Biotechnology Journal International
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Reproductive Performance of Carica papaya, Hibiscus rosa-sinensis and Ipomoea batatas on Female African Catfish (Clarias gariepinus)
This study investigated the reproductive performance of pawpaw (Carica papaya) seeds, hibiscus plant (Hibiscus rosa-sinensis) leaves and sweet potato (Ipomoea batatas) leaves on some reproductive parameters (ovary weight, mean egg diameter and egg fecundity) in female African catfish (Clarias gariepinus). One hundred and twenty (120) juveniles of C. gariepinus were collected from the University of Calabar fish farm. The 120 fish were randomly divided into 12 experimental tanks measuring 80x80x80 cm (L x W x H) using a completely randomized design (CRD). Three grams (3 g) of each test plant were incorporated into 1 kg of Coppens feed (3 g/kg) and reformulated into four experimental diets; Treatment A- Control, B- pawpaw seed meal, C- Hibiscus leaf meal and D- sweet potato leaf meal. The experiment was done in three replications. The fish were fed twice daily for 6 months. Data obtained were analyzed using a one way analysis of variance (ANOVA). Results obtained revealed that the different test substances significantly (p<0.05) negatively affected the different reproductive parameters studied. The ovary weight, gonadosomatic index (GSI), egg diameter, fecundity as well as total weight significantly (p<0.05) decreased in all the treated fish when compared with that of the control. Pawpaw seed meal (PSM) had the highest effect on the reproductive parameters of the fish studied (ovary weight, GSI, fecundity and egg diameter values of 14.89±5.51, 0.82±0.30, 19371±51.84 and 0.63±0.07, respectively) when compared to the other test plants. The findings of this study suggest that C. papaya seeds, hibiscus leaves and sweet potato leaves have the potential to impair reproductive performance in female African catfish. Therefore, holistic measures should always be taken when using these plants considering the effect it could exert on other aquatic inhabitants and systems
Effects of Hydrogen-Rich Water on the Growth and Photosynthetic Characteristics of Cucumber Seedlings
Aims: The aim of the paper was to study the effect and specific mechanism of hydrogen-rich water (HRW) improving cucumber (Cucumis sativus) seedlings growth.
Study Design: 14-day-old cucumber seedlings were treated with different saturation hydrogen-rich water in root for three times every three days. After one day of the last treatment, growth parameters were determined and plants tissues were sampled for test photosynthetic characteristics.
Place and Duration of Study: In 2017, cucumber (C. sativus ‘JinYan4’) were germinated in College of Bioscience and Biotechnology of Shenyang Agricultural University (Lab ChuangXin).
Methodology: 20 cucumbers and 3 replicates were determined for growth parameters. The determination of total soluble sugar and soluble protein content are the methods of Anthrone colorimetry and Coomassic Brilliant Blue. Chlorophyll content was tested by using ethanol immersion extraction method. Gas exchange parameters were measured with LI-6400XT portable photosynthesis system. Chlorophyll fluorescence parameters of leaves were measured by a Handy PEA Fluorometer.
Results: Our results showed that 50% saturation HRW significantly enhanced the growth and development of cucumber seedlings, including the improvement of fresh weight, plant height, stem diameter and leaf area. These responses were consistent with a significant increase of leaf water content, total soluble sugar content and soluble protein content, which was further confirmed by the determination of photosynthetic related parameters. Also, research results illustrated that HRW up-regulated chlorophyll content and changed chlorophyll fluorescence parameters of leaves. The increase of chlorophyll content promoted the absorption of light and enhanced plant photosynthesis. Furthermore, HRW changed the leaf stomata conductance and decreased transpiration so as to improve the photosynthetic rate.
Conclusion: Taken together, these results suggest that the effect of HRW on cucumber seedling was associated with plant photosynthesis. Therefore, the application of HRW may be a promising strategy to improve cucumber growth
Bacterial Community Profile and Phylogenetic Diversity of Water and Surface Sediments in Iko River Estuary, Akwa Ibom State- Nigeria
A large amount of crude oil is spilled annually into the terrestrial and aquatic environments in Nigeria. This tends to upset the natural equilibrium of the marine environment as a microbial habitat. Thus, there is a need to evaluate the effect of exposure to crude oil on bacterial load and diversity using Iko River and Odoro Ikot pond as the study and pristine sites respectively. The bacterial isolates were molecularly identified using the 16S rRNA sequencing protocol. The total heterotrophic bacterial count (THBC) in the surface water (SW), sub-surface water (SSW) and sediment segments of Iko River ranged from 2.23±0.87 to 9.67 ± 0.43 x 106 CFU/ML while the THBC in the SW, SSW and sediment segments of the pristine site (Odoro Ikot pond) ranged from 1.87±0.53 to 4.8± 0.04x106 CFU/ML. The sediment had a significantly higher (P<0.05) THBC than the water segments (SW and SSW) in both Iko River and Odoro Ikot pond. The hydrocarbon utilizing bacteria (HUB) count in Odoro Ikot pond ranged from 0.40 ± 0.01 to 1.10 ± 0.03 x 106 CFU/ML while the HUB count in Iko River ranged from 0.53 ± 0.02 to 0.93 ± 0.04 x 106 CFU/ML, making Iko River have a higher number of total heterotrophic bacteria and hydrocarbon utilizing bacteria than Odoro Ikot pond. The Proteobacteria isolates had the highest bacterial diversity (77%) while members of the Firmicutes phylum had a 23% bacterial diversity. However, higher bacterial count and diversity were obtained from the sediment segment than from the water segments in both Iko River and Odoro Ikot pond implying that the sediment is more favourable for bacterial growth. Although the bacterial profile was affected by exposure to crude oil, there were variations in the phylogenetic diversities obtained from the different water segments attributable to crude oil concentration. 
Screening of Thermotolerant Acetic Acid Bacteria Involved in Cocoa Fermentation in Six Major Cocoa Producing Regions in Côte d’Ivoire
Aims: This work intends to screen and to identify thermotolerant acetic acid bacteria with acetic acid production capacity at high temperature in cocoa beans fermentation from six cocoa producing regions of Côte d’Ivoire.
Study Design: Thermotolerant acetic acid bacteria were isolated from cocoa fermentation. These thermotolerant strains were biochemically characterized and tested for the production of acetic acid in culture medium.
Place and Duration of Study: This study was performed in Biotechnology Laboratory, University Félix Houphouët-Boigny (Côte d’Ivoire) from January to November 2017.
Methodology: Several strains of acetic acid bacteria were isolated from the traditional cocoa beans heap fermentation process occurred in six major cocoa producing regions of Côte d’Ivoire. These isolates were screened to select thermotolerant strains that were able to produce a good amount of acetic acid. Biochemical identification of thermotolerant acetic acid bacteria was carried out on the basis of biochemical characteristics analysis as acid production from ethanol, oxidation of acetate and lactate, ketogenesis from glycerol or mannitol, formation of water-soluble brown pigment, growth on different carbon sources and acid production from sugars and sugar alcohols.
Results: A total of 821 acetic acid bacteria strains were isolated from the cocoa beans heap fermentation of these six regions. Among them, 26 (31.15%) showed growth capacity at 45°C and six (6) grown at 50°C. These 26 strains displayed also acid production capacity at 35°C and at 45°C with acid amount ranged from 1.2 to 24.63 and 0.80 to 1.70 respectively. Biochemical analyses of these thermotolerant strains revealed that the isolates belong to three genera notably Acetobacter, Gluconacetobacter or Gluconobacter. Moreover, all strains were able to grow in medium containing 10% ethanol and to produce acid from various carbohydrates sources. In addition, strain T6HS14 displayed acetoin production capacity while 8 strains were able to produce brown pigment on Yeast extract-Ethanol-Peptone-Glucose medium.
Conclusion: This study highlighted the presence of thermotolerant acetic acid bacteria strains involved in Ivorian cocoa fermentation. Furthermore, some isolates displayed a diversity of technological properties which could be used for the improvement of cocoa fermentation process. These predictors, however, need further work to validate reliability
Effect of Herbal Supplement on the Laying Performance and Antioxidant Status of Serum and Egg Yolk of Laying Birds
Aims: The experiment was carried out to investigate the effect of herbal supplement on laying performance and antioxidant status of serum and egg yolk of laying birds.
Study Design: The experiment employed a completely randomised design, all data generated were subjected to analysis of variance, P =0.05.
Methodology: A total of ninety-six (26-week old laying birds) of Isa-brown laying birds were used in a 120-day feeding trial. The birds were assigned to 4 dietary treatments replicated eight times with 3 hens per replicate and were fed the control diet and diets supplemented with 5 g/kg garlic powder, 5 g/kg moringa leafmeal and 5 g/kg ginger powder as diets 2,3,4, respectively. The birds were fed with experimental diets for the 120-day period during which data were obtained on laying performance, internal and external egg qualities and antioxidant status of serum and egg yolk.
Results: There was no significant difference in hen day production, feed intake in birds fed control diet and diets supplemented with herbs. Final live-weight and weight change decreased (p<0.05) with dietary herbal supplementation. Supplementation of diets with herbs resulted in a significant increase in egg weight, eggshell weight, eggshell thickness, eggshell surface area compared to the control diet. The yolk weight and albumen weight were significantly (p<0.05) influenced by the dietary herbal supplement. Serum superoxide dismutase (SOD) activity increased in the dietary herbal supplement and was maximised (288 µml-1) at diet 3 likewise, the egg yolk superoxide dismutase (SOD) activity increased (p<0.05) in the dietary herbal supplement and was maximised (9.21 µml-1) at diet 3.
Conclusion: Diet supplemented with 5 g/kg moringa leafmeal gave optimum performance, 5g/kg moringa leafmeal may be used as feed additive to improve performance, egg qualities characteristics and antioxidant status of serum and egg yolk of laying birds
Decolorization of Synthetic Dyes by Ficus carica Latex Peroxidase Isoenzymes
Aims: The current study aims to elucidate the potential of Ficus carica latex peroxidase isoenzymes for decolorizing different synthetic dyes in comparison to the commercial horseradish peroxidase.
Study Design: The decolorization of 20 dyes was investigated using the purified F. carica latex peroxidase isoenzymes (purified FP1 and partially purified FP2, and FP3), and horseradish peroxidase (HRP) as a control.
Place and Duration of Study: Molecular Biology Department, Genetic Engineering and Biotechnology Research Division, National Research Centre, Egypt, between January 2017 and March 2018.
Methodology: The purified and partially fractions of peroxidase isolated from latex of F. carica were used for the present study. Stock solutions of the dyes were prepared in 0.05 M sodium acetate buffer (pH 5.5) and diluted to the requested concentrations ranged from 12 to 330 µM in order to get maximum absorbance does not exceed 1.5 as initial reading. The efficiency of decolorization was expressed in terms of percentage. All experiments were performed in triplicate.
Results: F. carica latex peroxidase isoenzymes and commercial horseradish peroxidase were able to decolorize some of tested dyes and the extent of decolorization achieved with different dyes classes were varied according to different chemical structure of each dye. The decolorization efficiency after 3 h of incubation at 40°C using 6.4 U/ml of peroxidase activity of FP1, FP2, FP3 and HRP, was found to be extremely efficient in decolorizing some dyes and relatively low in other dyes.
Conclusion: The efficiency of F. carica latex peroxidase isoenzymes toward different synthetic dyes meet the prerequisites needed for environmental and industrial applications
Immobilised Phytase Production from Aspergillus foetidus MTCC 11682 Using an Optimized Media
Aim: Immobilised fungal phytase production from the novel strain Aspergillus foetidus MTTC 11682 and optimisation of cultural conditions for a better and continuous economic yield.
Study Design: The study was designed based on the classical method of changing one independent variable while fixing all other at a certain level- one factor at a time, a close ended system for the optimisation of fermentation process.
Methodology: Physical and nutritional parameters were optimised for phytase production and subjected to statistical analysis. Adsorption and Entrapment techniques were employed to immobilise the production strain.
Results: The optimum physical conditions for augmenting the yield up to 6 days incubation period were as follows: pH of 3.5, 30ºC temperatures and 5% inoculum size. Amongst the nutritional parameters, lactose and sodium nitrate were found to be the best carbon and nitrogen sources. K++, Mg++, Mn++ and Fe++ ions supported the phytase production. TritonX 100 and tween 80 showed an inducing effect on the secretion of phytase enzyme. Immobilised fungal phytase production resulted in an increased yield of 32.5% with poly urethane foam (PUF) as the matrix. A scale up fermentation resulted in an activity of 52.7 FTU/mL for immobilised cells as compared to 25.5 FTU/mL by its free counterpart.
Conclusion: Phytase produced in an optimised media employing immobilised Aspergillus foetidus 11682 on poly urethane foam cubes exhibited better phytase activity, improved stability and long shelf life
Production of β-Mannanase by Penicillium italicum Subjected to Different Growth Conditions
Aims: The process parameters affecting enzyme production were optimized to ascertain the best optimal conditions for β-mannanase production by Penicillium italicum in solid state fermentation.
Study Design: Four stages of experimental processes were designed for this study. The first experiment, samples were withdrawn after 24, 48, 72, 96, 120, 144,168 and 192 h incubation. In second experiment, the fermentation media were incubated at different temperatures. In third experiment, the effect of different pH values on β-mannanase production was evaluated, while the fourth experiment described the supplementation of surfactants in mineral salt solution for β-mannanase production.
Place and Duration of Study: Microbiology Research Laboratory, Federal University of Technology, Akure Nigeria between September 2011 and March 2012.
Methodology: β-mannanase production was conducted using Locust Bean Gum (LBG) as the sole carbon source; moisten with mineral salt solution, and enzyme activity determined by dinitrosalicylic acid method, while protein content was determined by Lowry method.
Results: Maximum enzyme activity (146.389 U/ml) was observed after 72 h of incubation. Different surfactants were supplemented in the basal medium, and Sodium Dodecyl Sulfate (SDS) was observed to give the highest β-mannanase activity of 53.335 U/ml. Initial pH of the culture medium was optimized and a pH of 6.0 was found to support maximum enzyme activity (173.241 U/mg protein). The optimum incubation temperature was achieved at 35°C.
Conclusion: The results obtained provide information on optimal process parameters that might improve the yield of β-mannanase by P. italicum for better fish feed formulation, especially in the larval stages of fish fingerlings when the enzyme system is not efficient
Gene Action of Cassava Resistance Metabolites to Whitefly (Bemisia tabaci)
Several metabolites are linked to cassava resistance to whitefly. There is limited information however, on the mode of gene action of the metabolites associated with cassava resistance to whitefly (Bemisia tabaci). The objective of the study was to determine the combining abilities and mode of gene action of salicylic acid, antioxidative capacity, total phenolic content, flavonoid, tannin, peroxidase and protein of selected cassava genotypes. Ten genotypes were crossed in half diallel and the parental and 45 S1 progenies evaluated for nymph count, whitefly count, leaf damage and sooty mold at Namulonge in season two of 2016. In season one and two of 2017, the parental genotypes and their corresponding forty-five S1 progenies were evaluated in randomized complete blocks with two replications for Bemisia tabaci population and leaf damage and leaf metabolite content assayed. The results indicated highly significant (P<.001) differences among genotypes for general combining ability (GCA) to antioxidative capacity; (P<.05) for total phenolic content, peroxidase and protein and (P<.01) for salicylic acid, tannin and flavonoid. The specific combining ability (SCA)showed significant (P<.001) differences for salicylic acid; (P<.01) for antioxidative capacity and total phenolic content. The additive variance was significant (P< .05) for flavonoid, protein; (P<.01) for antioxidative capacity, total phenolic content, tannin and peroxidase. The dominance variance had high significance (P<.001) for salicylic acid; (P<.05) for flavonoid and (P<.01) for antioxidative capacity and total phenolic content. Although additive gene action was higher than non-additive, both were influencing most metabolites indicating complexity of inheritance. A critical evaluation is necessary when exploiting metabolite related traits in breeding for resistance to Bemisia tabaci
Mycorrhizal Diversity Associated to Liparis japonica (Miq.) Maxim. in China
Orchidaceae (Orchidaceae) is the second largest family of angiosperms. It’s the "flagship" group in plant protection. The existence of orchid plant is closely related to mycorrhizal fungi. The relationship between orchids and their symbiotic mycorrhizal fungi is a benefit to the protection and population restoration of orchids.
Aims: The research was aimed to study molecular identification about 15 strains of mycorrhizal fungi from 6 plots by rDNA ITS technology in order to understand and utilise the mycorrhizal fungi of Liparis japonica (Miq.) Maxim.
Study Design: The mycorrhizal fungi collected from different geographical locations were isolated and purified from the mycorrhizal fungi symbiotic with Liparis japonica in Northeast China, which were identified by rDNA ITS, meanwhile computed evolutionary distance and constructed the phylogenetic tree.
Place and Duration of Study: In 2017, the root segments of Liparis japonica were collected separately from Qianshan, Changbaishan, Gaoguan, Guanmenshan, Dongling, Daqinggou.
Methodology: Fifteen strains of mycorrhizal fungi collected from six plots were identified by rDNA ITS. Using DNAMAN software to analyse, the pairwise homology was compared by using the optimal global sequencing option. The evolutionary distances of fifteen strains were calculated by MEGA (Molecular Evolutionary Genetics Analysis) software package and their phylogenetic trees were constructed by neighbour-joining method.
Results: With primers ITS1 and ITS4, the 15 mycorrhizal fungi strains of rDNA ITS got about 600 bp length. ITS length was about 582-613 bp, in which ITS1 length was about 177-190 bp, and ITS2 length was 246-273 bp. The mycorrhizal fungi strains were highly homology separated from one plot, mostly above 90%. The plots from the south to the north were as follows: Qianshan, Guanmenshan, Gaoguan, Dongling, Changbaishan, Daqinggou in China. Fifteen strains after separated and purified were identified to be the Epulorhiza of Orchid Rhizoctonia blasted with Genbank. The homology of the strains gradually decreased affected by the difference of the north and the south, namely there was an increasing trend of diversity from south to north.
Conclusion: The homology of mycorrhizal fungi from one plot was higher because of the same soil environment and climate environment and so on, and strain type was single. Under the influence of microclimate in Northeast China, the homology of strains decreased gradually in the sample area, that is, the diversity gradually increased from the south to the north