Biotechnology Journal International
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Screening and Characterisation of Keratin-Degrading Bacillus sp. from Feather Waste
No full textAim: This study focuses on the screening and characterisation of keratin-degrading Bacillus species from feather waste.
Methods: Nine bacteria were isolated from feather waste obtained from a poultry layout at Egbeda local government secretariat, Ibadan, Nigeria. These bacteria were grown in basal medium with feather as primary source of carbon, nitrogen, sulfur and energy. Feather degrading bacteria were screened for both proteolytic activity and keratin degradation on skimmed milk agar and keratin azure medium respectively. They were also screened for their ability to degrade other keratin substrates such as hair and nail.
Results: Three of the isolates with higher feather degradation levels also showed high proteolytic activity and release of azure dye. They were selected and identified phenotypically and genotypically using 16S rRNA sequencing as Bacillus licheniformis-K51, Bacillus subtilis-K50 and Bacillus sp.-K53. The bacteria were capable of degrading other keratin-containing substrates such as nail and hair. Bacillus subtilis-K50 and Bacillus licheniformis-K51 showed significant difference (P) in degradation among the three different keratin sources used yielding higher degradation with feather as keratin source with respective optical densities of 0.07 and 0.11 followed by hair and least in nails with optical densities of 0.05 and 0.07 respectively. Highest degradation of all the three keratin substrates was observed in Bacillus licheniformis-K51.
Conclusion: The three isolated bacteria possess the ability to degrade keratin and utilize feather as keratin substrate. As a result, these can be considered as potential candidates for degradation and utilization of feather keratin
Evaluation of Antimicrobial Activities of Fractions of Plant Parts of Pterocarpus santalinoides
No full textAims: The study aims to investigate the antimicrobial activities of the leaves, seeds, bark, and root of Pterocarpus santalinoides plant.
Study Design: Agar well diffusion and Agar well dilution methods were used to test the preliminary antimicrobial and minimum inhibitory/bactericidal/fungicidal concentrations respectively of Pterocarpus santalinoides plants.
Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Agulu Campus, Nigeria, between February – October, 2017.
Methodology: Primary extraction and fractionation of the plant parts were undertaken with methanol, butanol, ethyl acetate, and n-hexane. Agar diffusion method for the primary antimicrobial screening on Muller-Hinton agar (bacteria) and Sabouraud dextrose agar (fungi) were used to assess the antimicrobial activities of the sixteen (16) samples on some microbial isolates namely Salmonella typhi, Escherichia coli, Candida albicans, Aspergillus niger, Microsporon canis, and Trichophyton rubrum. The minimum Inhibitory concentration (MIC), minimum bactericidal/fungicidal concentration (MBC/MFC) and percentage inhibition diameter growth (PIDG) of the samples that yielded positive activity were also evaluated.
Results: Twelve (12) samples exhibited inhibitory activity on at least one or more of the test isolates. The MIC range observed for the extracts and fractions that yielded positive activity was 12.5 – 100 mg/ml. The n-hexane fraction of the plant root indicated the best value of 12.5 mg/ml against M. canis. The best MBC/MFC value of 25 mg/ml was observed with the ethyl acetate fraction of the bark (against E. coli and M. canis) and the n-hexane fraction of the root (against M. canis). The result showed S. typhi to be the most sensitive organism to the metabolites of P. santalinoides. Extended-spectrum activity was observed with the ethyl acetate fraction of the bark against three (3) of the test isolates namely S. typhi, E. coli and M. canis. The determination of PIDG values for the test organisms against the plants’ extracts/fractions showed that crude methanol extract (28.57%) and ethyl-acetate fraction (0.14%) of the leaves, butanol fraction (0.14%) of the root (all against S. typhi) were the most potent test samples.
Conclusion: The results indicated that the plant parts may have potential medicinal values and confirmed its use in traditional medicine
Preliminary Studies on Isolation, Bile Tolerance and Antibiogram of Potential Probiotics (Probionts) from Locally Fermented Food Products at Beach Market, Nsukka Metropolis, Enugu State, Nigeria
No full textAims: The study aims to isolate, evaluate bile tolerance and antibiogram studies of potential probiotics (Lactobacillus spp) from locally fermented Food Products (Akamu, Aqua Rafa® Yoghurt, Ogiri, Okpeye) and Kunu at Beach Market, Nsukka.
Study Design: A ten - fold serial dilution and spread plate method using De Man, Rogosa and Sharpe (MRS) medium was adopted for isolation of potential Probionts.
Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, University of Nigeria Nsukka, between March - September, 2018.
Methodology: Only catalase negative and Gram positive isolates characteristic of lactobacilli were used. Bile tolerance test was performed by monitoring the bacterial growth at different Bile salt concentrations (0.2%, 0.3% and 2%). The antibiogram of the isolates was assessed using the Kirby-Bauer disc diffusion method against commercial antibiotic discs of ampicillin, vancomycin, gentamycin, ciprofloxacin, methicillin and erythromycin.
Results: All the 18 screened isolates were tolerant to bile salt at 0.2 % and 0.3 % with inhibition of growth at 2 % bile concentration. All isolates were observed to be resistant to methicillin (100 %) but very sensitive to gentamycin (11%) and ciprofloxacin (22%) respectively. The isolates showed intermediate resistance to other antibiotics: vancomycin (33%), erythromycin (33%) and ampicillin (44%). The decreasing pattern of resistance was thus: methicillin > ampicillin > vancomycin and erythromycin > ciprofloxacin > gentamycin. Isolates from Yoghurt (66.67%) and Ogiri (53.33%) provided most of the resistant isolates. Methicillin would provide best antagonist potential as all the isolates exhibited very high level of resistance (100 %).
Conclusion: These results suggest that all the eighteen potential Lactobacillus spp strain show potential for probiotic applications and the locally fermented food products are rich sources of probionts
Biosurfactant Production by Fusarium oxysporum LM 5634 Using Peels from the fruit of Bactris gasipaes (Kunth) as Substrate
No full textAim: The aim of this work was to find new fungal sources of biosurfactants via bioprospecting and to produce biosurfactants from fungal isolates using inexpensive culture media.
Methodology: Bioprocess tests were conducted with five fungal species isolated from soil from the Amazon region in order to investigate which of them presented the highest productivity in a kinetics experiment. The possibility of using the peels from the fruit of Astrocaryum aculeatum Meyer (PFAC), Bactris gasipaes Kunth (PFBG), Theobroma grandiflorum Schumann (PFTG), Musa paradisiaca (PFMP) as substrates was also evaluated.
Results: Fusarium oxysporum LM 5634 had the highest productivity, and PFBG was the best substrate for biosurfactant production.
Conclusion: Thus, the present study showed the potential of fungi and wastes (fruit peels) from the Amazon region for biosurfactant production
Antioxidant Activity and Hepatoprotective Potential of Flavonoids from Arbutus pavarii against CCl4 Induced Hepatic Damage
No full textFlavonoids have been shown to have antioxidant factors and effective against hepatotoxicity. This in vivo study aimed to evaluate the efficacy of flavonoids rich extracts in a model of chemical-induced liver cell injury.
Materials and Methods: Flavonoids were extracted from leaves and flowers of Arbutus pavarii using Microwave assisted extraction method. Different concentrations of extracted flavonoids (200, 500, 1000, 2000 and 5000mg/kg bw) were evaluated up to two weeks on mice model. The hepatoprotective effects of the extracts were examined using mice pretreated orally with 200 and 400 mg/kg bw of flavonoids extracted from leaves and flowers as well as their combination (200 mg/kg; 1:1) for 28 days. At day 28, the mice were received orally a single dose of 1ml/kg CCl4 in corn oil. Forty-eight hours after Carbon tetrachloride (CCl4) treatment, the animals were sacrificed and their liver and blood samples were collected for determination of biochemical parameters (Alkaline phosphatase (ALT), Aspartate-aminotransferase (AST) and Alanine-aminotransferase (ALP)), histopathological investigation and antioxidant status.
Results: Treatment of the mice with a daily dose of flavonoids extracts up to 5 g/kg bw did not cause mortality and did not show hepatotoxicity. Pretreatment with extracts decreased the increased serum levels of ALT, AST, and ALP, decreased lipid peroxidation and maintained the levels of glutathione and antioxidant enzymes status in the CCl4 treated mice, especially in the group treated with combined extracts. The hepato-protcitve effects were confirmed by histopathological examinations.
Conclusion: The results shown by the extracted flavonoids need further investigation
Effect of Ronstar® on the Hematological Parameters of the Freshwater Catfish Clarias albopunctatus Fingerlings
No full textClarias albopunctatus juvenile (mean weight 85.40±2.67 g) was exposed to sublethal concentrations of Ronstar (0, 0.2, 0.6 and 1.2 mg/l) in a static renewal bioassay system for 15 days. The hematological parameters of the fish were determined every 5 days. When compared with the control, the erythrocyte count (RBC), hemoglobin (Hb) and the hematocrit (Hct) were significantly reduced (P<.05) between the treatment groups. These values also differed (P<.05) within the treatment groups. The leucocyte counts increased with increasing Ronstar concentration and during the exposure. Compared with the control, significant (P<05) lymphocytosis, monocytopenia, neutropenia and reduced eosinophil were all evident in the treatment groups. The reduction in the erythrocyte count and hemoglobin are indications of anemia in the fish exposed to Ronstar. The assay of these parameters could be of immense value in establishing safe limits for pesticides in Nigerian waters as well as in the monitoring of fish health
Application of Metagenomic Study for the Detection of Diphtheria Toxin Gene of Corynebacterium diphtheriae in Patients Displaying Symptoms of Pharyngitis in Kaduna State
No full textThis study was set to detect the A and B subunit of Diphtheria toxin gene among patients with pharyngitis and to identify which of the possible biovars is endemic in Kaduna State. The result of standard collection of twenty-two (22) throat swab samples of patients using standard technique of purposive sampling, packing in sterile water at 4°C and DNA extraction using the phenol/ chloroform extraction method gave two 2 (9%) positive PCR samples out of the twenty-two, (22) throat swabs analysed; both A and B subunits of the gene’s amplicon were identified at 248bp and 297bp respectively. Sanger’s (dye-terminator) sequencing of one of the genes’ two, 2 subunits yielding two sequences obtained in the fasta format for bioinformatics analysis using the blast tool at the NCBI website. The 2 subunits of the gene were homologous to the biovars gravis and mitis complete coding sequence gene of Corynebacterium diphtheriae biovar mitis and gravis strains with percentage identity of 97 to both strains. Mutation in the 2 subunits of genes was also noticed with a total of 8 gaps that signify indels
Non-target Host Immune Gene Modulation in Transgenic Silkworm Bombyx mori Endowed with RNAi Silence BmNPV Genes
No full textAim: To reveal differential expression of host- response genes activated after nuclear polyhedro virus infection in transgenic silkworm Bombyx mori larva and to show an influence of ‘BmNPV transgenes’ on expression pattern of host- response genes.
Study Design: Relative expression profile of immune genes was analysed after BmNPV infection in transgenic and non-transgenic larvae by real-time PCR.
Place and Duration of Study: Genomics Division, Seribiotech Research Laboratory, Bangalore, India; 2014 January – 2016 December.
Methodology: Expression of immune genes encoding components of Toll and melanisation pathways was analysed in third instar larvae of transgenic B. mori line mff118B by quantitative PCR at 0, 6, 9 and 24 h after BmNPV infection and compared with infected non-transgenic larvae. A significant difference in relative expression was analyzed by Students\u27t – test or ANOVA and correlation in expression pattern, by linear regression in the probability distribution of Y as a function of X, at significance level P < 0.05.
Results: In transgenic larvae, survival rate after NPV infection was up to 70% compared to 30% in non-transgenic larvae. Immune genes encoding NF-kappa B inhibition factor, Cactus, NF kappa B transcription factors, Dorsal and Relish, Toll- activating cytokine Spätzle, melanization pathway components, prophenol oxidase activating enzyme and prophenol oxidase1 showed significantly lower expression in BmNPV infected- transgenic larva whereas enhanced expression in infected non-transgenic larvae. In infected transgenic larvae, selected gene pairs Cactus – Dorsal, Cactus – Relish, Spatzle - Dorsal, Spatzle - Cactus, Relish – Dorsal showed positively correlated expression whereas the correlation derailed in infected non-transgenic larvae.
Conclusion: RNA interference-mediated inhibition of BmNPV multiplication was engineered previously in B. mori. In infected transgenic silkworm, NPV multiplication rate is low and host-response genes showed low expression level. Under the influence of transgenes, host response genes showed correlated expression thus transgenes preserve specific host- gene interactions after NPV infection. Notably influence of ‘BmNPV transgenes\u27 on expression of host response gene is a crucial revelation in the field of transgenesis to develop better antiviral resistance in silkworms
DNA Polymorphism and Genetic Variation among Bivoltine Silkworm (Bombyx mori L.) Genotypes Revealed by RAPD Markers
No full textBackground: In breeding programs, the selection of parental lines based on their phenotypic traits along with genetic variation at their DNA level is the better option in directional breeding as specific DNA profiles would be useful in producing reliable estimates of genetic identity and diversity at their genomic levels. In this context, Random amplified polymorphic DNA (RAPD) marker approach was used to ascertain genetic diversity and relationship among 12 bivoltine silkworms (Bombyx mori L.) genotypes for the identification of diverse genotypes to be used as parents for the improvement of quantitative traits in silkworms.
Results: RAPD analysis with 12 silkworm genotypes revealed high polymorphism of 88.88%. Ten RAPD primers utilized in this study generated PCR products in the range of 202-2876 bp across 12 silkworm genotypes. The Polymorphism information content (PIC) ranged from 0.295 to 0.493%. Based on Nei’s similarity coefficients, the genetic distance among the studied silkworm genotypes ranged from 0.93 to 0.99. Of the pairwise combinations, CSR18 and SH6, showed the highest similarity (0.99) whereas, SKAU-R-6 and DUN6 showed the lowest similarity (0.93) among the genotypes. The dendrogram generated, using Unweighted pair group method using the arithmetic average (UPGMA) from this marker, grouped the 12 silkworm genotypes into five main clusters, which corroborates the differences in their phenotypic characters.
Conclusion: The study clearly indicated the potential use of RAPD markers in differentiating silkworm genotypes based on their characters. The information generated on genetically divergent genotypes identified in this study would be useful in future silkworm breeding programmes for modifying the yield potentials of silkworms so as to boost up bivoltine silk production in the interest of temperate sericulture industry
Microbial Degradation of Petroleum Hydrocarbons: Realities, Challenges and Prospects
No full textOne of the emerging environmental challenges today is hydrocarbon pollution arising from activities of petrochemical industries, natural sources of crude oil pollution and other anthropogenic activities. These petroleum products contain harmful, carcinogenic and mutagenic compounds which could have severe consequences on biotic and abiotic components of the ecosystem. Physicochemical and biological methods are employed for the remediation of hydrocarbon contaminated systems; however, the negative impacts of the physicochemical approach are presently directing greater attention to the exploitation of the biological alternatives. This article reviews basic concepts of petroleum hydrocarbons, hydrocarbon degrading microorganisms, factors that influence the biodegradation of contaminants and innovative technologies for the effective removal of these pollutants. This article also discusses the applications of relative advances in molecular biological techniques, such as the isolation of plasmid DNA of microbial communities and the use of genetically engineered microorganisms to increase the rates of biodegradation of hydrocarbon pollutants in the environment