ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-008, March, 2019. © International Scholars Journals
Full Length Research Paper
Enrichment and diversity analysis of the thermophilic microbes in a high temperature petroleum reservoir
Guihong Lan1,2, Zengting Li1, Hui zhang3, Changjun Zou2, Dairong Qiao1 and Yi Cao1*
1School of Life Science, Sichuan University, Chengdu 610065, China.
2School of Chemistry and Chemical Engineering, Southwest Petroleum University, Chengdu 610500, China.
3Biogas Institute of Ministry of Agriculture P. R. China, Chengdu 610041, China.
Accepted 13 January, 2019
Abstract
Thermophilic microbial diversity in production water from a high temperature, water-flooded petroleum reservoir of an offshore oilfield in China was characterized by enrichment and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Six different function enrichment cultures were cultivated one year, at 75°C DGGE and sequence analyses of 16S rRNA gene fragments were used to assess the thermophilic microbial diversity. A total of 27 bacterial and 9 archaeal DGGE bands were excised and sequenced. Phylogenetic analysis of these sequences indicated that 21 bacterial and 7 archaeal phylotypes were affiliated with thermophilic microbe. The bacterial sequences were mainly bonged to the genera Fervidobacterium, Thermotoga, Dictyoglomus, Symbiobacterium, Moorella, Thermoanaerobacter, Desulfotomaculum, Thermosyntropha, Coprothermobacter, Caloramator, Thermacetogenium, and the archaeal phylogypes were represented in the genera Geoglobus and Thermococcus, Methanomethylovorans, Methanothermobacter Methanoculleus and Methanosaeta. So many thermophiles were detected suggesting that they might be common habitants in high-temperature petroleum reservoirs. The results of this work provide further insight into the thermophilic composition of microbial communities in high temperature petroleum reservoirs.
Key words: Thermophilic microbial diversity, petroleum reservoir, denaturing gradient gel electrophoresis.
Hui zhang, Dairong Qiao and Yi Cao*, Changjun Zou, Guihong Lan, Zengting Li
Research Article
The effect of fertilizer amendment on diesel biodegradation in contaminated soils
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-011, March, 2019. © International Scholars Journals
Full Length Researc
The effect of fertilizer amendment on diesel biodegradation in contaminated soils
D. Padayachee and J. Lin*
School of Biochemistry, Genetics and Microbiology, University of KwaZulu-Natal (Westville), Private Bag X 54001, Durban, Republic of South Africa.
Accepted 10 January, 2019
Abstract
Diesel biodegradation and the effect of application frequency were investigated in artificially contaminated soils (loam soil and sea sand) amended with commercial fertilizers and the effect of application frequency. Two sets of four equal portions (2 kg each) of diesel-contaminated soils were amended with 10% (w/w) of 2 different commercial fertilizers at 4 different but regular intervals for up to four times during a 7- week study period. The microcosms were then incubated at 30°C and sampled weekly. One sterilized soil microcosm contaminated with 10% (v/w) diesel and supplemented with 10% (w/w) fertilizer and one naturally attenuated microcosm served as controls. The amount of diesel remaining in each sample was determined using the Soxhlet extraction method and compared to the amount of diesel present in the same sample at day 0. The quantities of hydrocarbon contents in the supplemented samples and the controls were determined by GC-MS. The total heterotrophic population of each sample was also monitored. The population increase was found to correspond to the higher diesel degradation percentage in the study on sea sand. A significant enhancement (p<0.05) of diesel degradation was observed after the supplementation of fertilizers especially in sea sand compared with those in both controls. There were no significant differences in diesel degradation with the fertilizer application frequency. In conclusion, supplementation of diesel contaminated soils with commercial fertilizer results in a significant increase on diesel biodegradation. Additional supplementation did not further stimulate the degradation under the same conditions.
Key words: Diesel, bioremediation, fertilizers, application frequency.
J. Lin*, D. Padayachee
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-006, March, 2019. © International Scholars Journals
Full Length Research Paper
Prevalence of dental caries bacterial pathogens and evaluation of inhibitory concentration effect on different tooth pastes against Streptococcus spp
A. Maripandi2,1*, Arun Kumar T.2 and Ali A. Al Salamah3
1Department of Microbiology, Government Arts College, Krishnagiri, Tamilnadu, India.
2Department of Microbiology, K. S. Rangasamy College of Arts and Science, Tiruchengode- 637 209, Namakkal, India.
3Unit of Medical Bacteriology, Department of Botany and Microbiology, College of Science, King Saudi University, P. O. Box 2455, Riyadh, Saudi Arabia.
Accepted 23 January, 2019
Abstract
Dental caries is a microbiologic infection of the tooth which results in localized dissolution and destruction of the calcified tissue and it is one of the most common chronic infectious diseases in humans throughout the world. People spent huge amounts of money and time in treating dental caries. Hence, it is essential to prevent and control dental caries in public health. In our studies, a total of 70 clinical specimens, 37 from male and 33 from female dental caries patient’s samples were collected. All the clinical samples showed positive results with 87 bacterial isolates from which 63 were facultative anaerobes and 24 were anaerobe, among facultative anaerobe, Streptococcus mutants (22.98%) S. salivarius (13.79%) and Candida spp (14.97%), in anaerobes, Prevotella spp (12.64%) and Fusobacterium spp (5.74%). The antibiotic susceptibility test was carried out against Streptococcus spp. the results showed all isolates were resistance to penicillin and sensitive to tetracycline. The antibacterial activity of commercially available tooth paste was checked against Streptococcus spp., by agar well diffusion method. The results showed that pepsodent and colgate have good antibacterial activity against Streptococcus spp.
Key words: Dental caries, Streptococcus spp, Prevotella spp, antibiotics, tooth pastes.
Arun Kumar T and Ali A. Al Salamah, A. Maripandi*
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-006, March, 2019. © International Scholars Journals
Full Length Research Paper
The antifungal mechanism of Bacillus subtilis against Pestalotiopsis eugeniae and its development for commercial applications against wax apple infection
Hsiu Fen Lin1, Tsang Hai Chen2, and Shan Da Liu3*
1Graduate Institute of Bioresources, National Pingtung University of Science and Technology, Pingtung 912, Taiwan, Republic of China.
2Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung 912, Taiwan, Republic of China.
3Graduate Institute of Health and Biotechnology Industry, Meiho University, Pingtung 912, Taiwan, Republic of China.
Accepted 16 February, 2019
Abstract
The inhibition zones of three Bacillus subtilis isolates, BS-99, BS-23857 and BS-33608 against Pestalotiopsis eugeniae were, 13.5, 0, and 0 mm, respectively. BS-99 showed the strongest inhibitory activity, whereas, no inhibitory activity was reported for BS-23857 or BS-33608. The spore germination rates of P. eugeniae were not significantly inhibited by n-hexane, ethyl acetate (EtOAc), or methanol (MeOH) extracts of the fermentation broths of BS-99, BS-23857, and BS-33608. However, the hyphae of P. eugeniae became swollen and malformed after 12 h and stopped growing after 30 h of treatment with the MeOH extract of the BS-99 fermentation broth. Analysis by polymerase chain reaction (PCR), blood agar test plates, and high performance liquid chromatography (HPLC) indicated that, BS-99 produced the antibiotics, iturin A and surfactin, whereas, BS-23857 and BS-33608 only produced surfactin. These data suggest that, the antifungal activity of the B. subtilis BS-99 isolate against P. eugeniae is activated only when both iturin A and surfactin are present and not by surfactin alone. Unveiling the antifungal mechanism of B. subtilis, BS-99 could promote its commercial development and application as a biofungicide for controlling P. eugeniae infections of the wax apple, which is a highly valued fruit in Taiwan.
Key words: Bacillus subtilis, iturin A, solvent extract, surfactin.
Hsiu Fen Lin, Tsang Hai Chen and Shan Da Liu*
Research Article
Determination of an economical medium for growth of Clostridium butyricum TK2 using orthogonal test
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-005, March, 2019. © International Scholars Journals
Full Length Research Paper
Determination of an economical medium for growth of Clostridium butyricum TK2 using orthogonal test
HaiKuan Wang, AnDong Li , FeiFei Liu and Wei Qi*
Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology,
Tianjin University of Science and Technology, Tianjin, P. O. 300457, P. R. China.
Accepted 07 February, 2019
Abstract
Clostridium butyricum is a widely utilized probiotics, also used as an alternative to antibiotics for humans and growth promotion in a wide variety of livestock species. The objective of this study is to develop an economical and practical fermentation medium for the growth of C. butyricum TK2 using orthogonal test. The optimal fermentation medium was obtained by the single factor and the orthogonal test of carbon source, nitrogen source and growth factors design. The fermentation medium was estimated to be the most economical formula (per liter): 8 g glucose, 20 g soybean meal (hydrolyzed for 3 h by neutral protease), 5 g brewer’s yeast powder. After incubating for 48 h in the optimum fermentation medium, the populations of C. butyricum TK2 were estimated to be 8 × 108 CFU ml-1, while the cost was 90% lower than ever. The optimized medium is not only more economical but also good for the growth of C. butyricum TK2. The low cost medium developed in this study can be used for large-scale commercial application where economics are quite likely to be important.
Key words: Clostridium butyricum, soybean meal, brewer’s yeast powder, medium optimization.
FeiFei Liu and Wei Qi*, HaiKuan Wang, AnDong Li
Research Article
Single multiplex PCR assay to identify the shiga toxin
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-007, March, 2019. © International Scholars Journals
Full Length Research Paper
Single multiplex PCR assay to identify the shiga toxin
Eisa Tahmasbpour Marzony1, Mahdi Kamali2, Mojtaba Saadati3, Amir Homayoun Keihan2*, Abbas Ali Imani Fooladi 4, and Sharareh Sajjadi5
1Young Research Bashgah, Islamic Azad University, Sari Branch, Tehran, Sari Iran.
2Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
3Department of Biology, Imam Hossein University, Tehran, Iran.
4Microbial Products Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
5Department of biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran.
Accepted 19 February, 2019
Abstract
Strains of shiga toxin–producing Escherichia coli (STEC) and Shigella dysenteriae type1 have been associated with outbreaks of diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS) in humans. Most clinical signs of disease arise as consequence of the production of shiga toxin/shiga toxin 1 (Stx/Stx1), shiga toxin 2 (Stx2) or combination of these toxins. Here, we designed a Multiplex PCR technique to identify stx/stx1 and stx2 genes with the incorporation of mdh gene of E. coli and Shigella. A total of six primers were used: SFI and SRI, which produce a 199bp product that serves as an internal positive control; Ka2F and Ka2R, which yield a 381bp fragment of stx2 gene, and Ka1F and Ka1R, which amplify a 622bp fragment of stx/stx1. The thermal profile, which was preceded by a 5 min incubation at 95°C for 20 to 25 cycles with the following parameters: 95°C at 1 min, 60°C at 1 min, 72°C at 1 min, and 5 min incubation at 72°C as final extension. PCR amplification products identifying the stx/stx1 and stx2 gene sequences were observed only in E. coli 0157:H7 and Shigella dysenteriae type1. Template nucleic acid extracted from other Gram-negative bacteria was found to be negative. The sensitivity of the PCR procedure for detection of shiga toxin genes was determined to be 2.1 pg/µl of total nucleic acid and 320 cfu/µl.
Key words: Shiga toxin, multiplex PCR, diagnosis.
Mojtaba Saadati, Mahdi Kamali, and Sharareh Sajjadi, Amir Homayoun Keihan*, Eisa Tahmasbpour Marzony, Abbas Ali Imani Fooladi