AJMR | Table of Contents

Table of Contents 2016

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

Fingerprinting of repetitive DNA sequences in the genus Anabaena using PCR- based techniques

Anbalagan Ezhilarasi* and Narayanaswamy Anand

Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai–600025, India.

Accepted 23 March, 2015

Abstract

In this study, ten species of Anabaena were used to test the congruence between the traditional morphological classification system and the present molecular classification system. The electrophoretic patterns for 10 cyanobacterial strains belonging to the genera (Anabaena) were used for molecular analysis using the repeated PCR technique. Genetic diversity was assessed using the banding patterns of repetitive DNA sequences including the HIP1 sequences, enterobacterial repetitive intergenic consensus [ERIC] sequences and REP sequences that are present in the cyanobacterial genome. PCR with different sets of repetitive oligonucleotides as primers were used to evaluate the various repetitive DNA sequences. The dendrograms based on the HIP1, ERIC and REP sequences did not show a clear correlation with morphological dendrogram based on traditional classification.

Key words: Anabaena, cyanobacteria, highly iterated palindromic sequences, enterobacterial repetitive intergenic consensus sequences, repetitive extragenic palindromic sequences, polymerase chain reaction.

Anbalagan Ezhilarasi*, , Narayanaswamy An

Page: 1 - 10

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

The effect of cultivation conditions on the mycelial growth of a dark-septate endophytic isolate

Ya-li Lv, Li-hua Sun, Fu-sheng Zhang, Yue Zhao and Shun-xing Guo*

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, P. R. China.

Accepted 9 March, 2016

Abstract

The EF-37 isolate, one of DSE fungi, is beneficial to the growth and development of its host plant, Saussurea involucrata Kar. et Kir. The cultivation requirements including basic culture medium, temperature, light, pH, carbon source and nitrogen compounds were studied for their effects on mycelial growth of a dark-septate endophytic (DSE) fungus EF-37 by using one-factor-at-a- time method. Potato dextrose agar (PDA) was the best medium for the growth of endophyte EF-37. Our studies showed that 20°C, 24 h dark cultivation and pH 7 significantly influenced the growth of endophyte EF-37 on PDA medium. Moreover, glucose and calcium nitrate were found to be the best nutrients for EF- 37 growth. Under the optimal cultivation conditions, DSE fungus EF-37 isolate could grow actively. This is the first study about the effect of cultivation conditions on the growth of this strain, which provides the preparatory knowledge for the biological characteristics of DSE fungus EF-37.

Key words: Cultural conditions, dark-septate endophytic (DSE) fungus EF-37, mycelial growth, optimization, Saussurea involucrata Kar. et Kir.

Yue Zhao and Shun-xing Guo*, Fu-sheng Zhang, Li-hua Sun, Ya-li Lv

Page: 1 - 10

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

The identification of the first isolate of influenza B virus using a duplex RT- PCR DNA sequencing in Saudi Arabia (B/Riyadh/01/2007)

Fahad N. Almajhdi

Department of Botany and Microbiology, Center of Excellence in Biotechnology Research, College of Sciences, King Saudi University, Saudi Arabia. E-mail: [email protected]. Tel: +966-4677778. Fax: +966-4675833.

Accepted 24 February, 2016

Abstract

Although Influenza virus infections threaten thousands of lives each year worldwide and bear the risk of been an epidemic, little is known about the circulating strains in Saudi Arabia. The availability of a rapid and accurate diagnostic approach is essential for effective treatment and disease control. In the current study, we optimized a duplex RT-PCR assay for the simultaneous detection of influenza A and B viruses in clinical samples. The developed assay was utilized for testing 100 nasopharyngeal aspirates collected from young children hospitalized with acute respiratory tract infection in Riyadh. Influenza B not A viruses, were identified as possible causes of the disease syndrome. Recovery of influenza B virus from clinical samples was achieved by serial passage in chicken embryos and the virus isolate was designated as Influenza B/Riyadh/01/2007. The identity of B/Riyadh/01/2007 was confirmed by sequencing the RT-PCR product. The sequenced data was submitted to the Gene Bank under the accession number GU135839. An almost complete homology was recognized with all Influenza B virus strains available on Gene Bank. Although several studies suggested the presence of influenza B virus in respiratory tract infection of Saudi children, this is the first report that describes the isolation and partial identification of the circulating virus. Further studies that fully characterize the isolated strain are ongoing.

Key words: Influenza B virus, isolation, RT-PCR, DNA sequencing, Saudi Arabia.

Fahad N. Almajhdi

Page: 1 - 10

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

Development and evaluation of colloidal gold immunochromatographic strip for detection of Escherichia coli O157

Xihong Zhao¹, Xiaowei He¹*, Wenmei Li ^1,3, Yao Liu², Liansheng Yang¹ and Jihua Wang³

¹College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China.

²Zhongshan Supervision Testing Institute of Quality and Metrology, Zhongshan 528403, China.

³Guangzhou Wondfo Biotech CO., LTD, 510530, Guangzhou, China.

Accepted 24 February, 2016

Abstract

Escherichia coli O157:H7 is a serious and common human pathogen that can cause diarrhoea, haemorrhagic colitis, and haemolytic uraemic syndrome (HUS). In this study, the synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates probe specific to E. coli O157:H7 were performed, and the preparation of colloidal gold immunochromatographic strip based the biotin-streptavidin system was developed for detection of E. coli O157:H7.Monodispersional nanogold colloid was synthesized and preparation of nanogold-labeled polyclonal antibody probe to E. coli O157:H7 by citrate method. Combination of antibody with nanogold particles was also characterized by UV-visible (UV-vis) light absorption spectra and transmission electron microscopy (TEM). Furthermore, nanogold-labeled probe was used to develop an immunochromatographic (IC) strip for E. coli O157:H7 analysis. With this method, analysis could be completed in less than 10 min. Examination of the 65 known strains (36 E. coli O157 strains and 29 serotypes other than E. coli) showed 98.5% specifity and 100% sensitivity, only yield a false-positive reaction with Salmonella choleraesuis . The sensitivity of the IC strip was tested using 10-fold dilution E. coli O157 in foods, could be detected at a minimum of 2.3 × 10³ CFU/ml without enrichment and 2.3 CFU/ml after enrichment. Application of IC strip test were performed on 265 water samples, 340 beef samples, 208 milk samples and 120 cake samples after enrichment, the specificity of the strip was 99.2, 97.9, 94.6 and 94.9%, respectively. The sensitivity of the strip was 100% agreement with tradition culture method. The established method is very useful for monitoring E. coli O157 containment in food samples.

Key words: Escherichia coli O157, colloidal gold immunochromatographic assay, rapid test.

Liansheng Yang and Jihua Wang, Wenmei Li, Xihong Zhao, Yao Liu, Xiaowei He*

Page: 1 - 10

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

Preliminary in-vitro antibacterial activities of ethanolic extracts of Ficus sycomorus Linn. and Ficus platyphylla Del. (Moraceae)

Adeshina Gbonjubola Olusesan*, Okeke Claire-Lorentz Ebele, Osuagwu Nonyelum Onwuegbuchulam and Ehinmidu Joseph Olorunmola

Department of Pharmaceutics and Pharmaceutical Microbiology, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Kaduna State, Nigeria.

Accepted 23 March, 2016

Abstract

Antibacterial therapeutic failure due to emergence of resistant bacterial strain is a worldwide phenomenon. The search for effective antibacterial substances from sources such as plants has become a necessity to overcome emergent of bacterial resistant in clinical practice. The dried leaves and stem barks of Ficus sycomorus and Ficus platyphylla were collected in Samaru-Zaria, Nigeria in July 2006 and extracted with 70% aqueous ethanol at room temperature. The antibacterial activities such as susceptibility, Minimum inhibitory concentrations (M.I.C.) and the minimum bactericidal concentrations (M.B.C.) were determined using appropriate methods. Using the same concentration of the two test plants extracts, the zones of inhibition showed by F. sycomorus ranged between 11.5 - 21.5 mm while that of F. platyphylla was from 17.0 - 22.0 mm. The values of the M.I.C and M.B.C of F. sycomorus were 1.95, 31.3 and 3.91, 250 mg/ml, respectively. Similarly, F. platyphylla displayed 1.95 and 7.81 mg/ml M.I.C. values and 3.91 to 62.5 mg/ml M.B.C. values against the test organisms. The observed antibacterial activities in this study proved that the leaves and stem bark extracts of Ficus spp. obtained in Zaria support the forcloric claims of the use of Ficus plants in the treatment of ailment such as wound dressing.

Key words: Antibacterial activity, medicinal plants, bacterial resistance, Ficus spp.

Okeke Claire-Lorentz Ebele, Osuagwu Nonyelum Onwuegbuchulam and Ehinmidu Joseph Olorunmola, Adeshina Gbonjubola Olusesan*

Page: 1 - 10

Research Article

Advanced Journal of Microbiology Research Vol. 2016

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

Full Length Research Paper

Prevalence rate and antibiotic resistant pattern of methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis

Muhammad Arfat Yameen², Hina Nasim², Naeem Akhtar¹, Saira Iram¹, Imran Javed² and Abdul Hameed^2*

¹Microbiology Department, Holy Family Hospital, Rawalpindi, Pakistan 46000, Pakistan.

²Microbiology Research Laboratory, Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan 45320, Pakistan.

Accepted 17 December, 2015

Abstract

The aim of this study was to evaluate the prevalence rate and antibiotic resistant pattern of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE). A prospective study was conducted at Holy Family Hospital Rawalpindi, Pakistan and Microbiology Research Laboratory, Quaid-i-Azam University, Islamabad, Pakistan during the period from December 2007 to August 2008. The antibiotic resistance pattern was studied for MRSA and MRSE isolated from nasal samples from patients admitted in medical and surgical intensive care units. The study was conducted on 283 isolates. The results depicted that 25% isolates of S. aureus were MRSA and 29.78% isolates of S. epidermidis were MRSE. All MRSA and MRSE were susceptible to vancomycin and quinopristin/dalfopristin while all isolates of MRSE were susceptible to teicoplanin. All the isolates of MRSA and MRSE were multidrug-resistant. The susceptibility of the isolates to the drugs varied greatly. The resistance rate of MRSA to various antibiotics was found to be as follow: cephalaxin (90%), cephalothin (58%), cephradine (86%), ciprofloxacin (80%), gentamicin (34%), imipenum (42%), levofloxacin (75%), tetracycline (49%), rifampicin (14%) and teicoplanin (3%). The resistance rate of MRSE to various antibiotics was found to be as follow: cephalaxin (64%), cephalothin (29%), cephradine (64%), ciprofloxacin (50%), gentamicin (21%), imipenum (7%), levofloxacin (21%), tetracycline (21%) and rifampicin (29%). The minimum inhibitory concentration (MIC) value for MRSA and MRSE in case of vancomycin ranged 1-4 g/ml, for tetracycline 4-128 g/ml, for rifampicin 0.5-32 g/ml and for gentamicin 0.5 – 64 g/ml. Both MRSA and MRSE showed variable susceptibility with different antibiotic groups but high susceptibility with streptogramin and glycopeptide antibiotics.

Key words: Antibiotic resistance, staphylococcus aureus, staphylococcus epidermidis, minimum inhibitory concentration.

Hina Nasim, Imran Javed and Abdul Hameed*, Muhammad Arfat Yameen, Saira Iram, Naeem Akhtar

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