Characterization of selected phages for biocontrol of food-spoilage pseudomonads.

Pseudomonas spp., such as P. fluorescens group, P. fragi, and P. putida, are the major psychrophilic spoilage bacteria in the food industry. Bacteriophages (phages) are a promising tool for controlling food-spoilage and food-poisoning bacteria; however, there are few reports on phages effective on food-spoilage bacteria such as Pseudomonas spp. In this study, 12 Pseudomonas phages were isolated from chicken and soil samples. Based on the host range and lytic activity at 30 °C and 4 °C and various combinations of phages, phages vB_PflP-PCS4 and vB_PflP-PCW2 were selected to prepare phage cocktails to control Pseudomonas spp. The phage cocktail consisting of vB_PflP-PCS4 and vB_PflP-PCW2 showed the strongest lytic activity and retarded regrowth of P. fluorescens and P. putida at 30 °C, 8 °C, and 4 °C at a multiplicity of infection of 100. Nucleotide sequence analysis of the genomic DNA indicated that vB_PflP-PCS4 and vB_PflP-PCW2 phages were lytic phages of the Podoviridae family and lacked tRNA, toxin, or virulence genes. A novel endolysin gene was found in the genomic DNA of phage vB_PflP-PCS4. The results of this study suggest that the phage cocktail consisting of vB_PflP-PCS4 and vB_PflP-PCW2 is a promising tool for the biocontrol of psychrophilic food-spoilage pseudomonads during cold storage and distribution.

Potential application of phage vB_EfKS5 to control Enterococcus faecalis and its biofilm in food.

Contaminated food with antibiotic-resistant Enterococcus spp. could be the vehicle for transmitting Enterococcus to humans and accordingly cause a public health problem. The accumulation of biogenic amines produced by Enterococcus faecalis (E. faecalis) in food may have cytological effects. Bacteriophages (phage in short) are natural antimicrobial agents and can be used alone or in combination with other food preservatives to reduce food microbial contaminants. The aim of this study was to isolate a novel phage against E. faecalis and determine its host range to evaluate its potential application. Bacteriophage, vB_EfKS5, with a broad host range, was isolated to control the growth of E. faecalis. The vB_EfKS5 genome is 59,246 bp in length and has a GC content of 39.7%. The computational analysis of phage vB_EfKS5 genome confirmed that it does not contain any lysogenic, toxic, or virulent genes. Phage vB_EfKS5 exhibited lytic activity against most E. faecalis isolates with different multiplicities of infections and it infected 75.5% (22/29) of E. faecalis isolates and 42.3% (3/7) of E. faecium isolates. It was also able to destroy the biofilm formed by E. faecalis with different MOIs. Phage vB_EfKS5 alone or in combination with nisin could control the growth of E. faecalis in broth and milk. Based on its high productivity, stability, short latent period, and large burst size, phage vB_EfKS5 has a high potential for applications both in food and medical applications.

The Fabrication of Halogen-Doped FeWO4 Heterostructure Anchored over Graphene Oxide Nanosheets for the Sunlight-Driven Photocatalytic Degradation of Methylene Blue Dye.

Rapid industrialization and urbanization are the two significant issues causing environmental pollution. The polluted water from various industries contains refractory organic materials such as dyes. Heterogeneous photocatalysis using semiconductor metal oxides is an effective remediation technique for wastewater treatment. In this research, we used a co-precipitation-assisted hydrothermal method to synthesize a novel I-FeWO4/GO sunlight-active nanocomposite. Introducing dopant reductive iodine species improved the catalytic activity of FeWO4/GO. I- ions improved the catalytic performance of H2O2 by doping into FeWO4/GO composite. Due to I- doping and the introduction of graphene as a support medium, enhanced charge separation and transfer were observed, which is crucial for efficient heterogeneous surface reactions. Various techniques, like FTIR, SEM-EDX, XRD, and UV-Vis spectroscopy, were used to characterize composites. The Tauc plot method was used to calculate pristine and iodine-doped FeWO4/GO bandgap. Iodine doping reduced the bandgap from 2.8 eV to 2.6 eV. The degradation of methylene blue (MB) was evaluated by optimizing various parameters like catalyst concentration, oxidant dose, pH, and time. The optimum conditions for photocatalysts where maximum degradation occurred were pH = 7 for both FeWO4/GO and I-FeWO4/GO; oxidant dose = 9 mM and 7 mM for FeWO4/GO and I-FeWO4/GO; and catalyst concentration = 30 mg and 35 mg/100 mL for FeWO4/GO and I-FeWO4/GO; the optimum time was 120 min. Under these optimum conditions, FeWO4/GO and I-FeWO4/GO showed 92.0% and 97.0% degradation of MB dye.

Comparison of prevalence, characterization, antimicrobial resistance and pathogenicity of foodborne Listeria monocytogenes in recent 5 years in Japan.

This study investigated the prevalence, serotype, antimicrobial resistance (AMR), virulence potential, and biofilm formation of Listeria monocytogenes isolated in 2022 in Japan and compared their profiles with those of isolates in 2012 and 2017. A total of 85 chicken samples were randomly collected from different supermarkets in Fukuoka in 2022. L. monocytogenes were isolated by conventional method and characterized by MALDI-TOF MS. Among 85 samples tested in 2022, 9 (10.6%) were positive for L. monocytogenes and 17 strains were isolated from the positive samples. The isolates were serotyped as 1/2b (41.2%), 3a (29.4%), 3b (23.5%) and 1/2a (5.9%). Antimicrobial susceptibility tests of the 2022 isolates showed susceptibility to majority of the antibiotics, except cefoxitin, oxacillin, and fosfomycin. Compared to the previous surveillance results, the prevalence of L. monocytogenes in 2022 (10.6%) was significantly lower (p < 0.05) than those of the isolates in 2017 (24%) and 2012 (52.9%). The distribution of serotypes 1/2a and 1/2b decreased over time, and serotype 4b was not detected in the 2022 isolates. The proportion of multidrug resistant strains in 2022 (16.7%) was significantly lower than those in 2012 (46.7%) and 2017 (82.6%). Moreover, a total of 36 isolates (12 isolates/ year) were used to detect the virulence genes (hlyA, plcA, clpC, and inlA) and biofilm-forming capacity. Most of the isolates from different years harboured four virulence genes. The biofilm formation of the 2022 isolates was significantly weaker (p < 0.05) than those of the 2012 and 2017 isolates. Thus, despite the low rates of contamination in chicken meat and AMR of the isolates, virulent L. monocytogenes contamination in food should still be acknowledged.

Investigation of boron-doped graphene oxide anchored with copper sulphide flowers as visible light active photocatalyst for methylene blue degradation.

The non-biodegradable nature of waste emitted from the agriculture and industrial sector contaminates freshwater reserves. Fabrication of highly effective and low-cost heterogeneous photocatalysts is crucial for sustainable wastewater treatment. The present research study aims to construct a novel photocatalyst using a facile ultrasonication-assisted hydrothermal method. Metal sulphides and doped carbon support materials work well to fabricate hybrid sunlight active systems that efficiently harness green energy and are eco-friendly. Boron-doped graphene oxide-supported copper sulphide nanocomposite was synthesized hydrothermally and was assessed for sunlight-assisted photocatalytic degradation of methylene blue dye. BGO/CuS was characterized through various techniques such as SEM-EDS, XRD, XPS, FTIR, BET, PL, and UV-Vis DRS spectroscopy. The bandgap of BGO-CuS was found to be 2.51 eV as evaluated through the tauc plot method. The enhanced dye degradation was obtained at optimum conditions of pH = 8, catalyst concentration (20 mg/100 mL for BGO-CuS), oxidant dose (10 mM for BGO-CuS), and optimum time of irradiation was 60 min. The novel boron-doped nanocomposite effectively degraded methylene blue up to 95% under sunlight. Holes and hydroxyl radicals were the key reactive species. Response surface methodology was used to analyze the interaction among several interacting parameters to remove dye methylene blue effectively.

Biocontrol of Salmonella Typhimurium in milk, lettuce, raw pork meat and ready-to-eat steamed-chicken breast by using a novel bacteriophage with broad host range.

Salmonella spp., one of the most frequently reported bacteria, causes foodborne illness and economic losses. Due to the threat of increasing antibiotic resistant foodborne pathogens, application of bacteriophages as novel antibacterial agents in food matrices has become an emerging strategy. In this study, a novel Salmonella phage PS3-1 with high lytic activity against Salmonella Typhimurium was identified from previously isolated phages. PS3-1 belonged to the class Caudoviricetes with a broad host range, and had relatively short latent period (15 min), large burst size (92 PFU/cell), high pH stability (pH 3.0-11.0) and thermal tolerance (4-60 °C). Genome sequencing analysis showed that PS3-1 genome consisted of 107,110 bp DNA, without antibiotic resistance and virulence related genes. The results of growth curve and time-kill assay showed that PS3-1 not only inhibited the growth of S. Typhimurium, but also effectively decreased the viable cell counts (0.30-4.72 log) after 24-h incubation at 7, 25 and 37 °C (P < 0.05). Moreover, >1.28 log of established biofilm cells were effectively removed after 24-h treatment with PS3-1. Besides, PS3-1 significantly reduced the viability of S. Typhimurium in milk, lettuce, raw pork meat and ready-to-eat steamed-chicken breast at different temperatures (P < 0.05). These results demonstrated that PS3-1 may be an excellent antibacterial agent for controlling S. Typhimurium in food industry.

Impact of alternate Mn doping in ternary nanocomposites on their structural, optical and antimicrobial properties: Comparative analysis of photocatalytic degradation and antibacterial activity.

The present study was done to investigate and compare the photocatalytic and antibacterial activity of two in situ Manganese doped ternary nanocomposites. The dual ternary hybrid systems comprised Mn-doped Ag2WO4 coupled with MoS2-GO and Mn-doped MoS2 coupled with Ag2WO4-GO. Both hierarchical alternate Mn-doped ternary heterojunctions formed efficient plasmonic catalysts for wastewater treatment. The novel nanocomposites were well-characterized using XRD, FTIR, SEM-EDS, HR-TEM, XPS, UV-VIS DRS, and PL techniques confirming the successful insertion of Mn+2 ions in respective host substrates. The bandgap of the ternary nanocomposites evaluated by the tauc plot showed them visible light-active nanocomposites. The photocatalytic ability of both Mn-doped coupled nanocomposites was investigated against the dye methylene blue. Both ternary nanocomposites showed excellent sunlight harvesting ability for dye degradation in 60 min. The maximum catalytic efficiency of both photocatalysts was obtained at a solution pH value of 8, photocatalyst dose and oxidant dose of 30 mg/100 mL and 1 mM for Mn-Ag2WO4/MoS2-GO, 50 mg/100 mL, 3 mM for Mn-MoS2/Ag2WO4-GO keeping IDC of 10 ppm for all photocatalysts. The nanocomposites showed excellent photocatalytic stability after five successive cycles. The response surface methodology was used as a statistical tool for the evaluation of the photocatalytic response of several interacting parameters for dye degradation by ternary composites. The antibacterial activity was determined by the inactivation of gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria by support-based doped ternary hybrids.

Characterization of novel antimicrobial peptides designed on the basis of amino acid sequence of peptides from egg white hydrolysate.

Salmonella enterica subsp. enterica serotype Typhimurium (S. Typhimurium) is one of the most prevalent foodborne pathogens responsible for food poisoning and is spread through the consumption of contaminated poultry products. In this study, four antimicrobial peptides (AMPs) with varying hydrophobicity and helical structure-forming tendencies were designed and synthesized based on the amino acid sequences of peptides from egg white hydrolysate. Two of these AMPs, P1R3 (KSWKKHVVSGFFLR) and P1C (KSWKKHVVSGFFLRLWVHKK), exhibited inhibitory activity against S. Typhimurium and compromised its biofilm-forming ability. Investigation of their modes of action revealed that P1R3 and P1C interact with and permeabilize the cytoplasmic membrane of bacteria, leading to membrane potential dissipation, damage to membrane integrity, and consequent bacterial death. P1R3 also bound to S. Typhimurium DNA, resulting in DNA aggregation or precipitation. Moreover, both peptides showed negligible cytotoxicity to Vero cells, and P1C displayed significant antimicrobial activity in chicken meat. Peptides P1R3 and P1C, therefore, have the potential to be developed as promising food preservatives, especially against pathogenic S. Typhimurium.

Fabrication of visible light active Mn-doped Bi2WO6-GO/MoS2 heterostructure for enhanced photocatalytic degradation of methylene blue.

The increase in environmental pollution has led to an increased investigation in the development of novel ternary photocatalytic systems for remediation. These photocatalytic systems exhibit superior photocatalytic action for the removal of pollutants because of their visible light active bandgaps. A highly effective visible light active ternary heterojunction was fabricated using a hydrothermal method assisted by ultrasonication. Herein, we report the in situ hydrothermal synthesis of Mn-doped Bi2WO6-GO/ MoS2 photocatalyst, efficiently exhibiting greater photocatalytic activity for the wastewater treatment under solar light. The binary metal sulphide (MoS2) used as a co-catalyst, acted as an electron collector and graphene oxide (GO) as a support material for interfacial electron transfer to and from bismuth tungstate and MoS2. The as-prepared samples were characterized using SEM-EDX, FT-IR, XRD, XPS, BET, PL, and UV-Vis techniques. The bandgap of the novel photocatalyst was found in the visible region (2.2 eV) which helped in suppressing photoinduced electron-hole pairs recombination. The ternary Mn-doped Bi2WO6-GO/MoS2 showed 99% methylene blue removal after 60 minutes of sunlight irradiation at the optimum conditions of pH 8, catalyst dose 50 mg/100ml, and initial MB concentration of 10ppm under sunlight irradiation. The doped ternary heterostructure has proved to be an effective sunlight-active photocatalyst that can be reused without substantial loss in photocatalytic efficiency.

Comparative analysis of statistical tools for oil palm phytochemical research.

Recent advances in phytochemical analysis have allowed the accumulation of data for crop researchers due to its capacity to footprint and distinguish metabolites that are present within an organisms, tissues or cells. Apart from genotypic traits, slight changes either by biotic or abiotic stimuli will have significant impact on the metabolite abundances and will eventually be observed through physicochemical characteristics. Apposite data mining to interpret the mounds of phytochemical information from such a dynamic system is thus incumbent. In this investigation, several statistical software platforms ranging from exploratory and confirmatory technique of multivariate data analysis from four different statistical tools of COVAIN, SIMCA-P+, MetaboAnalyst and RIKEN Excel Macro were appraised using an oil palm phytochemical data set. As different software tool encompasses its own advantages and limitations, the insights gained from this assessment were documented to enlighten several aspects of functions and suitability for the adaptation of the tools into the oil palm phytochemistry pipeline. This comparative analysis will certainly provide scientists with salient notes on data assessment and data mining that will later allow the depiction of the overall oil palm status in-situ and ex-situ.

Sustainable Palm Oil-The Role of Screening and Advanced Analytical Techniques for Geographical Traceability and Authenticity Verification.

Palm oil production from oil palm (Elaeis guineensis Jacq.) is vital for the economy of Malaysia. As of late, sustainable production of palm oil has been a key focus due to demand by consumer groups, and important progress has been made in establishing standards that promote good agricultural practices that minimize impact on the environment. In line with the industrial goal to build a traceable supply chain, several measures have been implemented to ensure that traceability can be monitored. Although the palm oil supply chain can be highly complex, and achieving full traceability is not an easy task, the industry has to be proactive in developing improved systems that support the existing methods, which rely on recorded information in the supply chain. The Malaysian Palm Oil Board (MPOB) as the custodian of the palm oil industry in Malaysia has taken the initiative to assess and develop technologies that can ensure authenticity and traceability of palm oil in the major supply chains from the point of harvesting all the way to key downstream applications. This review describes the underlying framework related to palm oil geographical traceability using various state-of-the-art analytical techniques, which are also being explored to address adulteration in the global palm oil supply chain.

Biosynthesis of Saxitoxin in Marine Dinoflagellates: An Omics Perspective.

Saxitoxin is an alkaloid neurotoxin originally isolated from the clam Saxidomus giganteus in 1957. This group of neurotoxins is produced by several species of freshwater cyanobacteria and marine dinoflagellates. The saxitoxin biosynthesis pathway was described for the first time in the 1980s and, since then, it was studied in more than seven cyanobacterial genera, comprising 26 genes that form a cluster ranging from 25.7 kb to 35 kb in sequence length. Due to the complexity of the genomic landscape, saxitoxin biosynthesis in dinoflagellates remains unknown. In order to reveal and understand the dynamics of the activity in such impressive unicellular organisms with a complex genome, a strategy that can carefully engage them in a systems view is necessary. Advances in omics technology (the collective tools of biological sciences) facilitated high-throughput studies of the genome, transcriptome, proteome, and metabolome of dinoflagellates. The omics approach was utilized to address saxitoxin-producing dinoflagellates in response to environmental stresses to improve understanding of dinoflagellates gene-environment interactions. Therefore, in this review, the progress in understanding dinoflagellate saxitoxin biosynthesis using an omics approach is emphasized. Further potential applications of metabolomics and genomics to unravel novel insights into saxitoxin biosynthesis in dinoflagellates are also reviewed.

Antimicrobial resistance profiles of Listeria monocytogenes isolated from chicken meat in Fukuoka, Japan.

In this study, the antimicrobial resistance profiles of L. monocytogenes isolated from chicken meat in Fukuoka in 2017 were compared with the isolates of 2012. A total of 85 and 50 chicken meat samples, including different body parts, were collected from different supermarkets in Fukuoka in 2012 and 2017, respectively. Detection, isolation, identification, and characterization of L. monocytogenes were performed according to the conventional methods. Forty-five among 85 samples (53%) were positive for L. monocytogenes in 2012, while 12 among 50 samples in 2017 (24%) tested positive. One hundred fifty-three and 29 L. monocytogenes strains were isolated in 2012 and 2017, respectively. The serotypes of isolates in 2012 were 1/2a (21.5%), 1/2b (73.9%), 1/2c (1.5%), and 4b/4e (3.1%). In contrast, the 2017 isolates showed 1/2a (48.3%) and 1/2b (51.7%) serotypes. While all isolates in 2012 were positive for hlyA (listeriolysin O) in the PCR assay with hlyA primer set 7, only 17 hlyA positive isolates were seen in 2017. Moreover, 75 isolates with different ribotypes in 2012 and 29 isolates in 2017, respectively, were tested for antimicrobial susceptibility by broth microdilution for 18 different antimicrobial agents. Most of the 2012 and 2017 isolates displayed antimicrobial susceptibility. However, among the 2012 and 2017 isolates, 98.7% and 100% of the isolates were resistant to cefoxitin, 57.3% and 95.7% to fosfomycin, 72.0% and 82.6% to oxacillin, 8.0% and 17.4% to clindamycin, respectively. In addition, 2.7% of the isolates in 2012 were resistant to flomoxef and 4.3% of the isolates in 2017 to linezolid. Multidrug resistance (MDR) to 3 or more antimicrobials was observed in 35/75 (46.7%) isolates of 2012 and 19/23 (82.6%) in 2017. Detection of antimicrobial resistance (AMR) genes by PCR showed that the resistant isolates of 2012 were positive for mecA (96.3%) and ermC (83.3%), whereas the resistant isolates in 2017 screened positive for mecA (94.7%) and mefA (25.0%). Other cfxA, ermA, ermB, fosA, fosB, and fosC genes were absent in the PCR assay for any of the isolates. This study investigated for the first time the change in the L. monocytogenes contamination of chicken meat and antibiotic resistance of the isolated L. monocytogenes strains in Fukuoka, Japan, in the course of 5 years. Although the contamination rate of L. monocytogenes in 2017 was found to be lower than that in 2012, AMR of the isolates in 2017 was higher.

Use of headspace-gas chromatography-ion mobility spectrometry to detect volatile fingerprints of palm fibre oil and sludge palm oil in samples of crude palm oil.

OBJECTIVE: The addition of residual oils such as palm fibre oil (PFO) and sludge palm oil (SPO) to crude palm oil (CPO) can be problematic within supply chains. PFO is thought to aggravate the accumulation of monochloropropanediols (MCPDs) in CPO, whilst SPO is an acidic by-product of CPO milling and is not fit for human consumption. Traditional targeted techniques to detect such additives are costly, time-consuming and require highly trained operators. Therefore, we seek to assess the use of gas chromatography-ion mobility spectrometry (GC-IMS) for rapid, cost-effective screening of CPO for the presence of characteristic PFO and SPO volatile organic compound (VOC) fingerprints. RESULTS: Lab-pressed CPO and commercial dispatch tank (DT) CPO were spiked with PFO and SPO, respectively. Both additives were detectable at concentrations of 1% and 10% (w/w) in spiked lab-pressed CPO, via seven PFO-associated VOCs and 21 SPO-associated VOCs. DT controls could not be distinguished from PFO-spiked DT CPO, suggesting these samples may have already contained low levels of PFO. DT controls were free of SPO. SPO was detected in all SPO-spiked dispatch tank samples by all 21 of the previously distinguished VOCs and had a significant fingerprint consisting of four spectral regions.

Decision making process and factors contributing to research participation among general practitioners: A grounded theory study.

INTRODUCTION: The participation of general practitioners (GPs) in primary care research is variable and often poor. We aimed to develop a substantive and empirical theoretical framework to explain GPs' decision-making process to participate in research. METHODS: We used the grounded theory approach to construct a substantive theory to explain the decision-making process of GPs to participate in research activities. Five in-depth interviews and four focus group discussions were conducted among 21 GPs. Purposeful sampling followed by theoretical sampling were used to attempt saturation of the core category. Data were collected using semi-structured open-ended questions. Interviews were recorded, transcribed verbatim and checked prior to analysis. Open line-by-line coding followed by focus coding were used to arrive at a substantive theory. Memoing was used to help bring concepts to higher abstract levels. RESULTS: The GPs' decision to participate in research was attributed to their inner drive and appreciation for primary care research and their confidence in managing their social and research environments. The drive and appreciation for research motivated the GPs to undergo research training to enhance their research knowledge, skills and confidence. However, the critical step in the GPs' decision to participate in research was their ability to align their research agenda with priorities in their social environment, which included personal life goals, clinical practice and organisational culture. Perceived support for research, such as funding and technical expertise, facilitated the GPs' participation in research. In addition, prior experiences participating in research also influenced the GPs' confidence in taking part in future research. CONCLUSIONS: The key to GPs deciding to participate in research is whether the research agenda aligns with the priorities in their social environment. Therefore, research training is important, but should be included in further measures and should comply with GPs' social environments and research support.

Biopolymers composites with peanut hull waste biomass and application for Crystal Violet adsorption.

Composites of polyaniline, starch, polypyrrole, chitosan aniline and chitosan pyrrole using peanut waste were prepared and employed for the adsorption of Crystal Violet (CV) dye from aqueous media. The process variables i.e., composite doses, pH, contact time, CV initial concentration and temperature were optimized. Thermodynamic, equilibrium modelling and kinetics models were fitted to the CV adsorption data in order to understand the mechanism and nature of CV adsorption onto native and composite adsorbents. Maximum CV adsorption of 100.6mg/g was achieved (onto chitosan aniline composite) using 150mg/L dye initial concentration, 50°C temperature, 60min contact time, 0.05g adsorbent dose and>7pH. Effect of composites pre-treatments with salts, surfactants and co-metals ions were also studied. The CV adsorption efficiencies of adsorbents were found in following order; chitosan aniline composite>starch composite>chitosan pyrrole composite>polyaniline composite>polypyrrole composite>native peanut biomass. The pseudo-second-order kinetic model and Freundlich isotherm fitted well to the CV equilibrium adsorption data and intraparticle diffusion was the rate limiting step. Composites showed endothermic and energetically stable nature for CV adsorption. Composites also showed good desorption properties, which revealed the recycling ability of prepared composites.

Characterization of apigenin and luteolin derivatives from oil palm (Elaeis guineensis Jacq.) leaf using LC-ESI-MS/MS.

The palm oil industry generates several byproducts, and more than half of the dry weight of the waste is of oil palm leaf whereby the tissue is underutilized. Recently, several research studies found promising potential of oil palm fronds as a source of nutraceutical due to its bioactive properties. However, the chemical composition of the tissue is still not deciphered. Using reversed-phase liquid chromatography (LC) electrospray mass spectrometry (ESI-MS), glycosylated apigenin and luteolin were separated and identified from oil palm (Elaeis guineensis Jacq.) leaf and structures of the constituents were elucidated by collision-induced dissociation (CID) tandem MS. From 28 derivatives of the flavones, 9 compounds were conjugated with hydroxymethylglutaric (HMG) acid. Improved knowledge on oil palm especially on bioactive component of the leaf tissue will allow correlation of its beneficial effects and further promotes efficient utilization of this agriculture byproduct.