Displaying publications 21 - 40 of 91 in total

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  1. Comparison of microplate- and bottle-based methods to age yeast for chronological life span assays
    Kwong MMY, Lee JW, Samian MR, Watanabe N, Osada H, Ong EBB
    J Microbiol Methods, 2019 12;167:105743.
    PMID: 31629019 DOI: 10.1016/j.mimet.2019.105743
    This study compared the chronological life span and survival of Saccharomyces cerevisiae aged in a microplate or bottle, under different aeration and calorie restriction conditions. Our data shows that limited aeration in the microplate-aged culture contributed to slower outgrowth but extended yeast CLS compared to the bottle-aged culture.
    Matched MeSH terms: Microbial Viability
  2. Microencapsulation of Lactobacillus rhamnosus GG with flaxseed mucilage using co-extrusion technique
    Lai K, How Y, Pui L
    J Microencapsul, 2021 Mar;38(2):134-148.
    PMID: 33306440 DOI: 10.1080/02652048.2020.1863490
    AIM: This study aimed to evaluate the protective effect of flaxseed mucilage on the co-extrusion microencapsulation of Lactobacillus rhamnosus GG.

    METHODS: Core flow rate, chitosan coating, and flaxseed mucilage concentration were optimised for the microencapsulation of L. rhamnosus. The microbeads were characterised and evaluated on microencapsulation efficiency and cell released after 6 h of sequential digestion.

    RESULTS: The optimised parameters for the L. rhamnosus microencapsulation were 1.0 mL/min core flow rate, 0.4% (w/v) chitosan coating, and 0.4% (w/v) flaxseed mucilage. The L. rhamnosus microbeads with flaxseed mucilage in core and wall materials had a smooth surface with 781.3 µm diameter, the highest microencapsulation efficiency (98.8% w/w), lowest swelling (5196.7% w/w) and erosion ratio (515.5% w/w), and least cell release (<40% w/w) with 9.31 log10 CFU mL-1 after sequential digestion.

    CONCLUSIONS: This study showed the protective capacity of flaxseed mucilage towards the L. rhamnosus GG during microencapsulation and gastrointestinal environment.

    Matched MeSH terms: Microbial Viability
  3. Fulltext Nematode Peptides with Host-Directed Anti-inflammatory Activity Rescue Caenorhabditis elegans from a Burkholderia pseudomallei Infection
    Lim MP, Firdaus-Raih M, Nathan S
    Front Microbiol, 2016;7:1436.
    PMID: 27672387 DOI: 10.3389/fmicb.2016.01436
    Burkholderia pseudomallei, the causative agent of melioidosis, is among a growing number of bacterial pathogens that are increasingly antibiotic resistant. Antimicrobial peptides (AMPs) have been investigated as an alternative approach to treat microbial infections, as generally, there is a lower likelihood that a pathogen will develop resistance to AMPs. In this study, 36 candidate Caenorhabditis elegans genes that encode secreted peptides of <150 amino acids and previously shown to be overexpressed during infection by B. pseudomallei were identified from the expression profile of infected nematodes. RNA interference (RNAi)-based knockdown of 12/34 peptide-encoding genes resulted in enhanced nematode susceptibility to B. pseudomallei without affecting worm fitness. A microdilution test demonstrated that two peptides, NLP-31 and Y43C5A.3, exhibited anti-B. pseudomallei activity in a dose dependent manner on different pathogens. Time kill analysis proposed that these peptides were bacteriostatic against B. pseudomallei at concentrations up to 8× MIC90. The SYTOX green assay demonstrated that NLP-31 and Y43C5A.3 did not disrupt the B. pseudomallei membrane. Instead, gel retardation assays revealed that both peptides were able to bind to DNA and interfere with bacterial viability. In parallel, microscopic examination showed induction of cellular filamentation, a hallmark of DNA synthesis inhibition, of NLP-31 and Y43C5A.3 treated cells. In addition, the peptides also regulated the expression of inflammatory cytokines in B. pseudomallei infected macrophage cells. Collectively, these findings demonstrate the potential of NLP-31 and Y43C5A.3 as anti-B. pseudomallei peptides based on their function as immune modulators.
    Matched MeSH terms: Microbial Viability
  4. Fulltext Does Campylobacter jejuni form biofilms in food-related environments?
    Teh AH, Lee SM, Dykes GA
    Appl Environ Microbiol, 2014 Sep;80(17):5154-60.
    PMID: 24928882 DOI: 10.1128/AEM.01493-14
    Campylobacter jejuni is one of the most frequent causes of bacterial gastrointestinal food-borne infection worldwide. This species is part of the normal flora of the gastrointestinal tracts of animals used for food production, including poultry, which is regarded as the primary source of human Campylobacter infections. The survival and persistence of C. jejuni in food processing environments, especially in poultry processing plants, represent significant risk factors that contribute to the spread of this pathogen through the food chain. Compared to other food-borne pathogens, C. jejuni is more fastidious in its growth requirements and is very susceptible to various environmental stressors. Biofilm formation is suggested to play a significant role in the survival of C. jejuni in the food production and processing environment. The aims of this minireview were (i) to examine the evidence that C. jejuni forms biofilms and (ii) to establish the extent to which reported and largely laboratory-based studies of C. jejuni biofilms provide evidence for biofilm formation by this pathogen in food processing environments. Overall existing studies do not provide strong evidence for biofilm formation (as usually defined) by most C. jejuni strains in food-related environments under the combined conditions of atmosphere, temperature, and shear that they are likely to encounter. Simple attachment to and survival on surfaces and in existing biofilms of other species are far more likely to contribute to C. jejuni survival in food-related environments based on our current understanding of this species.
    Matched MeSH terms: Microbial Viability
  5. Improved production of live cells of Lactobacillus rhamnosus by continuous cultivation using glucose-yeast extract medium
    Ling LS, Mohamad R, Rahim RA, Wan HY, Ariff AB
    J Microbiol, 2006 Aug;44(4):439-46.
    PMID: 16953180
    In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates (0.05 h(-1) to 0.40 h(-1)) using a 2 L stirred tank fermenter with a working volume of 600 ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, mu-max, was estimated at 0.40 h(-1), and the Monod cell growth saturation constant, Ks, at approximately 0.25 g/L. Maximum cell viability (1.3 x 10(10) CFU/ml) was achieved in the dilution rate range of D = 0.28 h(-1) to 0.35 h(-1). Both maximum viable cell yield and productivity were achieved at D = 0.35 h(-1). The continuous cultivation of L. rhamnosus at D = 0.35 h(-1) resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.
    Matched MeSH terms: Microbial Viability
  6. Fulltext Altered Proteome of Burkholderia pseudomallei Colony Variants Induced by Exposure to Human Lung Epithelial Cells
    Al-Maleki AR, Mariappan V, Vellasamy KM, Tay ST, Vadivelu J
    PLoS One, 2015;10(5):e0127398.
    PMID: 25996927 DOI: 10.1371/journal.pone.0127398
    Burkholderia pseudomallei primary diagnostic cultures demonstrate colony morphology variation associated with expression of virulence and adaptation proteins. This study aims to examine the ability of B. pseudomallei colony variants (wild type [WT] and small colony variant [SCV]) to survive and replicate intracellularly in A549 cells and to identify the alterations in the protein expression of these variants, post-exposure to the A549 cells. Intracellular survival and cytotoxicity assays were performed followed by proteomics analysis using two-dimensional gel electrophoresis. B. pseudomallei SCV survive longer than the WT. During post-exposure, among 259 and 260 protein spots of SCV and WT, respectively, 19 were differentially expressed. Among SCV post-exposure up-regulated proteins, glyceraldehyde 3-phosphate dehydrogenase, fructose-bisphosphate aldolase (CbbA) and betaine aldehyde dehydrogenase were associated with adhesion and virulence. Among the down-regulated proteins, enolase (Eno) is implicated in adhesion and virulence. Additionally, post-exposure expression profiles of both variants were compared with pre-exposure. In WT pre- vs post-exposure, 36 proteins were differentially expressed. Of the up-regulated proteins, translocator protein, Eno, nucleoside diphosphate kinase (Ndk), ferritin Dps-family DNA binding protein and peptidyl-prolyl cis-trans isomerase B were implicated in invasion and virulence. In SCV pre- vs post-exposure, 27 proteins were differentially expressed. Among the up-regulated proteins, flagellin, Eno, CbbA, Ndk and phenylacetate-coenzyme A ligase have similarly been implicated in adhesion, invasion. Protein profiles differences post-exposure provide insights into association between morphotypic and phenotypic characteristics of colony variants, strengthening the role of B. pseudomallei morphotypes in pathogenesis of melioidosis.
    Matched MeSH terms: Microbial Viability
  7. TolC facilitates the intracellular survival and immunomodulation of Salmonella Typhi in human host cells
    Hussain A, Ong EBB, Balaram P, Ismail A, Kien PK
    Virulence, 2024 Dec;15(1):2395831.
    PMID: 39185619 DOI: 10.1080/21505594.2024.2395831
    Salmonella enterica serovar Typhi (S. Typhi) causes typhoid fever, a systemic infection that affects millions of people worldwide. S. Typhi can invade and survive within host cells, such as intestinal epithelial cells and macrophages, by modulating their immune responses. However, the immunomodulatory capability of S. Typhi in relation to TolC-facilitated efflux pump function remains unclear. The role of TolC, an outer membrane protein that facilitates efflux pump function, in the invasion and immunomodulation of S. Typhi, was studied in human intestinal epithelial cells and macrophages. The tolC deletion mutant of S. Typhi was compared with the wild-type and its complemented strain in terms of their ability to invade epithelial cells, survive and induce cytotoxicity in macrophages, and elicit proinflammatory cytokine production in macrophages. The tolC mutant, which has a defective outer membrane, was impaired in invading epithelial cells compared to the wild-type strain, but the intracellular presence of the tolC mutant exhibited greater cytotoxicity and induced higher levels of proinflammatory cytokines (IL-1β and IL-8) in macrophages compared to the wild-type strain. These effects were reversed by complementing the tolC mutant with a functional tolC gene. Our results suggest that TolC plays a role in S. Typhi to efficiently invade epithelial cells and suppress host immune responses during infection. TolC may be a potential target for the development of novel therapeutics against typhoid fever.
    Matched MeSH terms: Microbial Viability
  8. Improvement of ɣ-Aminobutyric Acid Production and Cell Viability of Lactiplantibacillus plantarum B7 via Whole-Cell Immobilisation in Repeated Batch Fermentation System
    Pannerchelvan S, Muhamad FN, Wasoh H, Mohamed MS, Wong FWF, Mohamad R, et al.
    Probiotics Antimicrob Proteins, 2024 Dec;16(6):1907-1924.
    PMID: 38085437 DOI: 10.1007/s12602-023-10200-4
    Whole-cell immobilisation technology involving ℽ-aminobutyric acid GABA biosynthesis using lactic acid bacteria (LAB) has been extensively studied owing to its numerous benefits over free-living bacteria, including enhanced productivity, improved cell viability, ability to prevent cell lysis and protect cells against bacteriophages and other stressful conditions. Therefore, a novel LAB biocatalyst was developed using various fruit and fruit waste, immobilising a potential probiotic strain, Lactiplantibacillus plantarum B7, via an adsorption method to improve GABA and cell viability. Apple and watermelon rind have been known to be the ideal natural supports for L. plantarum B7 owing to higher GABA and lactic acid production and improved cell viability among the other natural supports tested and selected to be used in repeated batch fermentation (RBF) to improve GABA production and cell viability. In general, immobilisation of L. plantarum B7 on natural support has better GABA and lactic acid production with improved cell viability via RBF compared to free cells. Watermelon rind-supported cells and apple-supported cells could produce nine and eight successful GABA cycles, respectively, within RBF, whereas free cells could only produce up to four cycles. When using watermelon rind-supported cells and apple-supported cells in RBF, the GABA titer may be raised by up to 6.7 (218.480 ± 0.280 g/L) and 6 (195.439 ± 0.042 g/L) times, respectively, in comparison to GABA synthesis by free cells in single batch fermentation (32.65 ± 0.029 g/L). Additionally, natural support immobilised L. plantarum B7 could retain half of its cell viability even after the 12th cycle of RBF, while no cell was observed in control.
    Matched MeSH terms: Microbial Viability
  9. Comparative Study of the Effects of Fluconazole and Voriconazole on Candida glabrata, Candida parapsilosis and Candida rugosa Biofilms
    Madhavan P, Jamal F, Pei CP, Othman F, Karunanidhi A, Ng KP
    Mycopathologia, 2018 Jun;183(3):499-511.
    PMID: 29380188 DOI: 10.1007/s11046-018-0243-z
    Infections by non-albicans Candida species are a life-threatening condition, and formation of biofilms can lead to treatment failure in a clinical setting. This study was aimed to demonstrate the in vitro antibiofilm activity of fluconazole (FLU) and voriconazole (VOR) against C. glabrata, C. parapsilosis and C. rugosa with diverse antifungal susceptibilities to FLU and VOR. The antibiofilm activities of FLU and VOR in the form of suspension as well as pre-coatings were assessed by XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction assay. Morphological and intracellular changes exerted by the antifungal drugs on Candida cells were examined by scanning electron microscope (SEM) and transmission electron microscope (TEM). The results of the antibiofilm activities showed that FLU drug suspension was capable of killing C. parapsilosis and C. rugosa at minimum inhibitory concentrations (MICs) of 4× MIC FLU and 256× MIC FLU, respectively. While VOR MICs ranging from 2× to 32× were capable of killing the biofilms of all Candida spp tested. The antibiofilm activities of pre-coated FLU were able to kill the biofilms at ¼× MIC FLU and ½× MIC FLU for C. parapsilosis and C. rugosa strains, respectively. While pre-coated VOR was able to kill the biofilms, all three Candida sp at ½× MIC VOR. SEM and TEM examinations showed that FLU and VOR treatments exerted significant impact on Candida cell with various degrees of morphological changes. In conclusion, a fourfold reduction in MIC50 of FLU and VOR towards ATCC strains of C. glabrata, C. rugosa and C. rugosa clinical strain was observed in this study.
    Matched MeSH terms: Microbial Viability/drug effects
  10. The antifungal agent of silver nanoparticles activated by diode laser as light source to reduce C. albicans biofilms: an in vitro study
    Astuti SD, Puspita PS, Putra AP, Zaidan AH, Fahmi MZ, Syahrom A, et al.
    Lasers Med Sci, 2019 Jul;34(5):929-937.
    PMID: 30413898 DOI: 10.1007/s10103-018-2677-4
    Candida albicans is a normal flora caused fungal infections and has the ability to form biofilms. The aim of this study was to improve the antifungal effect of silver nanoparticles (AgNPs) and the light source for reducing the biofilm survival of C. albicans. AgNPs were prepared by silver nitrate (AgNO3) and trisodium citrate (Na3C6H5O7). To determine the antifungal effect of treatments on C. albicans biofilm, samples were distributed into four groups; L + P+ was treatment with laser irradiation and AgNPs; L + P- was treatment with laser irradiation only; L - P+ was treatment with AgNPs only (control positive); L - P- was no treatment with laser irradiation or AgNPs (control negative). The growth of fungi had been monitored by measuring the optical density at 405 nm with ELISA reader. The particle size of AgNPs was measured by using (particle size analyzer) and the zeta potential of AgNPs was measured by using Malvern zetasizer. The PSA test showed that the particle size of AgNPs was distributed between 7.531-5559.644 nm. The zeta potentials were found lower than - 30 mV with pH of 7, 9 or 11. The reduction percentage was analyzed by ANOVA test. The highest reduction difference was given at a lower level irradiation because irradiation with a density energy of 6.13 ± 0.002 J/cm2 resulted in the biofilm reduction of 7.07 ± 0.23% for the sample without AgNPs compared to the sample with AgNPs that increased the biofilm reduction of 64.48 ± 0.07%. The irradiation with a 450-nm light source had a significant fungicidal effect on C. albicans biofilm. The combination of light source and AgNPs provides an increase of biofilm reduction compared to the light source itself.
    Matched MeSH terms: Microbial Viability/drug effects
  11. Survival of antibiotic resistant Escherichia coli in vacuumpacked keropok lekor: Food safety alert among SME keropok lekor producers
    Wan-Hamat H, Lani MN, Hamzah Y, Alias R, Hassan Z, Mahat NA
    Trop Biomed, 2020 Mar 01;37(1):103-115.
    PMID: 33612722
    The microbiological quality of thirty ready-to-eat (RTE) keropok lekor (a sausage shape Malaysian fish product) was evaluated in comparison to microbiological guidelines for ready to eat foods. The two E. coli isolates were subjected to DNA sequencing, identified and tested for their resistance towards fifteen different antibiotics. The survival and growth of the isolated E. coli strains inoculated in keropok lekor at atmospheric air and vacuum packaging were also evaluated. Results revealed that four samples (13.33%) contained Enterobacteriaceae counts that exceeded the recommended allowable counts of 4.0 log10 CFU/g. Unsatisfactory level of coliforms (< 1.7 log10 CFU/g) was also observed in ten of the samples; two of which contained E. coli (2.1 ± 0.17 and 3.7 ± 0.02 log10 CFU/g), suggesting of poor hygiene and sanitation practices. While the 'Possible E10' E. coli strain was observably resistant towards Nalidixic acid (30µg) alone, B10 E. coli isolate was worryingly resistant towards Ampicillin (10µg), Ceftazidime (30µg), Ciprofloxacin (5µg), Ceftriaxone (30µg), Nalidixic acid (30µg) and Tetracycline (30µg). This study also revealed that the growth and survival of the 'Possible E10' and B10 E. coli strains were not significantly affected by vacuum packaging when stored at both 4°C and 28°C. Therefore, intervention programmes to alert and educate smallmedium enterprisers (SMEs) of keropok lekor producers on food safety as well as potential health risks that can be associated due to inappropriate handling procedures of such product, merits consideration.
    Matched MeSH terms: Microbial Viability*
  12. Enhancement of anti-candidal activity of endophytic fungus Phomopsis sp. ED2, isolated from Orthosiphon stamineus Benth, by incorporation of host plant extract in culture medium
    Yenn TW, Lee CC, Ibrahim D, Zakaria L
    J Microbiol, 2012 Aug;50(4):581-5.
    PMID: 22923105 DOI: 10.1007/s12275-012-2083-8
    This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anti-candidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.
    Matched MeSH terms: Microbial Viability/drug effects
  13. Profiling of recovery efficiencies for three standard protocols (FDA-BAM, ISO-11290, and modified USDA) on temperature-injured Listeria monocytogenes
    Lee HY, Chai LC, Pui CF, Wong WC, Mustafa S, Cheah YK, et al.
    J Microbiol Biotechnol, 2011 Sep;21(9):954-9.
    PMID: 21952372
    There have been a number of studies conducted in order to compare the efficiencies of recovery rates, utilizing different protocols, for the isolation of L. monocytogenes. However, the severity of multiple cell injury has not been included in these studies. In the current study, L. monocytogenes ATCC 19112 was injured by exposure to extreme temperatures (60°C and -20°C) for a one-step injury, and for a two-step injury the cells were transferred directly from a heat treatment to frozen state to induce a severe cell injury (up to 100% injury). The injured cells were then subjected to the US Food and Drug Administration (FDA), the ISO-11290, and the modified United States Department of Agriculture (mUSDA) protocols, and plated on TSAyeast (0.6% yeast), PALCAM agar, and CHROMAgar Listeria for 24 h or 48 h. The evaluation of the total recovery of injured cells was also calculated based on the costs involved in the preparation of media for each protocol. Results indicate that the mUSDA method is best able to aid the recovery of heat-injured, freeze-injured, and heat-freeze-injured cells and was shown to be the most cost effective for heat-freeze-injured cells.
    Matched MeSH terms: Microbial Viability*
  14. Synthesis and anti-microbial potencies of 1-(2-hydroxyethyl)-3-alkylimidazolium chloride ionic liquids: microbial viabilities at different ionic liquids concentrations
    Hossain MI, El-Harbawi M, Alitheen NB, Noaman YA, Lévêque JM, Yin CY
    Ecotoxicol Environ Saf, 2013 Jan;87:65-9.
    PMID: 23107478 DOI: 10.1016/j.ecoenv.2012.09.020
    Three 1-(2-hydroxyethyl)-3-alkylimidazolium chloride room temperature ionic liquids (ILs) [2OHimC(n)][Cl]; (n=0, 1, 4) have been synthesized from the appropriate imidazole precursors and characterized by IR and NMR spectroscopies and elemental analysis. Their anti-microbial activities were investigated using the well-diffusion method. The viabilities of Escherichia coli, Aeromonas hydrophila, Listeria monocytogenes and Salmonella enterica as a function of IL concentrations were studied. The minimal inhibitory concentrations (MICs) and EC₅₀ values for the present ILs were within the concentration range from 60 to 125 mM and 23 to 73 mM. The anti-microbial potencies of the present ILs were compared to a standard antibiotic, gentamicin. The finding affords additional perspective on the level of ILs toxicity to aquatic lifeforms and yet, this characteristic can be readily harnessed to detect microbial growth and activity.
    Matched MeSH terms: Microbial Viability/drug effects*
  15. The production and characterisation of trichotoxin peptaibols, by Trichoderma asperellum
    Chutrakul C, Alcocer M, Bailey K, Peberdy JF
    Chem Biodivers, 2008 Sep;5(9):1694-706.
    PMID: 18816522 DOI: 10.1002/cbdv.200890158
    Trichoderma spp. are regularly found as a constituent of the mycoflora of many soils and are noted for their antagonistic activity against bacteria and other fungi. This latter property is the basis for the widespread interest in their use in the biological control of soil-borne fungal plant pathogens. This antagonism is partly based on their ability to produce an impressive inventory of secondary metabolites. An important group of bioactive metabolites produced by Trichoderma spp. are the non-ribosomal peptides (NRPs), especially the peptaibols. A virulent antagonistic strain, T. asperellum, which had been used in biological control strategies in Malaysia and previously examined for mycolytic enzyme production, has been studied for its potential for peptaibol production. The present research demonstrated the ability of T. asperellum to produce at least two metabolites which were identified as acid trichotoxin 1704E (Ac-Aib-Gly-Aib-Leu-Aib-Gln-Aib-Aib-Aib-Ala-Ala-Aib-Pro-Leu-Aib-Iva-Glu-Vol) and neutral trichotoxin 1717A (Ac-Aib-Gly-Aib-Leu-Aib-Gln-Aib-Aib-Aib-Ala-Aib-Aib-Pro-Leu-Aib-Iva-Gln-Vol). Addition of free Aib to the culture medium enhanced the production of trichotoxins. Biological activity of these substances was investigated against Bacillus stearothermophilus. The general characteristics of peptaibols, also found in the trichotoxins, include the presence of high proportions of the uncommon amino acid Aib, the protection of the N- and C-termini by an acetyl group and reduction of the C-terminus to 2-amino alcohols, respectively, amphipathy and microheterogeneity.
    Matched MeSH terms: Microbial Viability/drug effects
  16. A mutant L-asparaginase II signal peptide improves the secretion of recombinant cyclodextrin glucanotransferase and the viability of Escherichia coli
    Ismail NF, Hamdan S, Mahadi NM, Murad AM, Rabu A, Bakar FD, et al.
    Biotechnol Lett, 2011 May;33(5):999-1005.
    PMID: 21234789 DOI: 10.1007/s10529-011-0517-8
    L-Asparaginase II signal peptide was used for the secretion of recombinant cyclodextrin glucanotransferase (CGTase) into the periplasmic space of E. coli. Despite its predominant localisation in the periplasm, CGTase activity was also detected in the extracellular medium, followed by cell lysis. Five mutant signal peptides were constructed to improve the periplasmic levels of CGTase. N1R3 is a mutated signal peptide with the number of positively charged amino acid residues in the n-region increased to a net charge of +5. This mutant peptide produced a 1.7-fold enhancement of CGTase activity in the periplasm and significantly decreased cell lysis to 7.8% of the wild-type level. The formation of intracellular inclusion bodies was also reduced when this mutated signal peptide was used as judged by SDS-PAGE. Therefore, these results provide evidence of a cost-effective means of expression of recombinant proteins in E. coli.
    Matched MeSH terms: Microbial Viability*
  17. Isoprenyl caffeate, a major compound in manuka propolis, is a quorum-sensing inhibitor in Chromobacterium violaceum
    Gemiarto AT, Ninyio NN, Lee SW, Logis J, Fatima A, Chan EW, et al.
    Antonie Van Leeuwenhoek, 2015 Aug;108(2):491-504.
    PMID: 26059863 DOI: 10.1007/s10482-015-0503-6
    The emergence of antibiotic-resistant bacterial pathogens, especially Gram-negative bacteria, has driven investigations into suppressing bacterial virulence via quorum sensing (QS) inhibition strategies instead of bactericidal and bacteriostatic approaches. Here, we investigated several bee products for potential compound(s) that exhibit significant QS inhibitory (QSI) properties at the phenotypic and molecular levels in Chromobacterium violaceum ATCC 12472 as a model organism. Manuka propolis produced the strongest violacein inhibition on C. violaceum lawn agar, while bee pollen had no detectable QSI activity and honey had bactericidal activity. Fractionated manuka propolis (pooled fraction 5 or PF5) exhibited the largest violacein inhibition zone (24.5 ± 2.5 mm) at 1 mg dry weight per disc. In C. violaceum liquid cultures, at least 450 µg/ml of manuka propolis PF5 completely inhibited violacein production. Gene expression studies of the vioABCDE operon, involved in violacein biosynthesis, showed significant (≥two-fold) down-regulation of vioA, vioD and vioE in response to manuka propolis PF5. A potential QSI compound identified in manuka propolis PF5 is a hydroxycinnamic acid-derivative, isoprenyl caffeate, with a [M-H] of 247. Complete violacein inhibition in C. violaceum liquid cultures was achieved with at least 50 µg/ml of commercial isoprenyl caffeate. In silico docking experiments suggest that isoprenyl caffeate may act as an inhibitor of the violacein biosynthetic pathway by acting as a competitor for the FAD-binding pockets of VioD and VioA. Further studies on these compounds are warranted toward the development of anti-pathogenic drugs as adjuvants to conventional antibiotic treatments, especially in antibiotic-resistant bacterial infections.
    Matched MeSH terms: Microbial Viability/drug effects
  18. Effects of ultraviolet light (UV-C) and heat treatment on the quality of fresh-cut Chokanan mango and Josephine pineapple
    George DS, Razali Z, Santhirasegaram V, Somasundram C
    J Food Sci, 2015 Feb;80(2):S426-34.
    PMID: 25586772 DOI: 10.1111/1750-3841.12762
    The effects of ultraviolet (UV-C) and medium heat (70 °C) treatments on the quality of fresh-cut Chokanan mango and Josephine pineapple were investigated. Quality attributes included physicochemical properties (pH, titratable acidity, and total soluble solids), ascorbic acid content (vitamin C), antioxidant activity, as well as microbial inactivation. Consumers' acceptance was also investigated through sensory evaluation of the attributes (appearance, texture, aroma and taste). Furthermore, shelf-life study of samples stored at 4 ± 1 °C was conducted for 15 d. The fresh-cut fruits were exposed to UV-C for 0, 15, 30, and 60 min while heat treatments were carried out at 70 °C for 0, 5, 10 and 20 min. Both UV-C and medium heat treatments resulted in no significant changes to the physicochemical attributes of both fruits. The ascorbic acid content of UV-C treated fruits was unaffected; however, medium heat treatment resulted in deterioration of ascorbic acids in both fruits. The antioxidants were enhanced with UV-C treatment which could prove invaluable to consumers. Heat treatments on the other hand resulted in decreased antioxidant activities. Microbial count in both fruits was significantly reduced by both treatments. The shelf life of the fresh-cut fruits were also successfully extended to a maximum of 15 d following treatments. As for consumers' acceptance, UV-C treated fruits were the most accepted as compared to their heat-treated counterparts. The results obtained through this study support the use of UV-C treatment for better retention of quality, effective microbial inactivation and enhancement of health promoting compounds for the benefit of consumers.
    Matched MeSH terms: Microbial Viability/radiation effects
  19. Fulltext Toxic effect of high concentration of sonochemically synthesized polyvinylpyrrolidone-coated silver nanoparticles on Citrobacter sp. A1 and Enterococcus sp. C1
    Lau CP, Abdul-Wahab MF, Jaafar J, Chan GF, Abdul Rashid NA
    J Microbiol Immunol Infect, 2017 Aug;50(4):427-434.
    PMID: 26427880 DOI: 10.1016/j.jmii.2015.08.004
    BACKGROUND/PURPOSE: Currently, silver nanoparticles (AgNPs) have gained importance in various industrial applications. However, their impact upon release into the environment on microorganisms remains unclear. The aim of this study was to analyze the effect of polyvinylpyrrolidone-capped AgNPs synthesized in this laboratory on two bacterial strains isolated from the environment, Gram-negative Citrobacter sp. A1 and Gram-positive Enterococcus sp. C1.

    METHODS: Polyvinylpyrrolidone-capped AgNPs were synthesized by ultrasound-assisted chemical reduction. Characterization of the AgNPs involved UV-visible spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, and energy dispersive X-ray spectroscopy. Citrobacter sp. A1 and Enterococcus sp. C1 were exposed to varying concentrations of AgNPs, and cell viability was determined. Scanning electron microscopy was performed to evaluate the morphological alteration of both species upon exposure to AgNPs at 1000 mg/L.

    RESULTS: The synthesized AgNPs were spherical in shape, with an average particle size of 15 nm. The AgNPs had different but prominent effects on either Citrobacter sp. A1 or Enterococcus sp. C1. At an AgNP concentration of 1000 mg/L, Citrobacter sp. A1 retained viability for 6 hours, while Enterococcus sp. C1 retained viability only for 3 hours. Citrobacter sp. A1 appeared to be more resistant to AgNPs than Enterococcus sp. C1. The cell wall of both strains was found to be morphologically altered at that concentration.

    CONCLUSION: Minute and spherical AgNPs significantly affected the viability of the two bacterial strains selected from the environment. Enterococcus sp. C1 was more vulnerable to AgNPs, probably due to its cell wall architecture and the absence of silver resistance-related genes.

    Matched MeSH terms: Microbial Viability/drug effects*
  20. Growth, biofilm formation, antifungal susceptibility and oxidative stress resistance of Candida glabrata are affected by different glucose concentrations
    Ng TS, Desa MNM, Sandai D, Chong PP, Than LTL
    Infect Genet Evol, 2016 06;40:331-338.
    PMID: 26358577 DOI: 10.1016/j.meegid.2015.09.004
    Glucose is an important fuel source to support many living organisms. Its importance in the physiological fitness and pathogenicity of Candida glabrata, an emerging human fungal pathogen has not been extensively studied. The present study aimed to investigate the effects of glucose on the growth, biofilm formation, antifungal susceptibility and oxidative stress resistance of C. glabrata. In addition, its effect on the expression of a putative high affinity glucose sensor gene, SNF3 was also investigated. Glucose concentrations were found to exert effects on the physiological responses of C. glabrata. The growth rate of the species correlated positively to the amount of glucose. In addition, low glucose environments were found to induce C. glabrata to form biofilm and resist amphotericin B. Conversely, high glucose environments promoted oxidative stress resistance of C. glabrata. The expression of CgSNF3 was found to be significantly up-regulated in low glucose environments. The expression of SNF3 gene in clinical isolates was found to be higher compared to ATCC laboratory strains in low glucose concentrations, which may explain the better survivability of clinical isolates in the low glucose environment. These observations demonstrated the impact of glucose in directing the physiology and virulence fitness of C. glabrata through the possible modulation by SNF3 as a glucose sensor, which in turn aids the species to adapt, survive and thrive in hostile host environment.
    Matched MeSH terms: Microbial Viability/drug effects
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