2-ethylhexyl Phthalate Research Articles

Antibacterial Activity, GC MS Analysis and In Silico Validation of Bioactive Compound from Endophytic Fungus Lasiodiplodia pseudotheobromae EF-9.

Secondary metabolites synthesized by endophytic fungi have garnered significant interest for their broad applications in treating various ailments. In this study involving 20 plant samples, 11 endophytic fungi were isolated and cultured, and Lasiodiplodia pseudotheobromae EF-9, derived from Hibiscus rosa-sinensis, demonstrated greater antibacterial efficacy than the other isolated endophytes. Phylogenetic analyses using 18S rRNA gene confirmed the EF-9 identity as L. pseudotheobromae. Following mass production, the active compound was partially purified using column chromatography. The fraction collected at the 60th min exhibited good antibacterial activity against Bacillus coagulans (MTCC 6735) and Shigella flexneri (ATCC 12022), with an inhibition zone of approximately 20 mm in diameter. UV spectral studies revealed a wide absorption band at 430 nm. High Performance Liquid Chromatography (HPLC) of the active fraction showed a distinct peak with a retention time of 4.216 min at 430 nm absorbance. Gas Chromatography-Mass Spectrometry (GC-MS) identified the active compound in the L. pseudotheobromae EF-9 culture broth extract as Bis(2-ethylhexyl) phthalate, which displayed a peak at 16.856 min and covered 66.69% of the area in the spectral analysis.

Characteristics, prediction, and risk assessment of phthalates, organophosphate esters, and polycyclic aromatic hydrocarbons in vegetables from plastic greenhouses of Northeast China.

We investigated the contaminations of phthalates (PAEs), organophosphate esters (OPEs), and polycyclic aromatic hydrocarbons (PAHs) in the vegetables and their corresponding soils from 26 plastic greenhouses of Northeast China. PAEs, OPEs, and PAHs in the edible portion of vegetables were in the range of 2620-21800, 115-852, and 32.4-602 ng/g, while the levels of these chemicals in the greenhouse soils were 5770-18800, 196-935, and 109-1600 ng/g, respectively. PAEs are the main organic pollutants in greenhouses, which were 1-2 orders of magnitude higher than that of OPEs and PAHs. Leafy vegetables showed the highest contamination level, which is ∼1-3 times that of root and fruit vegetables. Bioaccumulation factors (BAFs) of chemicals are significantly negatively correlated with their physicochemical properties, e.g., octanol-water partition coefficient and organic carbon partition coefficient. The partition-limited model can accurately predict the contamination level of greenhouse vegetables to a certain extent based on the chemical's concentration in the corresponding soil. We assessed the hazard quotients of target compounds through daily intake of greenhouse vegetables, and found a low risk for di(2-ethylhexyl) phthalate. This research emphasized the potential dietary exposure risks caused by greenhouse leafy vegetables, and proposed a method for evaluating the risk of greenhouse vegetables through soil monitoring.

The role of hydroponic solution of Ulva fasciata in modulating the antibiotic resistance of Vibrio alginolyticus.

This study aimed to ascertain the active substances in the hydroponic solution of Ulva (U.) fasciata (HSUF) and their effects on antibiotics resistance of pathogenic bacteria Vibrio alginolyticus (Va.) at environmental levels. Analytical results of HSUF by solid-phase extraction and gas chromatography-mass spectrometry showed that HSUF contained rich active substances and dominated by 2,4,6-tribromophenol, acetal, mono (2-ethylhexyl) phthalate and 2- (4-hydroxybenzene) ethanol, etc. Thereinto, 2,4,6-tribromophenol had the lowest minimal inhibitory concentrations (MICs) of 64-128μg mL-1 for Va. strains. The rich antibacterial substances of HSUF inhibited 47%-63% of isolated Va. strains. After 20 generations of subculture for Va. strains in three dilutes (1/2 (HT), 1/20 (MT) and 1/50 (LT)) of HSUF (20g L-1), the resistance to streptomycin of Va. S1 and Va. M3 changed from intermediate resistance ability to susceptible level. The large promotion of reactive oxygen species was observed in different HSUF levels, but the biofilm formation of Va. S1 did not changed significantly. Transcriptome sequencing of Va. S1 demonstrated that antibiotic resistance gene lpxA was down-regulated at different HSUF levels. Live U. fasciata excreted a variety of active secondary metabolites (SMs) in HSUF, which exhibited strong inhibitory effects on three isolated Va. strains. HSUF changed the antibiotic resistance of Va. Strain, especially at a higher HSUF concentrations.

Dietary exposure to di (2-ethylhexyl) phthalate (DEHP) for 6months alters markers of female reproductive aging in mice.

The female reproductive system ages before any other physiological system, making it a sensitive indicator of aging. Early reproductive aging is associated with the early onset of infertility and an increased risk of several diseases. During aging, systemic and reproductive oxidative stress and inflammation levels increase through inflammasome activation, leading to ovarian follicle loss. Other markers of reproductive aging include increased fibrosis and shortening of telomeres in ovarian cells. The factors that accelerate reproductive aging are unclear, but likely involve exposure to endocrine-disrupting chemicals such as phthalates. Di(2-ethylhexyl) phthalate (DEHP) is a widely used phthalate and humans are exposed to it daily. Several studies show that DEHP induces reproductive toxicity by affecting estrous cyclicity, follicle numbers, and hormone levels. However, little is known about the mechanisms underlying DEHP-induced early onset of reproductive aging. Thus, this study tested the hypothesis that dietary exposure to DEHP induces early reproductive aging by affecting inflammation, fibrosis, and the expression of telomere regulators and antioxidant enzymes. Adult CD-1 female mice were exposed to vehicle (corn oil) or DEHP (0.5, 1.5, or 1500ppm) via the chow for six months. Exposure to DEHP increased the expression of antioxidant enzymes and Casp3, increased expression of telomere-associated genes, and increased fibrosis levels in the ovary. In addition, DEHP exposure for 6months altered ovarian and systemic inflammatory status. Collectively, our novel data suggest that 6-month dietary exposure to DEHP may accelerate reproductive aging by affecting several reproductive aging markers in female mice.

Simulation study on heavy metals, phthalate esters, and organic halogens: Content and distribution characteristics during waste paper recycling.

Imported waste paper and recycled pulp may contain pollutants, posing potential environmental risks to the ecosystem of China. This study examined the presence and distribution patterns of heavy metals, phthalate esters (PAEs), and adsorbable organic halogens (AOX) in recycled pulp and production wastewater after various recycling processes of three typical restricted types of imported waste paper. The results indicated that the concentration ranges of heavy metals, PAEs, and AOX in the three types of imported recycled waste paper were 21.61-40.38mg/kg, 15.35-27.88mg/kg, and 19.21-57.72mg/kg, respectively. The comparative analysis with the initial waste paper demonstrated a reduction in heavy metal content in the recycled pulp by 17.80-49.75%, PAEs by 65.42-90.55%, and AOX by 32.80-42.34%. The average concentrations of these pollutants in wastewater were 0.85-1.66mg/L, 27.28-59.86mg/L, and 1.15-3.34mg/L, respectively. Chromium and lead were identified as the primary heavy metals present in the waste paper. Following pulping, No. 1 and No. 2 met the arsenic and lead levels specified in the "Reuse Fiber Pulp" standard (GB/T24320-2021), whereas No. 3 met these criteria after de-inking only. The main PAEs detected in the waste paper were dibutyl phthalate and di(2-ethylhexyl) phthalate, most of which were removed during the pulping stage. Significantly higher levels of AOX were observed in No. 2 and No. 3 than in No. 1, with a minimal impact on AOX removal from the pulp during the recycling process.

Comprehensive utilization of in silico approach and in vitro experiment to unveil the molecular mechanisms of mono (2-ethylhexyl) phthalate-induced lung adenocarcinoma

Mono (2-ethylhexyl) phthalate (MEHP), the main bioactive metabolite of commonly used plasticizer Di (2-ethylhexyl) phthalate, has received increasing attention due to its carcinogenic toxicity. This study aims to systematically explore the molecular mechanisms underlying MEHP-induced lung adenocarcinoma (LUAD). Firstly, network toxicology was employed to construct the interaction network of MEHP-targeted LUAD-related proteins and identify core proteins. Subsequently, functional analyses were used to determine the key pathways of these proteins enriched. Next, expression and survival analyses of multiple public datasets were conducted to emphasize the importance of core genes, and an optimized prognostic model was constructed based on independent prognostic genes to explore the relationship of gene risk with immune infiltration and immunotherapy. Ultimately, molecular docking and dynamics simulation were used to predict the binding modes and affinities of MEHP with core proteins, and surface plasmon resonance experiments were utilized to further validate their direct interactions. The findings demonstrated that MEHP targets 167 LUAD-related proteins, including 28 core target proteins. These proteins form the critical networks that regulate cancer and immune-associated pathways to induce the occurrence and development of LUAD, and further coordinate patient prognosis and treatment by altering the immune microenvironment. Most importantly, their direct interactions (especially PTGS2) lay the structural foundation of MEHP regulating core proteins, greatly supporting its LUAD toxicity. In conclusion, this study introduces a novel approach for evaluating the safety of plasticizers and elucidates the molecular mechanisms behind MEHP-induced LUAD, thus offering new and effective targets and strategies for cancer prevention and treatment.

Photoinduced Energy and Electron Transfer in Nitrogen-Doped Graphene/ZnO Nanorod Composites for Self-Powered Photoelectrochemical Aptasensing of Bis(2-ethylhexyl)phthalate

Photoinduced Energy and Electron Transfer in Nitrogen-Doped Graphene/ZnO Nanorod Composites for Self-Powered Photoelectrochemical Aptasensing of Bis(2-ethylhexyl)phthalate

New Antimicrobial Materials Based on Plasticized Polyvinyl Chloride for Urinary Catheters: Preparation and Testing.

Given the constant increased number of nosocomial infections in hospitals, especially associated with prolonged usage of inserted medical devices, our work aims to ameliorate clinical experience and promote faster healing of patients undergoing urinary catheterization by improving the properties of medical devices materials. Within this research, nine different composites were prepared based on polyvinyl chloride, using three different plasticizers (di-(2-ethylhexyl) phthalate, Proviplast 2646, and Proviplast 2755), and two different antimicrobial additives containing silver nanoparticles. The prepared materials were analyzed, and their physicochemical properties were determined: water absorption, relative density, plasticizer migration, hydrophobicity/hydrophilicity by contact angle measurement, Shore A hardness, tensile strength, and elongation at break. Structure and morphology were also investigated by means of FTIR, SEM, and EDX analyses, and thermal (TG-DSC) and biological properties were evaluated. The most important aspects of obtained results are showing that plasticizer migration was significantly reduced (to almost zero) and that the usage of antimicrobial additives improved the materials' biocompatibility. Thus, based on the concluded favorable properties, the obtained materials can be further used for catheter development. Pressure-flow studies for different sizes and configurations are the next steps toward advanced in vivo and clinical trials.

Associations of Phthalate Exposure with Adiposity and Metabolic Syndrome in US Adolescents and Adults, NHANES 2013-2018

Abstract Background/Objective Phthalates are ubiquitous endocrine-disrupting chemicals and suspected obesogens. However, the associations with fat distribution and associated cardiometabolic complications remain unclear. Therefore, we examined the associations between phthalate exposure, body fat (total and distribution patterns), and metabolic syndrome (MetS) among US adolescents and adults. Methods We analyzed cross-sectional data from 829 adolescents and 3905 adults in the 2013-2018 National Health and Nutrition Examination Survey. Total percent body fat (%BF), visceral adipose tissue (VAT) mass, and android to gynoid (A/G) ratio were determined using dual-energy x-ray absorptiometry. Associations between molar sums of low molecular weight (∑LMW), high molecular weight (∑HMW), and di(2-ethylhexyl) phthalate (∑DEHP) metabolites, and adiposity indicators and MetS were analyzed with multivariable linear and logistic regressions. Models included sex interaction terms, were stratified by age group, and adjusted for relevant covariates. Results ∑HMW and ∑DEHP exposures were positively associated with %BF in males, and all phthalate groups were associated with greater VAT mass and A/G ratio in adolescent males. Five-fold increases in ∑HMW and ∑DEHP metabolites were associated with 21.7% (95% CI: 10.5-33.9) and 18.0% (95% CI: 7.72-29.2) greater VAT mass among adolescent males, respectively. Sex modified the relationship between ∑HMW exposure and A/G ratio among adolescents (interaction p-value = 0.0185). Phthalates were not associated with odds of MetS. When assessing individual MetS components, phthalates were associated with hyperglycemia in adult males. Conclusion Greater exposure to phthalates was associated with greater %BF in all males, and with fat distribution in adolescent males; however, phthalates were not linked to MetS.

Hydroxyl radical oxidation of chemical contaminants on indoor surfaces and dust

Humans are widely exposed to semivolatile organic contaminants in indoor environments. Many contaminants have long lifetimes following partitioning to the large surface reservoirs present indoors, which leads to long exposure times to gas-phase oxidants and multiphase chemistry. Studies have shown selective multiphase oxidation of organics on indoor surfaces, but the presence of hydroxyl radicals with nonselective reactivity and evidence of multiphase OH radical reactivity toward common indoor contaminants indicates that there may be additional unknown transformation chemistry indoors. We screened genuine indoor samples for 60 OH radical oxidation products of the common plasticizer and endocrine-disrupting contaminant bis(2-ethylhexyl) phthalate (DEHP) identified in laboratory experiments using nontargeted high-resolution mass spectrometry. At least 30 and 10 of these products are observed in indoor dust and DEHP films exposed to ambient indoor conditions, respectively, indicating that multiphase OH reactions occur indoors. Using the PROTEX model and a multimedia indoor chemical fate model, we demonstrate that these products have long indoor lifetimes and cause a higher potential for human exposure than DEHP. Some of these products are more active endocrine disruptors than DEHP itself, but most have unknown toxicities. Coexposure to all oxidation products will likely have an additive effect, leading to higher human health risks from indoor organic contaminants than previously thought. Due to the nonselective reactivity of OH radicals, it is likely that most indoor contaminants follow similar chemistry, and further study is needed to understand the prevalence and human health implications of such multiphase chemistry.

Associations between pre- and post-natal exposure to phthalate and DINCH metabolites and gut microbiota in one-year old children

The gut microbiota is a collection of symbiotic microorganisms in the gastrointestinal tract. Its sensitivity to chemicals with widespread exposure, such as phthalates, is little known. We aimed to investigate the impact of perinatal exposure to phthalates on the infant gut microbiota at 12 months of age. Within SEPAGES cohort (Suivi de l’Exposition à la Pollution Atmosphérique durant la Grossesse et Effet sur la Santé), we assessed 13 phthalate metabolites and 2 di(isononyl) cyclohexane-1,2-dicarboxylate (DINCH) metabolites in repeated urine samples collected in pregnant women and their offspring. We obtained stool samples from 356 children at 12 months of age and sequenced the V3-V4 region of the 16S rRNA gene, allowing gut bacterial profiling. We used single-chemical (linear regressions) and mixture (BKMR, Bayesian Kernel Machine Regression) models to examine associations of phthalates and DINCH metabolites, with gut microbiota indices of α-diversity (specific richness and Shannon diversity) and the relative abundances of the most abundant microbiota phyla and genera. After correction for multiple testing, di(2-ethylhexyl) phthalate (ΣDEHP), diethyl phthalate (DEP) and bis(2-propylheptyl) phthalate (DPHP) metabolites 12-month urinary concentrations were associated with higher Shannon α-diversity of the child gut microbiota in single-chemical models. The multiple-chemical model (BKMR) suggested higher α-diversity with exposure to the phthalate mixture at 12 months, driven by the same phthalates. There were no associations between phthalate and DINCH exposure biomarkers at other time points and α-diversity after correction for multiple testing. ΣDEHP metabolites concentration at 12 months was associated with higher Coprococcus genus. Finally, ΣDEHP exposure at 12 months tended to be associated with higher phylum Firmicutes, an association not maintained after correction for multiple testing. Infancy exposure to phthalate might disrupt children's gut microbiota. The observed associations were cross-sectional, so that reverse causality cannot be excluded.

Environmental Stress-Induced Alterations in Embryo Developmental Morphokinetics.

The association between embryo morphokinetics and its developmental competence is well documented. For instance, early cleaved embryos are more competent in developing to blastocysts, whereas the proportion of abnormally cleaved embryos that further developed to blastocysts is low. Numerous factors, such as the parental age, lifestyle, health, and smoking habits have been reported to affect the embryo morphokinetics and, consequently, its development. However, less is known about the effect of environmental stressors on embryo morphokinetics. The current review discusses the effect of the most concerning environmental stressors on embryo morphokinetics. These stresses include heat stress and human-made chemicals such as phthalates (e.g., bis-(2-ethylhexyl phthalate, dibutyl phthalate, dimethyl phthalate, and their primary metabolites), herbicides (e.g., diaminochlorotriazine, the primary metabolite of atrazine), pharmaceutical compounds (e.g., carbamazepine, nocodazole) and pro-oxidant agents (cumene hydroperoxide, Triton X-100), as well as naturally occurring toxins such as mycotoxin (e.g., aflatoxin B1 and its metabolite, and ochratoxin A). In addition, this review discusses the effect of ionizing or non-ionizing radiation and viral infections (e.g., SARS-CoV-2, papillomavirus). Finally, it points out some potential mechanisms that underlie the impairment of embryo morphokinetics, and it suggests protective compounds, mainly the supplementation of antioxidants to improve the morphokinetics, and consequently, the embryo developmental competence.

Environmental persistence, bioaccumulation, and hazards of chemicals in e-cigarette e-liquids: short-listing chemicals for risk assessments

Background/MethodsIncreased use and sales of e-cigarettes raises concerns about the potential environmental impacts throughout their life-cycle. However, few available research studies focus on the environmental impacts and ecotoxicity of e-cigarettes....

Associations of 1.5- and 3-year phthalate exposure levels with early adiposity rebound and overweight/obesity in Japanese children: An adjunct study of the Japan Environment and Children's Study

The relationship between early childhood phthalate exposure and early adiposity rebound (EAR) is unclear. This study aimed to investigate the association between 1.5- and 3-year phthalate exposure and EAR and overweight/obesity in 7.5-year-old Japanese children. A total of 452 mother–child pairs were enrolled from the Aichi Regional Cohort of the Japan Environment and Children's Study. The children were followed up at birth and at 1.5, 2, 3, 4, 5, 6, and 7.5 years of age for physical examination. Human biomonitoring of 16 urinary metabolites of eight phthalates was performed at 1.5 and 3 years of age. Latent class mixed models, binary logistic regression, and quantile g-computation were performed to identify body mass index (BMI) trajectories and investigate the relationships of single or mixed phthalate exposure with EAR and overweight/obesity. A one-unit increase in log10-transformed 3-year-old Σdi(2-ethylhexyl) phthalate (ΣDEHP) exposure levels was significantly associated with 6-year-old BMI in girls. The 1.5-year mono-iso-butyl phthalate and 3-year Σdi-isodecyl phthalate exposure levels were significantly associated with the repeated measures of longitudinal BMIs in girls. Single phthalate exposure showed null associations with EAR or overweight/obesity in the 7.5-year-old children. Σdi-isononyl phthalate, ΣDEHP, and mono-n-butyl phthalate exhibited the highest proportion of partial positive weights of being in the EAR trajectory after confounder adjustment. Phthalate mixture exposure in 1.5- and 3-year-old children was not significantly associated with EAR. Early childhood phthalate exposure was not related to EAR or overweight/obesity in 7.5-year-old Japanese children. However, few phthalates were positively associated with longitudinal BMIs in girls.

Phytochemical Profiling and Nutraceutical Benefits of Methanol Leaf Extracts from Datura alba ness

Introduction: Species of the Datura plant have demonstrated significant pharmacological benefits in combating various diseases over the past decade. However, little information is available on Datura alba ness (DAN), and no studies have yet reported its proximate composition or characterized its phytochemical components. This study aims to investigate the nutritional, phytochemical and bioactive properties of Datura alba ness. Materials and Methods: Fresh leaves of Datura alba ness were collected from Abua Odua local government area of Rivers State, Nigeria and properly identified and authenticated. The fresh leaves were subjected to proximate analysis using standard protocols while the dried leaves were extracted using methanol by the method of cold maceration to obtain methanolic extract of Datura alba ness (MEDAN). The MEDAN was further subjected to phytochemical screening to obtain the various phytochemicals and Gas chromatography and mass spectrometry (GC-MS) analysis for detailed identification and characterization of the various bioactive compounds. Results: Proximate analysis of fresh leaves of DAN showed the presence of moisture (60.4%), crude protein (21.9%), carbohydrate (7.6%), crude fibre (4.77%), ash (4.17%) and fat (1.19%) while the phytochemical screening revealed the presence of flavonoids (49.14%), alkaloids (19.16%), tannins 1(12.19%), saponins (11.72%), phenols (5.33%), cardiac glycosides (1.91%), oxalates (0.43%) and phytates (0.43%). Also, the GC-MS analysis of MEDAN identified twenty-nine (29) active compounds with four (4) observed as the most abundant (with area concentration >10%). They include Bis (2-ethylhexyl) phthalate (15.97%), 13-hexyloxacyclotridec-10-en-2-one (14.26%), n-hexadecanoic acid (12.87%) and Gamma.-Sitosterol (10.77). Conclusion: The identification of various nutrients, compounds, and bioactive substances through proximate, phytochemical, and GC-MS analysis of fresh leaves and methanol extract of Datura alba ness highlights its potential as a supplement and therapeutic agent. These findings suggest that Datura alba ness contains numerous bioactive compounds that may offer effective treatments for conditions related to oxidative stress, nervous system disorders, tissue damage, inflammation, and bacterial infections.

Revealing contaminants in China's recycled PET: Enabling safe food contact applications

The lack of data on chemical contamination in recycled polyethylene terephthalate (rPET) in food contact applications, impedes the legalization of rPET in food contact applications in China. This study offers a comprehensive analysis of contaminants in hot-washed rPET flakes, identifying 1011 compounds. While most substances were detected infrequently, some i.e., bis(2-ethylhexyl) phthalate and mineral oil aromatic hydrocarbons stood out for their relatively high average concentrations. Limonene emerged as the most frequently detected flavorants. Rare genotoxic compounds like safrole and diethyl sulfate were detected at low levels, emphasizing the importance of adopting a conservative evaluation approach. Molecular descriptor analysis guided the proposal of four new surrogates, i.e., dimethyl pentanedioate, 2-tridecanone, bis(2-ethylhexyl) terephthalate, and 2,2-dimethoxy-1,2-diphenylethanone, for challenge tests, aiming to better capture the chemical diversity of rPET contaminants. This research unveils the contamination profile of Chinese rPET and suggests adjustments to evaluation protocols, potentially facilitating its high-value applications and minimizing potential harms.

Novel Insights into Ethanol-Soluble Oyster Peptide-Zinc-Chelating Agents: Structural Characterization, Chelation Mechanism, and Potential Protection on MEHP-Induced Leydig Cells.

Numerous studies have reported that mono-(2-ethylhexyl) phthalate (MEHP) (bioactive metabolite of Di(2-ethylhexyl) phthalate) has inhibitory effects on Leydig cells. This study aims to prepare an oyster peptide-zinc complex (PEP-Zn) to alleviate MEHP-induced damage in Leydig cells. Zinc-binding peptides were obtained through the following processes: zinc-immobilized affinity chromatography (IMAC-Zn2+), liquid chromatography-mass spectrometry technology (LC-MS/MS) analysis, molecular docking, molecular dynamic simulation, and structural characterization. Then, the Zn-binding peptide (PEP) named Glu-His-Ala-Pro-Asn-His-Asp-Asn-Pro-Gly-Asp-Leu (EHAPNHDNPGDL) was identified. EHAPNHDNPGDL showed the highest zinc-chelating ability of 49.74 ± 1.44%, which was higher than that of the ethanol-soluble oyster peptides (27.50 ± 0.41%). In the EHAPNHDNPGDL-Zn complex, Asn-5, Asp-7, Asn-8, His-2, and Asp-11 played an important role in binding to the zinc ion. Additionally, EHAPNHDNPGDL-Zn was found to increase the cell viability, significantly increase the relative activity of antioxidant enzymes and testosterone content, and decrease malondialdehyde (MDA) content in MEHP-induced TM3 cells. The results also indicated that EHAPNHDNPGDL-Zn could alleviate MEHP-induced apoptosis by reducing the protein level of p53, p21, and Bax, and increasing the protein level of Bcl-2. These results indicate that the zinc-chelating peptides derived from oyster peptides could be used as a potential dietary zinc supplement.

In vitro antimicrobial and antioxidant activities of essential oil extracts from underutilized Zanthoxylum ovalifolium tutcher fruits: Prospective applications in the food industry

Zanthoxylum ovalifolium fruits and its oils are being used in traditional foods as flavoring agent. So, the present investigation was taken to evaluate the potential of essential oils extracted from fruits activity against food borne spoilage microbes. Phytochemical screening revealed the presence of flavonoids and phytosterols. GC-MS results showed the presence of a total of 20 compounds, including Oleoyl chloride in the oil extracted from young fruit and Bis(2-ethylhexyl) phthalate in both oils. The LC-MS results revealed a total of 48 compounds, highlighting variations in composition between ZEY and ZEM, likely due to the fruit's maturity stage. DPPH free radical scavenging potential of young fruit essential oils extract showed highest percentage of scavenging activity and IC50 values. The antimicrobial activities of the essential oils were evaluated against bacterial and fungal strains by well diffusion method. S. aureus showed highest zone of inhibition and all fungal strains showed excellent inhibition compared to standard. These essential oils have appreciated capability of suppressing growth of wide variety of food borne microbes in turn scavenging the free radicals released by them. Derived from natural sources, this essential oil offers a promising alternative for extending the shelf life of food products.

Synergistic action of bacteriophage and metabolites of Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM against Enterotoxigenic Escherichia coli and Bacillus cereus and their biofilm

BackgroundFoodborne disease and food spoilage are the prime public health issue and food security round the globe. Significant disease outbreaks mostly linked to the existence of pathogenic bacteria that extremely challenging due to the persistence of biofilm-forming. Proteins and bacterial metabolites have been shown to have good antibacterial activity and effectively removal bacterial biofilm. Recently, bacteriophage and their encoded lytic proteins such as lysin have attracted attention as potential alternative agent to control undesirable pathogens in human body infection, increasing food safety as advance preservations and medical treatment such as phage therapy. For these reasons, the efficacy of bacteriophage and their potential in combination with bacterial metabolites from Phyllosphere and Actinomycetes bacteria (Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM crude extracts) was the aim of this present study.ResultsIn this study, bacteriophage BC-VP (1.28 ± 0.29 × 1011 PFU/ml) and ETEC-phage-TG (8.9 ± 2.19 × 108 PFU/ml) isolated from artificial lake water from previous study showed potential activity to control Bacillus cereus (BC) and Enterotoxigenic Escherichia coli (ETEC) population. The combination of BC-VP with metabolite (P. fluorescens JB3B and S. thermocarboxydus 18PM) which were known from previous study had antibiofilm activities were able to inhibit (86.1%; 83.3%) and destruct (41%; 45.5%) biofilm formation of B. cereus respectively. Likewise, the synergy of bacteriophage ETEC-phage-TG with the same crude extract also showed promising activity against biofilm of ETEC with percentage of inhibition (81.9%; 76.4%) and percentage of destruction (54.1%; 44.4%). Application in various food, combination of BC-VP and bacterial metabolite extract (P. fluorescens JB3B; S. thermocarboxydus 18PM) were able to reduce Bacillus cereus population in mashed potato (99.6%; 99.4%) at cold temperature (4 °C) and (68.9%; 56.6%) at room temperature (28 °C), boiled pasta (99.5%; 99.4%) and (84.7%; 75.7%), also soymilk (96.9%; 96.7%) and (42.4%; 39.4%) respectively. Likewise, combination of ETEC-phage-TG and bacterial metabolite (P. fluorescens JB3B; S. thermocarboxydus 18PM) potentially reduced ETEC population after two different temperatures (4 °C and 28 °C) incubation in bean sprouts (TFTC; TFTC) and (47.5%; 49.1%), chicken meat (TFTC; TFTC) and (58.1%; 54%), also minced beef (99.5%; 99.4%) and (41.1%; 28%). GC-MS determination performed, oxalic acid, phenol, phenylethyl alcohol, N-hexadecanoic acid, and pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl was the most active compound in P. fluorescens JB3B. 2,4-Di-tert-butylphenol, phenyl acetic acid, N-Hexadecanoic acid, pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl, and Bis(2-ethylhexyl) phthalate was most active compound in the S. thermocarboxydus 18PM isolates.ConclusionsThe combination of isolated bacteriophages and bacterial metabolite showed promising results to be used as biocontrol candidate to overcome biofilm formed by foodborne and food spoilage bacteria using their ability to produce antibiofilm compounds and lytic activity. In addition, this combination also potentially reduces the use or replace the drawbacks of common application such as antibiotic treatment.

GC-MS SCRUTINY OF PHYTOCONSTITUENTS, IN VITRO EVALUATION OF ANTIHYPERGLYCEMIC, ANTIADIPOGENIC ACTIVITIES, AND CYTOTOXIC EFFECT USING 3T3 L1 ADIPOCYTE CELL LINE AND MOLECULAR DOCKING STUDIES OF PREMNA CORYMBOSA

Objective: The present study was intended to list out the phytochemical multiples and to investigate the antihyperglycemic effect of Premna corymbosa using in vitro assays and in silico molecular docking methods. Methods: The phytochemical multiples of methanol proportion of P. corymbosa leaves were appraised by Gas Chromatography-Mass Spectrometry (GC-MS) scrutiny to illustrate the attendance of phytochemical composites. Moreover, the in vitro antihyperglycemic, antiadipogenic activities, and cytotoxic effects of the extract were elucidated using a 3T3 L1 adipocyte cell line. Mode of action of phytochemical composites in methanol leaf extract of P. corymbosa was probed by Western blotting with IRS1, IRS2, mTOR, and glucose transporter type 4 (GLUT 4) receptors. At present, to probe the consequence of the aboriginal drugs, it is necessary to perform in silico docking on the diabetic receptor which could be useful for the progress of enhanced formulation for the psychoanalysis of diabetes. Results: The GC-MS scrutiny depicted the being there of thirty-five phytochemical multipart. Amid the thirty-five multipart’s recognized, focal composites were Phytol, acetate (RT-16.78), n-Hexadecanoic acid (RT-18.16), Phytol (RT-19.51), 9,12,15-octadecatrienoic acid (Z,Z,Z) (RT-19.85), octadecanoic acid (RT-20.04), and Bis (2-ethylhexyl) phthalate (RT-23.09). The results of the glucose conception assay, adipocyte differentiation assay, and MTT assay showed potent in vitro antihyperglycemic activity with methanol leaf extract of P. corymbosa in 3T3l1Cell line. The results attained from western blotting revealed good antihyperglycemic activity of P. corymbosa. The in silico molecular docking results illustrated that the selected herbal lead compound is an effective target against the receptors. The compound showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as an antihyperglycemic compound. Conclusion: The outcome of the current study substantiates the antihyperglycemic prospective of the methanol leaf extract of P. corymbosa on the hyperglycemic causal agents and its activity against diabetes by a molecular approach.