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  1. Secondary metabolites produced by marine streptomyces as antibiofilm and quorum-sensing inhibitor of uropathogen Proteus mirabilis
    Younis KM, Usup G, Ahmad A
    Environ Sci Pollut Res Int, 2016 Mar;23(5):4756-67.
    PMID: 26538254 DOI: 10.1007/s11356-015-5687-9
    Quorum-sensing regulates bacterial biofilm formation and virulence factors, thereby making it an interesting target for attenuating pathogens. In this study, we investigated anti-biofilm and anti-quorum-sensing compounds from secondary metabolites of halophiles marine streptomyces against urinary catheter biofilm forming Proteus mirabilis without effect on growth viability. A total of 40 actinomycetes were isolated from samples collected from different places in Iraq including marine sediments and soil samples. Fifteen isolates identified as streptomyces and their supernatant screened as anti-quorum-sensing by inhibiting quorum-sensing regulated prodigiosin biosynthesis of Serratia marcescens strain Smj-11 as a reporter strain. Isolate Sediment Lake Iraq (sdLi) showed potential anti-quorum-sensing activity. Out of 35 clinical isolates obtained from Urinary catheter used by patient at the Universiti Kebangsaan Malaysia Medical Center, 22 isolates were characterized and identified as Proteus mirabilis. Isolate Urinary Catheter B4 (UCB4) showed the highest biofilm formation with highest resistance to used antibiotic and was chosen for further studies. Ethyl acetate secondary metabolites extract was produced from sdLi isolate. First, we determined the Minimum Inhibitory Concentration (MIC) of sdLi crude extract against UCB4 isolate, and all further experiments used concentrations below the MIC. Tests of subinhibitory concentrations of sdLi crude extract showed good inhibition against UCB4 isolate biofilm formation on urinary catheter and cover glass using Scanning electron microscopy and light microscopy respectively. The influence of sub-MIC of sdLi crude extract was also found to attenuate the quorum sensing (QS)-dependent factors such as hemolysin activity, urease activity, pH value, and motility of UCB4 isolate. Evidence is presented that these nontoxic secondary metabolites may act as antagonists of bacterial quorum sensing by competing with quorum-sensing signals for receptor binding.
  2. Fulltext Isolation, detection of virulence genes, antibiotic resistance genes, plasmid profile, and molecular typing among Vibrio parahaemolyticus isolated in Malaysian seawater from recreational beaches and fish
    Faja OM, Sharad AA, Younis KM, Alwan MG, Mohammed BJ, Ahmad A
    Vet World, 2019 Jul;12(7):1140-1149.
    PMID: 31528045 DOI: 10.14202/vetworld.2019.1140-1149
    BACKGROUND AND AIM: Despite the importance of the global emergence of Vibrio parahaemolyticus infections worldwide, there has been scanty information on its occurrence in Malaysian seawaters and fish. This study aimed to determine the occurrence of V. parahaemolyticus isolates using polymerase chain reaction targeted at toxin operon gene, thermostable direct hemolysin (tdh), and tdh-related hemolysin genes and to determine antibiotic resistance pattern, genes, and plasmid profile of V. parahaemolyticus from Malaysian seawaters and fish.

    MATERIALS AND METHODS: Samples were collected from four recreational beaches in Malaysia (Port Klang; Bachok; Port Dickson; and Mersing). Thiosulfate-citrate-bile salts-sucrose (TCBS) agar and chromogenic Vibrio agar were used for isolation and identification. Colonies with yellow color on TCBS and green color on chromogenic vibrio (CV) agar were considered to be V. parahaemolyticus and they were subjected to biochemical tests. All V. parahaemolyticus isolates were further subjected to identification using seven specific gene markers.

    RESULTS: Seventy-three Vibrio isolates were recovered. Only one gene thermostable direct hemolysin (tdh) from seawater isolates of Vibrio has high virulence gene percentage (95.23%). Two genes alkaline serine protease (asp) and (tdh) had high percentage of virulence (83.87% and 80.64%, respectively) from fish. Comparatively, fish isolates have a higher virulence percentage compared to seawater isolates. Only gene streptomycin resistance B (strB) from seawater had 100% of the resistance genes. All isolates were multi-antibiotic resistant. Seventeen antibiotic resistance patterns were observed. The isolates had plasmids of varying sizes ranging from 2.7 kb to 42.4 kb. Dendrogram based on antibiotic resistance patterns of V. parahaemolyticus isolates discriminated the isolates into three clusters.

    CONCLUSION: This study demonstrated the occurrence of pathogenic, multi-antibiotic-resistant V. parahaemolyticus strains in Malaysian coastal waters and fish, and this could constitute potential public health risks.

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