Open Access Short Research Article

A Rapid Microwave Method for Isolation of Genomic DNA and Identification of White Rot Fungi

G. Rao Ramya, A. Ravichandran, A. Dhali, A. P. Kolte, K. Giridhar, Sridhar Manpal

Biotechnology Journal International, Page 1-7
DOI: 10.9734/BJI/2018/42191

A 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.

Open Access Original Research Article

Low-dose Sekikaic Acid Modulates Host Immunity and Protects Cells from Respiratory Syncytial Virus Infection

Damian C. Odimegwu

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BJI/2018/40802

Introduction: Investigations of Ramalina farinaceae and its isolated compounds had reportedly shown appreciable anti-RSV activity albeit within a slim therapeutic window. Sekikaic acid represent one of such compounds. In this present study, sekikaic acid in very low dose was evaluated  as a cell restorative and immunomodulatory factor.

Methodology: The effect of sekikaic acid on the cell viability of RSV-infected HEp2 cells was investigated. Further assay for cellular immune response in primed mouse splenocytes was established by cell culture and flow cytometry.  Splenocytes were treated with graded concentrations of sekikaic acid (6.25-25 µg/ml) and screened for their effect on the expression of IFNγ and IL-2 and T-lymphocytes markers CD4+ and CD8+ using intracellular cytokine staining and Fluorescence-activated cell sorting (FACS) analysis.

Results: Results reveal that sekikaic acid protects RSV-infected Hep 2 cells from infection-induced cytopathology. Moreover, sekikaic acid displayed some degree of immune-regulatory activity in the primed mouse splenocytes increasing the proportion of CD8+/IFNγ+ (70.95-73.8%) and CD4+/ IFNγ+ (3.44-4.13%) T lymphocytes when compared to the cells in untreated controls 2.75% (CD4+) and 69.35% (CD8+) respectively. Interleukin-2 (IL-2) expression and signalling by T-Lymphocytes was selective but pronounced for CD4+ cells activation (sekikaic acid 2.67-3.6%, control 0.44%). Similar scenario was recorded for Intracellular IL-2 secretion by the T-lymphocytes.

Conclusion: Low-dose sekikaic acid protects RSV infected cells and lead to immune lymphocytes proliferation. This recorded T-lymphocyte-specific immune-modulatory property may contribute to explain in part the dynamics associated with the overall antiviral effect of Sekikaic acid, and may also find relevance as a necessary cellular immune response precursor to infection-associated disease management.

Open Access Original Research Article

GIS-Based Estimate of GHG Emission from Livestock in Anambra State of Nigeria

E. C. Chukwuma, C. O. Nwajinka, L. C. Orakwe, C. C. Odoh

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2018/40814

The 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.

Open Access Original Research Article

Screening of Thermotolerant Acetic Acid Bacteria Involved in Cocoa Fermentation in Six Major Cocoa Producing Regions in Côte d’Ivoire

Pelepo M. Coulibaly, Bernadette G. Goualié, Lamine Samagaci, Honoré G. Ouattara, Sébastien L. Niamké

Biotechnology Journal International, Page 1-15
DOI: 10.9734/BJI/2018/41147

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.

Open Access Original Research Article

Reproductive Performance of Carica papaya, Hibiscus rosa-sinensis and Ipomoea batatas on Female African Catfish (Clarias gariepinus)

P. B. Ekpo, U. U. Uno, C. M. Adilieje, A. J. Umoyen, F. O. Okey

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2018/42026

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.