Preparation of phosphorus containing products by co-incineration of sludge ash and calcium-based additives: Focusing one-step and multi-step method.

Due to the irreplaceable nature of phosphorus (P) in biological growth and the shortage of P rock, it is necessary to recover P from waste, such as sludge ash. P-containing products were prepared using sludge ash and calcium-based additives (CaCO3 and eggshell). In addition, the effects of different incineration methods (one-step method (OSM) and multi-step method (MSM)), additive doses, and incineration temperature on the P content and species in the products were investigated. The results indicated that as the dose of calcium-based additives increased, total P (TP) content in P-containing products reduced, apatite P (AP) content increased, non-apatite P (NAIP) content declined, and P solubility in citric acid content decreased. The amount of AP increased, NAIP reduced, and P solubility in citric acid decreased as the incineration temperature climbed. Although P in P-containing products prepared by OSM and MSM changed in a similar way at different additive doses and temperatures, P-containing products prepared by MSM had at least a 6.1% increase in P solubility in citric acid. Compared with OSM, MSM could save 10% of calcium-based additives when reaching the maximum AP value. Additionally, pure materials were employed to investigate how P species changed during the incineration procedure. The advantage of the MSM-prepared product over the OSM-prepared product may be explained by the high concentration of Ca3(PO4)2 and low concentration of amorphous calcium bound P (Ca-P). Overall, MSM is an effective method to reduce the dose of calcium-based additives and increase the bioavailability of P in P-containing products.

Phosphorus recovery from incinerated sewage sludge ash using electrodialysis coupled with plant extractant enhancement technology.

Phosphorus (P) is an integral mineral nutrient for the growth of plants and animals. As the increasing population worldwide, the demand for P resources keeps increasing. Therefore, it is necessary to recover P from secondary resources. Unlike conventional P recovery processes, this work focused on the recovery of P from incinerated sewage sludge ash (ISSA) using electrodialysis as the main technology coupled with plant extractants. In this study, Amaranthus and hydrolyzed polymaleic anhydride (HPMA) were used as P extractants, investigating the effects of HPMA concentration and pH of the compound agent on the migration of P and heavy metals from ISSA. The results showed that the concentration of HPMA and pH of the compound agent had a significant influence on the mobility of P and heavy metals. Meanwhile, the impacts of eggshell additions and voltage on the recovery efficiency of P was also studied by using waste eggshells as calcium sources. We found that when eggshells were added at 10 g/L and the voltage was 10 V, the recovery efficiency of P reached 96.05%. Moreover, XRD patterns revealed that the mineral phase of recovered P-containing products was predominantly hydroxyapatite, which had good environmental benefits. Generally, the favorable results have been achieved in the recovery efficiency of P and has practical implications for P recovery from ISSA.

YY1-induced long non-coding RNA PSMA3 antisense RNA 1 functions as a competing endogenous RNA for microRNA 214-5p to expedite the viability and restrict the apoptosis of bladder cancer cells via regulating programmed cell death-ligand 1.

Bladder cancer (BC) is one of the most common malignant tumors in the urinary system. Our research aimed to explore the function and underlying mechanisms of long noncoding RNA (lncRNA) PSMA3-AS1 in BC. RT-qPCR was utilized to detect the levels of PSMA3-AS1, miR-214-5p, and PD-L1. ChIP assay was employed to confirm the transcription factor of PSMA3-AS1. Luciferase reporter assay was carried out to demonstrate the relationships between miR-214-5p and PSMA3-AS1 or PD-L1. The diagnostic value of PSMA3-AS1 was evaluated by the ROC curve. CCK-8, wound healing, transwell, and flow cytometry assays were applied to analyze cell viability, migration, invasion, and apoptosis. Western blotting was used to confirm the expression of cleaved caspase-3. The present study revealed that BC tissues and cells exhibited an increased expression in PSMA3-AS1. High expression of PSMA3-AS1 was related to poor prognosis in BC patients. Then, the area under the ROC curve for PSMA3-AS1 was up to 0.8954. Moreover, ChIP assay elaborated that YY1 could bind to the PSMA3-AS1 promoter region. Furthermore, it was found that that PSMA3-AS1 knockdown repressed BC cell viability and metastasis, and promoted apoptosis. In addition, miR-214-5p was inversely correlated with PSMA3-AS1 or PD-L1 levels. MiR-214-5p deletion reversed the impacts of PSMA3-AS1 deletion on BC progression, and PD-L1 inhibition also abrogated the influence of miR-214-5p deletion in BC development. In conclusion, YY1-induced PSMA3-AS1 exerted an oncogenic function in BC cells via targeting miR-214-5p and enhancing PD-L1, providing potential biomarkers for BC therapy.

Transformation of phosphorus by MgCl2 and CaCl2 during sewage sludge incineration.

Phosphorus (P) recovery from sewage sludge (SS) have been regarded as an effective method of P recycling. The effects of incineration temperature, incineration time, and chlorine additives on the distribution of P speciation during sludge incineration were studied. Moreover, the reactions between model compounds AlPO4 and additives (MgCl2 and CaCl2) were investigated by thermogravimetric differential thermal analysis and X-ray diffraction measurements. The results demonstrated that the increase in temperature and time stimulated the volatilization of non-apatite inorganic phosphorus (NAIP) instead of apatite phosphorus (AP). MgCl2 and CaCl2 can greatly promote the conversion of NAIP to AP. Additionally, AlPO4 reacted with MgCl2 are incinerated at 500-600 °C to form Mg3(PO4)2, which is mainly due to the reaction of the intermediate product MgO and AlPO4. Reactions between AlPO4 and CaCl2 occurred at 700-750 °C and produced Ca2PO4Cl, which can be directly used with high bioavailability. These findings suggested that chlorine additives in the SS incineration process can obtain phosphorus-containing minerals with higher bioavailability to realize the resource utilization of P in sludge.

Improvement of phosphorus release from sludge by combined electrochemical-EDTA treatment.

In this paper, combined with the addition of ethylenediaminetetraacetic acid (EDTA), the electrochemical treatment of waste activated sludge (WAS) was investigated to explore its effect on the release of phosphorus (P) from WAS. The results showed that during the electrochemical treatment, the addition of EDTA could significantly promote the release of P from the WAS to the supernatant, the optimal amount of EDTA was 0.4 g/g total suspended solids (TSS), when the release of total dissolved phosphorus (TDP), organic phosphorus (OP) and molybdate reactive phosphorus (PO43--P) were 187.30, 173.84 and 13.46 mg/L, respectively. OP was the most likely form of P to be released during this process. Moreover, combined electrochemical-EDTA treatment could promote the release of P and metal ions from extracellular polymeric substances (EPSs) to the supernatant, and increase the solubility and disintegration of sludge. EDTA chelated the metal ions of sludge flocs and phosphate precipitates to cause sludge floc decomposition, thereby promoting the release of P from WAS.

Behavior of fast and slow phosphorus release from sewage sludge-derived biochar amended with CaO.

The pyrolyzation of sewage sludge (SS) could efficiently transform inherent phosphorus (P) into bioavailable phosphate forms, which endows SS-derived biochar (SSB) the potential as a soil fertilizer. However, the details about the release behavior of P in SSB have not been systematically investigated. This study evaluated the fast and slow P releasing behaviors from SSB and CaO-amended SSB prepared under different pyrolysis temperature. The higher pyrolysis temperature and CaO addition could enhance the conversion of non-apatite inorganic phosphorus (NAIP) into more bioavailable apatite inorganic phosphorous (AP). Acidic and alkaline conditions were favorable for the fast release of P from SSB. Higher ionic strength condition gave greater releasing amounts of TP and the SO42- facilitating a rapid release of TP than those for Cl- and NO3-. SSBs with CaO addition showed a much slower TP release than those without CaO both in fast release (24 h, with CaO: 0.05~0.4 mg TP g-1 SSB, e.g., without CaO 0.5~5 mg TP g-1 SSB) and slow release tests (21 days, with CaO: 1.2~4.1 mg TP g-1 SSB, e.g., without CaO 1.8~5.7 mg TP g-1 SSB). Ortho-P release was more remarkable for the SSB amended with CaO (~54% of TP), which was likely due to the formation of orthophosphate. The results of this study suggested that SSB prepared by high pyrolysis temperature and CaO addition had high potential as a slow P-releasing fertilizer for the soil.

An efficient strategy of phosphorus recovery: Electrochemical pretreatment enhanced the anaerobic fermentation of waste activated sludge.

The anaerobic fermentation (AF) of waste activated sludge (WAS) with an electrochemical pretreatment (EPT) was investigated to determine its correlation with the release of phosphorus and the disintegration of WAS. The sludge was pretreated by holding under 4.5 V for 60 min, followed by AF for 9 days. Untreated sludge was used as the control group (no-EPT). Results showed that, with pretreatment, the total dissolved P (TDP), orthophosphate (PO43--P) and organic P (OP) reached the maximum values of 7.30 mg/L, 4.77 mg/L and 2.35 mg/L on day 8, respectively, which were approximately 5.3, 9.2 and 2.7 times greater than that in the control group. The analysis of soluble chemical oxygen demand (SCOD), protein and polysaccharides showed that the EPT promoted the disintegration of sludge, thereby enhancing the P release. The SCOD reached 1625 mg/L on day 6 in pretreatment experiment, which was about 9.8 times greater than that in control group. Additionally, the EPT contributed to fewer metal ions in sludge supernatant. This mechanism might have been due to the anions accumulating in the supernatant from the greater degree of sludge collapse after EPT, which caused the released metal ions to combine with anions to form insoluble compounds. In conclusion, EPT could be a promising method for the dissolution of sludge and the recovery of phosphorus from WAS under AF. Besides, the economic benefit evaluation showed the potential value of EPT for P recovery.

Enhancement of phosphorus release from waste activated sludge by electrochemical treatment.

The enhancement of phosphorus (P) released from waste activated sludge (WAS) by electrochemical treatment was investigated in this study. Results showed that the concentration of orthophosphate (ortho-P) and organic phosphorus (OP) in liquid both increased after electrochemical treatment. The ortho-P and OP concentration reached a maximum of 5.020 and 1.888 mg/L under the optimal condition respectively (voltage of 4.5 V and time of 60 min), which were 2.86 and 4.93 times higher than that in raw sludge. Meanwhile, the role of extracellular polymeric substances (EPS) in this process was also studied. The variation trends of P-release in tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) were different. In TB-EPS, the concentration of total phosphorus (TP) and ortho-P decreased when the voltage increased. In contrast, the concentration of TP and ortho-P in LB-EPS increased and reached the maximum under the optimal condition. Released metal ions (Ca, Mg, Fe, and Al) had some effects on P-release both in liquid and EPS, which indicated that EPS played an important role. SCOD and TSS revealed that the disintegration of sludge was also enhanced by electrochemical treatment. Additionally, the P fractions in sludge phase suggested that OP was more likely to be released in liquid phase.

Phosphorus recovery from sewage sludge via incineration with chlorine-based additives.

Phosphorus (P) is an essential nutrient for all organisms, and the recovery of P from sewage sludge (SS) has been the focus of recent research. The effect of temperature and chlorine-based additives (MgCl2 and CaCl2) on P recovery and heavy metal (Cu, Zn, and Cr) removal during SS incineration were investigated. The results showed that temperature and chlorine-based additives increased the P fixation and improved the P-bioavailability. The fixation rate of P reached a maximum of 98.5% in SS using 3% Mg (MgCl2) at 900 °C and 97.8% in SS using 5%Ca (CaCl2) at 800 °C. Additionally, the mechanism of P associated with aluminum oxides/hydroxides (Al-P) transferring to that of magnesium oxides/hydroxides (Mg-P) and calcium oxides/hydroxides (Ca-P) was also investigated. The results indicated that Mg3(PO4)2 was formed via the reaction between AlPO4 and MgCl2 during incineration, which increased the P solubility in 2% citric acid. When AlPO4 was incinerated with CaCl2, Ca2PO4Cl which has a higher P-bioavailability than AlPO4 was formed. In addition, the mass fraction and leaching concentration of Cu and Zn in treated SS greatly decreased during SS incineration, while no reduction in Cr. These findings suggest that chlorine-based additives can be used to treat SS at high temperatures to obtain an ideal material for P-fertilizer production.

Phosphorus speciation and bioavailability of sewage sludge derived biochar amended with CaO.

In this study, biochar samples were prepared from the co-pyrolysis of sewage sludge (SS) and CaO to explore the transformation of P speciation in sample. The potential of these biochar as a fertilizer to promote the growth of the plant was also evaluated. The result indicated that CaO addition can greatly facilitate the conversion of non-apatite inorganic phosphorus (NAIP) to apatite inorganic phosphorus (AP, mainly Ca3(PO4)2 and Ca3Mg3(PO4)4). The addition of 10% CaO in feedstock is sufficient to convert SS inherent P into more bioavailable AP. Under such a dosage, AP content in biochar increased by 21.2-33.6% in contrast to CaO free sample at pyrolysis temperature of 500-800 °C, and water-soluble phosphorus (WSP) content decreased to less than 1% of TP. The addition of CaO can also apparently reduce Zn, Mn leaching from biochar. Additionally, hydroponics assay showed that CaO amended SS biochar can promote the growth of rice seedling. The results of this study indicate that preparing CaO amended SS biochar is a technically feasible strategy to utilize P resource in SS.

Olive oil polyphenol extract inhibits vegetative cells of Bacillus cereus isolated from raw milk.

This study was conducted to analyze the antibacterial effect of olive oil polyphenol extract (OOPE) against vegetative cells of Bacillus cereus isolated from raw milk and reveal the possible antibacterial mechanism. The diameter of inhibition zone, minimum inhibitory concentration, minimum bactericidal concentration, and survival counts of bacterial cells in sterile normal saline and pasteurized milk were used to evaluate the antibacterial activity of OOPE against B. cereus vegetative cells. The changes in intracellular ATP concentration, cell membrane potential, content of bacterial protein, and cell morphology were analyzed to reveal possible mechanisms of action. Our results showed the diameter of inhibition zone, minimum inhibitory concentration, and minimum bactericidal concentration of OOPE against B. cereus vegetative cells were 18.44 ± 0.55 mm, 0.625 mg/mL, and 1.25 mg/mL, respectively. Bacillus cereus GF-1 vegetative cells were decreased to undetectable levels from about 8 log cfu/mL after treatments with 0.625 mg/mL of OOPE in normal saline at 30°C for 3 h and in pasteurized milk at 30°C for 10 h. The antibacterial mechanisms of OOPE against B. cereus GF-1 vegetative cells may be due to the reduction of intracellular ATP concentrations, cell membrane depolarization, decrease of bacterial protein content, and leakage from cytoplasm. These findings illustrated that OOPE could be used to prevent the growth of contaminating B. cereus cells in dairy products.

Standardized Ginkgo biloba extract EGb 761® attenuates early brain injury following subarachnoid hemorrhage via suppressing neuronal apoptosis through the activation of Akt signaling.

BACKGROUND: Early brain injury (EBI) plays a critical role in determining the outcome of subarachnoid hemorrhage (SAH). The present study was designed to investigate the role of EGb 761, a standardized extract of Ginkgo biloba, in SAH-induced EBI and to explore its potential mechanism of action. METHOD: A rat SAH model was established by the endovascular perforation process. Doses of 10, 50 and 100 mg/kg EGb 761 were injected intraperitoneally 2 h after SAH was induced. Mortality, SAH grade, neurological score and brain water content were measured 24 h after SAH was induced. A Western blot assay was performed to assess the expression of the apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3, Akt and phosphorylated Akt (p-Akt). Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and neuronal nuclei (NeuN) double immunofluorescence staining were used to detect apoptotic neurons. RESULTS: Animals suffered from serious neurological deficits and increased brain water content after induction of SAH. Rats treated with EGb 761 experienced dose-dependent attenuation of neurological dysfunction and decreased brain water content. In addition, EGb 761 significantly activated Akt signaling accompanied by increased Bcl-2 levels and decreased expression of Bax and cleaved caspase-3. Moreover, EGb 761 decreased the number of TUNEL/NeuN-positive cells in a dose-dependent manner. However, all the beneficial effects of EGb 761 for SAH were abolished by the Akt inhibitor MK2206. CONCLUSION: Our results indicated that EGb 761 could ameliorate SAH-induced EBI and that the neuroprotective effects of EGb 761 against SAH were exerted via suppression of neuronal apoptosis through activation of the Akt pathway.

Remediation of chromium-contaminated soil by electrokinetics and electrokinetics coupled with CaAl-LDH permeable reaction barrier.

The remediation of Cr(VI)-contaminated soil was investigated by electrokinetic (EK) and permeable-reactive-barrier assisted electrokinetic (EK-PRB). The medium of PRB was hydrocalumite (CaAl-LDH). The results showed that removal efficiency of hexavalent chromium (Cr(VI)) in EK-PRB and EK system was 96.49 and 85.50%, respectively. Simultaneously, the removal efficiency of total chromium (TCr) was 69.34 and 40.97% after 120-h treatment. The XRD, FTIR, and XPS analyses indicated that the reactive barrier media of CaAl-LDH successfully captured the chromium. Besides, the migration rate of chromium in EK-PRB was relatively faster than EK, since the media of PRB captured chromium in-time and reduced the influence of chromium accumulation on the migration of chromium. Moreover, the trivalent chromium (Cr(III)) was generated in EK/EK-PRB, and the chromium was stabilized in soil with the chemical speciations of oxidizable and residual fractions. Therefore, the treatment of EK-PRB and EK both increased the removal of chromium and decreased its environmental risks.

Antimicrobial Activity of Punicalagin Against Staphylococcus aureus and Its Effect on Biofilm Formation.

Punicalagin, one of the main active compounds in pomegranate peel, has been reported to possess many properties, including antioxidant, antimicrobial, antiviral, and immunosuppressive activities. The aim of this study was to investigate the antibacterial effect of punicalagin against Staphylococcus aureus and possible mode of action. Growth inhibition activity was examined by the agar diffusion method. Then agar dilution method was adopted to determine the minimum inhibitory concentration (MIC). The effects of punicalagin on cell membrane were assessed by measuring potassium efflux. Morphological changes of S. aureus were assessed by scanning and transmission electron microscopy. Crystal violet assay was applied to investigate antibiofilm activity of punicalagin. Punicalagin exhibited good antistaphylococcal effect with an MIC of 0.25 mg/mL. An increase of potassium efflux was observed when cells were treated with punicalagin at 2 × MIC. Punicalagin induced morphological damages to the cell membrane. Moreover, punicalagin exerted a remarkable inhibitory effect on biofilm formation of S. aureus. These findings suggest that punicalagin has antimicrobial and antibiofilm activities against S. aureus and may have potential application to control S. aureus contamination in food industry.

The anti-infective activity of punicalagin against Salmonella enterica subsp. enterica serovar typhimurium in mice.

Punicalagin, a major bioactive component of pomegranate peel, has been proven to have antioxidant, antiviral, anti-apoptosis, and hepatoprotective properties. The aim of this study was to investigate the anti-infective activity of punicalagin in a mouse model. C57BL/6 mice were initially challenged with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and then treated with punicalagin. Food and water consumption and body weight were recorded daily. On day 8 post infection, the mice were sacrificed to examine pathogen counts in tissues, hematological parameters, cytokine levels, and histological changes. Compared to mice only infected with S. typhimurium, punicalagin-treated mice had more food consumption and less weight loss. A higher survival rate and lower counts of viable S. typhimurium in feces, liver, spleen, and kidney were found in the punicalagin-treated mice. The enzyme linked immunosorbent assay showed that the levels of IL-6, IL-10, and IFN-γ in serum and the spleen and TNF-α in serum, the spleen and the liver were reduced by punicalagin. Moreover, more neutrophils and higher neutrophil-to-mononuclear cell ratios in the punicalagin-treated mice were observed. Histological examination showed that punicalagin protected cells in the liver and spleen from hemorrhagic necrosis. It is concluded that punicalagin has a beneficial effect against S. typhimurium infection in mice. The anti-infective properties, together with other nutritionally beneficial effects, make punicalagin a promising supplement in human food or animal feeds to prevent disease associated with S. typhimurium.

Aqueous Cr (VI) removal by Friedel's salt adsorbent prepared from calcium aluminate-rich cementitious materials.

This research paper investigated a novel absorbent of calcium aluminate-rich cementitious materials (Friedel's salt adsorbent, FA) for aqueous hexavalent chromium (VI) removal. The adsorption kinetics showed that the maximum adsorption capacities of FA were 3.36, 14.66, and 26.17 mg/g when the initial Cr(VI) concentration was 10, 50, and 100 mg/L, respectively. The adsorption fitted with the pseudo-second-order kinetic model, suggesting the important roles of intercalation in the adsorption process with increasing Cr(VI) concentrations. This Friedel's salt adsorbent is suggested as an adaptive and effective adsorbent for Cr(VI) removal in contaminated groundwater.

CaFeAl mixed oxide derived heterogeneous catalysts for transesterification of soybean oil to biodiesel.

CaAl layered double oxides (LDO) were prepared by co-precipitation and calcined at 750°C, and then applied to biodiesel production by transesterification reaction between methanol and soybean oil. Compared with characteristics of CaFe/LDO and CaAl/LDO, CaFeAl/LDO had the best performance based on prominent catalytic activity and stability, and achieved over 90% biodiesel yield, which stayed stable (over 85%) even after 8 cycles of reaction. The optimal catalytic reaction condition was 12:1M-ratio of methanol/oil, reaction temperatures of 60°C, 270rpm stirring rate, 60min reaction time, and 6% weight-ratio of catalyst/oil. In addition, the CaFeAl/LDO catalyst is insoluble in both methanol and methyl esters and can be easily separated for further reaction, turning it into an excellent alternative for biodiesel synthesis.

Hepatocyte nuclear factor 4 alpha is a key factor related to depression and physiological homeostasis in the mouse brain.

Major depressive disorder (MDD) is a common psychiatric disorder that involves marked disabilities in global functioning, anorexia, and severe medical comorbidities. MDD is associated with not only psychological and sociocultural problems, but also pervasive physical dysfunctions such as metabolic, neurobiological and immunological abnormalities. Nevertheless, the mechanisms underlying the interactions between these factors have yet to be determined in detail. The aim of the present study was to identify the molecular mechanisms responsible for the interactions between MDD and dysregulation of physiological homeostasis, including immunological function as well as lipid metabolism, coagulation, and hormonal activity in the brain. We generated depression-like behavior in mice using chronic mild stress (CMS) as a model of depression. We compared the gene expression profiles in the prefrontal cortex (PFC) of CMS and control mice using microarrays. We subsequently categorized genes using two web-based bioinformatics applications: Ingenuity Pathway Analysis and The Database for Annotation, Visualization, and Integrated Discovery. We then confirmed significant group-differences by analyzing mRNA and protein expression levels not only in the PFC, but also in the thalamus and hippocampus. These web tools revealed that hepatocyte nuclear factor 4 alpha (Hnf4a) may exert direct effects on various genes specifically associated with amine synthesis, such as genes involved in serotonin metabolism and related immunological functions. Moreover, these genes may influence lipid metabolism, coagulation, and hormonal activity. We also confirmed the significant effects of Hnf4a on both mRNA and protein expression levels in the brain. These results suggest that Hnf4a may have a critical influence on physiological homeostasis under depressive states, and may be associated with the mechanisms responsible for the interactions between MDD and the dysregulation of physiological homeostasis in humans.

Tannin-rich pomegranate rind extracts reduce adhesion to and invasion of Caco-2 Cells by Listeria monocytogenes and decrease its expression of virulence genes.

Pomegranate rind is rich in tannins that have remarkable antimicrobial activities. This study aimed to evaluate the effects of a tannin-rich fraction from pomegranate rind (TFPR) on Listeria monocytogenes virulence gene expression and on the pathogen's interaction with human epithelial cells. Growth curves were monitored to determine the effect of TFPR on L. monocytogenes growth. The 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide and fluorescence staining assays were used to examine the cytotoxicity of TFPR. The effects of TFPR on L. monocytogenes adhesion to and invasion of epithelial cells were investigated using Caco-2 cells. Real-time quantitative PCR analysis was conducted to quantify mRNA levels of three virulence genes in L. monocytogenes. Results showed that a MIC of TFPR against L. monocytogenes was 5 mg/ml in this study. TFPR exhibited cytotoxicity against Caco-2 cells when the concentration was 2.5 mg/ml. Subinhibitory concentrations of TFPR significantly reduced, in a dose-dependent manner, adhesion to and invasion of Caco-2 cells by L. monocytogenes. When L. monocytogenes was grown in the presence of 2.5 mg/ml TFPR, the transcriptional levels of prfA, inlA, and hly decreased by 17-, 34-, and 28-fold, respectively.

Tannin-rich fraction from pomegranate rind damages membrane of Listeria monocytogenes.

Pomegranate rind has been reported to inhibit several foodborne pathogens, and its antimicrobial activity has been attributed mainly to its tannin fraction. This study aimed to investigate the antimicrobial activity of the tannin-rich fraction from pomegranate rind (TFPR) against Listeria monocytogenes and its mechanism of action. The tannin-related components of TFPR were analyzed by high-performance liquid chromatography and liquid chromatography-mass spectrometry, and the minimum inhibitory concentration (MIC) of TFPR was determined using the agar dilution method. Extracellular potassium concentration, the release of cell constituents, intra- and extracellular ATP concentrations, membrane potential, and intracellular pH (pHin) were measured to elucidate a possible antibacterial mechanism. Punicalagin (64.2%, g/g) and ellagic acid (3.1%, g/g) were detected in TFPR, and the MICs of TFPR were determined to be 1.25-5.0 mg/mL for different L. monocytogenes strains. Treatment with TFPR induced a decrease of the intracellular ATP concentration, an increase of the extracellular concentrations of potassium and ATP, and the release of cell constituents. A reduction of pHin and cell membrane hyperpolarization were observed after treatment. Electron microscopic observations showed that the cell membrane structures of L. monocytogenes were apparently impaired by TFPR. It is concluded that TFPR could destroy the integrity of the cell membrane of L. monocytogenes, leading to a loss of cell homeostasis. These findings indicate that TFPR has the potential to be used as a food preservative in order to control L. monocytogenes contamination in food and reduce the risk of listeriosis.