Biotechnology Journal International
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Actinobacteria: Versatile Microorganisms with Medical and Pharmaceutical Application
No full textThe Actinobacteria receive much attention, since they produce a variety of metabolites, including antibiotics and enzyme inhibitors. These bacteria are distributed in various habitats, including soil, ocean, extreme environments, lichen, plants, and animals. The classification of Actinobacteria based upon the morphological observation, physiological and biochemical characteristics were not adequate to differentiate the genera of this phylum. Following, another identification was available based in the distribution of specific constituents from the cellular wall, such as diaminopimelic acid and carbohydrates. With the advent of molecular biology, the identification of genera and species was more reliable. The screening of microbial natural products has become an important route to discover new bioactive compounds in order to develop new therapeutic agents. Actinobacteria remains one of the leading producers of biopharmaceuticals; endophytic Actinobacteria also yield secondary metabolites with wide range of biological activity. This review focus on gathering relevant information on identification, classification, chemical diversity of Actinobacteria, as well as reveals some biotechnological applications of these bacteria.  
Phytochemical Screening and Antimicrobial Activity of Bryophyllum pinnatum Extracts
No full textThe hexane, ethyl acetate and methanol extracts of the wood and stem bark of Bryophyllum pinnatum were investigated for their phytochemical constituents and activity against selected microorganisms. Phytochemicals found present were reducing sugars, saponins, steroids, tannins, alkaloids, flavonoids and phenols. The test microorganisms were Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Shigella dysenteriae, Salmonella typhi, Pseudomonas aeruginosa, Candida albicans, Aspergillus fumigatus, Aspergillus niger, Microsporum spp and Trichophyton rubrum. The Ethyl acetate extracts were the most effective against S. aureus, E. coli, P. aeruginosa and K. Pneumonia (MIC 5.0 mg/mL) and S. dysenteriae, C. albicans, Microsporum spp, and T. rubrum (MIC 10 mg/mL)
Effects of Input Variables on the Conversion of 5 Ton/h Processed Tomato Juice in a Triple-Effect Evaporator
No full textTomato is grown in many regions of the world; and processed tomato products account for 80% of total tomato consumption. A worldwide tomato processing industry has since evolved with a range of technologies which are able to convert tomatoes into products such as paste, sauces and ketchup. Tomato concentrate production can be classified as either Hot Break (HB) process or Cold Break (CB) process. CB process involves chopping the fresh tomato suitable for processing at lower temperatures, ranging from 65 to 75°C, resulting in a less viscous, but slightly better flavoured paste which is mainly used for triple concentrate paste, packed for domestic use. The heated tomato pulp is then conveyed via a special pump to an extraction unit. Two products, the refined juice for concentrate and the waste for disposal come out of the extraction unit. The yield from extractor (about 95%) varies according to the variety of tomatoes treated, pulp’s temperature, type of sieve fitted, the rotation speed, and shape of the rotor on each operating stations. The refined juice is collected and constantly feed to an evaporator from where it passes through different effect(s) where its concentration level is gradually increased until the required concentration is obtained.
This study is aimed at using Matrix Laboratory (MATLAB®) to model a triple-effect evaporator for a 5 ton/h processed tomato juice initially at 5% concentration to 35% tomato paste concentration. The steam economy (SE) was reduced from 3.7 at an interval of 0.4 to study its effect on the outlet tomato paste concentration. Based on some thermodynamic assumptions, the effect of varying some input variables on the output parameters was studied. Other values of SE were later assumed and the process repeated until the desired 35% tomato paste concentration was obtained. The desired concentration was obtained in the third effect when a steam economy of 2.1 was used. With this SE of 2.1, the concentration of tomato paste obtained in first effect is 11%; second effect is 7% while the target concentration of 35% is obtained in the third effect. It was concluded that most of the output parameters, such as tomato paste concentration; water removed; volumetric flow rate; and area decrease with increasing SE; only mass of flow rate increases with increasing SE. Also, the tomato paste concentration increases as the number of effect increases; with a tomato paste concentration of 35% obtained in the third effect. The choice of SE and the number of effects should be properly considered before the design of tomato paste production facility
Screen House and Field Resistance of Taro Cultivars to Taro Leaf Blight Disease (Phytophtora colocasiae)
No full textIntroduction: Taro leaf blight disease cause by Phytophtora colocasiae has become an economic disease in Cocoyam growing regions of Cameroon.
Aims: To screen for resistance 10 improved and 4 local cultivars of taro against taro leaf blight disease.
Study Design: A randomized complete block design study.
Place of Study: Studies were conducted at the International Institute of Tropical Agriculture (IITA) Yaounde Nkolbisson from July 2013 to January 2014.
Methodology: Taro cultivars from tissue culture were planted in the screen house conditions and tested for virulence and pathogenicity with 4 isolates of Phythophthora colocasiae at spore density of 3×104 spores /ml of distilled water. Plants were planted in the field to assess disease incidence and severity.
Results: The results obtained on the different taro cultivars, revealed that all the 4 isolates showed variable pathogenicity. They caused lesions on inoculated leaves. There was variability in pathogenicity based on the small lesion lengths produced on cultivars, these included BL/SM132 and Red petiole. Isolate 3 showed a stronger sensitivity to leaf collapse and defoliation irrespective of the cultivar tested. There was a significant difference (p = 0.05) in tissue collapse and leaf defoliation on exposure to the different fungal isolates. The result of field infection rates of P. colocasiae at 126 DAP-154 DAP on 10 improved and 4 local cultivars indicated that there was significant variability (p = 0.05) in incidence and disease severity, with high incidence and severity occurring at 154 DAP in all cultivars. Improved cultivar BL/SM132 showed no classic symptoms of P. colocasiae and therefore it was resistant to Phytophthora colocasiae.
Conclusion: The results obtained on virulence and pathogenicity of Phythophthora colocasiae on the different taro cultivars revealed that all the 4 isolates showed variable pathogenicity. They caused lesions, on inoculated leaves. Isolate 3 showed a stronger sensitivity to leaf collapse and defoliation irrespective of the cultivar tested. The result of field infection rates of P. colocasiae at 126 DAP-154 DAP on 10 improved and 4 local cultivars indicated that there was a significant variability (p = 0.05) in disease incidence and severity, with high incidence and severity occurring at 154 DAP in all cultivars. Improved cultivar BL/SM132 showed no classic symptoms of P. colocasiae and therefore it was resistant to Phytophthora colocasiae as compared to all the other cultivars which showed high severity rates of infection of the disease and thus were susceptible to the disease
Waste Oil Biodegradation Potential among Bacteria Isolates from Waste Oil and Pristine Soils
No full textThe research study was aimed at investigating waste oil biodegradation potential among bacteria isolates from waste oil and pristine soil samples. Auto-mechanic workshop and pristine soil environments were randomly sampled within Calabar Metropolis. The study was completed within a period of six months. Standard microbiological methods were used to isolate, characterize and identify bacteria from the collected soil samples, while waste oil biodegradation potential of the bacteria isolates were determined using screen test for used engine oil and hydraulic oil utilization by bacteria isolates from both waste oil polluted and pristine soil. The level of turbidity, degree of precipitation and colour of precipitate were used to assess the biodegradation ability of the bacteria isolates. The total heterotrophic bacteria counts from the waste oil polluted soil samples ranged from 3.16x106 to 4.15x106cfu/g while that of the pristine soil sample ranged from 1.91x105 to 9.12x105cfu/g. Used engine oil biodegraders from the waste oil polluted soil were identified as Serratia spp, Enterobacter spp while efficient used engine oil and hydraulic oil biodegraders were identified as Shigella spp, Corynecbacterium spp, Klebsiella spp, Yersinia spp, Serratia spp and Aeromonas spp, Bacillus spp and Pseudomonas spp, while Serratia spp and Yersinia spp from the pristine soil also showed used engine oil biodegradation potentials and Bacillus spp was identified as efficient used engine and hydraulic oil biodegraders. However, it was obvious from the study that waste oil degraders and their proportion within an area appear to be marginally higher in environment with waste oil exposure and pollution as confirmed by the biodegradation potentials of bacteria isolates from the auto-mechanic workshop investigated, this therefore calls for the optimization of the process as it could serve as a more eco-friendly approach for the bioremediation of waste oil contaminated sites. 
Current Opportunities and Challenges of Next Generation Sequencing (NGS) of DNA; Determining Health and Diseases
No full textMany publications have demonstrated the huge potential of NGS methods in terms of new species discovery, environment monitoring, ecological studies, etc. [24,35,92,97,103]. Undoubtedly, NGS will become one the major tools for species identification and for routine diagnostic use. While read lengths are still quite short for most existing systems ranging between 50 bp and 800 bp, they are likely to improve soon. This will enable easier, faster, and more reliable contig assembly and subsequent matching against reference databases. When data generation is no longer a bottleneck, the storage, speed of analysis, and interpretation of DNA sequence data are becoming the major challenges. Also, the integration or the use of data originating from diverse datasets and a variety of data providers are serious issues that need to be addressed. Poor sequence record annotations and species name assignments are known problems that should be instantly addressed and would allow the creation of reference databases used for routine diagnostics based on NGS. Samples with huge amounts of short DNA fragments need to be analyzed and compared against reference databases in an efficient and fast way. Although a number of solutions have been proposed by Industry; offering commercial software, there still remain hurdles to take. One of the challenges that we need to address is data upload from client’s computers to central or distributed data storage and analysis services. Another one is the efficient parallelization of analyses using cloud or grid solutions. The reliability and up-time of storage and analyses facilities is another important problem that need to be addressed if one wants to use it for routine diagnostics. Finally, the management, reporting and visualization of the analyses results are among the last issues, but not the least challenging ones. Considering the constant growth of computational power and storage capacity needed by different bioinformatics applications, working with single or a limited number of servers is no longer realistic. Using a cloud environment and grid computing is becoming a must. Even single cloud service provider can be restrictive for bioinformatics applications and working with more than one cloud can make the workflow more robust in the face of failures and always growing capacity needs. In this white paper we review the current state of the art in this field. We discuss the main limitations and challenges that we need to address such as; data upload from client’s computers to central or distributed data storage and analysis services; efficient parallelization of analyses using grid solutions; reliability and up-time of storage and analyses facilities for routine diagnostics; management, retrieving and visualization of the analyses results
Epidermal Growth Factor Receptor Regulates Matrix Metalloproteinase-2 Activity in MDA-MB-231 Human Breast Cancer Cells
No full textAims and Study Design: Overexpression of epidermal growth factor receptor (EGFR) and its phosphorylated form (p-EGFR) in breast carcinomas has been associated with an increase in invasive potential and decreased survival. EGFR mediated signal transduction has been reported to be involved in regulation of matrix metalloproteinases (MMPs). Elevated MMP-2 expression and activity shows significant correlation with increased invasive potential in breast cancer. As MMP-2 plays a crucial role in tumour invasion, the role of EGFR in regulation of MMP-2 expression and activity in breast cancer was studied using the human breast adenocarcinoma cell line MDA-MB-231 as a model.
Methodology: MDA-MB-231 cells were cultured on 1 µg/ml epidermal growth factor (EGF) coated culture dishes for 24 hours. Control cells were cultured without EGF. MMP-2 activity was assayed by gelatin zymography. Expression of EGFR, MMP-2, focal adhesion kinase (FAK) and mitogen activated protein kinase (p38MAPK) and phosphorylation of EGFR were assayed by Western blot. Results: When MDA-MB- 231 cells were cultured on EGF, increased activation of MMP-2 and an overall increase in MMP-2 expression and activity was observed. The observed upregulation of MMP-2 was appreciably inhibited if cells were pre-treated with anti-EGFR antibody, thus blocking EGFR. Phosphorylation of EGFR and expression of p38MAPK and FAK were appreciably increased upon culture of cells on EGF.
Conclusion: In MDA-MB-231 breast cancer cells, EGFR-EGF interactions promote activation of MMP-2 and an increase in MMP-2 expression and activity via EGFR mediated signal transduction cascades involving FAK and p38MAPK. As elevated MMP-2 expression and activity correlate with tumour aggressiveness, tumour cell EGF interaction via EGFR might increase the invasive potential of breast cancer cells
Biddulphia (Odontella) sp. Isolated from the Seawater of Kayalpattinam Coastal Area, Gulf of Mannar and Maintained in Tuticorin Research Centre of ICAR-CMFRI
No full textAim: Present study was aimed to isolate marine algae from the sea water samples of Kayalpattinam Coastal area, Gulf of Mannar.
Methodology: Seawater samples were collected for the isolation of phytoplankton from the Kayalpattinam Coastal area, Tuticorin on May 2016. The collected seawater was maintained in Walne medium with optimum conditions (light intensity of 1000 Lux, pH of 8.2-8.7, temperature of 18-25°C and salinity of 35 ppt) for obtaining maximum growth. After the visible change observed, microscopic observation was carried out.
Results: Microscopic observation revealed that the sample contained Biddulphia (Odontella) sp. which is also known as Chinese Diatom. There were significant numbers of the species. It is further confirmed by the morphological identification. It has been maintained in phytoplankton repository of Tuticorin Research Centre, ICAR-CMFRI for sale and research. Cells are observed either in isolated or in the colony which are organized in straight lines or in a zigzag pattern and they are yellow/brown in colour. The size is measured and varied from 25 to 40 µ.
Conclusion: Biddulphia (Odontella) sp. has been widely used as a feed for zooplanktons in aquaculture industry. This is the first report of method to isolate and cultivate this species. Further more research is needed in order to investigate the potential utilization of this species in medicinal, nutritional and other possible useful properties of Biddulphia (Odontella) sp
Isolation of a Major Antimicrobial Compound from Stem Bark of Glossonema boveanum (Decne)
No full textMedicinal plants have been used as traditional remedies for the treatments of different ailments, and large number of human population still depends on these medicinal plants for their preventive and curative properties Glossonema boveanum is a medicinal plant widely used in a folk medicine in both West and North Africa. Column chromatography of the ethyl acetate extract from stem bark of Glossonema boveanum yielded one pentacyclic triterpenoid (betulinic acid) which is reported for the first time in this plant. The structure of the isolated compound was identified on the basis of 1D NMR (1H, 13C and DEPT), 2D NMR (COSY, HSQC, HMBC and NOESY) and by comparison with reported data. The compound was then tested for its antibacterial activity against two Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and three Gram-negative bacteria (Salmonella typhi, Shigella dysenteriae and Escherichia coli) and was found to be effective against the bacteria. The result of the MIC revealed that the lowest concentration of betulinic acid that inhibited the growth of the bacteria was 3.125 µg/ml for S. aureus and S. dysenteriae. This study concluded that the stem bark of G. boveanum, used traditionally as a medicinal plant for the treatment of typhoid fever, bacillary dysentery, diarrhoea and stomach pain has antibacterial activities against the causative bacteria
Apoptotic Gene Expression in Sheep Hepatocytes during Fasciola hepatica Infection (Fascioliasis)
No full textAims: The main objective of this work was to investigate the apoptotic genes of sheep liver hepatocytes to elucidate the apoptosis pathway mechanisms during Fasciola hepatica infection using molecular and serological techniques.
Study Design: This is a laboratory based study whereby F. hepatica infected liver specimens were used.
Methodology: Total RNA was extracted from fresh-frozen liver tissue. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to investigate the genes encoding the mRNA for following proteins: 18S; Bax; Bcl-2; Caspase-3. Additionally, the concentration of total protein in each sample was estimated spectrophotometrically. Polyclonal antibodies were produced by immunizing rabbits and diluted with 5% skimmed milk in PBS containing 0.1% Tween 20 (1:1000). The detection of the reacted antigen/antibody products was performed using immunoblotting technique were used to assess the quantification of protein kinetic to apoptotic genes. Data obtained from this investigation were analyzed using Portable IBM SPSS 2006 Statistics software.
Results: We investigated the apoptotic gene expression in sheep liver hypatocytes during F. hepatica infection by mRNA expression of three genes involved in apoptosis; Bax, Bcl-2 and Caspase-3 and histopathological parameter after infection with F. hepatica in the liver cells. Quantitative real-time PCR, histological examination and immunoblotting were used to quantify apoptotic genes, histopathology and protein kinetic, respectively. F. hepatica infection induces apoptosis in the liver cells via Bax, Bcl-2 and Caspase-3 genes.
Conclusions: F. hepatica infection induces apoptosis in the liver cells via Bax, Bcl-2 and Caspase-3 genes and it precedes necrosis. Thus, this study suggests that the induced apoptotic gene expression was due to the outcome of F. hepatica