamin B compounds, which are thought to play an important role as antioxidants recently, have been used to see effect of vitamin B on pepper androgenesis. When the results of the study were evaluated, it was observed that not only differences were found in cultivars but also different vitamin B compounds added to the media were positively affecting androgenesis
Effect of Ad-libitum, Split and Restricted Feeding on Performance, Digestibility and Welfare of Broiler Chickens
No full textAn experiment was conducted to assess the performance, welfare and digestibility of broilers fed varying forms of feeding regimes. The objective of this work research was investigate the possibility of improving growth performances, digestibility and welfare of broilers using ad libitum, split and restricted feeding.
Formulated diet was given in four different forms Ad-libitum (Treatment 1) twice a day meal/split feeding (Treatment 2) thrice a day feeding (Treatment 3) 3 hours restriction feeding (Treatment 4). Experimental design was completely randomized design. 240 broilers were randomly allotted to four treatments with 6 replicates and 10 birds per replicate. Birds were weighed weekly to evaluate their performances, faeces was collected on day 42, to determine digestibility and rectal temperature was taken as a measure of bird’s wellbeing.
Results showed that there were no significant differences (p≥ 0.05) in performance of birds across treatments but values from birds fed Ad-libitum were higher. Rectal temperature (RT) differ at week 4, birds fed 3 hours restricted feeding had the highest RT (41.62°C) while the least RT were from birds fed Ad-libitum (41.20°C). Significant differences were obtained in digestibility and nutrient utilization of broilers with respect to feeding regimes. Crude protein (CP) digestibility was significantly higher in birds fed 3 hours restriction (81.91%) while birds fed Ad-libitum had the least (46.76%). The digestibility of crude fibre was higher in birds fed 3 hours restricted feeding regime (86.77%) the least was from birds fed thrice a day feeding regime (62.66%). A similar trend was obtained for digestibility of gross energy.
Ad-libitum feeding had the highest non-significant performance values but split feeding optimized feed and nutrient utilization thereby enhancing performance and minimizing nutrient waste at the same time
Enhancing the Performance of Three White-rot Fungi in the Mycoremediation of Crude Oil Contaminated Soil
No full textContamination of soils by petroleum hydrocarbon is on the increase, particularly in the oil producing areas of Nigeria. White-rot fungi have enzymes which are capable of turning these organic compounds into harmless substances. This work investigated the performance of three white-rot fungi (Pleurotus tuber-regium, P. ostreatus and P. pulmonarius) for the remediation of different concentrations (0, 1, 2.5, 5, 10 and 20% w/v) of crude oil contaminated soils with sawdust and poultry manure as bedding materials. Ten grammes of each of the white-rot fungi were separately inoculated in each bottle containing exhaustively cropped topsoil (200 g), rice straw (40 g) and wheat bran (20 g). Each treatment was conducted in three replicates and arranged in a 3 x 6 x 3 complete randomised design. The bottles (54) were incubated for zero, one and three months in a dark room, exposed to light, watered daily thereafter for twelve days for fruiting bodies to spring out and harvested. P. ostreatus had best agronomic performance and P. tuber-regium removed hydrocarbons and heavy metals more than either P. ostreatus or P. pulmonarius under similar experimental conditions. The yield and mycoremedial performance of the three tested white-rot fungi demonstrated potentials for cleaning-up petroleum hydrocarbon contaminated soil, but their performance reduced from 5.0% (w/v) crude oil substrate contamination
Production and Characterization of Halo Tolerant Alkaline Protease of Enterobacter aerogenes Strain ZB and Its Application
No full textAims: The present investigation was carried out to isolate a halo tolerant Enterobacter aerogenes strain ZB for the production of alkaline protease to degrade protein wastes from the tannery wastewater.
Study Design: Halo tolerant protease producing bacteria was isolated from the salt pan. It was used for the production of halo tolerant protease in submerged fermentation. The crude protease thus obtained was characterized and its protein removal efficacy was tested in tannery wastewater.
Place and Duration of Study: Smykon Biotech Private LtD, Nagercoil, Kanyakumari District, Tamilnadu, India between January 2017 and March 2017.
Methodology: The halo tolerant bacterial isolates were screened for the production of alkaline protease using skimmed milk agar plates. Then a potent bacterial isolate was selected for the production of halo tolerant protease in submerged fermentation. Crude protease was used treat tannery wastewater and the protein removal efficacy of the enzyme was evaluated.
Results: A halo tolerant strain able to grow at wide sodium chloride concentration was isolated from a salt pan and identified as E. aerogenes strain ZB. The isolate was the potential to produce protease which showed catalytic activity under high saline conditions. The E. aerogenes strain ZB protease activity was found to be maximum at pH 8.0 and enzyme activity was reduced at pH 10.0. The protease from E. aerogenes strain ZB was highly stable upto pH 8.0 and declined at higher pH. Enzyme activity was found to be maximum at 50°C and was stable upto 40°C. Enzyme was highly active in the presence of ions like Ca2+, Na+ and Mg2+ ions (0.005 M) and the enzyme activity was 101.3%, 109% and 102% respectively. This halo tolerant enzyme showed the protein degrading capacity in tannery wastewater at 1% and 5% enzyme concentrations.
Conclusion: This halo tolerant enzyme effectively removed proteinaceous waste from the tannery wastewater. A maximum protein removal was achieved at 5% enzyme concentration. The proteases produced by E. aerogenes strain ZB could have potential application in tannery wastewater treatment
Charge Transference Detection through Cyclic Voltammograms in the Interaction of Cratylia mollis Seed Lectin-carbohydrate Inhibitor
No full textAims: To evaluate the charge transference and the electrochemical potential of Cratylia mollis seed lectin, Cramoll 1,4, adsorbed on Nafion beads after the interaction of glucose ligand.
Study Design: Based on detection of electrochemical currents and potentials of Cramoll 1,4 by electrochemical techniques.
Place and Duration of Study: Department of Chemistry, Catholic University of Pernambuco, between April 2014 and November 2015.
Methodology: Cyclic Voltammetry (CV) was performed in an electrochemical cell containing three electrodes (a calomel electrode, a platinum wire counter electrode and a platinum electrode), connected to a potentiostat to obtain electrochemical currents related to the charge transference. An electrochemical cell containing a calomel electrode and a platinum electrode coupled to a multimeter was used to register the potentials. A saline solution was used as support to control the charge distribution inside of the cell. Cramoll 1,4-glucose interaction was evaluated in the concentration of 100, 200 and 300 mM of glucose.
Results: CV measurements showed significant charge transference after Cramoll 1,4-glucose interaction. Cathodic and anodic peaks paired near 100 mV were detected in the range 100-300 mM glucose, achieving a maximum current response of 1300 μA, approximately. Positive electrochemical potentials of Cramoll 1,4 adsorbed to Nafion-beads was achieved showing a linear behaviour with the increase of glucose concentration at 300 mM.
Conclusion: The system is useful for characterization of lectin-carbohydrate interactions and as a glucose sensor to estimate the activity of lectins
Attempt to Establish Direct Gene Transformation System to Seeds of Sweet Potato (Ipomoea batatas) Using Electroporation Method
No full textAims: The purpose of this study is to get transformants by using the seeds of sweet potato and the instrument of NEPA21, an electroporation method.
Study Design: As the first step, we used grafting method to get seeds from the combinations of Ipomoea crassicaulis cv. [Kidachiasagao] as stock and I. batatas (L.) Lam varieties as graft. Then, we used an electroporation method with instrument of NEPA21 to get the GUS transformants by using direct gene introduction into the seeds.
Place and Duration of Study: Faculty of Environmental and Horticultural Science, Minami Kyushu University, between September 2014 and December 2016.
Methodology: The materials for grafting between Ipomoea crassicaulis var. [Kidachiasagao] as stock and I. batatas (L.) varieties as graft to get seeds, were provided by Kyushu-Okinawa Agricultural Research Center (KOARC), and they were cultivated in growth chambers at Minami Kyusyu U. For the judgment of crossing incompatibility (CI) among the varieties of sweet potato, the standard of KOARC was adopted. For crossing experiment of sweet potato, the flowers of any two varieties were used for pollination, and the performance was stopped before 10 am. The seeds were harvested when they matured.
For the transformation of GUS into sweet potato, the seeds obtained from [Koganesengan] were used in this study. As the seeds were wrapped with hard skin, they were damaged with knife to absorb water easily. Then, the seeds were placed onto wetted filter papers laid in the dishes for 3~4 days. When the seeds germinated, they were used for transformation according to the procedures of NEPA21. After the process was finished, the container was placed in dark condition for 2 days at 25℃, and then, dyed with GUS dyeing solution.
Results: 1) According to the standard of selection for varieties having crossing ability based on the result of CI conducted by KOARC, we selected 3 varieties of [Koganesengan], [Narutokintoki] and [Beniazuma] out from 6 ones held; 2) According to the standard of selection suitable to be used as either for seed or pollen we have obtained mature seeds successfully from the 3 varieties; 3) Using the sweet potato seeds collected from [Koganesengan] and electroporation have made GUS transformant of [Koganesengan] with GUS gene expression, for the first time, detected by GUS dyeing with blue color which appeared on the surfaces of the seeds and young buds of the transformants.
Conclusion: The results obtained in this study, provide a practical sweet potato transformation system by two steps: 1) Using the grafting between [Kidachiasagao] and sweet potato can consistently obtain mature seeds of sweet potato according to the judgmental standard for CI provided by KOARC, and 2) Using the combination of seeds of sweet potato and electroporation instrument of NEPA21, can get GUS transformants of sweet potato by using the direct gene transformation system. The establishment of GUS direct gene transformation system in seeds of sweet potato can provide a powerful tool for the ASG-1 gene transformation into sweet potato, which is considered as a crop, difficult to be cultured for plant regeneration. Furthermore, the system can be expected to open the way for all of the seed-set crops with a simplified and practical method to produce transformants.  
Assessment of the Influence of Transgenic Cotton on Beneficial Soil Rhizosphere Microbes
No full textIn this study the impact of Bt cotton has been accessed on the population of beneficial microbes in the rhizosphere of transgenic Bt cotton line (RCH-2 BG II) and its counterpart (a non-Bt cotton line). The Bacillus thuringiensis (Bt) proteins i.e., Cry1Ac and Cry2Ab were released from root exudates of Bt cotton (RCH-2 BG II) was confirmed by qualitative Enzyme-Linked Immunosorbent Assay (ELISA). Rhizosphere soil samples were collected in fifteen days interval i.e., 15, 30, 45 and 60 days after sowing (DAS). Azospirillum brasilense, Bacillus megaterium, Pseudomonas fluorescens population was more on 15th day in non Bt rhizosphere than Bt except Trichoderma viride, where as in 45th and 60th day all four microbes were more in Bt cotton (RCH-2 BG II) rhizosphere than its counterpart RCH-2. The statistical analysis of the data obtained from the pot cuture showed no significant differences in the numbers of CFU of Azospirillum brasilense, Bacillus megaterium (biofertilizer), Pseudomonas fluorescens and Trichoderma viride (biocontrol agents) between rhizosphere soil of Bt and non-Bt cotton. Restriction enzyme analysis of 16S rRNA of Azospirillum brasilense, Bacillus megaterium (biofertilizer), Pseudomonas fluorescens and ITS region of Trichoderma viride obtained from Bt and non Bt cotton rhizosphere in different intervals showed similar banding pattern between them
Optimisation of Growth Factors for Effective Use of Phosphate Solubilizing Bacterial Strains and Its Use as Bioinoculants for the Growth of Groundnut (Arachis hypogaea) Plant
No full textPhosphorus (P) is one of the essential macronutrients for plant growth. Phosphate solubilizing bacteria (PSB) are organisms which are used as bioinoculants to enhance the plant growth. Plants take phosphate in the form of soluble orthophosphate ions but due to the presence of calcium, magnesium, potassium, sodium, aluminium and ferrous ions in soil, the soluble orthophosphate is converted in to insoluble form. Because of this process plants utilize very little amount of phosphate, even though phosphorus containing fertilizers are added to the plants. Phosphate-solubilizing bacteria (PSB) has been added as fertilizer to increase phosphorus uptake and plant growth. It is usually observed that the solubilization of phosphate by the phosphate solubilizing bacteria would drop the pH of the medium. Acidification of the medium may be due to production of organic acids by the bacterial strains. Such plant growth-promoting bacteria has the ability to produce enzymes such as Phosphatase and 1-aminocyclopropane-1-carboxylate (ACC) deaminase to lower plant ethylene levels, that often results in various stresses, which shows the efficacious functioning of these bacteria. This study aims in the optimization of growth factors of the isolated PSB strains for use as a Biofertilizers, specifically to study the effects of temperature, pH and different carbon, nitrogen sources and NaCl concentrations on phosphate solubilization ability. The results showed phosphate solubilization was expressed maximum at pH 6, temperature 25 °C, dextrose as carbon source, ammonium sulphate as Nitrogen source and (1.2%) NaCl concentration by the PSB strains. Further the PSB was applied as bioinoculants to enhance the root, shoot length and phosphorus accumulation content of ground nut (Arachis hypogaea) seedlings. Present study highlights the importance of these plants growth promoting bacterial strains and their uses for agriculture purposes as a Biofertilizer
Prokaryotic Diversity of the Alkaline Lake Acıgöl, Turkey by Using Culture-dependent and Culture-Independent Methods
No full textAims: The prokaryotic diversity of Acıgöl lake, Turkey, was studied in samples taken from soil, water and mud. Culture -dependent and -independent methods were used to analyse the results.
Methodology: Bacterial cultures were isolated by using six different growth media. Isolates were identified according to their 16S rDNA sequence analysis. Liquid media were used to test antibacterial substance production of the isolates. Bacillus cereus, Serratia marcescens, Pectobacterium carotovorum, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, Micrococcus luteus, Escherichia coli and Listeria innocua were used as indicator bacteria. The extracted DNA samples taken from soil, water and mud were used in PCR reactions to amplify 16S rDNA genes by using universal bacterial primers. Amplicons were cloned by TA cloning after which white colonies were selected and colony PCR were applied for the amplification of the fragments. Each amplicon obtained from clones was sequenced and similarities were compared to those of the other bacterial groups. The DNA extracted from water sample was also used to amplify archeal 16S rDNAs. For the amplification reaction, the primers used were specific to the archaeal domain and their amplicons were cloned by TA cloning. Randomly picked white colonies were used for M13 PCR. The sequences of amplicons were analyzed by BLAST to determine the highest similarities with the other archeal groups.
Results: In this study, fourty-nine strains were identified as belonging to Bacillus, Halomonas Enterococcus, Exiquobacterium, Piscibacillus, Haloalkalibacillus, Aerococcus, Acinetobacter, Lysinibacillus, Oceanobacillus, Planococcus, Micrococcus genera according to 16S rDNA analysis. Among these isolates, the GBA17, GBA34 and GBA36 weakly inhibit the growth of M. luteus. The analysis of 16S rDNA from different environmental samples suggests that the bacterial clones belong to the class of Gemmatimonadetes (n=1), Alphaproteobacteria (n=4), Flavobacteria (n=1), Opitutae (n=2), Gammaproteobacteria (n=1), Saprospira (n=1), Verrucomicrobia (n=1). All of the identified archeal clones were affiliated to the Class Halobacteria with 12 members belonging to the Halobacteriales order and two members belonging to the Haloferaceles order. No match was found between the taxa determined by culture-dependent method and those determined by independent method. Growth media used in the isolation experiments might not support the growth of extreme halo-alkaliphilic bacteria found as a result of cloning.
Haloalkaliphilic and alkaliphilic isolates obtained in our study may be ideal candidates as industrially important substance producers
Nutritional and Quality Characteristics of White Maize Ogi Flour Enriched with Moringa oleifera Seed
No full textAim: Aim is to investigate the effect of boiling and fermentation of Moringa oleifera (M. oleifera) seed on nutritional quality of enriched maize ogi flour.
Place and Duration: The study was carried out in the Department of Food Science and Technology, Obafemi Awolowo University, Ife, Nigeria, between May-December, 2014.
Methodology: Raw and treated M. oleifera seed were wet milled together with fermented maize in the ratio 80:20 and sieved to obtain enriched ogi slurry. The slurry was dewatered, dried and milled to obtain ogi flour. Ogi prepared from 100% maize was used as control.
Results: The results showed significant increase (P=0.05) in protein (9.73 - 10.77%), ash (0.67 - 0.91%), and fat (10.84 - 12.34%) contents of the enriched products as compared with control (6.58, 0.53, and 5.05% for protein, ash and fat respectively). Fermentation and boiling of the seed reduced the fat content but boiling indicated the highest protein content while fermentation resulted in highest ash content. Fermentation improved P, Ca, Na and Fe (195.60, 21.80, 31.35 and 4.57 mg/100g) content of the enriched ogi. Results of sensory evaluation showed that sample with fermented M. oleifera seed was not significantly different (P=0.05) from the control.
Conclusion: The study shows that enriching white maize ogi with M. oleifera seed improves nutrients quality while fermentation and boiling lower the antinutritional factors without adverse effect on sensory attributes