The effect of mesenchymal stem cell conditioned medium incorporated within chitosan nanostructure in clearance of common gastroenteritis bacteria in-vitro and in-vivo.

Gastroenteritis infection is a major public health concern worldwide, especially in developing countries due to the high annual mortality rate. The antimicrobial and antibiofilm activity of human mesenchymal stem cell-derived conditioned medium (hMSCsCM) encapsulated in chitosan nanoparticles (ChNPs) was studied in vitro and in vivo against common gastroenteritis bacteria. The synthesized ChNPs were characterized using Zeta potential, scanning electron microscopy (SEM), and dynamic light scattering (DLS) techniques. HMSC-derived conditioned medium incorporated into chitosan NPs (hMSCsCM-ChNPs) composite was fabricated by chitosan nanoparticles loaded with BM-MSCs (positive for CD73 and CD44 markers). The antimicrobial and antibiofilm activity of composite was investigated against four common gastroenteritis bacteria (Campylobacter jejuni ATCC29428, Salmonella enteritidis ATCC13076, Shigella dysenteriae PTCC1188, and E. coli ATCC25922) in-vitro and in-vivo. Majority of ChNPs (96%) had an average particle size of 329 nm with zeta potential 7.08 mV. The SEM images confirmed the synthesis of spherical shape for ChNPs and a near-spherical shape for hMSCsCM-ChNPs. Entrapment efficiency of hMSCsCM-ChNPs was 75%. Kinetic profiling revealed that the release rate of mesenchymal stem cells was reduced following the pH reduction. The antibacterial activity of hMSCsCM-ChNPs was significantly greater than that of hMSCsCM and ChNPs at dilutions of 1:2 to 1:8 (P < 0.05) against four common gastroenteritis bacteria. The number of bacteria present decreased more significantly in the group of mice treated with the hMSCsCM-ChNPs composite than in the groups treated with hMSCsCM and ChNPs. The antibacterial activity of hMSCsCM against common gastroenteritis bacteria in an in vivo assay decreased from > 106 CFU/ml to approximately (102 to 10) after 72 h. Both in vitro and in vivo assays demonstrated the antimicrobial and antibiofilm activities of ChNPs at a concentration of 0.1% and hMSCsCM at a concentration of 1000 µg/ml to be inferior to that of hMSCsCM-ChNPs (1000 µg/ml + 0.1%) composite. These results indicated the existence of a synergistic effect between ChNPs and hMSCsCM. The designed composite exhibited notable antibiofilm and antibacterial activities, demonstrating optimal release in simulated intestinal lumen conditions. The utilization of this composite is proposed as a novel treatment approach to combat gastroenteritis bacteria in the context of more challenging infections.

Battling Salmonella enteritidis infections: integrating proteomics and in vivo assessment of Galla Chinensis tannic acid.

Salmonella infections pose a significant threat to animal and human health. Phytochemicals present a potential alternative treatment. Galla chinensis tannic acid (GCTA), a hydrolyzable polyphenolic compound, inhibits bacterial growth and demonstrates potential as an alternative or supplement to antibiotics to prevent Salmonella infections. However, little is known about the antimicrobial mechanism of GCTA against Salmonella. Here, we revealed 456 differentially expressed proteins upon GCTA treatment, impacting pathways related to DNA replication, repair, genomic stability, cell wall biogenesis, and lipid metabolism using TMT-labeled proteomic analysis. TEM analysis suggested altered bacterial morphology and structure post-treatment. A Salmonella-infected-mouse model indicated that GCTA administration improved inflammatory markers, alleviated intestinal histopathological alterations, and reduced Salmonella enterica serovar Enteritidis (S. Enteritidis) colonization in the liver and spleen of Salmonella-infected mice. The LD50 of GCTA was 4100 mg/kg with an oral single dose, vastly exceeding the therapeutic dose. Thus, GCTA exhibited antibacterial and anti-infective activity against S. Enteritidis. Our results provided insight into the molecular mechanisms of these antibacterial effects, and highlights the potential of GCTA as an alternative to antibiotics.

Red osier dogwood extract vs. trimethoprim-sulfadiazine (Part 2). Pharmacodynamic effects on ileal and cecal microbiota of broiler chickens challenged orally with Salmonella Enteritidis.

With the subsisting restrictions on the use of antibiotics in poultry production, the use of plant extracts has shown some promising antimicrobial capacity similar to antibiotics; however, such capacity is largely dependent on their total polyphenol concentration and profile. Given the emerging antimicrobial potential of red osier dogwood (ROD) extract, the study aimed to investigate the pharmacodynamic effect of ROD extract on the ileal and cecal microbiota of broiler chickens challenged orally with Salmonella Enteritidis (SE). A 21 d 4 × 2 factorial experiment was conducted based on 2 main factors, including diets and SE challenge. A total of 384 one-day-old mixed-sex Cobb-500 broiler chicks were randomly allotted to 4 dietary treatments; Negative control (NC), NC + 0.075 mg trimethoprim-sulfadiazine (TMP/SDZ)/kg of diet, and NC containing either 0.3 or 0.5% ROD extract. On d 1, half of the birds were orally challenged with 0.5 mL of phosphate-buffered saline (Noninfected group) and the remaining half with 0.5 mL of 3.1 × 105 CFU/mL SE (Infected group). Dietary treatments were randomly assigned to 8 replicate cages at 6 birds/cage. On d 21, 10 birds/treatment were euthanized and eviscerated to collect ileal and cecal digesta for gut microbiota analysis. The ileal and cecal microbiota was dominated by phyla Firmicutes, Proteobacteria, and Actinobacteriota. The SE infection decreased (P < 0.05) the relative abundance of Proteobacteria and Actinobacteriota in the ileum and ceca, respectively, however, it increased (P < 0.05) Proteobacteria in the ceca. Both 0.3 and 0.5% ROD extracts (P < 0.05) depressed the relative abundance of Actinobacteriota in the ileum but marginally improved (P < 0.05) it in the ceca compared to the TMP/SDZ treatment. Dietary TMP/SDZ increased (P < 0.05) genus Bifidobacterium at the ileal and cecal segments compared to other treatments. Dietary 0.3 and 0.5% marginally improved (P < 0.05) Bifidobacterium in the ceca and depressed (P < 0.05) Weissella and was comparably similar to TMP/SDZ in the ileum. Regardless of the dietary treatments and SE infection, alpha diversity differed (P < 0.05) between ileal and cecal microbiota. Beta diversity was distinct (P < 0.05) in both ileal and cecal digesta along the SE infection model. Conclusively, both ROD extract levels yielded a pharmacodynamic effect similar to antibiotics on ileal and cecal microbiota.

Variations in the Composition, Antioxidant and Antimicrobial Activities of Cystoseira compressa during Seasonal Growth.

The underexplored biodiversity of seaweeds has recently drawn great attention from researchers to find the bioactive compounds that might contribute to the growth of the blue economy. In this study, we aimed to explore the effect of seasonal growth (from May to September) on the in vitro antioxidant (FRAP, DPPH, and ORAC) and antimicrobial effects (MIC and MBC) of Cystoseira compressa collected in the Central Adriatic Sea. Algal compounds were analyzed by UPLC-PDA-ESI-QTOF, and TPC and TTC were determined. Fatty acids, among which oleic acid, palmitoleic acid, and palmitic acid were the dominant compounds in samples. The highest TPC, TTC and FRAP were obtained for June extract, 83.4 ± 4.0 mg GAE/g, 8.8 ± 0.8 mg CE/g and 2.7 ± 0.1 mM TE, respectively. The highest ORAC value of 72.1 ± 1.2 µM TE was obtained for the August samples, and all samples showed extremely high free radical scavenging activity and DPPH inhibition (>80%). The MIC and MBC results showed the best antibacterial activity for the June, July and August samples, when sea temperature was the highest, against Listeria monocytogenes, Staphylococcus aureus, and Salmonella enteritidis. The results show C. compressa as a potential species for the industrial production of nutraceuticals or functional food ingredients.

A combined approach using slightly acidic electrolyzed water and tea polyphenols to inhibit lipid oxidation and ensure microbiological safety during beef preservation.

Slightly acidic electrolyzed water (SAEW) is often used as a disinfectant in beef preservation to ensure microbiological safety. However, it ineffectively inhibit lipid oxidation. Therefore, the combination of SAEW and tea polyphenols (TPs) was tested to inhibit lipid oxidation and microbial growth in beef preservation. SAEW and TPs were selected as the optimum sanitizer and antioxidant, respectively. Then, the inactivation efficacies of different combination treatments of SAEW and TPs of Salmonella enteritidis in beef were compared and treatment of SAEW-TPs (SAEW immersion at an available chlorine concentration of 30 mg/L for 2.5 min, followed by the TPs immersion at a 0.1% concentration for 2.5 min) was selected. Finally, the effectiveness of SAEW-TPs on the microbiological and physicochemical properties of beef during storage was evaluated. The results revealed that the required quality standard of beef treated with SAEW-TPs was prolonged by approximately 9 d at 4 °C, and this treatment had greater antimicrobial and antioxidant effects than did the single treatment.

Chlorine-stressed Salmonella cells are significantly more engulfed by Acanthamoeba trophozoites and have a longer intracystic survival than the non-stressed cells.

In the present study, the effect of sublethal chlorine-induced oxidative stress on the subsequent interaction of Salmonella enterica serovars Enteritidis and Typhimurium with Acanthamoeba castellanii and A. polyphaga was evaluated. Sublethal chlorine concentration was determined using the lag phase extension information and used to prepare chlorine-stressed Salmonella cells. Coculture experiments of Acanthamoeba and Salmonella cells were performed in Page's amoeba saline (PAS) at 25 °C for 2 h. The results showed that the chlorine-stressed Salmonella cells were significantly more engulfed by A. castellanii and A. polyphaga trophozoites than the non-stressed cells. The uptake rates of the chlorine-stressed and non-stressed Salmonella cells were in the range of 14.17-27.34 and 6.51-11.52% for A. castellanii, and in the range of 8.32-17.76 and 2.28-6.12% for A. polyphaga trophozoites, respectively. Moreover, intracystic survival time of chlorine-stressed cells of S. Enteritidis and S. Typhimurium was significantly longer than that of non-stressed cells. While, non-stressed Salmonella cells survived within A. castellanii and A. polyphaga cysts for 13-20 and 8-15 days, chlorine-stressed cells were recovered from A. castellanii and A. polyphaga cysts after 22-32 and 15-24 days, respectively. These results underscore the importance of bacterial exposure to sublethal chlorine concentrations in their interaction with free-living amoebae, and may lead to a better understanding of the parameters affecting the persistence of Salmonella enterica serovars in food-related environments.

Immobilization and Release Studies of Triazole Derivatives from Grafted Copolymer Based on Gellan-Carrying Betaine Units.

New polymer-bioactive compound systems were obtained by immobilization of triazole derivatives onto grafted copolymers and grafted copolymers carrying betaine units based on gellan and N-vinylimidazole. For preparation of bioactive compound, two new types of heterocyclic thio-derivatives with different substituents were combined in a single molecule to increase the selectivity of the biological action. The 5-aryl-amino-1,3,4 thiadiazole and 5-mercapto-1,2,4-triazole derivatives, each containing 2-mercapto-benzoxazole nucleus, were prepared by an intramolecular cyclization of thiosemicarbazides-1,4 disubstituted in acidic and basic medium. The structures of the new bioactive compounds were confirmed by elemental and spectral analysis (FT-IR and 1H-NMR). The antimicrobial activity of 1,3,4 thiadiazoles and 1,2,4 triazoles was tested on gram-positive and gram-negative bacteria. The triazole compound was chosen to be immobilized onto polymeric particles by adsorption. The Langmuir, Freundlich, and Dubinin-Radushkevich adsorption isotherm were used to describe the adsorption equilibrium. Also, the pseudo-first and pseudo-second models were used to elucidate the adsorption mechanism of triazole onto grafted copolymer based on N-vinylimidazole and gellan (PG copolymer) and grafted copolymers carrying betaine units (PGB1 copolymer). In vitro release studies have shown that the release mechanism of triazole from PG and PGB1 copolymers is characteristic of an anomalous transport mechanism.

Antimicrobial activity of ClO2 gas against Salmonella Enteritidis on almonds.

Nuts, including almonds, are occasionally contaminated with Salmonella spp. In this study, we used chlorine dioxide (ClO2) gas to inactivate S. enterica subsp. Enterica serovar Enteritidis on almonds. Almonds inoculated with a single strain of S. Enteritidis (8.95 log cfu/mL) were exposed to ClO2 gas generated from 1.0 or 1.5 mL ClO2 solution in a sealed container at 50 or 60 °C (43% relative humidity) for up to 10 h. The concentration of ClO2 gas peaked at 354-510 and 750-786 ppm within 0.5 h upon deposition of 1.0 and 1.5 mL of aqueous ClO2, respectively, and gradually decreased thereafter. Population of S. Enteritidis on almonds treated at 50 °C decreased to 1.70-2.32 log cfu/sample within 1 h of exposure to ClO2 gas and decreased to below the detection limit (1.7 log cfu/sample) at all ClO2 concentrations after 8 h. At 60 °C, the microbial population fell below the detection limit within 1 h, regardless of the volume of ClO2 solution supplied. Microbial survival on almonds treated with ClO2 gas and stored at 12 or 25 °C was observed for up to 8 weeks and the organism was not recovered from the almonds treated for 10 h and stored at 12 °C for 2-8 weeks. The lightness (L value) and redness (a value) of almonds treated for 10 h were not changed by ClO2 gas treatment, but yellowness (b value) increased. Results showed that Salmonella on almonds was successfully inactivated by ClO2 gas treatment and the microbial survival did not occur during storage.

Cyperus esculentus L. Tubers (Tiger Nuts) Protect Epithelial Barrier Function in Caco-2 Cells Infected by Salmonella Enteritidis and Promote Lactobacillus plantarum Growth.

Cyperus esculentus L. tubers (tiger nuts) contain different compounds with several intestinal health-promoting properties. Here, we studied the capacity of tiger nuts from Valencia, Spain, to prevent epithelial barrier function disruption induced by Salmonella enteritidis in Caco-2 cell cultures. Paracellular permeability was assessed by transepithelial electrical resistance (TER) and tight junction protein immunolocalization. Moreover, the effect of tiger nuts on S. enteritidis agglutination, oxidative stress, and Lactobacillus plantarum growth was tested. Compared to controls, tiger nuts partially restored TER in S. enteritidis-infected cultures, an effect confirmed by immunolocalization of tight junction proteins ZO-1 and occludin. The results also revealed that this protective effect may be associated with the capacity to agglutinate the pathogen, restore TER in TNFα-stimulated cultures, and reduce reactive oxygen species in H2O2-stimulated cultures. Moreover, they favor L. plantarum growth. In conclusion, this study demonstrates that the tiger nut protects epithelial barrier function by reducing bacterial invasion, along with counteracting TNFα and H2O2 effects, thus giving an additional value to this tuber as a potential functional food.

Case Report: Ceftriaxone-Resistant Invasive Salmonella Enteritidis Infection with Secondary Hemophagocytic Lymphohistiocytosis: A Contrast with Enteric Fever.

In contrast to enteric fever, reports of secondary hemophagocytic lymphohistiocytosis (HLH) in invasive non-typhoidal salmonellosis are scarce. We report a child with ceftriaxone-resistant invasive Salmonella Enteritidis infection with secondary HLH, who was successfully managed with intravenous meropenem. Secondary HLH in the context of S. Enteritidis has not been described before.

Walnut (Juglans regia L.) Septum: Assessment of Bioactive Molecules and In Vitro Biological Effects.

Walnut (Juglans regia L.) septum represents an interesting bioactive compound source by-product. In our study, a rich phenolic walnut septum extract, previously selected, was further examined. The tocopherol content determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed higher amounts of α-tocopherol compared to γ- and δ-tocopherols. Moreover, several biological activities were investigated. The in vitro inhibiting assessment against acetylcholinesterase, α-glucosidase, or lipase attested a real management potential in diabetes or obesity. The extract demonstrated very strong antimicrobial potential against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enteritidis. It also revealed moderate (36.08%) and strong (43.27%) antimutagenic inhibitory effects against TA 98 and TA 100 strains. The cytotoxicity of the extract was assessed on cancerous (A549, T47D-KBluc, MCF-7) and normal (human gingival fibroblasts (HGF)) cell lines. Flow cytometry measurements confirmed the cytotoxicity of the extract in the cancerous cell lines. Additionally, the extract demonstrated antioxidant activity on all four cell types, as well as anti-inflammatory activity by lowering the inflammatory cytokines (interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1 ß (IL-1ß)) evaluated in HGF cells. To the best of our knowledge, most of the cellular model analyses were performed for the first time in this matrix. The results prove that walnut septum may be a potential phytochemical source for pharmaceutical and food industry.

Characteristics of New Peptides GQLGEHGGAGMG, GEHGGAGMGGGQFQPV, EQGFLPGPEESGR, RLARAGLAQ, YGNPVGGVGH, and GNPVGGVGHGTTGT as Inhibitors of Enzymes Involved in Metabolic Syndrome and Antimicrobial Potential.

The aim of this study was to determine the cytotoxic properties, influence on enzyme activity involved in metabolic syndrome, and antimicrobial activity of synthetic peptides with GQLGEHGGAGMG, GEHGGAGMGGGQFQPV, EQGFLPGPEESGR, RLARAGLAQ, YGNPVGGVGH, and GNPVGGVGHGTTGT sequences. Peptides have no cytotoxic effect on cells. The highest inhibitory effect on angiotensin converting enzyme I was noted for peptide GT-14 (IC50 = 525.63 µg/mL). None of the tested peptides had an influence on α-glucosidase. The highest α-amylase and lipase inhibitory activity was noted for GG-12 (IC50 = 56.72 and 60.62 µg/mL, respectively). The highest lipoxidase inhibitory activity was determined for peptide ER-13 (IC50 = 84.35 µg/mL). Peptide RQ-9 was characterized by the highest COX inhibitory activity (0.31 and 4.77 µg/mL for COX-1 and COX-2, respectively). Only peptide RQ-9 inhibited S. enteritidis ATCC 4931 growth (42%-48%) in all tested concentrations (15.62-250 mg/mL).

Resistencia a nitrofuranos en Salmonella enterica aisladas de carne para consumo humano / Nitrofuran resistance in Salmonella enterica isolated from meat for human consumption

RESUMEN En el presente estudio, se analizaron los mecanismos de resistencia a nitrofuranos en 18 muestras cár nicas con Salmonella enterica (15 de pollo, 2 de ternera y 1 de cerdo) de mercados de Lima (Perú). Determinaron los serotipos de los aislamientos y la sensibilidad a furazolidona y nitrofurantoina (con y sin el inhibidor de bombas de expulsión Phenyl-Arginine-β-Naphthylamide [PAβN]), las mutaciones en los genes snrA y cnr por PCR y la transferabilidad de la resistencia por conjugación. Se identificaron 15 muestras con S. infantis (13 muestras de pollo), 2 con S. enteritidis y 1 con S. anatum. Todos los aisla mientos, excepto S. anatum, fueron resistentes a ambos nitrofuranos (concentración mínima inhibidora [CMI] a furazolidona: 32-64 µg/mL, CMI a nitrofurantoina: 128-256 µg/mL), sin diferencias al adicio narse PAβN. Todos los aislamientos resistentes a nitrofuranos presentaron sustituciones en snrA y cnr (S. infantis: snrA STOP-151; cnr STOP-137; S. enteritidis: snrA STOP-180; cnr STOP-179). No se detectaron mecanismos transferibles de resistencia a nitrofuranos.

ABSTRACT The mechanisms of resistance to nitrofurans from 18 meat samples with Salmonella enterica (chicken: 15; beef: 2; pork: 1) collected in Lima (Peru) were analyzed. The isolates were serotyped and the susceptibility levels to furazolidone and nitrofurantoin [with and without the efflux pump inhibitor Phenyl-Arginine- β-naphthylamide (PAβN)], the presence of mutations in the snrA and cnr genes and the transferability of resistance by conjugation were established. Fifteen samples with S. infantis (13 from chicken samples), 2 with S. enteritidis and 1 with S. anatum were identified. All isolates except the S. anatum were resistant to both nitrofurans showing MICs (minimum inhibitory concentration) of furazolidone and nitrofurantoin of 32-64 μg/mL and 128-256 μg/mL, respectively. The addition of PAßN had no effect on the MIC levels. All nitrofuran-resistant isolates showed amino acid codon alterations at both snrA and cnr (S. infantis: snrA STOP-151; cnr STOP-137; S. enteritidis: snrA STOP-180; cnr STOP-179). No transferable mecha nisms of nitrofuran resistance were detected.

Contribution of the colicin receptor CirA to biofilm formation, antibotic resistance, and pathogenicity of Salmonella Enteritidis.

Salmonella Enteritidis is an important foodborne pathogen that can infect a wide range of animal species including human beings, resulting in great losses to commercial husbandry and human health. CirA is an outer membrane receptor involved in iron uptake and colicin1A/B-mediated competitive killing. Although iron uptake is crucial to bacterial virulence, limited literature is available about the role of CirA in infection. In the present work, we aimed to evaluate the role of CirA during S. Enteritidis infection. For this purpose, we generated a CirA-deficient mutant of the S. Enteritidis strain C50336 and examined its biological characteristics. The results showed that cirA gene inactivation caused sharply decreased biofilm formation and apparently impaired antibiotic resistance. Furthermore, the cirA gene deletion mutant showed markedly reduced adhesion and invasion to human epithelial cell line Caco-2 cells and decreased proliferation in mouse macrophage cell line RAW264.7 cells. Moreover, attenuated virulence was determined by a mouse model, with an LD50 increase of approximately 1,000-fold. These data indicated that CirA plays critical roles in the S. Enteritidis infection process.

Preparation and characterization of functional sodium caseinate/guar gum/TiO2/cumin essential oil composite film.

Eco-friendly functional bionanocomposite films were prepared using sodium caseinate (SC) and guar gum (GG) as the polymer matrix and TiO2 and cumin essential oil (CEO) as functional fillers. 0.2 vol% GG selected for the preparation of SC/GG composite film and various amount of TiO2 and CEO (1 and 2 wt% based on SC) were incorporated into the SC/GG film either individually or in combination. The addition of TiO2 and CEO significantly improved the water permeability and sensitivity properties and mechanical characteristics such as the strength (TS), stiffness (YM), and flexibility (EB) of the composite films. Also, the SC/GG films incorporated with TiO2 and CEO exhibited remarkable antibacterial activity against both Gram-positive (L. monocytogenes and S. aureus) and Gram-negative (E. coli O157: H7 and S. enteritidis) bacteria. All the film properties were varied not only on the concentration of TiO2 and CEO, but also increased synergistically when they were added together.

Efficacy of oregano and rosemary essential oils to affect morphology and membrane functions of noncultivable sessile cells of Salmonella Enteritidis 86 in biofilms formed on stainless steel.

AIMS: This study evaluated the efficacy of essential oil from Origanum vulgare L. (oregano; OVEO) and Rosmarinus officinalis L. (rosemary; ROEO) to inactivate sessile cells of Salmonella enterica serovar Enteritidis 86 (SE86) in young and mature biofilms formed on stainless steel. METHODS AND RESULTS: Ultrastructural alterations and damage in different physiological functions caused by OVEO and ROEO in noncultivable sessile cells of SE86 were investigated using scanning electron microscopy and flow cytometry. OVEO (2·5 µl ml-1 ) and ROEO (40 µl ml-1 ) were effective to eradicate young and mature biofilms formed by SE86 sessile cells on stainless steel surfaces; however, the efficacy varied with exposure time. OVEO and ROEO caused alterations in morphology of SE86 sessile cells, inducing the occurrence of bubbles or spots on cell surface. OVEO and ROEO compromised membrane polarization, permeability and efflux activity in noncultivable SE86 sessile cells. These findings show that OVEO and ROEO act by a multitarget mechanism on SE86 membrane functions. CONCLUSIONS: ROEO and OVEO showed efficacy to eradicate SE86 sessile cells in preformed biofilms on stainless steel, displaying a time-dependent effect and multitarget action mode on bacterial cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides for the first time the effects of OVEO and ROEO on morphology and physiological functions of noncultivable sessile cells of S. Enteritidis biofilms preformed on stainless steel surfaces.

The synergistic effects of slightly acidic electrolyzed water and UV-C light on the inactivation of Salmonella enteritidis on contaminated eggshells.

Salmonella enteritidis (S. enteritidis) infection has been recognized as one of the most common bacterial causes of human gastroenteritis worldwide and is closely associated with eggs. Slightly acidic electrolyzed water (SAEW) is an emerging environmentally friendly technology for disinfecting eggshell surfaces to remove dirt and pathogenic microorganisms. However, the efficiency of SAEW could be affected by the presence of manure. UV-based advanced oxidation processes have been studied to improve the microorganism's inactivation effect of disinfection. Therefore, in this study, the synergistic bactericidal efficacy of SAEW and UV-C light (ultraviolet lamp, λ = 254 nm) for inactivation of S. enteritidis on artificially inoculated eggshells with or without manure was evaluated, and the bactericidal efficacy of different combination treatments of SAEW and UV-C light was compared. Without manure interference, complete inactivation (reduction of 6.54 log10 CFU/g) of S. enteritidis on the surface of eggshells was achieved following a 4-min treatment with SAEW+UV at an available chlorine concentration (ACC) of 20 mg/L. In the presence of manure, a 3.02 log reduction was achieved following a 4-min treatment with SAEW+UV at an ACC of 30 mg/L. Simultaneous treatment with SAEW and UV light exhibits higher bactericidal activity for eggshells than other combination process methods with UV and SAEW. The results suggest that the combined treatment of SAEW+UV is a novel method to enhance the microbial safety of eggshells.

The dietary combination of essential oils and organic acids reduces Salmonella enteritidis in challenged chicks.

This study was conducted to determine the effects of essential oils and organic acids (EOA) on Salmonella Enteritidis (S. Enteritidis) challenged chickens. One-day-old specific pathogen-free (SPF) chicks (250) were randomly assigned to 5 groups, with 50 birds in each group. The treatment groups were as follows: 1) basal diet, negative control group (NC); 2) basal diet + S. Enteritidis, positive control group (PC); 3) PC + 4,000 g/t of enrofloxacin (5%), antibiotic group (ENR); 4) PC + 800 g/t of EOA1, thymol-benzoic acid group (TBA); and 5) PC + 800 g/t of EOA2, cinnamylaldehyde-caproic acid group (CCA). At 7 D of age, each bird, except those in NC, was orally gavaged with 0.4 mL of a suspension of 4.4 × 109 cfu S. Enteritidis/mL. Results revealed that ENR reduced bacterial counts in the liver and spleen on days 3, 5, and 7 post-challenge more (P < 0.05) than any other treatments. However, bacterial counts in cecal contents among ENR, TBA, and CCA were similar at 5 and 7 D post-challenge but lower than those of PC. Additionally, the bacterial counts in liver, spleen, and cecum contents in TBA were lower (P < 0.05) than in PC at 3, 5, and 7 D post-challenge; the bacterial counts in spleen contents in TBA were lower (P < 0.05) than in CCA at 7 D post-challenge. Tumor necrosis factor-α contents in TBA and CCA were lower (P < 0.05) than those in PC. Also, the ratio of villus height to crypt depth in the ileum of CCA was higher (P < 0.05) than that of PC and ENR; however, there was no difference in the secretory IgA content of the jejunum among the groups. In conclusion, EOA had a bacteriostatic effect on S. Enteritidis, and the effect of the thymol-benzoic acid complex surpassed that of the cinnamaldehyde-caproic acid complex. Therefore, EOA may act as an effective antibiotic substitute for animals in the prevention and treatment of Salmonella.

Electrospun essential oil-doped chitosan/poly(ε-caprolactone) hybrid nanofibrous mats for antimicrobial food biopackaging exploits.

Development of antimicrobial electrospun nanofibrous mats is one of the most recent trends in the field of food biopackaging. Herein, novel antimicrobial fibrous mats consisting of blend electrospun chitosan (CS)/poly(ε-caprolactone) (PCL) embedded with oregano essential oil (OEO) were fabricated via electrospinning approach and investigated. FE-SEM manifested that as-spun fibers were well-oriented with uniform diameters along their lengths, and AFM displayed the dense nature of the fibrous mats, with surface roughness in the range of 62.9-84.6 nm. The physical interactions and hydrogen bonding between CS/PCL and OEO were verified by FTIR. This novel hybrid mat demonstrated excellent mechanical strength of 14.0 MPa and achieved good water vapor barrier performance. In vitro release assay also revealed that most of plant essential oil loaded fiber mats was remained (56.4-81.7%) after 96 h, demonstrating its durability. Furthermore, the CS/OEO(5%)/PCL nanofiber mats exhibited distinctive antibacterial activity towards Gram-positive (Staphylococcus aureus, Listeria monocytogenes) and Gram-negative (Salmonella enteritidis, Escherichia coli) bacteria. This study gives insights to design new fiber-based antimicrobial nanomaterials of interest in food packaging exploits.

Transcriptional profiling of Salmonella enterica serovar Enteritidis exposed to ethanolic extract of organic cranberry pomace.

Non-typhoidal Salmonella enterica serovars continue to be an important food safety issue worldwide. Cranberry (Vaccinium macrocarpon Ait) fruits possess antimicrobial properties due to their various acids and phenolic compounds; however, the underlying mechanism of actions is poorly understood. We evaluated the effects of cranberry extracts on the growth rate of Salmonella enterica serovars Typhimurium, Enteritidis and Heidelberg and on the transcriptomic profile of Salmonella Enteritidis to gain insight into phenotypic and transcriptional changes induced by cranberry extracts on this pathogen. An ethanolic extract from cranberry pomaces (KCOH) and two of its sub-fractions, anthocyanins (CRFa20) and non-anthocyanin polyphenols (CRFp85), were used. The minimum inhibitory (MICs) and bactericidal (MBCs) concentrations of these fractions against tested pathogens were obtained using the broth micro-dilution method according to the Clinical Laboratory Standard Institute's guidelines. Transcriptional profiles of S. Enteritidis grown in cation-adjusted Mueller-Hinton broth supplemented with or without 2 or 4 mg/ml of KCOH were compared by RNASeq to reveal gene modulations serving as markers for biological activity. The MIC and MBC values of KCOH were 8 and 16 mg/mL, respectively, against all tested S. enterica isolates. The MIC value was 4 mg/mL for both CRFa20 and CRFp85 sub-fractions, and a reduced MBC value was obtained for CRFp85 (4 mg/ml). Treatment of S. Enteritidis with KCOH revealed a concentration-dependent transcriptional signature. Compared to the control, 2 mg/ml of KCOH exposure resulted in 89 differentially expressed genes (DEGs), of which 53 and 36 were downregulated and upregulated, respectively. The upregulated genes included those involved in citrate metabolism, enterobactin synthesis and transport, and virulence. Exposure to 4 mg/ml KCOH led to the modulated expression of 376 genes, of which 233 were downregulated and 143 upregulated, which is 4.2 times more DEGs than from exposure to 2 mg/ml KCOH. The downregulated genes were related to flagellar motility, Salmonella Pathogenicity Island-1 (SPI-1), cell wall/membrane biogenesis, and transcription. Moreover, genes involved in energy production and conversion, carbohydrate transport and metabolism, and coenzyme transport and metabolism were upregulated during exposure to 4 mg/ml KCOH. Overall, 57 genes were differentially expressed (48 downregulated and 9 upregulated) in response to both concentrations. Both concentrations of KCOH downregulated expression of hilA, which is a major SPI-1 transcriptional regulator. This study provides information on the response of Salmonella exposed to cranberry extracts, which could be used in the control of this important foodborne pathogen.