ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-006, January, 2019. © International Scholars Journals
Full Length Research Paper
Sensitive and specific detection of Agrobacterium tumefaciens in soil using a rapid polymerase chain reaction (PCR)
Wei Yang1,2, Lei Ji3, Li-Rong Tan2, Shi-Mo Li1, Yan Wang3, Hong-Xia Liu2*, and Yu-Ming Luo1*
1Jiangsu Key Laboratory of Eco-Agricultural Biotechnology around Hongze Lake, Huaiyin Teacher’s College, No. 111, Changjiang West, Huai’an, 223001, Jiangsu Province, China.
2Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
3Shanghai Forestry Station, Shanghai, 200072, China.
Accepted 13 January, 2019
Abstract
One pair of primers was designed based on the sequence of tmr locus for specific and sensitive detection of Agrobacterium tumefaciens. Only the A. tumefaciens strain can produce the 236bp target fragment among the fourteen bacterial species that tested. The sensitivity of the specific PCR system was determined by a nested-PCR amplification which can numbered the copies of the template DNA. According to the results, it can give positive band when only 100 copies were in the template. The protocol was carried out for detection A. tumefaciens of twelve soil samples collected from six different gardens in Shanghai where crown gall happened. Two of the samples which collected from symptomless gardens also give the positive band. Based on the results we can make a conclusion that this pair of primers can be a useful tool in detecting A. tumefaciens, especially in detecting latent infection of this devastating pathogen.
Key words: Agrobacterium tumefaciens, detection, polymerase chain reaction (PCR).
and Yu-Ming Luo*, Li-Rong Tan, Wei Yang, Shi-Mo Li, Lei Ji, Hong-Xia Liu*, Yan Wang
Page: 1 - 6
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-004, January, 2019. © International Scholars Journals
Full Length Research Paper
E-test versus disk diffusion method in determining multidrug resistant strains of Escherichia coli in urinary tract infection
Y. Erfani1, A. Rasti2*, A. Mirsalehian3, S. M. Mirafshar3, V. Ownegh3
1Department of Laboratory medicine, School of Allied health, Tehran University of Medical Sciences, P. O. Box: 13185-1678, Tehran, Iran.
2School of Nursing and Midwifery, Tehran University of Medical Sciences, Tehran, Iran.
3School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Accepted 15 January, 2019
Abstract
Multidrug resistant strains of Escherichia coli are becoming a major challenge in treatment of patients with urinary tract infection. Developing effective screening methods for selection of antimicrobial resistant strains is necessary. In this study, we have compared descriptively results of E-test with Disk diffusion agar method in selection of multidrug resistant strains of E. coli among patients with urinary tract infection. In disk diffusion test, 19 of 256 urine collections were resistant to 5 antibiotics used in the study. Later E-test was performed on these 19 collections with the same 5 antibiotics. In E-test, 10.5% of the specimens were sensitive to Bacterim, 21.1% to gentamicin, 47.7% to nitrofurantoin, 10.5% to ciprofloxacin and 10.5% to Ceftazidime. Results indicate that performing E-test on strains that were resistant in disk diffusion test may increase specificity in determination of multidrug resistant strains of E. coli in patients with urinary tract infection. This antibiotic susceptibility study showed difference between E-test and disk diffusion agar in assessing the antibiotic susceptibility and found E-test accuracy and its superiority to disk diffusion in detecting multidrug resistance.
Key words: Escherichia coli, resistance, E-test, disk diffusion.
S. M. Mirafshar and V. Ownegh, A. Rasti*, A. Mirsalehian, Y. Erfani
Page: 1 - 4
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-006, January, 2019. © International Scholars Journals
Full Length Research Paper
Effects of arbuscular mycorrhizal (AM) fungi Glomus mosseae on characteristics of leaf development of Paeonia suffruticosa under salt stress
Guo Shaoxia1,2* Han Tingting2 and Liu Runjin1
1Institute of Mycorrhizal Biotechnology, Qingdao Agricultural University, Qingdao Shandong Province 266109, China.
2College of Garden and Horticulture, Qingdao Agricultural University, Qingdao Shandong Province 266109, China.
Accepted 08 January, 2019
Abstract
The effects of Arbuscular mycorrhizal (AM) fungi on the characteristics of leaf development of Paeonia suffruticosa at different levels of salt stress (0, 8, 16 and 24%) were studied. Potted ‘Feng Dan’ seedlings were inoculated with Glomus mosseae, and the non-inoculated was used as the control. The results showed that under salt stress leaf relative water content, sclerophyllous index of leaves and leaf succulence level of P. suffruticosa seedlings inoculated with G. mosseae were significantly higher than those of non-inoculated seedling, and specific leaf area was significantly lower than that of the control. Leaf water loss rate of the non-inoculated peony was faster, the percentage of water loss in the total amount of water each time point is the highest. These results suggest that G. mosseae may play an important role in the leaf traits and enhanced salt tolerance of tree peony seedlings.
Key words: Arbuscular mycorrhizal fungi, characteristics of leaf development, Paeonia suffruticosa, salt stress.
Guo Shaoxia* Han Tingting, Liu Runjin
Page: 1 - 6
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-008, January, 2019. © International Scholars Journals
Full Length Research Paper
Application of phytodisinfectants in water purification in rural Cameroon
K. A. Yongabi1*, D. M. Lewis1 and P. L. Harris2
1School of Chemical Engineering, The University of Adelaide, South Australia.
2School of Agriculture, Food and Wine, The University of Adelaide, South Australia.
Accepted 11 November, 2018
Abstract
Findings from a preliminary lab-scale study show strong potentials of phytodisinfectants as a low-cost, appropriate and ecological alternative technology in purifying water in rural Cameroon. A survey of plants used in water purification in Bamenda, Cameroon, indicated that there are many plants used in water treatment. A rapid screening on the coagulative and disinfection potential of four most frequently used plants was carried out on; Moringa oleifera, Jatropha curcas, calyx of Hibiscus sabdarifa, sclerotium of Pleurotus tuberregium against their crude methanol extracts, aluminum sulphate and sodium hypochlorite controls on turbid surface water samples. A beaker experiment with varying weights (0.5 to 5 g) of dried pulverized plant materials and alum (control) were placed in 200 ml each of the three-turbid water samples and left for thirty minutes retention time. A 95% reduction in bacterial loads of the water samples by M. oleifera in fifteen minutes residence time was observed. J. curcas seeds, as well as H. sabdarifa calyx also reduced the bacterial loads between 75 to 90%. All the plant extracts except P. tuberregium inhibited an Escherichia coli isolate from the turbid water with highest zone of inhibition (15 mm) recorded for M. oleifera seed extract. The inhibition zones produced by three of the plant extracts were comparable to aluminum sulphate (6 mm) and sodium hypochlorite (17 mm). Crude methanol extracts from M. oleifera seeds, J. curcas seeds and H. sabdariffa calyx used directly on turbid water drastically reduced the total aerobic mesophilic bacterial counts far more than the unextracted plant powders. The turbidity of both phytodisinfectant and alum treated water samples drastically reduced while no turbidity reduction was observed with sodium hypochlorite treated water samples. The pH of alum treated water was observed to decrease from neutral to 5.0 while pH of phytocoagulant treated water was 7.0. This report suggest that M. oleifera seeds, J. curcas seeds and calyx of Hibiscus sabdariffa posses both phytodisinfectant and phytocoagulant property in water purification. Scerotium of P. tuberregium poses only phytocoagulant (mycocoagulant) activity. Plant materials can be used as phytocoagulants and phytodisinfectants in treating turbid water and can be applied in wastewater treatment. Further studies on the application of Phytodisinfectants in domestic water purification, especially the phytodisinfection potentials of M. oleifera are exigent.
Key words: Phytodisinfectants, phytobiotechnology, phytocoagulants, bacteria turbid water, plants, Moringa oleifera, Cameroon.
K. A. Yongabi*, D. M. Lewis and P. L. Harris
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Short Communication
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-003, January, 2019. © International Scholars Journals
Short Communication
Anthrax in animals of the Eastern Mediterranean region of Turkey
Ekrem Kirecci1*, Suleyman Aslan2 and Nevin Turut2
1School of Health, Kahramanmaras Sutcuimam University, 46050, Kahramanmaras, Turkey.
2Veterinary Control and Research Institute, Adana, Turkey.
Accepted 21 December, 2018
Abstract
In this study, we aim to perform a microbiological evaluation on the anthrax cases encountered in small and large ruminants as well as in equidae in the Eastern Mediterranean region of Turkey between March 2005 and July 2010. Blood specimens from 44 animals raised in the Mediterranean region of Turkey and suspected of having anthrax in light of the clinical examination results between 2005 and 2010, were subjected to bacteriological analysis. Conventional methods such as Giemsa and Gram staining as well as culture and motility tests were carried out. Smears prepared from blood specimens were treated with Giemsa and Bacillus anthracis pathogens with typical capsules were investigated. During culture test, specimens were cultivated in nutrient broth, blood agar, and MacConkey agar after which they were incubated at 37°C for 24 - 48 h in an aerobic environment. Colony morphologies and microscopic appearances of the bacteria that grew in the nutrient broth and blood agar were evaluated. Thus, bacteria displaying an appearance of typical hair-like shape and a configuration of long chains similar to bamboo sticks, with nonmotile and R-form like colony view, were diagnosed as B. anthracis. The animals with suspected anthrax in this study were comprised of 19 cattle, 14 goats, 10 sheeps and 1 mule. In 12 (27.3%) of 44 specimens, B. anthracis was isolated and identified. B. anthracis was isolated from the animals with suspected anthrax. Anthrax, recognized as a dangerous zoonosis across the entire world, is still of endemic status in our country and preventive measures should include prevention of illegal animal slaughter by performing efficient supervision, applying routine vaccination programs and raising awareness among the livestock owners.
Key words: Anthrax, animals, Bacillus anthracis, pathogens.
Ekrem Kirecci*, Suleyman Aslan and Nevin Turut
Page: 1 - 3
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-010, January, 2019. © International Scholars Journals
Full Length Research Paper
Cloning, codon-optimized expression and homology modeling of structural protein VP1 from foot and mouth disease virus
Xin-sheng Liu, Yong-lu Wang*, Yong-guang Zhang, Yu-zhen Fang, Li Pan, Jian-liang Lü, Peng Zhou, Zhong-wang Zhang, Cheng Qi-wei, Gang Wang, Ji-wei Wang, Hui Lou and Shou-tian Jiang
State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046,China.
Accepted 17 January, 2019
Abstract
Structural protein VP1 of foot-and-mouth disease virus (FMDV) is the most frequently studied protein due to its significant roles in virus attachment, protective immunity, and serotype specificity. The coding sequence of VP1was amplified and then identified by polymerase chain reaction (PCR) and sequencing. To achieve high-level expression of VP1 protein, we optimized VP1 gene base on Escherichia coli preferred codons and synthesized the optimized gene. The synthetical gene was cloned into the fusion expression vector pET-28a and expressed in E. coli BL21(DE3). After induced with Isopropyl -D-1-Thiogalactopyranoside (IPTG) and optimized the conditions of expression, the VP1 fusion protein was highly expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. Based on the primary and secondary structure analysis of VP1, Three-dimensional structure of VP1 was developed by homology modeling methods. The validation of 3-D structure was done with the help of PROCHECK encompassing amino acid residues in the most favored region of almost all strains. Potential epitopes of VP1 was predicted with different methods. In this study, the VP1 protein was expressed in E. coli efficiently and highly purified VP1 was obtained, which laid a foundation of refolding and further study on activity of the protein. The VP1 model in the productive conformation can now be used for structure-based design purposes as well as structure-function relation of VP1 protein.
Key words: Foot-and-mouth disease virus, VP1 protein, codon optimization, homology modeling.
Yong-guang Zhang, Peng Zhou, Yong-lu Wang*, Zhong-wang Zhang, Gang Wang, Ji-wei Wang, Xin-sheng Liu, Cheng Qi-wei, Hui Lou and Shou-tian Jiang, Yu-zhen Fang, Li Pan, Jian-liang Lü
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