ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-005, March, 2019. © International Scholars Journals
Full Length Research Paper
Development and application of a novel multiplex polymerase chain reaction (PCR) assay for rapid detection of various types of staphylococci strains
Zhenbo Xu1,2, Lin Li1, Xihong Zhao3*, Jin Chu4, Bing Li1, Lei Shi1, Jianyu Su1* and Mark E. Shirtliff2,5
1College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China.
2Department of Microbial Pathogenesis, Dental School, University of Maryland, Baltimore, MD 21201, USA.
3Key Laboratory for Green Chemical Process of Ministry of Education, School of Chemical Engineering and Pharmacy,
Wuhan Institute of Technology, Wuhan 430073, Hubei, China.
4School of Food Science and Nutrition, Faculty of Mathematics and Physical Sciences, University of Leeds, Leeds LS2 9JT, United Kingdom.
5Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA.
Accepted 15 January, 2019
Abstract
In this study, a novel multiplex- polymerase chain reaction (PCR) for rapid detection of various staphylococci strains, including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-sensitive Staphylococcus aureus, (MSSA) methicillin-resistant coagulase-negative staphylococci (MRCNS), methicillin-sensitive coagulase-negative staphylococci (MSCNS) and non-staphylococci strains, had been developed and applied. Six primers were specially designed on three target genes, which were mecA, 16S Ribosomal ribonucleic acid (rRNA) and femA. The specific amplification generated 3 bands on agarose gel, with sizes 374 bp for mecA, 542 bp for 16S rRNA and 823 bp for femA, respectively. The PCR product showed highest levels of resolution of DNA when 250 M of dNTP, primer concentration of mecA, 16S rRNA and femA reaching 1, 1 and 3 M respectively. No false positive amplification was observed, indicating the high specificity of the established multiplex PCR assay. Application of this multiplex-PCR had been further performed on detection for 262 MRSA and MRCNS strains with primers pairs M1 with M2 and F1 and F2. According to the results, multiplex-PCR results showed expected products for either MRSA or MRCNS strains, demonstrating the multiplex-PCR assays established in this study to be useful and powerful methods for differentiation of MRSA, MSSA, MRCNS, MSCNS and non-staphylococci strains.
Key words: Staphylococcus, multiplex-PCR, rapid detection.
Jianyu Su* and Mark E. Shirtliff, Bing Li, Jin Chu4, Xihong Zhao*, Lei Shi, Zhenbo Xu, Lin Li
Research Article
Prevalence of Leptospira hardjo antibody in bulk tank milk in some dairy herds in Mashhad suburb
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-005, March, 2019. © International Scholars Journals
Full Length Research Paper
Prevalence of Leptospira hardjo antibody in bulk tank milk in some dairy herds in Mashhad suburb
Elias Tabatabaeizadeh1,2, Gholamreza Hashemi Tabar1,2*, Nima Farzaneh3 and Hesam A. Seifi3
1Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
2The Research Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
3Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Accepted 19 January, 2019
Abstract
Bulk tank milk samples were collected from 47 dairy herds in Mashhad suburb. From 19 herds, one sample was collected at each season in 2009 and from the rest of the 28 dairy herds one sample was collected only at autumn, 2009 and samples was tested by ELISA method for detecting antibodies to Leptospira interrogans serovar hardjo. These herds, which had not been vaccinated against leptospirosis within the previous five years, were categorised according to the herds’ regions. Nine herds (19.15%) out of 47 had a positive ELISA result. The antibody level in the herd milk samples were affected by the herd size (P<0.05). Region and season had no significant effect on antibody level (P>0.05). Larger herds were significantly more likely to have higher mean concentrations of antibody. It was concluded that a number of unvaccinated dairy herds in Mashhad suburb are exposed to infection with Leptospira hardjo.
Key words: Cattle, Leptospira, antibodies, ELISA, hardjo.
Nima Farzaneh and Hesam A. Seifi, Elias Tabatabaeizadeh, Gholamreza Hashemi Tabar*
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-007, March, 2019. © International Scholars Journals
Full Length Research Paper
Virus infection disturbs cyclin expression, leading to cell cycle arrest in the unicellular marine algae Emiliania huxleyi and Chrysochromulina ericina
Jingwen Liu1, Tianling Zheng2*, Gunnar Bratbak3 and Runar Thyrhaug2
1Bioengineering College of Jimei University, Xiamen 361021, China.
2Key Laboratory of MOE for Coast and Wetland Ecosystem, Xiamen University, Xiamen, 361005, China.
3Department of Microbiology, University of Bergen, Postbox 7800, N-5020, Bergen, Norway.
Accepted 10 February, 2019
Abstract
To investigate the influences of viral infection on cell cycle of marine algae, we examined the changes of expression and activity of G 2/M-phase cell cycle regulators in two unicellular marine algae Emiliania huxleyi and Chrysochromulina ericina after viral infection by immunoblot. The results revealed that the expression of cyclins could be altered by viral infection, thus inhibiting the progression of cell cycle. E. huxleyi virus (EhV) repressed the activity of p34cdc2/cyclinB complex by inactivating p34cdc2 kinase through phosphorylation of Cdc2, and block the life cycle of host cells at G2/M checkpoint. C. ericina virus (CeV) interfere with the activity of p34cdc2/cyclin B complex by the downregulation of cyclinB combined with the overexpression of kinase-active p34cdc2, resulting in a G2 cell cycle. The G 2/M cell cycle arrest caused by EhV infection indicated that EhV may transcribe its own genes without relying on the host cell cycle control. C. ericina cells blocked in a delayed G2 phase may partly turn into the second S phase to synthesize proteins useful for viral nucleic acid metabolism, suggesting that the replication of CeV viruses was dependent on cell cycle control. Our results indicated that algal virus infection selectively activates/inactivates certain components of the cell cycle with the aim to establish a more efficient environment for their gene expression and DNA replication.
Key words: Marine algal virus, cyclins expression, cell cycle arrest.
Jingwen Liu, Gunnar Bratbak and Runar Thyrhaug, Tianling Zheng*
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
Agricultural utilisation of dairy sewage sludge: Its effect on enzymatic activity and microorganisms of the soil environment
Magdalena Frąc1* and Stefania Jezierska-Tys2
1Department of Plant and Soil System, Institute of Agrophysics Polish Academy of Sciences, ul.Doswiadczalna 4, 20-290 Lublin, Poland.
2Department of Agricultural Microbiology, University of Life Sciences in Lublin, ul. Leszczynskiego 7, 20-069 Lublin, Poland.
Accepted 09 February, 2019
Abstract
Biochemical indices related with the transformations of C, N and P are frequently used in the diagnosis of soil quality. Such an assessment usually includes analysis of the following enzymes – the activity of dehydrogenases and of hydrolytic enzymes such as phosphatases, urease and protease. The objective of the study presented here was evaluation of dairy sewage sludge as an agent causing an improvement of soil quality or its degradation. Estimation of soil quality was performed on the basis of a comprehensive study comprising analyses of microbiological and biochemical parameters. The results of the study indicate a stimulating effect of dairy sewage sludge on the soil microorganisms, which was apparent also as an increase in the activity of the enzymes studied. That effect should be attributed to soil enrichment in organic matter, total nitrogen and minerals.
Key words: Dairy sewage sludge, enzymatic activity, microorganisms, soil.
Stefania Jezierska-Tys, Magdalena FrÄ…c*
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-007, March, 2019. © International Scholars Journals
Full Length Research Paper
Effect of neem (Azadiracta indica) aqueous extract on Casuarina equisetifolia seed germination, growth of Frankia and some rhizospheric microorganisms
Sayed W. F.*, Salem W. M. and Ali M. S.
Department of Botany, Faculty of Science, Qena, South Valley University, 83523 Qena, Egypt.
Accepted 10 January, 2019
Abstract
Aqueous neem (Azadiracta indica) leaf extract was tested for its effect on Casuarina equisetifolia seed germination capacity, growth of Frankia and on some dominant rhizosphere microorganisms under casuarinas grown in soil contaminated with sewage water. Analysis was carried out to determine the main micro- and macroelement content of different extract amounts. The addition of extract amount between 10- 20 µl per 10 ml medium (1 - 2 µl ml-1) selectively stimulated Frankia growth and inhibited other non- beneficial bacteria and fungi in the rhizosphere such as Bacillus subtilis, Drechslera sp and Curvularia lunata. Slight increase was observed in Casuarina seed germination capacity with 1- 2 µ ml-1 of the extract, but with no significant differences, indicating at least no negative effect of the extract on seed germination. According to the results and as the roots and leaves of both Casuarina and neem release their active components in soil, and according to previous recommendations, we suggest a significant synergistic effect between Casuarina, rhizosphere microorganisms and neem trees for the benefits of Casuarina nodulation, growth performance and nitrogen fixation.
Key words: Neem, Casuarina, aqueous extract, Frankia, rhizospheric microorganisms.
Salem W. M. and Ali M. S., Sayed W. F.*
Research Article
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-005, March, 2019. © International Scholars Journals
Full Length Research Paper
Direct detection of Escherichia coli O157 and its major virulence factor genes in animal faeces at slaughter using multiplex polymerase chain reaction (PCR)
Khandaghi Jalil1*, Razavilar Vadood2 and Barzegari Abolfazl3
1Food Hygiene, Islamic Azad University, Sarab Branch, Sarab, Iran.
2Food Hygiene, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
3Research Center of Pharmaceutical Biotechnology (RCPN), Tabriz University of Medical Science, Tabriz, Iran.
Accepted 10 January, 2019
Abstract
Although more than 100 serotypes of shigatoxigenic Escherichia coli (STEC) have been implicated in cases of human diseases, E. coli O157 is the most common serogroup connected with sporadic cases and large outbreaks of diseases in many countries. Rapid and sensitive identification of this dangerous pathogen is important for patient management and for prompt epidemiological investigations. PCR has become a very rapid and reliable tool for the molecular diagnosis of E. coli O157. PCR assays are usually aimed at detecting the shiga toxins, the intimin protein and enterohaemolysin. In the present study, a mPCR-based protocol is described as that which uses one primer set to detect the gene responsible for the production of the O- antigen synthesis (rfb O157) and four primer set to detect the major virulence factor genes including the Shigatoxin type 1 and 2 ( stx1 and stx2), intimin (eaeA) and enterohemolysin (EHEC hlyA) directly from 190 samples of animal faeces at the time of slaughter after overnight incubation of stool specimens in BPW. In this research, we use one primer set for detection of the gene responsible for the production of the O- antigen synthesis (rfb O157) and four primer set for detection of the Shigatoxin type 1 and 2 (stx1 and stx2), intimin (eaeA ) and enterohemolysin (EHEC hlyA) producing genes directly from 190 samples of animal faeces at the time of slaughter after overnight incubation of stool specimens in BPW. This study has established the presence of rather high prevalence of E. coli O157-positive animals at abattoirs (These consisted of 4.2% of cattle and 2.1% of sheep), providing an increased risk of transmission of E. coli O157 to the food chain and contamination of human.
Key words: Shigatoxigenic Escherichia coli (STEC), E. coli O157, shiga toxins, intimin, enterohaemolysin, mPCR.
Razavilar Vadood and Barzegari Abolfazl, Khandaghi Jalil*