Current trends in polymerase chain reaction based detection of three major human pathogenic vibrios.

Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus are the most significant aquatic pathogens of the genera Vibrio, account for most Vibrio-associated outbreaks worldwide. Rapid identification of these pathogens is of great importance for disease surveillance, outbreak investigations and food safety maintenance. Traditional culture dependent methods are time-consuming and labor-intensive whereas culture-independent polymerase chain reaction (PCR) based assays are reliable, consistent, rapid and reproducible. This review covers the recent development and applications of PCR based techniques, which have accelerated advances in the analysis of nucleic acids to identify three major pathogenic vibrios. Emphasis has been given to analytical approaches as well as advantages and limits of the available methods. Overall, this review article possesses the substantial merit to be used as a reference guide for the researchers to develop improved PCR based techniques for the differential detection and quantification of Vibrio species.

Rapid and sensitive detection of Salmonella with reduced graphene oxide-carbon nanotube based electrochemical aptasensor.

Rapid detection of foodborne pathogens is crucial as ingestion of contaminated food products may endanger human health. Thus, the objective of this study was to develop a biosensor using reduced graphene oxide-carbon nanotubes (rGO-CNT) nanocomposite via the hydrothermal method for accurate and rapid label-free electrochemical detection of pathogenic bacteria such as Salmonella enterica. The rGO-CNT nanocomposite was characterized using Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray diffraction and transmission electron microscopy. The nanocomposite was dropped cast on the glassy carbon electrode and further modified with amino-modified DNA aptamer. The resultant ssDNA/rGO-CNT/GCE aptasensor was then used to detect bacteria by using differential pulse voltammetry (DPV) technique. Synergistic effects of aptasensor was evident through the combination of enhanced electrical properties and facile chemical functionality of both rGO and CNT for the stable interface. Under optimal experimental conditions, the aptasensor could detect S. Typhimurium in a wide linear dynamic range from 101 until 108 cfu mL-1 with a 101 cfu mL-1 of the limit of detection. This aptasensor also showed good sensitivity, selectivity and specificity for the detection of microorganisms. Furthermore, we have successfully applied the aptasensor for S. Typhimurium detection in real food samples.

Carbon nanotube-based aptasensor for sensitive electrochemical detection of whole-cell Salmonella.

In this study, an amino-modified aptasensor using multi-walled carbon nanotubes (MWCNTs)-deposited ITO electrode was prepared and evaluated for the detection of pathogenic Salmonella bacteria. An amino-modified aptamer (ssDNA) which binds selectively to whole-cell Salmonella was immobilised on the COOH-rich MWCNTs to produce the ssDNA/MWCNT/ITO electrode. The morphology of the MWCNT before and after interaction with the aptamers were observed using scanning electron microscopy (SEM). Cyclic voltammetry and electrochemical impedance spectroscopy techniques were used to investigate the electrochemical properties and conductivity of the aptasensor. The results showed that the impedance measured at the ssDNA/MWCNT/ITO electrode surface increased after exposure to Salmonella cells, which indicated successful binding of Salmonella on the aptamer-functionalised surface. The developed ssDNA/MWCNT/ITO aptasensor was stable and maintained linearity when the scan rate was increased from 10 mV s-1 to 90 mV s-1. The detection limit of the ssDNA/MWCNT/ITO aptasensor, determined from the sensitivity analysis, was found to be 5.5 × 101 cfu mL-1 and 6.7 × 101 cfu mL-1 for S. Enteritidis and S. Typhimurium, respectively. The specificity test demonstrated that Salmonella bound specifically to the ssDNA/MWCNT/ITO aptasensor surface, when compared with non-Salmonella spp. The prepared aptasensor was successfully applied for the detection of Salmonella in food samples.

TiO2 loaded on glycidol functionalized poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) nanobiocomposite film for photocatalytic and antibacterial activities.

The rapid acceleration of industrialization and urbanization has exacerbated water pollution, which is primarily caused by the presence of highly toxic, non-biodegradable contaminants in industrial waste and effluents. In response to this urgent issue, a novel nanobiocomposite film with titanium dioxide (TiO2) loaded onto a poly(3-hydroxybutyrate-co-18 mol% 3-hydroxyhexanoate) (18PHBH) matrix was developed to serve as an effective dual-function material with photocatalytic and antibacterial properties. Through Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR), Diffuse reflectance ultraviolet-visible (DRUV-Vis), Scanning Electron Microscope (SEM), and X-ray diffraction (XRD) analyses, the physicochemical properties of the TiO2/Gly/18PHBH nanobiocomposite film were exhaustively characterized, revealing effective TiO2 loading and uniform distribution on the film's surface. The film exhibited extraordinary photocatalytic degradation of methylene blue (MB) dye, with the 5TiO2/Gly/18PHBH film demonstrating the greatest efficiency. In addition, antibacterial testing revealed that the film was effective against 99.8 % of Staphylococcus aureus and 96.9 % of Pseudomonas aeruginosa. These results demonstrate the potential of polyhydroxyalkanoate-based films as exceptional nanoparticle matrices and position the 5TiO2/Gly/18PHBH film as a versatile candidate for applications in photocatalysis and antibacterial interventions, providing innovative solutions to critical environmental challenges.

Sustainable oil palm trunk fibre based activated carbon for the adsorption of methylene blue.

Activated carbon (AC) is becoming the limelight due to its widespread application as an adsorbent for wastewater treatment, gases, and catalysis. However, its high consumption and price have drawn more attention to the sustainable use of natural resources as precursor for AC production. This study focuses on synthesising AC from two types of oil palm trunk (OPT) fibres, a significant agricultural waste products produced by Malaysia's thriving palm oil industries. The BET surface area of about 2057.9 m2 g-1 was achieved by chemical activation with phosphoric acid (H3PO4). The efficiency of the synthesised AC was critically analysed based on the adsorption experiments with methylene blue (MB) by varying several parameters (dosage of adsorbent, pH, initial dye concentration, and temperature of the solution) to elucidate the adsorption mechanism(s). A maximum adsorption capacity of 320.4 mg g-1 at 50 °C was achieved, and the Temkin (r2 = 0.98, 0.95, 0.95) and Langmuir (r2 = 0.94, 0.93, 0.95) isotherm models fitted the adsorption process better than the Freundlich (r2 = 0.95, 0.90, 0.86) model. Besides, the pseudo-second-order model (r2 > 0.90) best described the adsorption process, favouring chemisorption over physisorption. Thermodynamics showed MB adsorption on AC was spontaneous except at the highest dye concentration. It was exothermic at lower dye concentrations (50 and 100 mg L-1) and endothermic at higher ones (300, 500, and 700 mg L-1). In a nutshell, this study reveals that OPT fibre is a promising precursor for synthesising highly porous AC for the adsorption of MB dye.

Exploring Various Techniques for the Chemical and Biological Synthesis of Polymeric Nanoparticles.

Nanoparticles (NPs) have remarkable properties for delivering therapeutic drugs to the body's targeted cells. NPs have shown to be significantly more efficient as drug delivery carriers than micron-sized particles, which are quickly eliminated by the immune system. Biopolymer-based polymeric nanoparticles (PNPs) are colloidal systems composed of either natural or synthetic polymers and can be synthesized by the direct polymerization of monomers (e.g., emulsion polymerization, surfactant-free emulsion polymerization, mini-emulsion polymerization, micro-emulsion polymerization, and microbial polymerization) or by the dispersion of preformed polymers (e.g., nanoprecipitation, emulsification solvent evaporation, emulsification solvent diffusion, and salting-out). The desired characteristics of NPs and their target applications are determining factors in the choice of method used for their production. This review article aims to shed light on the different methods employed for the production of PNPs and to discuss the effect of experimental parameters on the physicochemical properties of PNPs. Thus, this review highlights specific properties of PNPs that can be tailored to be employed as drug carriers, especially in hospitals for point-of-care diagnostics for targeted therapies.

Activated carbon from biomass waste precursors: Factors affecting production and adsorption mechanism.

The use of activated carbon is evidenced by the increased scope of carbon-based applications in various industrial applications including pharmaceutical antidotes, wastewater remediation, aquaculture and toxin removal. Activated carbon produced from biomass waste by various processing methods and conditions is emerging as a promising adsorbent for remediation of the ecosystem due to extensive discharge of pollutants. Methods of producing activated carbon, nature of lignocellulosic biomass waste, and interaction of adsorbent-adsorbate are some of the crucial factors that need to be scrutinized to produce an effective adsorbent. However, these factors have not been thoroughly discussed in the literature. Activated carbon needs to go through continuous and rigorous research and development through optimization of key parameters such as type of activation (physical/chemical) and processing conditions, especially for large-scale production. It is imperative to have a detailed understanding of the preeminent characteristics of the activated carbon such as pore size distribution, total pore volume, surface area, and yield of activated carbon that control the extents of adsorptions and production of activated carbon. To further clarify the involved mechanism, studies should focus on all the possible variables that influence the system. Therefore, this review provides a better understanding of factors that affect the production of an efficient activated carbon, important properties to be used as an adsorbent, and the involved mechanisms during the adsorption process followed by increasing demand for activated carbon in various fields.

Biomedical Applications of Polyhydroxyalkanoate in Tissue Engineering.

Tissue engineering technology aids in the regeneration of new tissue to replace damaged or wounded tissue. Three-dimensional biodegradable and porous scaffolds are often utilized in this area to mimic the structure and function of the extracellular matrix. Scaffold material and design are significant areas of biomaterial research and the most favorable material for seeding of in vitro and in vivo cells. Polyhydroxyalkanoates (PHAs) are biopolyesters (thermoplastic) that are appropriate for this application due to their biodegradability, thermo-processability, enhanced biocompatibility, mechanical properties, non-toxicity, and environmental origin. Additionally, they offer enormous potential for modification through biological, chemical and physical alteration, including blending with various other materials. PHAs are produced by bacterial fermentation under nutrient-limiting circumstances and have been reported to offer new perspectives for devices in biological applications. The present review discusses PHAs in the applications of conventional medical devices, especially for soft tissue (sutures, wound dressings, cardiac patches and blood vessels) and hard tissue (bone and cartilage scaffolds) regeneration applications. The paper also addresses a recent advance highlighting the usage of PHAs in implantable devices, such as heart valves, stents, nerve guidance conduits and nanoparticles, including drug delivery. This review summarizes the in vivo and in vitro biodegradability of PHAs and conducts an overview of current scientific research and achievements in the development of PHAs in the biomedical sector. In the future, PHAs may replace synthetic plastics as the material of choice for medical researchers and practitioners.

Antimicrobial resistance: Prevalence, economic burden, mechanisms of resistance and strategies to overcome.

Antibiotic resistance is a major health concern globally and has been estimated to cause 10 million deaths worldwide by year 2050 if the current trend of inappropriate and excessive use of antibiotics continues. Although, the discovery of antibiotics has saved countless of lives for the past 80 years, increasing levels of bacterial resistance to antibiotics would jeopardize the progress in clinical and agricultural sectors and may cause life-threatening situations even for previously treatable bacterial infections. Antibiotic resistance would increase the levels of poverty of low-middle income countries mostly due to extended hospital stays, higher cost of treatment and untimely deaths that directly affect the total productivity rate. Recent incidences of antibiotic resistance have been gradually increasing globally and this may potentiate horizontal transmission of the resistant gene and have been linked with cross-resistance to other antibiotic families as well. This review summarizes the global burden of antibiotic resistance from the economic viewpoint, highlights the recent incidences of antibiotic resistance mainly related to Escherichia coli, Acinetobacter baumannii, Klebsiella pneumoniae, Salmonella spp. and Staphylococcus aureus, describes the common mechanistic actions of antibiotic resistance and potential strategies to overcome antibiotic resistance.

Mechanistic actions and contributing factors affecting the antibacterial property and cytotoxicity of graphene oxide.

Recent advances in the field of nanotechnology contributed to the increasing use of nanomaterials in the engineering, health and biological sectors. Graphene oxide (GO) has great potentials as it could be fine-tuned to be adapted into various applications, especially in the electrical, electronic, industrial and clinical fields. One of the important applications of GO is its use as an antibacterial material due to its promising activity against a broad range of bacteria. However, our understanding of the mechanism of action of GO towards bacteria is still lacking and is often less described. Therefore, a comprehensive overview of bactericidal mechanistic actions of GO and the roles of physicochemical factors including size, aggregation, functionalization and adsorption behavior contributing to its antibacterial activities are described in this review. As the use of GO is expected to increase exponentially in the health sector, the cytotoxicity of GO among the cell lines is also discussed. Thus, this review emphasizes the physicochemical characteristics of GO that can be tailored for optimal antibacterial properties that is of importance to the health industry.

Antibacterial potential of Malaysian ethnomedicinal plants against methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA).

This study aimed to evaluate the antibacterial activities of 61 plant extracts from 49 Malaysian ethnomedicinal plants and to investigate the interaction of the active plant extracts in combination with synthetic antibiotics against the MSSA and MRSA strains. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the plant extracts were determined using a microdilution method against MSSA and MRSA strains. The interaction between active plant extracts and the antibiotics was assessed using the checkerboard method. The total fractional inhibitory concentration (∑FIC) indices from the combination were calculated to determine the nature of the interaction. Out of the 61 plant extracts tested against the MSSA strain, 7 plant extracts (~ 11%) showed MIC values of less than 200 µg/mL, 17 extracts (~ 28%) showed MIC between 200 and 800 µg/mL and seed extracts of Areca catechu showed MBC values of 400 µg/mL. The seed extract of A. catechu showed MIC and MBC of 400 µg/mL against the MRSA strains while leaf extract of Cocos nucifera showed MIC of 400 µg/mL against MRSA NCTC 12493. When the active plant extracts (MIC ≤ 200 µg/mL for MSSA, and ≤ 400 µg/mL for MRSA) were tested in combination with vancomycin and ciprofloxacin, they showed no interaction against both MSSA and MRSA with ∑FIC between 1.06 and 2.03. These findings provide a preliminary overview of the anti-MSSA and anti-MRSA properties of Malaysian ethnobotanical plants to combat Staphylococcal infections. Further research is needed to establish an antibacterial profile of the tested plant extracts.

Synergistic antibacterial actions of graphene oxide and antibiotics towards bacteria and the toxicological effects of graphene oxide on human epidermal keratinocytes.

Graphene oxide (GO) has displayed antibacterial activity that has been investigated in the past, however, information on synergistic activity of GO with conventional antibiotics is still lacking. The objectives of the study were to determine the combinatorial actions of GO and antibiotics against Gram-positive and Gram-negative bacteria and the toxicological effects of GO towards human epidermal keratinocytes (HaCaT). Interactions at molecular level between GO and antibiotics were analyzed using Attenuated Total Reflectance-Fourier-transform infrared spectroscopy (ATR-FTIR). Changes in the antibacterial activity of antibiotics towards bacteria through the addition of GO was investigated. Toxicity of GO towards HaCaT cells were examined as skin cells play a role as the first line of defense of the human body. The ATR-FTIR characterizations of GO and antibiotics showed adsorption of tested antibiotics onto GO. The combinatorial antibacterial activity of GO and antibiotics were found to increase when compared to GO or antibiotic alone. This was attributed to the ability of GO to disrupt bacterial membrane to allow for better adsorption of antibiotics. Cytotoxicity of GO was found to be dose-dependent towards HaCaT cell line, it is found to impose negligible toxic effects against the skin cells at concentration below 100 µg/mL.

Graphene oxide exhibits differential mechanistic action towards Gram-positive and Gram-negative bacteria.

The antibacterial nature of graphene oxide (GO) has stimulated wide interest in the medical field. Although the antibacterial activity of GO towards bacteria has been well studied, a deeper understanding of the mechanism of action of GO is still lacking. The objective of the study was to elucidate the difference in the interactions of GO towards Gram-positive and Gram-negative bacteria. The synthesized GO was characterized by Ultraviolet-visible spectroscopy (UV-vis), Raman and Attenuated Total Reflectance-Fourier-transform infrared spectroscopy (ATR-FTIR). Viability, time-kill and Lactose Dehydrogenase (LDH) release assays were carried out along with FESEM, TEM and ATR-FTIR analysis of GO treated bacterial cells. Characterizations of synthesized GO confirmed the transition of graphene to GO and the antibacterial activity of GO was concentration and time-dependent. Loss of membrane integrity in bacteria was enhanced with increasing GO concentrations and this corresponded to the elevated release of LDH in the reaction medium. Surface morphology of GO treated bacterial culture showed apparent differences in the mechanism of action of GO towards Gram-positive and Gram-negative bacteria where cell entrapment was mainly observed for Gram-positive Staphylococcus aureus and Enterococcus faecalis whereas membrane disruption due to physical contact was noted for Gram-negative Escherichia coli and Pseudomonas aeruginosa. ATR-FTIR characterizations of the GO treated bacterial cells showed changes in the fatty acids, amide I and amide II of proteins, peptides and amino acid regions compared to untreated bacterial cells. Therefore, the data generated further enhance our understanding of the antibacterial activity of GO towards bacteria.