Partial genomic characterization on potentially new bacteriophage: New addition to ICTV database?

A small infectious micro-organism that able to replicate only inside the living cells is named as viruses, which are also the most abundant life form on the earth. These subcellular organisms can be classified into four groups ; animal, plant, archaea and bacteria viruses. Among them, bacteria viruses or also known as bacteriophages have the highest numbers with the  total estimation of 1031 .  The fact is that, until today only ~ 6,000 types of phages have been discovered since 1959 (ICTV report, 2011). Therefore, there are lots of new phages waiting to be discovered. Partial genomic studies have been conducted on a potential new bacteriophage isolated from goat faeces. Thus, to fulfil the objective of this study genomic characterization was carried out to discover more genomic details in order to reveal the true identity of the newly isolated phage . As first part of the analysis, phage genome was extracted to identify the phage's genome type and to estimate the genome size. Along with that, the extracted genome was used to proceed with the genome profiling and also for  partial genome sequencing analysis. Based on the genomic analysis, the results showed the isolated phage carries DNA as it genetic material with the size of genome around 30-40kbp. Besides that, this phage produced different genomic profile in comparison with other known phages such as Lambda, T4 and T7 phages when digest with the same restriction enzymes. Besides that, based on partial genome sequencing and results run against NCBI database, this newly isolated phage might be related with another newly isolated phage known as Sodalis phage SO-1. As far as the genomic data collected, an assumption can be made that this isolated phage could be a new addition to the list of viruses in International Committee on Taxonomy of Viruses (ICTV) database

Exploring phage diversity and potential: Development of phage therapy for bacterial infection

The recent increase in drug-resistance bacteria has become a very serious threat to the treatment of infectious diseases. Over recent decades, a growing numbers of literatures have validated the application of bacteriophages for therapy against antibiotic-resistance bacteria. With rapid dissemination of these resistant bacteria pathogens, the interest in alternative remedies to antibiotics using bacteriophages therapy is gaining new ground. Based on the recent studies of bacteriophage applications against bacterial infections in countries where this alternative therapy has been approved, many scientists and companies believed that phages have the ability to treat and prevent diseases caused by bacteria. Malaysia, being well known as one of the mega diverse countries could promise the potential new phages that could be applied in phage therapy. The overall objectives of this research including isolation and purification of potentially new phages from environments, characterization of the isolated phages based on morphological study, physicochemical attributes, genomic and proteomic analysis. Lastly, the isolated phages would be developed into phage therapy against bacterial infection through in vitro and in vivo test. Preliminary results presented here, show a total of four phages were successfully isolated from human waste. Two of the phages were successfully isolated infecting Proteus mirabilis that caused urinary tract infection in human whereas another two phages infecting Escherichia coli 0157:H7 (caused food-borne diseases) and Escherichia coli ATCC 13706, respectively

Isolation and characterization of phage specific to E. coli O157:H7

. Phage therapy is not new because it has been practiced since the early 1900s to cure diseases such as shigellosis, typhus and dysentery that caused by bacteria. However, when antibiotics were discovered in 1941, this antibacterial therapy was quickly abandoned by most western scientists. Antibiotic treatment is usually prescribed to treat pathogenic bacteria E. coliO157:H7. However, since 1950s, the widespread resistance to antibiotics has raised public concern on development of alternatives to antibiotics. As a result of this resistance, there becomes a need for an alternative treatment to work synergistically with antibiotic to deal with bacterial infections. This has led to a new approach by the use of bacteriophage which is phage therapy. Once again, this treatment has received attention as an alternative method against bacterial infection. Phage therapy is the treatment on pathogenic bacterial infections by using bacteriophage which capable of invading bacterial cells and disrupting bacterial metabolism before lysing it. Phages are extremely host-specific and the natural enemies of bacteria. It soon becomes clear that there are huge numbers of bacteriophages existed and waiting to be isolated. Phage specific to E. coliO157:H7 was successfully isolated from sewage treatment sample. The appearance of plaques on lawn of E. coliO157:H7 plate indicates phage capabilities to infect and lyses the host cells. The isolated phage was further characterized based on its morphology, genomic profile, physicochemical attributes, and host specificity to assess their potentials to be developed as phage therapy against E. coliO157:H7.

Collaboration between Asian countries in materials, chemistry, life and food science, information technology, electric, civil engineering, architecture and mechanical engineering.

A bacteriophage infecting Pseudomonas aeruginosa USM AR2 was successfully isolated from hydrocarbon-contaminated muddy soil. Morphological study revealed a phage with icosahedral capsid head and long non-contractile tail. The genome of the phage was determined as double stranded DNA. Thus, the phage could be a member of the Siphoviridae family

Kaedah Kolorimetri untuk Analisis Kuantitatif Kapsaisin Secara Pencaman Corak Menggunakan Jaringan Neural Tiruan

A quantitative study for capsaicin based on the use of 2,6-dichloro-pbenzoquinone- 4-chlorimide (Gibbs reagent) and artificial neural network (ANN) has been carried out. The characterization include pH optimization, effect of reagent concentration, dynamic range of capsaicin concentration, photo stability, limit of detection and reproducibility. The optimum response was obtained at pH 11.0 and Gibbs reagent concentration of 2.96 x 104 M. The reproducibility of the method was very satisfactory with RSD values of 3.55%, 2.44% and 4.52% for capsaicin concentration of 200 ppm, 500 ppm and 800 ppm, respectively. Photostability test showed that the reagent was very stable with RSD value of 0.013% for the duration of 38 hours. A three layer feed-forward neural network was used and network training was performed by using back propagation algorithm. For the determination of capsaicin, a neural network with 20 hidden neurons, 0.00001% learning rate and trained over 47,738 cycles produced the best result. This network was able to extend the narrow dynamic range of capsaicin from 0 - 200 ppm to 0-600 ppm. The average interpolation error produced by this network was approximately 0.06

Molecular characterization of peroxisome proliferator activated receptor gamma2 (PPARγ2) promoter from bovine

Peroxisome proliferator activated receptor gamma (PPARγ) is a member of the PPAR subfamily of nuclear hormone receptors (Schoonjans et al., 1996a). So far, two PPARγ isoforms; γ1 and γ2, have been cloned in bovine and produced by alternative promoter usage and differential splicing of a single gene (Sundvold et al., 1997). PPARγ functions by forming heterodimeric complexes with 9-cis-retinoic acid X receptors (RXRs). This heterodimer regulates transcription by binding to PPRE, known also as DR-1 response element in the regulatory regions of target genes (Tontonoz et al., 1994a)

Surface modification of cellulose nanomaterial for urea biosensor application

Cellulose nanomaterial with rod-like structure and highly crystalline order, usually formed by elimination of the amorphous region from cellulose during acid hydrolysis. Cellulose nanomaterial with the property of biocompatibility and nontoxicity can be used for enzyme immobilization. In this work, urease enzyme was used as a model enzyme to study the surface modification of cellulose nanomaterial and its potential for biosensor application. The cellulose nanocrystal (CNC) surface was modified using 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation to introduce the carboxyl group at C6 primary alcohol. The success of enzyme immobilization and surface modification was confirmed using chemical tests and measured using UV-Visible spectrophotometer. The immobilization strategy was then applied for biosensor application for urea detection. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques were used for electroanalytical characterization of the urea biosensor

Prevalence of Salmonella spp. from Catfish (Clarias gariepinus) by using improvement isolation methods

Abstract. Catfish is an important freshwater fish used in food supply may be infected by pathogenic bacteria such as Salmonella spp. Due to its public health implications; an enhanced method of isolation of Salmonella spp. is desirable. A total of 60 samples (15 of whole-body of catfish, 15 of gills, 15 of intestines and 15 of water samples) collected from wet market in Penang (Malaysia) were examined. The aim of this study was to compare a pellet method to the conventional method (non-pellet method) of isolation of Salmonella spp. from different parts of catfish (Clarias Gariepinus) and water samples. In this study, the pellet method was assessed and compared to conventional method (non-pellet method). Three selective agars, Xylose Lysine Deoxycholate Agar (XLD), Xylose-Lysine-Tergitol 4 (XLT4), Bismuth Sulfite Agar (BSA), were used. Pellet method found to show significant difference (p<0.05) on a total of 15 catfish samples. Higher prevalence obtained in pellet method was 53.33%, 80%, 40% growth on XLD, XLT4 and BSA, respectively. By using non-pellet method, the prevalence was 46.67%, 53.33%, 13.33% on XLD, XLT4 and BSA, respectively. Salmonella spp. presented on whole-body of catfish, gills, intestine and water for 80%, 40% 20% and 6.67%, respectively. This result indicates that the pellet method can isolate Salmonella sp. higher compared to non-pellet method

Biosynthetic production of human growth hormone gene in methylotrophic yeast, Pichia pastoris

Human growth hormone (hGH) is secreted from the anterior pituitary gland and exerts a wide variety of functions such as, IGF-1 production, protein synthesis, glucose metabolism, lipolysis, lipogenesis, and cell proliferation/differentiation (Isaksson et al., 1985; Press, 1988; Thorner and Vance, 1988; Strobl and Thomas, 1994)