ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Use of RT-PCR in diagnosis of infectious hematopoietic necrosis in rainbow trout hatcheries, Iran
Mehdi Raissy1,2*, Hasan Momtaz1, Mahsa Ansari2 and Manouchehr Moumeni3
1Faculty of Veterinary Medicine, Islamic Azad University-Shahrekord Branch, Shahrekord, Iran.
2Young Researchers Club, Islamic Azad University-Shahrekord Branch, Shahrekord, Iran.
3Fishery Research Center, Islamic Azad University-Shahrekord Branch, Shahrekord, Iran.
Accepted 7 April, 2017
Abstract
Distribution of Infectious Hematopoietic Necrosis (IHN) was studied in rainbow trout fish farms in Chaharmahal and Bakhtyari Province which is a major trout culture region in Iran. Overall 150 suspected rainbow trout fry with less than 3 g body weight from 30 fish farms in Chaharmahal and Bakhtyari Province, Iran, were studied and fishes of 10 hatcheries (33.3% of all studied farms) found to be infected with IHNV. Reverse transcriptase polymerase chain reaction (RT-PCR) was applied to the detection of infectious hematopoietic necrosis virus (IHNV). The product of 371 bp was amplified from the RNA extracts of infected organs of suspected fish. DNA sequencing of the PCR products demonstrated specificity of the amplification. The RT-PCR was found to be a simple, highly specific and sensitive method allowing diagnosis of IHN in a short time.
Key words: Reverse transcription-polymerase chain reaction (RT-PCR), IHN, rainbow trout.
Mehdi Raissy*, Hasan Momtaz1, Mahsa Ansari and Manouchehr Moumeni
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full length Research Paper
Association among the Rv2629 gene, rpoB gene, RFP resistance and Beijing genotype in M. tuberculosis clinical isolates
Junxian Zhang1, Xueqiong Wu1*, Yourong Yang1, Yan Zhuang1, Bo Wang1, Jianqin Liang1, Yan Liang1, Hongmin Li 1 and Chuihuan Li2
1Institute of Tuberculosis Research, the 309th Hospital of PLA, Beijing 100091, China.
2Thorax Disease Hospital of Hebei province, Shijiazhuang 050041, China.
Accepted 2 May, 2017
Abstract
Rifampin (RFP) is a major first-line anti-tuberculosis drug. The molecular assay currently used is the detection of rpoB gene mutations in M. tuberculosis. Recently, the Rv2629 191C allele was found to have a correlation with RFP resistance, and might become a valuable marker for the detection of RFP resistance or the Beijing genotype. We studied the association among the Rv2629 gene, rpoB gene, RFP resistance and Beijing genotype in 69 M. tuberculosis clinical isolates using DNA sequencing, conventional drug susceptibility and spoligotyping. The 191C allele was present in 92.8% (64/69) isolates. Of 29 RFP-sensitive strains, none exhibited any mutations in rpoB genes, only one strain (3.4%) was found to carry the 191A allele and 28 strains (96.6%) had the 191C alleles. Of 40 RFP-resistant strains, 30 (75%) strains had rpoB gene mutations, only 4 strains (10%) carried 191A alleles, and 36 strains (90%) exhibited 191C alleles. The 191C allele was also present in INH-sensitive, SM -sensitive or EMB-sensitive isolates. Spoligotyping analysis showed 8 distinct spoligotyping patterns. 81.1% (30/37) strains were divided into one big cluster, which had a characteristic of the Beijing genotype. Rv2629 191C allele was present in 93.3% (28/30) Beijing genotype strains, but also in non- Beijing genotype strains. These results indicate that high association is present between the ropB gene and RFP resistance. No association is present between the Rv2629 191C allele and RFP resistance, or between the Beijing genotype and RFP resistance.
Key words: M. tuberculosis, Rv2629, rpoB, rifampin resistance, spoligotyping.
Bo Wang, Xueqiong Wu*, Jianqin Liang, Yan Zhuang, Junxian Zhang, Yan Liang, Yourong Yang, Hongmin Li and Chuihuan Li
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Validation of RNA integrity from low yield experiments with Mycobacterium tuberculosis for downstream application in real time PCR
Prathna Ramchandra and A. Willem Sturm*
Medical Microbiology Research Laboratory, Nelson R Mandela School of Medicine, University of KwaZulu-Natal, South Africa.
Accepted 22 June, 2017
Abstract
RNA extraction from mycobacterial cells is more challenging than from any other cell type. We used the Trizol Reagent® with silica beads to disrupt cells of Mycobacterium tuberculosis H37Rv. This method requires minimum reagents and handling and therefore aids in maintaining RNA integrity. The efficiency of cell disruption by this method was verified by microscopic analysis of the lysate. The yield and purity were determined using the Nanodrop-1000. A total volume of 50 µl containing 224.7 ng/µL of RNA was obtained from 3.1 x 108 cells, grown under sub-optimal conditions. RNA was visualized using the gene genius optical system. A 6 week old undisturbed culture of M. tuberculosis H37Rv, gave an expression ratio of fdxA/16s of -2.094. This extraction method is suitable for use of RNA in quantitative experiments, even if the cell numbers from which the RNA is extracted are low.
Key words: Mycobacteria, RNA extraction, RNA purification, quantitation.
A. Willem Sturm*, ra , Prathna Ramch
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Antibacterial effect of chlorine dioxide and hyaluronate on dental biofilm
F. Al-bayaty1,4*, T. Taiyeb-ali2, M. A. Abdulla3 and F. Hashim3
1Department of Restorative Dentistry, Faculty of Dentistry, University Teknologi MARA, Malaysia.
2Department of Oral Medicine and Periodontology, Faculty of Dentistry, University Malaya, Malaysia.
3Department of Molecular Medicine, Faculty of Medicine, University Malaya, Malaysia.
4Faculty of Dentistry, Level 19, Tower 2, Science and Technology Complex 40450, Shah Alam, Malaysia.
Accepted 8 April, 2017
Abstract
The objective of this study is to investigate antimicrobial action of chlorine dioxide (ClO2) gel and hyaluronate gel (Gengigel®) on dental biofilm. Pooled supra and subgingival dental biofilm were obtained from healthy individuals and incubated aerobically and anaerobically. Plaque bacteria investigated including Streptococcus constellatus, Streptococcus mitis, Eikenella corrodens, Fusobacterium nucleatum, dental plaque pool samples (aerobic and anaerobic) and Staphylococcus aureus and Escherichia coli as internal control microorganisms. All bacteria were grown in Brain Heart Infusion (BHI) broth and the clinical isolate strains were sub-cultured on BHI agar. Single pure colonies of bacteria were transferred into fresh BHI broth and incubated overnight. Bacterial counting was done using hemocytometer. Antibacterial activities were determined using bacteria grown on Mueller Hinton II agar and antimicrobial disc diffusion susceptibility testing with paper discs impregnated with ClO2 and Hyaluronate gels as well as by minimum inhibition concentration (MIC) test. Bacterial morphological alterations following treatment with ClO2 and Hyaluronate gels were viewed under Scanning Electron Microscope (SEM) at 3500x, 10000 x and 20000x magnification. Positive results were obtained with disc diffusion technique whereby both agents exhibited antibacterial action against the microorganisms tested. ClO2 gel produced large diameter inhibition zones while Hyaluronate gel resulted in smaller diameter inhibition zones. In MIC test the lowest MIC value of ClO2 gel (0.02% w/v) was obtained for S. aureus, S. mitis and S. constellatus. The other bacteria and pool samples of dental biofilm indicated slightly higher MIC values (0.2% w/v) for ClO2 gel. However, MIC values for Hyaluronate gel could not be determined. Under SEM, ClO2 gel produced obvious alterations to the bacterial morphology while no changes were observed after treatment with hyaluronate gel. Chlorine dioxide gel demonstrated stronger and obvious antibacterial activity.
Key words: Chlorine dioxide gel, Gengigel®, dental biofilm.
M. A. Abdulla and F. Hashim, F. Al-bayaty*, T. Taiyeb-ali
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full length Research Paper
Use of Scopulariopsis acremonium for the production of cellulase and xylanase through submerged fermentation
Loveleen Kaur Sarao*, Maninder Arora and V. K. Sehgal
Department of Processing and Food Engineering, Punjab Agricultural University, Ludhiana 141004, Punjab, India.
Accepted 5 May, 2017
Abstract
Production of cellulase and xylanase by Scopulariopsis acremonium through submerged fermentation was studied using shake flask cultivation media. The effect of process parameters such as effect of temperature, pH and inoculum size was investigated. The enzymes were partially purified using isopropanol and used for enzyme assays. The results indicated that the maximum cellulase and xylanase having an enzyme activity of 694.45 and 931.25 IU, respectively, were produced at 30°C incubation temperature. The pH optimum to achieve these enzyme activities was 5.5 with an inoculum size of 1 x 105 spores ml-1 of tween - 80. Cellulase was deactivated at 80°C while xylanase was deactivated at 70°C.
Key words: Scopulariopsis acremonium, cellulase, xylanase, submerged fermentation.
Maninder Arora and V. K. Sehgal, Loveleen Kaur Sarao*
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
The prevalence of group A rotavirus infection and some risk factors in pediatric diarrhea in Zaria, North central Nigeria
Pennap Grace1* and Umoh Jerald2
1Microbiology Unit, Nasarawa State University Keffi, P. M. B. 1022, Keffi, Nigeria.
2Department of Public Health, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria.
Accepted 8 June, 2017
Abstract
Group A rotavirus cause infectious, severe and dehydrating diarrhea which have great impact on childhood morbidity and mortality worldwide. This study was designed to investigate the disease burden and risk factors of rotavirus infection among children 1 - 5 years in Zaria, Nigeria. Stool samples were collected from 666 diarrheic children and 170 matched non -diarrheic controls and screened for rotavirus antigen by ELISA. Their socio-demographic information and clinical presentations were also noted. There was a 15.6 and 7.6% prevalence of rotavirus infections among the diarrheic and non-diarrheic children, respectively, with the peak of infection occurring between 1 – 6 months of age. The most common clinical features included dehydration (59.1%), fever and vomiting (41.3%), vomiting (34.6%) and fever (19.2%). Age, feeding regimen, and the presence of another person in the house with diarrhea were found to be risk factors for rotavirus diarrhea while gender and socioeconomic status were not. In view of the peak of infection in infants in the study area, rotavirus vaccine will be best administered in early infancy.
Key words: Rotavirus, prevalence, risk factors, pediatric, diarrhea.
Pennap Grace*, Umoh Jerald
Page: 1 - 10