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  1. Fulltext Glyphosate Herbicide Induces Changes in the Growth Pattern and Somatic Indices of Crossbred Red Tilapia (O. niloticus × O. mossambicus)
    Muhammad UA, Yasid NA, Daud HM, Shukor MY
    Animals (Basel), 2021 Apr 22;11(5).
    PMID: 33922293 DOI: 10.3390/ani11051209
    The development of glyphosate-resistant genetically modified organisms (GMO) has increased the use of herbicide glyphosate by several magnitudes in recent years. It is now the most commonly used pesticide globally that affects aquatic habitats, especially fish. This study aims to add new knowledge on the effect of technical grade glyphosate on several toxicity parameters and to identify the most effective parameter in predicting technical grade glyphosate chronic toxicity (seven weeks) to fish, especially Malaysia's heavily farmed red tilapia. The results show that a relatively high concentration of technical grade glyphosate is needed to induce significant changes in all tested parameters. However, the results also indicate that the bodyweight index is the most sensitive toxicity parameter in that a reduction in body weight was observed at 25 mg/L of glyphosate. Negative correlations between the glyphosate concentration and toxicity parameters such as specific growth rate (SGR), hepato-somatic index (HIS), and gonado-somatic index (GSI) were observed. The fish condition factor and feed conversion ratio were found not to be affected at the highest glyphosate concentration tested (150 mg/L). To conclude, crossbred red tilapia (O. niloticus × O. mossambicus) is one potential species for evaluating the toxic effects of technical grade glyphosate on fish.
  2. Fulltext Effective production of keratinase by gellan gum-immobilised Alcaligenes sp. AQ05-001 using heavy metal-free and polluted feather wastes as substrates
    Yusuf I, Ahmad SA, Phang LY, Yasid NA, Shukor MY
    3 Biotech, 2019 Jan;9(1):32.
    PMID: 30622870 DOI: 10.1007/s13205-018-1555-x
    The ability of gellan gum-immobilised cells of the heavy metal-tolerant bacterium Alcaligenes sp. AQ05-001 to utilise both heavy metal-free and heavy metal-polluted feathers (HMPFs) as substrates to produce keratinase enzyme was studied. Optimisation of the media pH, incubation temperature and immobilisation parameters (bead size, bead number, gellan gum concentration) was determined for the best possible production of keratinase using the one-factor-at-a-time technique. The results showed that the immobilised cells could tolerate a broader range of heavy metal concentrations and produced higher keratinase activity at a gellan gum concentration of 0.8% (w/v), a bead size of 3 mm, bead number of 250, pH of 8 and temperature of 30 °C. The entrapped bacterium was used repeatedly for ten cycles to produce keratinase using feathers polluted with 25 ppm of Co, Cu and Ag as substrates without the need for desorption. However, its inability to tolerate/utilise feathers polluted with Hg, Pb, and Zn above 5 ppm, and Ag and Cd above 10 ppm resulted in a considerable decrease in keratinase production. Furthermore, the immobilised cells could retain approximately 95% of their keratinase production capacity when 5 ppm of Co, Cu, and Ag, and 10 ppm of As and Cd were used to pollute feathers. When the feathers containing a mixture of Ag, Co, and Cu at 25 ppm each and Hg, Ni, Pb, and Zn at 5 ppm each were used as substrates, the immobilised cells maintained their operational stability and biological activity (keratinase production) at the end of 3rd and 4th cycles, respectively. The study indicates that HMPF can be effectively utilised as a substrate by the immobilised-cell system of Alcaligenes sp. AQ05-001 for the semi-continuous production of keratinase enzyme.
  3. In vivo and in vitro effects on cholinesterase of blood of Oreochromis mossambicus by copper
    Basirun AA, Ahmad SA, Sabullah MK, Yasid NA, Daud HM, Khalid A, et al.
    3 Biotech, 2019 Feb;9(2):64.
    PMID: 30729088 DOI: 10.1007/s13205-019-1592-0
    The present study is aimed to evaluate the effects of sub-acute toxicity testing of copper sulphate (CuSO4), on behavioural, histological and biochemical changes of the Oreochromis mossambicus (black tilapia) blood tissues. The effects were assessed according to the previous results on sub-acute toxicity test after exposing fish to several concentrations (0.0, 2.5, 5.0, and 10.0 mg/L). The observations of scanning electron microscope, and transmission electron microscope studies revealed severe histopathological changes on the surface and the cellular changes in blood tissues, respectively. The morphological alterations in blood involved irregular structure of red blood cell and blood clot formation. CuSO4 affected the biochemical alteration of the blood cholinesterase also known as serum cholinesterase (ChE). Blood ChE inhibited up to 80% of activity when exposed to 10.0 mg/L CuSO4. The findings from this study can further improve the quality standards of aquaculture industry and the fundamental basis in selecting suitable strains among freshwater fish species to be used as bioindicator.
  4. Fulltext Batch growth kinetic studies of locally isolated cyanide-degrading Serratia marcescens strain AQ07
    Karamba KI, Ahmad SA, Zulkharnain A, Yasid NA, Ibrahim S, Shukor MY
    3 Biotech, 2018 Jan;8(1):11.
    PMID: 29259886 DOI: 10.1007/s13205-017-1025-x
    The evaluation of degradation and growth kinetics of Serratia marcescens strain AQ07 was carried out using three half-order models at all the initial concentrations of cyanide with the values of regression exceeding 0.97. The presence of varying cyanide concentrations reveals that the growth and degradation of bacteria were affected by the increase in cyanide concentration with a total halt at 700 ppm KCN after 72 h incubation. In this study, specific growth and degradation rates were found to trail the substrate inhibition kinetics. These two rates fitted well to the kinetic models of Teissier, Luong, Aiba and Heldane, while the performance of Monod model was found to be unsatisfactory. These models were used to clarify the substrate inhibition on the bacteria growth. The analyses of these models have shown that Luong model has fitted the experimental data with the highest coefficient of determination (R2) value of 0.9794 and 0.9582 with the lowest root mean square error (RMSE) value of 0.000204 and 0.001, respectively, for the specific rate of degradation and growth. It is the only model that illustrates the maximum substrate concentration (Sm) of 713.4 and empirical constant (n) of 1.516. Tessier and Aiba fitted the experimental data with a R2 value of 0.8002 and 0.7661 with low RMSE of 0.0006, respectively, for specific biodegradation rate, while having a R2 value of 0.9 and RMSE of 0.001, respectively, for specific growth rate. Haldane has the lowest R2 value of 0.67 and 0.78 for specific biodegradation and growth rate with RMSE of 0.0006 and 0.002, respectively. This indicates the level of the bacteria stability in varying concentrations of cyanide and the maximum cyanide concentration it can tolerate within a specific time period. The biokinetic constant predicted from this model demonstrates a good ability of the locally isolated bacteria in cyanide remediation in industrial effluents.
  5. Optimisation of culture composition for glyphosate degradation byBurkholderia vietnamiensisstrain AQ5-12
    Manogaran M, Shukor MY, Yasid NA, Khalil KA, Ahmad SA
    3 Biotech, 2018 Feb;8(2):108.
    PMID: 29430369 DOI: 10.1007/s13205-018-1123-4
    The herbicide glyphosate is often used to control weeds in agricultural lands. However, despite its ability to effectively kill weeds at low cost, health problems are still reported due to its toxicity level. The removal of glyphosate from the environment is usually done by microbiological process since chemical process of degradation is ineffective due to the presence of highly stable bonds. Therefore, finding glyphosate-degrading microorganisms in the soil of interest is crucial to remediate this glyphosate.Burkholderia vietnamiensisstrain AQ5-12 was found to have glyphosate-degrading ability. Optimisation of biodegradation condition was carried out utilising one factor at a time (OFAT) and response surface methodology (RSM). Five parameters including carbon and nitrogen source, pH, temperature and glyphosate concentration were optimised. Based on OFAT result, glyphosate degradation was observed to be optimum at fructose concentration of 6, 0.5 g/L ammonia sulphate, pH 6.5, temperature of 32 °C and glyphosate concentration at 100 ppm. Meanwhile, RSM resulted in a better degradation with 92.32% of 100 ppm glyphosate compared to OFAT. The bacterium was seen to tolerate up to 500 ppm glyphosate while increasing concentration results in reduced degradation and bacterial growth rate.
  6. Characterisation of the simultaneous molybdenum reduction and glyphosate degradation byBurkholderia vietnamiensisAQ5-12 andBurkholderiasp. AQ5-13
    Manogaran M, Ahmad SA, Yasid NA, Yakasai HM, Shukor MY
    3 Biotech, 2018 Feb;8(2):117.
    PMID: 29430378 DOI: 10.1007/s13205-018-1141-2
    In this novel study, we report on the use of two molybdenum-reducing bacteria with the ability to utilise the herbicide glyphosate as the phosphorus source. The bacteria reduced sodium molybdate to molybdenum blue (Mo-blue), a colloidal and insoluble product, which is less toxic. The characterisation of the molybdenum-reducing bacteria was carried out using resting cells immersed in low-phosphate molybdenum media. Two glyphosate-degrading bacteria, namelyBurkholderia vietnamiensisAQ5-12 andBurkholderiasp. AQ5-13, were able to use glyphosate as a phosphorous source to support molybdenum reduction to Mo-blue. The bacteria optimally reduced molybdenum between the pHs of 6.25 and 8. The optimum concentrations of molybdate for strainBurkholderia vietnamiensis strainAQ5-12 was observed to be between 40 and 60 mM, while forBurkholderiasp. AQ5-13, the optimum molybdate concentration occurred between 40 and 50 mM. Furthermore, 5 mM of phosphate was seen as the optimum concentration supporting molybdenum reduction for both bacteria. The optimum temperature aiding Mo-blue formation ranged from 30 to 40 °C forBurkholderia vietnamiensis strainAQ5-12, whereas forBurkholderiasp. AQ5-13, the range was from 35 to 40 °C. Glucose was the best electron donor for supporting molybdate reduction, followed by sucrose, fructose and galactose for both strains. Ammonium sulphate was the best nitrogen source in supporting molybdenum reduction. Interestingly, increasing the glyphosate concentrations beyond 100 and 300 ppm forBurkholderia vietnamiensis strainAQ5-12 andBurkholderiasp. AQ5-13, respectively, significantly inhibited molybdenum reduction. The ability of these bacteria to reduce molybdenum while degrading glyphosate is a useful process for the bioremediation of both toxicants.
  7. Fulltext Evaluation of conventional and response surface level optimisation of n-dodecane (n-C12) mineralisation by psychrotolerant strains isolated from pristine soil at Southern Victoria Island, Antarctica
    Habib S, Ahmad SA, Johari WLW, Shukor MYA, Alias SA, Khalil KA, et al.
    Microb Cell Fact, 2018 Mar 17;17(1):44.
    PMID: 29549881 DOI: 10.1186/s12934-018-0889-8
    BACKGROUND: Biodegradation of hydrocarbons in Antarctic soil has been reported to be achieved through the utilisation of indigenous cold-adapted microorganisms. Although numerous bacteria isolated from hydrocarbon-contaminated sites in Antarctica were able to demonstrate promising outcomes in utilising hydrocarbon components as their energy source, reports on the utilisation of hydrocarbons by strains isolated from pristine Antarctic soil are scarce. In the present work, two psychrotolerant strains isolated from Antarctic pristine soil with the competency to utilise diesel fuel as the sole carbon source were identified and optimised through conventional and response surface method.

    RESULTS: Two potent hydrocarbon-degraders (ADL15 and ADL36) were identified via partial 16S rRNA gene sequence analysis, and revealed to be closely related to the genus Pseudomonas and Rhodococcus sp., respectively. Factors affecting diesel degradation such as temperature, hydrocarbon concentration, pH and salt tolerance were studied. Although strain ADL36 was able to withstand a higher concentration of diesel than strain ADL15, both strains showed similar optimal condition for the cell's growth at pH 7.0 and 1.0% (w/v) NaCl at the conventional 'one-factor-at-a-time' level. Both strains were observed to be psychrotrophs with optimal temperatures of 20 °C. Qualitative and quantitative analysis were performed with a gas chromatograph equipped with a flame ionisation detector to measure the reduction of n-alkane components in diesel. In the pre-screening medium, strain ADL36 showed 83.75% of n-dodecane mineralisation while the reduction of n-dodecane by strain ADL15 was merely at 22.39%. The optimised condition for n-dodecane mineralisation predicted through response surface methodology enhanced the reduction of n-dodecane to 99.89 and 38.32% for strain ADL36 and strain ADL15, respectively.

    CONCLUSIONS: Strain ADL36 proves to be a better candidate for bioaugmentation operations on sites contaminated with aliphatic hydrocarbons especially in the Antarctic and other cold regions. The results obtained throughout strongly supports the use of RSM for medium optimisation.

  8. Fulltext In Silico Study and Effects of BDMC33 on TNBS-Induced BMP Gene Expressions in Zebrafish Gut Inflammation-Associated Arthritis
    Mostofa F, Yasid NA, Shamsi S, Ahmad SA, Mohd-Yusoff NF, Abas F, et al.
    Molecules, 2022 Nov 28;27(23).
    PMID: 36500396 DOI: 10.3390/molecules27238304
    The bone morphogenic protein (BMP) family is a member of the TGF-beta superfamily and plays a crucial role during the onset of gut inflammation and arthritis diseases. Recent studies have reported a connection with the gut-joint axis; however, the genetic players are still less explored. Meanwhile, BDMC33 is a newly synthesized anti-inflammatory drug candidate. Therefore, in our present study, we analysed the genome-wide features of the BMP family as well as the role of BMP members in gut-associated arthritis in an inflammatory state and the ability of BDMC33 to attenuate this inflammation. Firstly, genome-wide analyses were performed on the BMP family in the zebrafish genome, employing several in silico techniques. Afterwards, the effects of curcumin analogues on BMP gene expression in zebrafish larvae induced with TNBS (0.78 mg/mL) were determined using real time-qPCR. A total of 38 identified BMP proteins were revealed to be clustered in five major clades and contain TGF beta and TGF beta pro peptide domains. Furthermore, BDMC33 suppressed the expression of four selected BMP genes in the TNBS-induced larvae, where the highest gene suppression was in the BMP2a gene (an eight-fold decrement), followed by BMP7b (four-fold decrement), BMP4 (four-fold decrement), and BMP6 (three-fold decrement). Therefore, this study reveals the role of BMPs in gut-associated arthritis and proves the ability of BDMC33 to act as a potential anti-inflammatory drug for suppressing TNBS-induced BMP genes in zebrafish larvae.
  9. Toxic effects of copper on liver and cholinesterase of Clarias gariepinus
    Padrilah SN, Ahmad SA, Yasid NA, Sabullah MK, Daud HM, Khalid A, et al.
    Environ Sci Pollut Res Int, 2017 Oct;24(28):22510-22523.
    PMID: 28804856 DOI: 10.1007/s11356-017-9923-3
    The release of pollutants, especially heavy metals, into the aquatic environment is known to have detrimental effects on such an environment and on living organisms including humans when those pollutants are allowed to enter the food chain. The aim of this study is to analyse the damage to Clarias gariepinus' liver caused by exposure to different concentrations of copper. In the present study, samples of C. gariepinus were exposed to sub-lethal copper sulphate (CuSO4) concentrations (from 0.2 to 20.0 mg/L) for 96 h. Physiological and behavioural alterations were observed with respect to their swimming pattern, mucus secretion and skin colour. Mortality was also observed at high concentrations of copper. Histopathological alterations of the liver were analysed under light, transmission and scanning electron microscopies. The liver of the untreated group showed normal tissue structures, while histopathological abnormalities were observed in the treated fish under light and electron microscopes with increased copper concentrations. Histopathological abnormalities include necrosis, melanomacrophage, hepatic fibrosis and congested blood vessels. In addition, the enzyme activity of liver cholinesterase (ChE) was also found to be affected by copper sulphate, as 100% of cholinesterase activity was inhibited at 20.0 mg/L. Thus, liver enzyme activity and histopathological changes are proven to be alternative sources for biomarkers of metal toxicity.
  10. Fulltext Microbiological Reduction of Molybdenum to Molybdenum Blue as a Sustainable Remediation Tool for Molybdenum: A Comprehensive Review
    Yakasai HM, Rahman MF, Manogaran M, Yasid NA, Syed MA, Shamaan NA, et al.
    Int J Environ Res Public Health, 2021 May 27;18(11).
    PMID: 34071757 DOI: 10.3390/ijerph18115731
    Molybdenum (Mo) microbial bioreduction is a phenomenon that is beginning to be recognized globally as a tool for the remediation of molybdenum toxicity. Molybdenum toxicity continues to be demonstrated in many animal models of spermatogenesis and oogenesis, particularly those of ruminants. The phenomenon has been reported for more than 100 years without a clear understanding of the reduction mechanism, indicating a clear gap in the scientific knowledge. This knowledge is not just fundamentally important-it is specifically important in applications for bioremediation measures and the sustainable recovery of metal from industrial or mine effluent. To date, about 52 molybdenum-reducing bacteria have been isolated globally. An increasing number of reports have also been published regarding the assimilation of other xenobiotics. This phenomenon is likely to be observed in current and future events in which the remediation of xenobiotics requires microorganisms capable of degrading or transforming multi-xenobiotics. This review aimed to comprehensively catalogue all of the characterizations of molybdenum-reducing microorganisms to date and identify future opportunities and improvements.
  11. Fulltext Rhizodegradation of Petroleum Oily Sludge-contaminated Soil Using Cajanus cajan Increases the Diversity of Soil Microbial Community
    Allamin IA, Halmi MIE, Yasid NA, Ahmad SA, Abdullah SRS, Shukor Y
    Sci Rep, 2020 Mar 05;10(1):4094.
    PMID: 32139706 DOI: 10.1038/s41598-020-60668-1
    Most components of petroleum oily sludge (POS) are toxic, mutagenic and cancer-causing. Often bioremediation using microorganisms is hindered by the toxicity of POS. Under this circumstance, phytoremediation is the main option as it can overcome the toxicity of POS. Cajanus cajan a legume plant, was evaluated as a phyto-remediating agent for petroleum oily sludge-spiked soil. Culture dependent and independent methods were used to determine the rhizosphere microorganisms' composition. Degradation rates were estimated gravimetrically. The population of total heterotrophic bacteria (THRB) was significantly higher in the uncontaminated soil compared to the contaminated rhizosphere soil with C. cajan, but the population of hydrocarbon-utilizing bacteria (HUB) was higher in the contaminated rhizosphere soil. The results show that for 1 to 3% oily sludge concentrations, an increase in microbial counts for all treatments from day 0 to 90 d was observed with the contaminated rhizosphere CR showing the highest significant increase (p  
  12. Fulltext Biodecolourisation of Reactive Red 120 as a Sole Carbon Source by a Bacterial Consortium-Toxicity Assessment and Statistical Optimisation
    Manogaran M, Yasid NA, Othman AR, Gunasekaran B, Halmi MIE, Shukor MYA
    Int J Environ Res Public Health, 2021 Mar 02;18(5).
    PMID: 33801387 DOI: 10.3390/ijerph18052424
    The application of microorganisms in azo dye remediation has gained significant attention, leading to various published studies reporting different methods for obtaining the best dye decolouriser. This paper investigates and compares the role of methods and media used in obtaining a bacterial consortium capable of decolourising azo dye as the sole carbon source, which is extremely rare to find. It was demonstrated that a prolonged acclimation under low substrate availability successfully isolated a novel consortium capable of utilising Reactive Red 120 dye as a sole carbon source in aerobic conditions. This consortium, known as JR3, consists of Pseudomonas aeruginosa strain MM01, Enterobacter sp. strain MM05 and Serratia marcescens strain MM06. Decolourised metabolites of consortium JR3 showed an improvement in mung bean's seed germination and shoot and root length. One-factor-at-time optimisation characterisation showed maximal of 82.9% decolourisation at 0.7 g/L ammonium sulphate, pH 8, 35 °C, and RR120 concentrations of 200 ppm. Decolourisation modelling utilising response surface methodology (RSM) successfully improved decolourisation even more. RSM resulted in maximal decolourisation of 92.79% using 0.645 g/L ammonium sulphate, pH 8.29, 34.5 °C and 200 ppm RR120.
  13. Fulltext Advances in Aptamer-Based Biosensors and Cell-Internalizing SELEX Technology for Diagnostic and Therapeutic Application
    Gan Z, Roslan MAM, Abd Shukor MY, Halim M, Yasid NA, Abdullah J, et al.
    Biosensors (Basel), 2022 Oct 25;12(11).
    PMID: 36354431 DOI: 10.3390/bios12110922
    Aptamers are a group of synthetic single-stranded nucleic acids. They are generated from a random library of single-stranded DNA or RNA by a technology named systematic evolution of ligands by exponential enrichment (SELEX). SELEX is a repetitive process to select and identify suitable aptamers that show high affinity and specificity towards target cells. Great strides have been achieved in the design, construction, and use of aptamers up to this point. However, only a small number of aptamer-based applications have achieved widespread commercial and clinical acceptance. Additionally, finding more effective ways to acquire aptamers with high affinity remains a challenge. Therefore, it is crucial to thoroughly examine the existing dearth and advancement in aptamer-related technologies. This review focuses on aptamers that are generated by SELEX to detect pathogenic microorganisms and mammalian cells, as well as in cell-internalizing SELEX for diagnostic and therapeutic purposes. The development of novel aptamer-based biosensors using optical and electrical methods for microbial detection is reported. The applications and limitations of aptamers are also discussed.
  14. Fulltext Production of Lipopeptide Biosurfactant by a Hydrocarbon-Degrading Antarctic Rhodococcus
    Habib S, Ahmad SA, Wan Johari WL, Abd Shukor MY, Alias SA, Smykla J, et al.
    Int J Mol Sci, 2020 Aug 26;21(17).
    PMID: 32858859 DOI: 10.3390/ijms21176138
    Rhodococci are renowned for their great metabolic repertoire partly because of their numerous putative pathways for large number of specialized metabolites such as biosurfactant. Screening and genome-based assessment for the capacity to produce surface-active molecules was conducted on Rhodococcus sp. ADL36, a diesel-degrading Antarctic bacterium. The strain showed a positive bacterial adhesion to hydrocarbon (BATH) assay, drop collapse test, oil displacement activity, microplate assay, maximal emulsification index at 45% and ability to reduce water surface tension to < 30 mN/m. The evaluation of the cell-free supernatant demonstrated its high stability across the temperature, pH and salinity gradient although no correlation was found between the surface and emulsification activity. Based on the positive relationship between the assessment of macromolecules content and infrared analysis, the extracted biosurfactant synthesized was classified as a lipopeptide. Prediction of the secondary metabolites in the non-ribosomal peptide synthetase (NRPS) clusters suggested the likelihood of the surface-active lipopeptide production in the strain's genomic data. This is the third report of surface-active lipopeptide producers from this phylotype and the first from the polar region. The lipopeptide synthesized by ADL36 has the prospect to be an Antarctic remediation tool while furnishing a distinctive natural product for biotechnological application and research.
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