Regulation of quorum sensing activities by the stringent response gene rsh in sphingomonads is species-specific and culture condition dependent.

Stringent response and quorum sensing (QS) are two essential mechanisms that control bacterial global metabolism for better survival. Sphingomonads are a clade of bacteria that survive successfully in diverse ecosystems. In silico survey indicated that 36 out of 79 investigated sphingomonads strains contained more than one luxI homolog, the gene responsible for the biosynthesis of QS signal acyl homoserine lactones (AHLs). Investigation of the regulatory effects of the stringent response gene rsh on QS related bioactivities were carried out using rsh mutants of Sphingobium japonicum UT26 and Sphingobium sp. SYK-6, both had three luxI homologs. Results indicated that deletion of rsh upregulated the overall production of AHLs and extracellular polymeric substances (EPS) in both UT26 and SYK-6 in rich medium, but affected expressions of these luxI/luxR homologs in different ways. In the poor medium (1% LB), rsh mutant of SYK-6 significantly lost AHLs production in broth cultivation but not in biofilm cultivation. The regulatory effects of rsh on QS activities were growth phase dependent in UT26 and culture condition dependent in SYK-6. Our results demonstrated the negative regulatory effect of rsh on QS activities in sphingomonads, which were very different from the positive effect found in sphingomonads containing only one luxI/R circuit. This study extends the current knowledge on the intricate networks between stringent response and QS system in sphingomonads, which would help to understand their survival advantage.

Marine litter and microplastic pollution in mangrove sediments in the Sea of Oman.

Marine litter and microplastic pollution in mangroves pose significant threats. This study of litter in Omani mangroves revealed an average density of 0.83 to 21.92 items/m2. Quriyat lagoon emerged as the most contaminated area, hosting 133 items/m2, while Qurum Natural Reserve lagoon showed the least contamination at 10 items/m2. Plastics constituted 73-96 % of the litter, with microplastic levels in sediment ranging from 6 to 256 pieces/kg. Al-Sawadi's lagoon had the highest microplastic abundance (27.52 ± 5.32 pieces/kg), in contrast to Al Qurum's Marine Protected Area with the lowest (0.60 ± 1.12 pieces/kg). Microplastics, categorized as fragments, pellets, and fibers, were distributed across zones 40.05 % landward, 30.97 % seaward, and 28.98 % in the middle mangrove zones. Primary plastic polymers identified were Polyethylene (PE) at 40 % and High-Density Polyethylene (HDPE) at 28 %, along with others found in specific areas. Our findings provide essential baseline data for future monitoring efforts and management strategies in Oman and other countries.

Nanomaterial grafted polymorphous activated carbon cloth surface for antibacterial, capacitive deionization and oil spill cleaning applications.

This work reports the development of multifunctional or polymorphous surfaces using zinc oxide (ZnO) nanorods, silica (SiO2), and fluoropolymer functionalization in a sequential process. Firstly, zinc oxide nanorods were grown on activated carbon cloth (ACC) using a simple low-temperature synthesis process. ZnO nanorods-coated ACC substrate was applied to investigate the antimicrobial properties, and the results showed inhibition of 50% for Escherichia coli (E.coli) and 55% for Bacillus subtilis (B.subtilis) over 48 h of incubation time. Subsequent in-situ modification of silica nanoparticles like layer on ZnO nanorods-coated ACC surface was developed and used as an electrode for brackish water desalination in a capacitive deionization system. ZnO-SiO2 modified ACC surface enhanced the desalination efficiency by 1.6 times, the salt removal rate (SRR) by threefold, and the durability (fouling prevention) for long-term usage compared to pristine ACC. Further modification of the ZnO-SiO2-ACC surface using fluoropolymer rendered the surface superhydrophobic and oleophilic. Vegetable (1.4 g/g) and crude oil (1.6 g/g) adsorption capacities were achieved for modified surface which was 70% enhancement compared with pristine ACC. The dynamic oil spill adsorption test exhibited the complete removal of oil spills on water surfaces within a few seconds, suggesting a potential application in oil spill cleaning.

"Omics" Techniques Used in Marine Biofouling Studies.

Biofouling is the growth of organisms on wet surfaces. Biofouling includes micro- (bacteria and unicellular algae) and macrofouling (mussels, barnacles, tube worms, bryozoans, etc.) and is a major problem for industries. However, the settlement and growth of some biofouling species, like oysters and corals, can be desirable. Thus, it is important to understand the process of biofouling in detail. Modern "omic" techniques, such as metabolomics, metagenomics, transcriptomics, and proteomics, provide unique opportunities to study biofouling organisms and communities and investigate their metabolites and environmental interactions. In this review, we analyze the recent publications that employ metagenomic, metabolomic, and proteomic techniques for the investigation of biofouling and biofouling organisms. Specific emphasis is given to metagenomics, proteomics and publications using combinations of different "omics" techniques. Finally, this review presents the future outlook for the use of "omics" techniques in marine biofouling studies. Like all trans-disciplinary research, environmental "omics" is in its infancy and will advance rapidly as researchers develop the necessary expertise, theory, and technology.

Epibiotic bacteria on the carapace of hawksbill and green sea turtles.

In this study, epibiotic bacteria on the carapace of sea turtles at three different sites in the Persian Gulf were studied. Bacterial density counted using a Scanning Electron Microscope showed that the highest (9.4 × 106 ± 0.8 × 106 cm-2) and lowest (5.3 × 106 ± 0.4 × 106 cm-2) average bacterial densities were found on the green and hawksbill sea turtles, respectively. Bacterial community analysis using Illumina 16S rRNA gene sequencing showed that Gamma- and Alpha-proteobacteria were the dominant classes on all substrates. Some genera, such as Anaerolinea, were site- and substrate-specific. In general, bacterial communities on sea turtles differed from those on the non-living substrate, stones, and exhibited lower species richness and diversity compared to the latter. Despite some similarities, the majority of bacterial communities on the two sea turtles were different. This study provides baseline information about the epibiotic bacteria of sea turtles of different species.

Outer membrane vesicles induce the mussel plantigrade settlement via regulation of c-di-GMP.

Despite the importance of outer membrane vesicles (OMVs) in benthic animal settlement, the underlying molecular mechanism remains elusive. Here, the impact of OMVs and OMVs synthesis-related tolB gene in Mytilus coruscus plantigrade settlement was tested. The OMVs were extracted from Pseudoalteromonas marina through density gradient centrifugation, and a tolB knockout strain, achieved by homologous recombination, was utilized for the investigation. Our results demonstrated that OMVs could significantly enhance M. coruscus plantigrades settlement. Deleting the tolB resulted in downregulation of c-di-GMP, accompanied by a reduction of OMV production, a decline in bacterial motility and increasing biofilm-forming ability. Enzyme treatment resulted in a 61.11% reduction in OMV-inducing activity and a 94.87% reduction in LPS content. Thus, OMVs regulate mussel settlement via LPS, and c-di-GMP is responsible for the OMV-inducing capacity. These findings provide new insights into the interactions between bacteria and mussels.

Development of portable sensor for the detection of bacteria: effect of gold nanoparticle size, effective surface area, and interparticle spacing upon sensing interface.

In this study, systematic development of a portable sensor for the rapid detection of Escherichia coli (E. coli) and Exiguobacterium aurantiacum (E. aurantiacum) was reported. A conductive glass was utilized as a substrate and developed the electrode patterns on it. Trisodium citrate (TSC) and chitosan-stabilized gold nanoparticles (AuNPs) (CHI-AuNP-TSC) and chitosan-stabilized AuNPs (CHI-AuNP) were synthesized and utilized as a sensing interface. The morphology, crystallinity, optical properties, chemical structures, and surface properties of immobilized AuNPs on the sensing electrodes were investigated. The sensing performance of the fabricated sensor was evaluated by using an electrochemical method to observe the current changes in cyclic voltammetric responses. The CHI-AuNP-TSC electrode has higher sensitivity toward E. coli than CHI-AuNP with a limit of detection (LOD) of 1.07 CFU/mL. TSC in the AuNPs synthesis process played a vital role in the particle size, the interparticle spacing, the sensor's effective surface area, and the presence of CHI around AuNPs, thus enhancing the sensing performance. Moreover, post-analysis of the fabricated sensor surface exhibited the sensor stability and the interaction between bacteria and the sensor surface. The sensing results showed a promising potential for rapid detection using a portable sensor for various water and food-borne pathogenic diseases.

Biofouling and Antifouling: Interactions between Microbes and Larvae of Invertebrates.

The biofouling process refers to the undesirable accumulation of micro- and macro-organisms on manufactured surfaces [...].

Effect of UV-radiation on the physiology of the invasive green seaweed Codium fragile and its associated bacteria.

Invasive species such as seaweeds often have a broad tolerance, allowing them to colonize novel habitats. During invasion, also new epibacteria can be formed on seaweeds, which have important chemo-ecological effects. Since UV-radiation (UVR) is one of the main factors affecting seaweeds and their epibacteria, we tested its effect on intertidal and subtidal thalli of the invasive seaweed Codium fragile from three sites and monitored photosynthesis, antioxidant activity and epibacteria. Exposure to UV-radiation resulted in photoinhibition with a subsequent low recovery in subtidal thalli from 23°S compared to 27°S and 30°S, which both showed a higher and almost complete recovery. However, a high antioxidant activity was present in all thalli, permitting to explain its relatively high tolerance to new environments. UV-radiation modified the composition of the epibacteria community by reducing its diversity and evenness. Our results showed that C. fragile responds plastic to variable UV-radiation (depending on site and water depth), which contributes to its high invasion potential.

Addition of Gallic Acid Overcomes Resistance to Cisplatin in Ovarian Cancer Cell Lines.

OBJECTIVE: Ovarian cancer is one of the leading causes of cancer-related mortality in women, and is often associated with drug resistance. Therefore, finding effective drugs, including naturally derived compounds, is urgently needed. Herein, we aimed to test the anti-cancer potential of gallic acid monohydrate (GA) and its congeners on cisplatin-sensitive (A2780S), and resistant (A2780CP) ovarian cancer and normal ovarian (HOSE6-3) cell lines. METHODS: Cytotoxicity was assessed by AlamarBlue and CCK08 assays by exposing cells to different concentrations of cisplatin (0-21µg/mL), GA and its congeners (0-100µg/mL), and a combination of GA and cisplatin. Apoptosis was estimated by Hoechst stain and monitoring the relative RNA expression of the apoptotic effector caspase-3 using qRT-PCR. RESULTS: GA decreased cell viability in a concentration-dependent manner in all cell lines, with an IC50 of 19.39µg/mL (A2780S), 35.59 µg/mL (A2780CP), and 49.32µg/mL (HOSE6-3). GA displayed higher cytotoxicity than its congeners. An apoptotic rate estimation of approximately 20% and 30% was obtained in A2780S and A2780CP. While the cytotoxicity observed with cisplatin and GA was comparable, combining the two enhanced the cytotoxicity significantly, especially in the A2780CP cell line (p<0.05). CONCLUSION: These data suggest that GA may help overcome the resistance. Hence, the cytotoxic effects of GA, especially on chemo-resistant ovarian cancer cells merit further investigation.
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Autopsy of Used Reverse Osmosis Membranes from the Largest Seawater Desalination Plant in Oman.

The Barka desalination plant, commissioned in 2018, is the largest desalination plant in Oman. It has a capacity of 281 MLD with a reverse osmosis (RO) first-pass recovery rate of 46%. As part of the standard operator practice, a membrane autopsy was conducted to determine the cause of reductions in membrane performance. This study investigated fouled membranes (model No. SW30HRLE-440) from two different locations in the membrane rack. Various analytical methods were used to conduct the membrane autopsy. Field-emission scanning electron microscopy/energy-dispersive X-ray (FESEM/EDS) analyses of membrane samples showed major components of inorganic foulants. Moreover, black and salt-like crystals deposited on the membrane surface revealed significant carbon (C) components and oxygen (O), with a small amount of magnesium (Mg), chloride (Cl), sodium (Na), aluminium (Al), and calcium (Ca), respectively. A Fourier transform infrared (FTIR) analysis revealed the presence of long-chain hydrocarbons, carboxylic acids/esters, carbohydrates/polysaccharides, and inorganic foulants. Thermogravimetric analyses (TGA) of the membranes showed a high initial weight loss due to organic and inorganic fouling. X-ray photoelectron (XPS) analyses further confirmed the presence of inorganic and organic foulants on the membrane surfaces. Bacteria identification results showed the presence of Bacillus cereus and Bacillus marisflavi. This paper offers a detailed analysis of the foulants present on the reverse osmosis membrane surface and sub-surface before and after a cleaning process.

Bioluminescence of the tropical Indian Ocean: a multiple-scale variation.

Large-scale surveys represented by 5800 bathymetric casts in the western Indian Ocean (0-22o N, 54-58o E), elucidated the 10-fold variation of the bioluminescent potential (BP) in the upper mixed layer, during the winter (north-east) monsoon season. The mesoscale survey in February 2017 consisted of 26 drift stations (4o N-3o S, 65-68o E) on which 5-10 bathymetric casts were deployed down to 60 m. The maximal BP was associated with the periphery of a cyclonic eddy. The two-fold to three-fold variation of BP characterized the spatial heterogeneity modulated by a detected eddy. High-frequency casts on drift stations resembled the fine-scale heterogeneity in which the three-fold variation was observed within the BP maximum at a 37 ± 13 m depth. The latter one was located above the deep chlorophyll maximum at a 80 m depth. A general decline of the BP variance from the large scale through mesoscale to fine scale, fits that of the zooplankton biomass.

Hymenialdisine is Cytotoxic Against Cisplatin-Sensitive but Not Against Cisplatin-Resistant Cell Lines.

OBJECTIVES: New compounds are needed to overcome the resistance to commonly used cytotoxic chemotherapy for epithelial ovarian cancer. Marine sponges are a rich source of diverse chemical compounds and hymenialdisine has been found to have antiproliferative effects. This study aimed to investigate the cytotoxic effect of hymenialdisine in cisplatin-sensitive and cisplatin resistant ovarian cancer cell lines. METHODS: This study took place at Sultan Qaboos University, Muscat, Oman between August and November, 2019. The anti-cancer effects of hymenialdisine or cisplatin were assessed using treating cells with different concentrations of hymenialdisine and cisplatin. Cell viability was determined using the AlamarBlue® Assay. RESULTS: The half-maximal inhibitory concentration (IC50) of cisplatin was estimated at 31.4 µM for A2780S and 76.9 µM for A2780CP, whereas the IC50 of hymenialdisine was evaluated at 146.8 µM for A2780S cells. Despite the higher concentrations of hymenialdisine (up to 300 µM), IC50 could not be determined for the A2780CP cell line. CONCLUSION: When compared to cisplatin, hymenialdisine was less toxic against both A2780S and A2780CP ovarian cancer cell lines.

Reduction of mussel metamorphosis by inactivation of the bacterial thioesterase gene via alteration of the fatty acid composition.

The molecular mechanism underlying modulation of metamorphosis of the bivalve Mytilus coruscus by bacteria remains unclear. Here, the functional role of the thioesterase gene tesA of the bacterium Pseudoalteromonas marina in larval metamorphosis was examined. The aim was to determine whether inactivation of the tesA gene altered the biofilm-inducing capacity, bacterial cell motility, biopolymers, or the intracellular c-di-GMP levels. Complete inactivation of tesA increased the c-di-GMP content in P. marina, accompanied by a reduced fatty acid content, weaker motility, upregulation of bacterial aggregation, and biofilm formation. The metamorphosis rate of mussel larvae on ΔtesA biofilms was reduced by ∼ 80% compared with those settling on wild-type P. marina. Exogenous addition of a mixture of extracted fatty acids from P. marina into the ΔtesA biofilms promoted the biofilm-inducing capacity. This study suggests that the bacterial thioesterase gene tesA altered the fatty acid composition of ΔtesA P. marina biofilms (BF) through regulation of its c-di-GMP, subsequently impacting mussel metamorphosis.

Marine biofilms on different fouling control coating types reveal differences in microbial community composition and abundance.

Marine biofouling imposes serious environmental and economic impacts on marine applications, especially in the shipping industry. To combat biofouling, protective coatings are applied on vessel hulls which are divided into two major groups: biocidal and non-toxic fouling release. The current study aimed to explore the effect of coating type on microbial biofilm community profiles to better understand the differences between the communities developed on fouling control biocidal antifouling and biocidal-free coatings. Biocidal (Intersmooth® 7460HS SPC), fouling release (Intersleek® 900), and inert surfaces were deployed in the marine environment for 4 months, and the biofilms that developed on these surfaces were investigated using Illumina NGS sequencing, targeting the prokaryotic 16S rRNA gene. The results confirmed differences in the community profiles between coating types. The biocidal coating supported communities dominated by Alphaproteobacteria (Loktanella, Sphingorhabdus, Erythrobacter) and Bacteroidetes (Gilvibacter), while other taxa, such as Portibacter and Sva0996 marine group, proliferated on the fouling-release surface. Knowledge of these marine biofilm components on fouling control coatings will serve as a guide for future investigations of marine microfouling as well as informing the coatings industry of potential microbial targets for robust coating formulations.

Malformin-A1 (MA1) Sensitizes Chemoresistant Ovarian Cancer Cells to Cisplatin-Induced Apoptosis.

High-grade epithelial ovarian cancer is a fatal disease in women frequently associated with drug resistance and poor outcomes. We previously demonstrated that a marine-derived compound MalforminA1 (MA1) was cytotoxic for the breast cancer cell line MCF-7. In this study, we aimed to examine the effect of MA1 on human ovarian cancer cells. The potential cytotoxicity of MA1was tested on cisplatin-sensitive (A2780S) and cisplatin-resistant (A2780CP) ovarian cancer cell lines using AlamarBlue assay, Hoechst dye, flow cytometry, Western blot, and RT-qPCR. MA1 had higher cytotoxic activity on A2780S (IC50 = 0.23 µM) and A2780CP (IC50 = 0.34 µM) cell lines when compared to cisplatin (IC50 = 31.4 µM and 76.9 µM, respectively). Flow cytometry analysis confirmed the cytotoxic effect of MA1. The synergistic effect of the two drugs was obvious, since only 13% of A2780S and 7% of A2780CP cells remained alive after 24 h of treatment with both MA1 and cisplatin. Moreover, we examined the expression of bcl2, p53, caspase3/9 genes at RNA and protein levels using RT-qPCR and Western blot, respectively, to figure out the cell death mechanism induced by MA1. A significant down-regulation in bcl2 and p53 genes was observed in treated cells compared to non-treated cells (p < 0.05), suggesting that MA1 may not follow the canonical pathway to induce apoptosis in ovarian cancer cell lines. MalforminA1 showed promising anticancer activity by inducing cytotoxicity in cisplatin-sensitive and cisplatin-resistant cancer cell lines. Interestingly, a synergistic effect was observed when MA1 was combined with cisplatin, leading to it overcoming its resistance to cisplatin.

Chitosan Nanocomposite Coatings Containing Chemically Resistant ZnO-SnOx Core-shell Nanoparticles for Photocatalytic Antifouling.

Functional nanocomposites with biopolymers and zinc oxide (ZnO) nanoparticles is an emerging application of photocatalysis in antifouling coatings. The reduced chemical stability of ZnO in the acidic media in which chitosan is soluble affects the performance of chitosan nanocomposites in antifouling applications. In this study, a thin shell of amorphous tin dioxide (SnOx) was grown on the surface of ZnO to form ZnO-SnOx core-shell nanoparticles that improved the chemical stability of the photocatalyst nanoparticles, as examined at pH 3 and 6. The photocatalytic activity of ZnO-SnOx in the degradation of methylene blue (MB) dye under visible light showed a higher efficiency than that of ZnO nanoparticles due to the passivation of electronic defects. Chitosan-based antifouling coatings with varying percentages of ZnO or ZnO-SnOx nanoparticles, with or without the glutaraldehyde (GA) crosslinking of chitosan, were developed and studied. The incorporation of photocatalysts into the chitosan matrix enhanced the thermal stability of the coatings. Through a mesocosm study using running natural seawater, it was found that chitosan/ZnO-SnOx/GA coatings enabled better inhibition of bacterial growth compared to chitosan coatings alone. This study demonstrates the antifouling potential of chitosan nanocomposite coatings containing core-shell nanoparticles as an effective solution for the prevention of biofouling.

Disinfection of Bacteria in Water by Capacitive Deionization.

Clean water is one of the primary UN sustainable development goals for 2,030 and sustainable water deionization and disinfection is the backbone of that goal. Capacitive deionization (CDI) is an upcoming technique for water deionization and has shown substantial promise for large scale commercialization. In this study, activated carbon cloth (ACC) electrode based CDI devices are used to study the removal of ionic contaminants in water and the effect of ion concentrations on the electrosorption and disinfection functions of the CDI device for mixed microbial communities in groundwater and a model bacterial strain Escherichia coli. Up to 75 % of microbial cells could be removed in a single pass through the CDI unit for both synthetic and groundwater, while maintaining the salt removal activity. Mortality of the microbial cells were also observed during the CDI cell regeneration and correlated with the chloride ion concentrations. The power consumption and salt removal capacity in the presence and absence of salt were mapped and shown to be as low as 0.1 kWh m-3 and 9.5 mg g-1, respectively. The results indicate that CDI could be a viable option for single step deionization and microbial disinfection of brackish water.

Anticorrosion Coated Stainless Steel as Durable Support for C-N-TiO2 Photo Catalyst Layer.

The development of durable photocatalytic supports resistant in harsh environment has become challenging in advanced oxidation processes (AOPs) focusing on water and wastewater remediation. In this study, stainless steel (SS), SS/Ti (N,O) and SS/Cr-N/Cr (N,O) anticorrosion layers on SS meshes were dip-coated with sol gel synthesised C-N-TiO2 photo catalysts pyrolysed at 350 °C for 105 min, using a heating rate of 50 °C/min under N2 gas. The supported C-N-TiO2 films were characterised by scanning electron microscopy (SEM) coupled with energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) and Raman spectroscopy. The results showed that C-N-TiO2 was successfully deposited on anticorrosion coated SS supports and had different morphologies. The amorphous C and TiO2 were predominant in C-N-TiO2 over anatase and rutile phases on the surface of SS and anticorrosion supports. The C-N-TiO2 coated films showed enhanced photocatalytic activity for the decolouration of O.II dye under both solar and UV radiation. The fabricated C-N-TiO2 films showed significant antibacterial activities in the dark as well as in visible light. Herein, we demonstrate that SS/Ti(N,O) and SS/Cr-N/Cr(N,O) anticorrosion coatings are adequate photocatalytic and corrosion resistant supports. The C-N-TiO2 photo catalytic coatings can be used for water and wastewater decontamination of pollutants and microbes.

ZnO nanorod-chitosan composite coatings with enhanced antifouling properties.

The current study describes the fabrication of chitosan­zinc oxide nanorods composite (CHT/ZnO) on fiberglass panels (support substrate). ZnO nanorods (NRs) with size ranging from 20 to 100 nm and some microrods with an approximate size of 0.5-1 µm were grown on fiberglass panels. CHT 1%/ZnO composite had ZnO NRs incorporated into chitosan (CHT) coating while ZnO NRs were not visible in the CHT 2%/ZnO NRs composite. XRD and FTIR results showed the presence of the ZnO and chitosan. The water contact angle decreased from 80° ± 2° (control) to 65° ± 2° for CHT 1%/ZnO NRs and 42 ± 2° for CHT 2%/ZnO NRs composite coatings. The antimicrobial activities of the coated fiberglass panels were investigated using biofilm-forming bacteria Bacillus subtilis and Escherichia coli under both light and dark conditions. CHT/ZnO composite coated fiberglass panels showed the strongest antimicrobial activity compared to chitosan, ZnO NRs coatings, and Zn-based antifouling paint in the experiments with B. subtilis and E. coli under light conditions. The highest antifouling activity was observed for CHT 2%/ZnO composites. CHT/ZnO composites can be good alternatives to the toxic antifouling paints.