Advanced Journal of Microbiology Research

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Isolation and characterization of Streptomyces species with antifungal activity from selected national parks in Kenya

James Oluoch Nonoh^1,2, Wilber Lwande¹, Daniel Masiga¹, Rafi Herrmann⁴, James K. Presnail⁴, Eric Schepers⁴, Matilda Angela Okech¹, Richard Bagine³, Paul Mungai³, Aggrey Bernard Nyende² and Hamadi Iddi Boga²*

¹International Center of Insect Physiology and Ecology (ICIPE), P.O. Box 30772 –00100, Nairobi, Kenya.

²Botany Department, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000 (00200), Nairobi, Kenya.

³Kenya Wildlife Service (KWS), P.O. Box 40241-00100, Nairobi, Kenya.

⁴Pioneer, A DuPont Company, Crop Genetics Research and Development, Rte 141 and Henry clay rd. Wilmington, DE 19880.

Accepted 15 April, 2016

Abstract

Microorganisms and their natural products are potentially important for the biological control of crop diseases without detrimental effects to the environment. In this study, acetonitrile-methanol extracts of 361 actinobacterial isolates obtained from Aberdares, Arabuko Sokoke, Lake Bogoria, Mt Kenya, Kakamega, Ruma, Shimba Hills and Imenti forest national parks in Kenya were screened for antagonism against Fusarium oxysporum, Fusarium spp and Colletotrichum kahawae, which are important crop pathogens. Twenty-three isolates showed antagonistic activity to one or all of the test fungi. Five isolates that were antagonistic against all test fungi were investigated further and were also found to have antibacterial activity against Staphylococcus aureus and Escherichia coli. Morphological and physiological studies show that the isolates belong to streptomycetes. Phylogenetic analysis of amplified actinobacterial 16S rRNA gene confirmed that all the five antagonistic isolates formed close phylogenetic clusters with known members of Streptomyces species with a (97 - 100%) sequence identity. The results suggest that protected areas may be ideal habitats for isolation of antagonistic actinobacterial species which may have the potential for beneficial application in biological control of fungal pathogens. However, further investigation by characterization of the antifungal and antibacterial compounds produced will be necessary.

Key words: Protected areas, soil streptomyces, bio-prospecting, antimicrobial, phytopathogens.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Function analysis of the predicted protein-tyrosine phosphatases gene (DR2161) in Deinococcus radiodurans

Shu Haiyan* and Tian Baoming

Biology Department, Zhengzhou University, Zhengzhou, China.

Accepted 13 April, 2015

Abstract

Protein phosphorylation on tyrosine has been demonstrated to occur in a wide array of bacterial species and appears to be ubiquitous among prokaryotes. In Deinococcus radiodurans, after the predicted protein-tyrosine phosphatase (PTP) gene DR2161 was deleted, the radiation resistance of the bacterium changed a little. The natural resistance-associated macrophage protein gene DR1709 was the possible target of PTP. But the radio resistance of the double mutant (DR1709 and DR2161 that were deleted) was almost the same as that of M1709. When the strain protease secretion was measured, the clearing area of M2161 was smaller than that of the wild type. But the protease secretion of the double mutant was similar to that of M1709. The influence on the bacterium DR2161, that was deleted seemed to have been covered by DR1709 being disrupted. The four strains (the wild type, M1709, M2161 and the double mutant) sensitivity to high concentrate of Mn²⁺ and Fe²⁺ were very similar, showing that they had similar resistance to high concentrate of Mn²⁺ or Fe²⁺. In liquid defined minimal medium (DMM) with 200 nM Mn, M1709 and the double mutant almost can not grow. But in DMM with 200 nM Fe, the two strains grew as quickly as the wild type. M1709 reaction to low concentrate of Mn²⁺ and Fe²⁺ was not affected by DR2161. The expression of DR1709 was not regulated by DR2161.

Key words: Deinococcus radiodurans, protein-tyrosine phosphatase, natural resistance-associated macrophage protein, DR1709, DR2161.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Escherichia coli phoA gene inactivation, insertion and restoration based on the  red system

J. M. Wambale^1,2 Ni zhaohui ¹, J. N. Kimatu³ and Fan Li¹*

¹School of Basic Medical Sciences, Jilin University, Changchun, 1 Xinmin Street, Changchun 130021, China.

²Faculty of Pharmaceutical Sciences, Kinshasa University, PO. Box 212 kinXI, Democratic Republic of Congo.

³Key laboratory of plant molecular epigenetics, Northeast Normal University, Changchun, Renmin street 5268, Zip 130024, China.

Accepted 24 February, 2015

Abstract

The red recombinase system of bacteriophage lambda has been used to inactivate chromosomal genes in Escherichia coli K-12 through homologous recombination using linear PCR products. Here, E. coli BW 25113 mutants were created by changing the localization of genes on the genome using a method which contains both inactivation and reinsertion steps. The phoA gene, the structural gene for alkaline phosphatase, was inactivated by having the ORF and its regulator region replaced with a kanamycin cassette flanked by FLP recognition target sites. The site for insertion was selected based on the intergenic distance and non-coding regions, thus the new neighboring genes sequences were not affected. The Insertion was carried out by designing primers armed with homologous sequences to the insertion site; the primers contained the enzyme restriction sites in order to ligate the gene PCR product with the FRT-kan -FRT marker prior to the electroporation. The PCR reactions, cultures in the media supplemented with appropriate antibiotics and comparative gene expression study were carried out to confirm the knockout, the insertion and the expression of the gene in its new location. This procedure and the created E. coli mutants can be efficiently used for the study of bacterial genomics, especially in systems biology to understand the relation between the genes loci on the genome and their expression alongside new neighboring genes and others with the same functional group.

Key words: Gene, operon, knockout, insertion, systems biology, electroporation.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals 

Full Length Research Paper

Antimicrobial activity of the lichen Lecanora frustulosa and Parmeliopsis hyperopta and their divaricatic acid and zeorin constituents

Kosani Marijana¹*, Rankovi Branislav¹ and Sukdolak Slobodan²

¹Department of Biology, Faculty of Science, University of Kragujevac, 34000 Kragujevac, Radoja Domanovi a 12, Serbia.

²Department of Chemistry, Faculty of Science, University of Kragujevac, 34000 Kragujevac, Radoja Domanovi a 12, Serbia.

Accepted 25 March, 2015

Abstract

Antibacterial and antifungal activity of the acetone, methanol and aqueous extracts of the lichen Lecanora frustulosa and Parmeliopsis hyperopta and their divaricatic acid and zeorin constituents has been screened in vitro against the following species of microorganisms: Bacillus mycoides, Bacillus subtilis, Staphylococcus aureus, Enterobacter cloaceae, Escerichia coli, Klebsiella pneumoniae, Aspergillus flavus, Aspergillus fumigatus, Botrytis cinerea, Candida albicans, Fusarium oxysporum, Mucor mucedo, Paecilomyces variotii, Penicillium purpurescens, Penicillium verrucosum and Trichoderma harsianum. The antimicrobal activity was estimated by the disc-difusion method and determination of the minimal inhibitory concentration (MIC) by the Broth tube Dilution method. The bacteria were more sensitive related to the tested fungi. The smallest MIC values were 0.39 mg/mL against bacteria and 1.56 mg/mL against fungi. Acetone and methanol extracts of the investigated lichens showed relatively strong antimicrobial activity, whereas aqueous extracts were inactive. Divaricatic acid and zeorin also showed strong activity against bacteria and fungi. There was no antimicrobial activity against E. coli species. The lichen extracts maximum activity showed in the concentrations of 0.78 mg/mL and the lichen components demonstrated maximum activity in the concetrations of 0.39 mg/mL. Generally, the tested lichen extracts and lichen compounds demostrated a strong antimicrobial effect against the tested microorganisms. That suggest a possibility of their use in the treatment of various diseases caused by these and similar microorganisms.

Key words: Antimicrobial activity, lichen extracts, lichen components.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Rationalizing some medicinal plants used in treatment of skin diseases

¹Adebayo-Tayo B.C, ^2,3Adegoke A.A, ^2,*Okoh, A.I and ⁴Ajibesin K.K

¹Department of Botany and Microbiology, University of Ibadan, Ibadan, Oyo State, Nigeria

²Applied and Environmental Microbiology Research Group (AEMREG),Department of Biochemistry and Microbiology, Plant Science Building, University of Fort Hare,Private Bag X1314, Alice 5700, South Africa

³Department of Microbiology, University of Uyo, Uyo, Akwa Ibom State, Nigeria ⁴Ogun State University, Ago-Iwoye, Ogun State, Nigeria.

Accepted 11 June, 2015

Abstract 

The hydroethanolic extracts obtained from ten plant species being used as components of skin disease remedy in Southwest Nigeria were subjected to phytochemical analysis and screened for antimicrobial activity. The antimicrobial activity was determined against Escherichia coli NCIB 86, Staphylococcus aureus NCIB 8588, Klebsiella pneumoniae NCIB 418, Pseudomonas aeruginosa NCIB 950, Proteus vulgaris NCIB 67, Bacillus subtilis NCIB 3610, Candida albicans and Aspergillus flavus by agar diffusion method. Terpenes, flavonoids, tannins and saponin were detected in the active plants through phytochemical screening and confirmed by thin layer chromatography (TLC) . The plant extracts demonstrated antimicrobial effect against bacteria and the fungal cultures used in this study at different levels. The plant species Funtumia elastica, Raphyostylis beninensis, Butyrospermum paradoxum, Serataria caudula, Parkia biglobosa and Curculigo pilosa showed significant antimicrobial activities against the test organisms. Curculigo pilosa, F. elastic and P. biglobosa gave the highest zone of inhibition of 30 mm at 80 mg/ml against Aspergillus flavus, Bacilus subtilis and Proteus vulgaris while Curculigo pilosa also gave the least zone of inhibition of 2 mm at 80 mg/ml against Klebsiella pneumoniae. This relative high active index explains the antimicrobial effects of the remedy for skin diseases.

Key words: Phytochemical analysis, antimicrobial activity, TLC, skin disease, Funtumia elastica.