Malay. J. Biochem. Mol. Biol. (2018) 21 (2)
TABLE OF CONTENTS
M. A. Adibah, A. Siti Nor Akmar and M. M. Hanafi
EVALUATION OF EXPRESSION OF OsAMT1;1 UNDER DIFFERENT NITROGEN CONCENTRATION
The NH4+ and NO3− are N sources available for plant nutrition in most soils. The NH4+ role in plant nutrition has most likely been underestimated. This study provides basic information on the expression of OsAMT1;1 that is needed in understanding factors contributing to upland rice Nitrogen Use Efficiency (NUE) at the molecular level. The objective of this studies is to evaluate OsAMT1;1 expression on landraces with different NUE under different nitrogen level. To further understand the molecular basis of responsiveness of these two contrasting upland rice landraces to different N inputs at the N uptake level, a laboratory experiment was conducted to study how members of ammonium transporter gene family respond to different nitrogen conditions. Two upland varieties, Landrace III (high Nitrogen Use Efficiency) and Landrace I (low Nitrogen Use Efficiency) were grown in 4 nitrogen levels (0.05mM NH4NO3, 0.1mM NH4NO3, 1mM NH4NO3 and 2mM NH4NO3). Quantitative RT-PCR was used to study the expression of patterns of rice OsAMT1;1 genes. Results from this study showed that Landrace III has relatively higher expression of OsAMT1;1 compared to Landrace I for all treatments. Expression of OsAMT1;1 is clearly up-regulated in response to low-nitrogen treatment. Expression levels of OsAMT1;1 in high-NUE upland rice landrace was higher compared to low-NUE upland rice landrace. These results suggested, expression of OsAMT1;1 had certain degree of influenced in NUE of upland rice landraces.
Ahmad Norasidi Mohd Raffie , Navindra Kumari Palanisany, Jamal Houssaini, Norazah Ahmad and Zaini Mohd Zain
MOLECULAR CHARACTERISTICS OF ANTIBIOTIC RESISTANCE IN DIFFERENT Haemophilus influenzae SEROTYPES
Antibiotic resistance in Haemophilus influenzae is increasingly common and caused significant burden in patients’ management and treatment. Factors contributing to the resistance vary in some strains and depend on genetic elements they possessed. In this study, we aim to determine the antibiotic resistant determinants in 18 clinical strains of Haemophilus influenzae. Serological typing has identified four strains to be H. influenzae type a (Hia) while fourteen were non-typeable H. influenzae (NTHi). Antibiotic susceptibility testing using disk diffusion method was carried out to determine their susceptibility patterns against three types of antibiotics. Strains that exhibited resistance to ampicillin (AMP), tetracycline (TE) or cotrimoxazole (SXT) were selected and underwent plasmid detection by PCR. These strains also were tested for the presence of integrative and conjugative elements (ICEs) and antibiotic resistance genes. Multiple resistances were only seen in NTHi with two strains exhibited resistance to all three antibiotics. AMP resistant gene (blaTEM-1) was detected in 7/8 AMP resistant strains; TE resistant genes, tet(B) was detected in 2/4 TE resistant strains and tet(M) in another 1/4 strains. Both trimethoprim and sulfamethoxazole resistant genes (dfrA1 and sul2) present in only 2/6 SXT resistant strains and the other 4/6 strains lack either gene. ICEs were not detected in Hia strains but present in six antibiotic resistant NTHi which represented by int and ofr51 genes. In conclusion, antibiotic resistance in Haemophilus influenzae is independent to the presence of ICEs and the genetic determinants of antibiotic resistance may differ among strains of Haemophilus influenzae.
Aziyah Abdul-Aziz, Izdinur Izzat Anuar, Sharifah Aminah Syed Mohamad, Zaini Mohd-Zain and Mohd Faiz Foong Abdullah
DETECTION OF TYPE I-V SCCmec IN METHICILLIN-RESISTANT Staphylococcus capitis ISOLATES FROM MALAYSIA
Staphylococcal Cassette Chromosomes carrying the mecA gene (SCCmec) confers resistant to methicillin and a number of β-lactam antibiotics in the pathogen Staphylococcus aureus. The SCCmec can be transferred to other staphylococcal species which are commensals on the human body. Seventeen clinical isolates of Staphylococcus capitis were tested for the presence of SCCmec type I to V using Polymerase Chain Reaction (PCR) and their correlation to resistance towards Cefoxitin and Oxacillin. All isolates (100%) were found to be resistant to Cefoxitin and Oxacillin. A total of 11 out of 17 isolates carry all five types of SCCmec elements tested. About 94% of isolates were positive for the presence of the mecA gene and SCCmec type V, while 88% of isolates show positive results for SCCmec type II and III and 82% of isolates were positive for both SCCmec type I and IV. Only one isolate tested negative for SCCmec typing and mecA despite being resistant to both antibiotics. The results show that commensal staphylococci can carry multiple versions of SCCmec and raise the concern that they may serve as reservoirs for antibiotic resistance genes.
Amyra Nurfarhana Mohd Zarib, Aliana Najwa Hamsah Zollappi, Mardhiah Zainal Abidin, Mohd Badrin Hanizam Abdul Rahim, Shafinaz Abdul Gani, Muhajir Hamid and Mohd Ezuan Khayat
TOTAL PHENOLIC, TOTAL FLAVONOID AND ANTIOXIDANT ACTIVITY OF EXTRACT FROM RHIZOME OF Cibotium Barometz PREPARED BY VARIOUS SOLVENTS
The extract from rhizome of Cibotium barometz prepared using a number of various solvents extraction technique was previously shown to possess antioxidant activity. However, due to the structural complexity of bioactive compound in plants, thus, it is important in determining the most suitable solvents for the antioxidant extraction. In this study, the extracts of rhizome from C. barometz was prepared using various polarities of solvents; cyclohexane, dichloromethane, ethyl acetate and methanol. The total phenolic and total flavonoid contents of the solvent extracts were determined using Folin-Ciocalteu method and aluminium colorimetric assay, while the antioxidant activity was determined using DPPH radical scavenging assay, ABTS radical scavenging assay and FRAP assay. Among the solvent extracts, ethyl acetate extract has the highest total phenolic content followed by methanol, cyclohexane and dichloromethane extract. Similarly, the highest flavonoid content was also observed in ethyl acetate but no significant different was observed between the flavonoid contents of the remaining solvent extracts. The antioxidant activity of the solvent extracts was also found to be related to the phenolic content where the highest DPPH and ABTS scavenging activity, and ferric reducing power were found in ethyl acetate extract, followed by methanol, dichloromethane and cyclohexane extract. Therefore, ethyl acetate appears to be the most suitable extracting solvent for antioxidant from the rhizome of C. barometz. The information obtained from the study could be used as a reference to extract antioxidant from rhizome of C. barometz for various applications
Mizaton Hazizul Hasan, Maisarah Mohd Zin, Ibtisam Abd Wahab, Thellie Ponto and Aishah Adam
MYRMECODIA PLATYTYREA AQUEOUS TUBER EXTRACT INHIBITS PAIN
Myrmecodia platytyrea, locally known as Sarang semut is an epiphytic plant mainly found throughout the entire Indonesian Archipelago, the Philippines, Papuasia and its associated islands, Solomon Island and the Cape York Peninsula in Australia. Myrmecodia sp. has been used as a remedy to treat minor ailments to severe diseases such as hyperuricemia, tuberculosis and cancer by local people of Indonesia. In this study, the analgesic effect of Myrmecodia platytyrea tuber aqueous extract (MPAE) was investigated using carrageenan-induced rat paw oedema model and acetic acid-induced writhing test. In both studies, acetylsalicylic acid (ASA, a non-steroidal inflammatory drug) (200 mg/kg) was used as a standard reference. In the carrageenan-induced rat paw oedema model, oedema was quantified by measuring the hind rat paw thickness after 4th hour of carrageenan induction. Administration of MPAE (100, 200 and 400 mg/kg, p.o.), significantly reduced the oedema thickness in a time-dependent manner. The same was also observed in rats treated with ASA. MPAE (100 mg/kg) was also able to reduce secretion of TNF-α, a pro-inflammatory cytokine. As for acetic acid-induced writhing test on mice, significant inhibition of abdominal constriction induced by 0.6% of acetic acid was observed in mice treated with MPAE (200 and 400 mg/kg, p.o), when compared to control. These analgesic effects shown by MPAE may be due to the presence of bioactive compounds such as flavonoids, phenolics and phytosterol in this extract. Our study suggests that M. platytyrea tuber aqueous extract have potential to be developed as an antinociceptive agent.
Nur Syahira Mohammad, Muhd Fauzi Safian, Shahrul Hisham Zainal Ariffin and Zaidah Zainal Ariffin
BIOTRANSFORMATION OF NITROFURANS ANTIBIOTICS BY ASPERGILLUS SPECIES – RESIDUAL ANTIBACTERIAL ACTIVITY
Nitrofurans antibiotics are banned drug. Nitrofurans antibiotics that administered to the animal will be excreted out in parent or metabolite through faeces. The excretion causes soil contamination and also the emerging of bacteria that resistant the nitrofurans antibiotics. The aim of this study is to isolate potential soil fungi species that able to degrade nitrofurans antibiotics and determine the antibacterial activity of the nitrofurans antibiotic residues. Nine different colonies were successfully identified using molecular tools, polymerase chain reactions. ITS 1 (forward) and ITS 4 (reverse) primer were used in this study. A total of six species from Aspergillus namely Aspergillus sydowii strain SCAU066, Aspergillus tamarii isolate TN-7, Aspergillus candidus strain KUFA 0062, Aspergillus versicolor isolate BAB- 6580, Aspergillus protuberus strain KAS 6024 and the rest are Cephaliophora tropica strain xsd08001 and Lichtheimia ramosa strain R and two species from Penicillium which are Penicillium citrinum strain FIB SR4 and Penicillium citrinum isolate J33. Growth curve of Aspergillus species was plotted, most fungi showed exponential phase from day 2 to day 6. Aspergillus species were used in degrading nitrofurans antibiotics furaltadone, furazolidone and nitrofurazone. This study showed Aspergillus tamarii efficiently degraded 12-30 % of nitrofurans antibiotics after 96 h of incubation. Thus the isolated soil fungi, Aspergillus tamarii showed the ability to degrade nitrofurans antibiotics.
Nurul Athirah Adam, Norrizah Jaafar Sidik, Nurul Izzati Osman and Nur Inani Rezal
EFFECTS OF DIFFERENT CULTURE SYSTEM FOR MICROPROPAGATION OF in vitro PLANTLETS Melastoma decemfidum
The current work was done to study the micropropagation of in vitro plantlets Melastoma decemfidum via different culture system which involve gelled cultures, permanent liquid cultures and temporary immersion bioreactor. In vitro grown cultures which aged three weeks were used in this study. Each system contained nutrient MS medium supplemented with 0.3 mg/L of BAP hormone. In temporary immersion bioreactor, in vitro plantlets is temporary immerse by liquid nutrient medium. In vitro plantlets were immersed with nutrient medium for 30 minutes in every one hour. The in vitro plantlets cultured in temporary immersion bioreactor gave the highest growth rate with significant differences (p<0.05) on shoot multiplication (4.62 ± 0.39), shoot length (0.34 ± 0.03) and leaves number (10.67 ± 0.62) as compared to permanent liquid cultures and agar gelled cultures. In this study, in vitro propagation of M. decemfidum has been successfully optimized by using temporary immersion bioreactor.
Rizal Husaini Razali, Nurul Ain Khoruddin, Hisyam Jamari, Geik Yong Ang, Choo Yee Yu, Richard Johari James, Kok Hoi Teh, Hishamshah Mohd Ibrahim, Lay Kek Teh and Mohd Zaki Salleh
UNDERSTANDING THE GENETIC RISK OF CHILDHOOD LEUKEMIA AMONG THE ORANG ASLI AND MALAY IN MALAYSIA: MINING THE WHOLE GENOME SEQUENCE DATABASE
Leukemia is the seventh most common childhood malignancy in Malaysia. It is of great interest to determine the genetic factors that cause leukemia in order to strategise for prevention and treatment modalities. We aim to determine the genetic variability that increases Acute Lymphoblastic Leukemia (ALL) susceptibility of the Orang Asli and the Malays using existing genome database. For the Orang Asli, the genomic DNA were isolated from blood and the whole genome sequencing analysis were performed. Genomes were assembled, aligned and variants were called using GATKBest Practise workflow. Leukemic-associated variants were identified and analysed from 98 Orang Asli and 96 Malays in this study. Of the 126 codon changes identified, the number of variants predicted to be deleterious among the Orang Asli was 24. The single nucleotide polymorphism (SNP) with the highest allele frequency associated with ALL risk in the Orang Asli and the Malays was XRCC1rs25487-G with 72.4% and 68.7%, respectively. This study is believed to be the first to report on the prediction of the genetic risks of susceptibility against leukemia among the Orang Asli compared to the Malays.
Wan Razarinah WAR., Rizlan Ross, E.E., Nor Farinna Abdul Rahim, Batrisyia Syazana Faridon and Khairul Adzfa Radzun.
ANTIMICROBIAL ACTIVITY OF MARINE GREEN ALGAE EXTRACT AGAINST MICROBIAL PATHOGENS
Marine macroalgae (seaweeds) had been recently discovered to be a highly potential alternative to commercial drugs in terms of their antimicrobial properties. Thus, this provides new option in production of novel drugs in pharmaceutical industries. In the present study, extract of green algae from Halimeda sp. which was found abundance in the coastal area of Peninsular Malaysia was evaluated for their antimicrobial activity. Green algae collected from the intertidal zone of Pulau Singgah, Port Dickson were extracted using methanol and further screened for their antimicrobial activities against microbial pathogens which are Gram positive bacteria (B. subtilis, S. aureus and B. cereus), Gram negative bacteria (P. aeruginosa, E. coli), yeast (C. albicans) and mold (A. niger). by disc diffusion method. The results showed that methanol extract of Halimeda sp. could inhibit the growth of B. subtilis, S. aureus and B. cereus. The highest zone of inhibition against B. subtilis S. aureus and B. cereus were recorded at 350 mg/ml concentrations with 13.67 ± 0.58 mm, 12.00 ± 0.00 mm and 9.67 ± 0.58 mm, respectively. Further tested with Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentrations (MBC) demonstrated that both values were between 9.37 mg/ml to 75 mg/ml. As a conclusion, Halimeda sp. collected from Pulau Singgah, Port Dickson showed promising antimicrobial activity against S. aureus and B. cereus.
Siti Aqlima Ahmad, Gayathrie Asokan, Nur Adeela Yasid, Norazah Mohammad Nawawi, Kavilasni Subramaniam, Nur Nadhirah Zakaria and Mohd Yunus Shukor
EFFECT OF HEAVY METALS ON BIODEGRADATION OF PHENOL BY ANTARCTIC BACTERIUM: ARTHROBACTER BAMBUSAE STRAIN AQ5-003
Antarctica which was known as the most pristine ice dessert is largely influenced by anthropogenic contamination. Usually, persistent organic pollutants (POPs) contaminate Antarctic environment via Long Range Atmospheric Transport (LRAT) as well as via in situ activities like research, tourism and marine transportation. The extreme environment of Antarctica becomes the greatest challenge in degrading these pollutants where they trapped in the environment and unsurprisingly accumulate in higher concentration. Unprocessed petroleum is a natural substance mainly composed of hydrocarbon molecules comprising hydrogen and carbon and can be broken down over time by microbes using unconventional metabolic pathways in a process called bioremediation. Nevertheless, the ability of microbes to bioremediate phenol can be affected by heavy metals that can be found in the environment. Hence, the purpose of this study was to investigate the effects of heavy metals on biodegradation of phenol using psychrophilic Arthrobacter bambusae strain AQ5-003 which was identified as Gram-positive cocci, tested (-) for Oxidase test and (+) for Catalase test. As a result, Arthrobacter bambusae strain AQ5-003 which completely degraded phenol at the concentration 0.2 g/L within 192 h at 15˚C was supplemented with heavy metals Ag, As, Cd, Co, Cr, Cu, Ni, Pb and Zn with the concentration 1 ppm and only Ag and Cd proven to inhibit the whole degradation process (p<0.001). Phenol degradation was affected by 0.4 ppm Ag which had caused a 40% inhibition while Cd concentration with 0.1 ppm had caused a 10% inhibition within 192 h of incubation. The IC50 of Ag and Cd on phenol degradation are 0.53 and 0.04 ppm. This study suggests that this Arthrobacter bambusae strain AQ5-003 has the potential to tolerate As, Co, Cr, Cu, Ni, Pb and Zn but intolerant to Ag and Cd in the bioremediation of phenol.