Zein-based nisin-loaded electrospun nanofibers as active packaging mats for control of Listeria monocytogenes on peach.

Zein-based nanofibers (NFs) functionalized with nisin (NS), reinforced with montmorillonite nanoclay (nMMT) were fabricated by uniaxial electrospinning (ES) for the first time to preserve yellow peach. Spinnability/viscosity/conductivity optimizations generated porous (95.09%), bead-free, ultrathin (119 nm) NFs of low hydrophobicity (26.05°). Glutaraldehyde (GTA) crosslinking fostered positive outcomes of tensile strength (1.23 MPa), elongation (5.0%), hydrophobicity (99.46°), surface area (201.38 m2.g-1), pore size (2.88 nm), thermal stability (Tmax = 342 °C), antioxidant/cytotoxic activities in optimized NFs that released NS sustainably according to Korsmeyer-Peppas model indicating a Fickian diffusion mechanism with R2 = 0.9587. The novel NFs inhibited growth of Listeria monocytogenes/aerobic mesophilic populations in peach after 4 days of abusive storage, evincing their robustness in food contact applications. Simultaneously, quality parameters (moisture/texture/browning/total soluble solids/pH) and peach physical appearance were maintained for up to 8 days, endorsing the practical value of zein-based NFs as a non-thermal postharvest intervention for prolonging fruits storage life.

Halochromic smart packaging film based on montmorillonite/polyvinyl alcohol-high amylose starch nanocomposite for monitoring chicken meat freshness.

Polyvinyl alcohol (PVA) was blended with high amylose starch (HAS) at a ratio of 3:1, and reinforced with montmorillonite (MMT K10) at different concentrations (1, 2, 5, and 7 % w/w of polymers) and anthocyanins (ANT) to develop an active and smart packaging film. MMT addition enhanced the film's mechanical, barrier, thermal, and water resistance properties. Incorporating ANT extracted from roselle calyx into the optimal nanocomposite film (MMT/PVA-HAS II) increased the films' antioxidant, pH-response, and antibacterial properties. FTIR, XRD, and SEM confirmed the intermolecular interactions and even distribution of ANT and MMT in the film matrix. Release rate of ANT was dependent on type of simulant, with higher rate in aqueous solutions compared to alcoholic/fatty food simulants, and cytotoxicity evaluation proved safety of films for food packaging applications. Storage experiments confirmed the potential applicability of the novel halochromic smart film as a promising candidate for monitoring chicken spoilage under abusive storage conditions.

Novel pectin-based nanocomposite film for active food packaging applications.

Novel pectin-based films reinforced with crystalline nanocellulose (CNC) and activated with zinc oxide nanoparticles (ZnO NPs) were prepared by solvent-casting method. Film ingredients enhanced UV-blocking, thermal, and antibacterial properties of active films against well-known foodborne pathogens. Optimal active films exhibited higher mechanical, water vapor barrier properties compared to pristine pectin films. SEM confirmed the even distribution of CNC and ZnO NPs in pectin matrix and their interactions were proven using FTIR. Wrapping hard cheese samples artificially contaminated with Staphylococcus aureus and Salmonella enterica with the ternary nanocomposite film at 7 °C for 5 days significantly reduced the total population counts by at least 1.02 log CFU/g. Zn2+ migrating to wrapped cheese samples was below the specific limit (5 mg/kg), confirming their safety for food contact. Overall, ZnO/CNC/pectin nanocomposite films represent promising candidates for active food packaging as safe, eco-friendly alternatives for synthetic packaging materials.

Dual Therapeutic Targeting of Lung Infection and Carcinoma Using Lactoferrin-Based Green Nanomedicine.

In view of the promising applications of nanoparticles in drug delivery, this study highlights the fabrication of new bioactive green protein-polysaccharide nanocomplexes with significant antibacterial and antitumor efficacies. We preformulated the water-insoluble drugs Quercetin (Quer) and Resveratrol (Res) as water-soluble nanocrystals to facilitate their entrapment in the electrostatic lactoferrin-chondroitin (Lf-ChS) nanocomplex. Quer and Res were physically entrapped in the Lf-ChS nanomatrix with high encapsulation efficiencies (EE %) of 85.2 and 90.1% w/w for Quer and Res, respectively. The in vitro synergetic antibacterial effects of the studied compounds against all bacterial strains were confirmed. Res-Quer Lf-ChS NPs revealed an enhanced cytotoxic effect against A549 lung cancer cells. A new model of polymicrobial lung infection was designed, where treatment with Res-Quer Lf-ChS NPs (233.5 ± 6.59 nm) resulted in a marked decline of 3.2 log units in bacterial counts. In the lung tumor model, the potent antitumor efficacy of the developed Res-Quer Lf-ChS NPs was demonstrated by a noticeable decline in both lung weight and the biomarkers compared to the positive control group that did not receive any treatment. In conclusion, the green Res-Quer Lf-ChS NPs possess antibacterial and antitumor attributes for potential lung infection and tumor therapy.

Investigation of the Immunomodulatory effect of Berberis vulgaris on core-pulsed dendritic cell vaccine.

BACKGROUND: Virus-induced dendritic cells (DCs) functional deficiency leads to sub-optimal initiation of adaptive immune responses and consequently chronic infection establishment. The present study reports an advanced hepatitis C virus (HCV) therapeutic vaccine model based on In vivo enrichment of DCs with barberry ethanolic crude extract (BCE) then pulsing them with HCV core protein. METHODS: DCs were enriched by BCE intravenous injection in BALB/c mice. Vaccine efficiency was assessed by flow cytometric analysis of splenocytes of immunized mice, cytokine profiling, cytotoxic T lymphocyte assay, and humoral immune response assessment. RESULTS: There was no significant difference in surface phenotypic characterization of splenocytes from mice immunized with non-BCE-enriched-core-pulsed DCs (iDcs-core) compared to those from mice injected with RPMI-1640 medium. However, splenocytes from mice immunized with BCE-enriched-core-pulsed DCs showed 197 % increase in CD16+ population, 33 % increase in MHCII(+) population, and 43 % decrease in CD3(+) population. In iDCs-core group, 57.9 % greater anti-core cytotoxic T lymphocyte activity, up-regulation in interferon gamma and interleukin (IL) -12 expression, and down-regulation in IL-4 and IL-10 were recorded. Moreover, sustained specific anti-core antibodies were detected only in sera of the same group. CONCLUSIONS: results indicate that BCE-enriched-core-transduced DCs may serve as a new model for immunotherapy of HCV chronic infection.

New Monoclonal Antibodies against a Novel Subtype of Shiga Toxin 1 Produced by Enterobacter cloacae and Their Use in Analysis of Human Serum.

Shiga toxin (Stx) is a major virulence factor of several bacterial pathogens that cause potentially fatal illness, including Escherichia coli and Shigella spp. The continual emergence of new subtypes of Stxs presents challenges for the clinical diagnosis of infections caused by Stx-producing organisms. Here, we report the development of four new monoclonal antibodies (MAbs) against Stx1e, a novel subtype of Stx1 that was produced by an Enterobacter cloacae strain and had limited reactivity with existing anti-Stx1 antibodies. Western blot analysis indicates that these MAbs were Stx1 specific, bound to the A subunit, and had distinct preferences for subtypes of Stx1. Of the four MAbs, Stx1e-2 was capable of partially neutralizing cytotoxicities derived from Stx1e in Vero cells. Enzyme-linked immunosorbent assays assembled with these high-affinity MAbs detected Stx1e at concentrations as low as 4.8 pg/ml in phosphate-buffered saline and 53.6 pg/ml in spiked human serum samples and were also capable of distinguishing Stx1e-producing strains in enriched cultures. These assays may therefore have clinical value in diagnosing Stx1e-producing bacterial infection. Additionally, characteristics of Stx1e, such as the origin of stx1e genes, conditions for toxin expression, receptor binding, and cytotoxicity, were investigated with the new antibodies developed in this study. This information should be useful for further understanding the clinical significance and prevalence of Stx1e-harboring E. cloacae and other organisms. IMPORTANCE Stxs are among the most clinically important virulence factors of Shigella and enterohemorrhagic Escherichia coli. There are many varieties of Stx, and although Stx1a and Stx2a are the most common and widely distributed types of Stx, new variants of Stx are continually emerging. These new variants of Stx can be challenging to detect, since most Stx detection kits are optimized for the detection of Stx1a and Stx2a. Stx1e, recently discovered in an atypical host (Enterobacter cloacae), is undetectable by many Stx assays. To formulate new assays for the detection of Stx1e, we generated four new MAbs that recognize this Stx subtype. Using these antibodies, we generated an assay capable of detecting Stx1e at low picogram-per-milliliter concentrations. This assay is also compatible with a human serum matrix, suggesting that it may have utility for the clinical detection and diagnosis of Stx1e-associated infections.

Phage-mediated Shiga toxin (Stx) horizontal gene transfer and expression in non-Shiga toxigenic Enterobacter and Escherichia coli strains.

Enterobacter cloacae M12X01451 strain recently identified from a clinical specimen produces a new Stx1 subtype (Stx1e) that was not neutralized by existing anti-Stx1 monoclonal antibodies. Acquisition of stx by Ent. cloacae is rare and origin/stability of stx1e in M12X01451 is not known. In this study, we confirmed the ability of Stx1a- and Stx1e-converting phages from an Escherichia coli O157:H7 strain RM8530 and M12X01451 respectively to infect several E. coli and Ent. cloacae strains. stx1e was detected in 97.5% and 72.5% of progenies of strains lysogenized by stx1e phage after 10 (T10) and 20 (T20) subcultures, versus 65% and 17.5% for stx1a gene. Infection of M12X01451 and RM8530 with each other's phages generated double lysogens containing both phages. stx1a was lost after T10, whereas the stx1e was maintained even after T20 in M12X01451 lysogens. In RM8530 lysogens, the acquired stx1e was retained with no mutations, but 20% of stx1a was lost after T20 ELISA and western blot analyses demonstrated that Stx1e was produced in all strains lysogenized by stx1e phage; however, Stx1a was not detected in any lysogenized strain. The study results highlight the potential risks of emerging Stx-producing strains via bacteriophages either in the human gastrointestinal tract or in food production environments, which are matters of great concern and may have serious impacts on human health.

Detection of shiga-toxin producing E. coli (STEC) in leafy greens sold at local retail markets in Alexandria, Egypt.

Leafy green vegetables, a popular and an indispensable ingredient of the daily menus of Egyptians' diets, currently presents a great concern in terms of microbiological hazards. To the best of our knowledge, this is the first report that provides scientific evidence for prevalence of shiga-toxigenic Escherichia coli (STEC) in leafy greens sold at open air local retail markets and superstores in the Egyptian environment. A total of 486 conventional and organic leafy green samples that are eaten raw were collected from different areas in Alexandria, evaluated for total E. coli counts (ECCs), and screened for E. coli O157:H7 using conventional and molecular methods. Recovery of E. coli (≥10(2)CFU/g) from all studied types of leafy greens was indicative of fecal contamination. Total ECCs in conventional samples ranged from 5.47 to 2.56 log CFU/g. Based on their inability to ferment sorbitol on CT-SMAC media, 26 presumptive E. coli O157 isolates were detected in 71.4% (270/378) of the studied conventional samples. From all studied organic samples, only 2 types (organic cabbage and parsley, 16.7%) were contaminated with presumptive E. coli O157. All 28 isolates were further serotyped as E. coli O157 by latex agglutination test, and biochemically confirmed as E. coli. Multiplex PCR assays confirmed the ability of 21.4% (6/28) of the E. coli O157 strains to produce shiga-toxins (Stxs), and their virulence markers were as follows: stx1, 66.6% (4/6); stx2, 50% (3/6); stx1/stx2, 16.7% (1/6); eaeA, 83.3% (5/6); and hlyA, 16.7% (1/6). Only 2 strains recovered from conventional and organic parsley could possibly be classified as E. coli O157:H7 based on the presence of stx-genes (either stx1 or stx2 or both). Results of the present research highlight that high E. coli loads, together with recovery of STEC O157 isolates could pose serious health risks to the produce consumers. This emphasizes the urgent need for health authorities to value and utilize the existing knowledge to identify strategies that reduce microbiological risks due to fecal contamination of agricultural products, and implement control measures at all stages of the food chain to specifically eliminate the presence of STEC O157 on the leafy green category.

Evaluation of the microbiological quality of conventional and organic leafy greens at the time of purchase from retail markets in Alexandria, Egypt.

This is a pioneer study in Egypt that provides some assessment of the microbiological quality of conventional and organic leafy green vegetables that constitute an essential component of the Egyptians' daily diet. A total of 380 samples of unpackaged whole conventional and 84 packaged whole organic leafy greens were collected from retail markets in Alexandria, and analyzed for total aerobic mesophilic count (AMC) and total E. coli count (ECC) using the standard spread plate method. Mean AMC values for organic samples were statistically less (p < 0.05) than those of the corresponding conventional samples. Conventional radish and organic parsley samples had the highest AMC of 7.17 and 7.68 log CFU/g respectively, while conventional green cabbage and organic basil had the lowest AMC of 3.63 and 3.23 log CFU/g respectively. The presence of E. coli in 100% of the studied leafy greens was indicative of potential fecal contamination, in view of open and unhygienic environmental and unsanitary handling conditions, as leafy green items are available for sale by street-vendors. Unsatisfactory AMC and ECC levels encountered in the studied samples, warrant future investigations to determine the potential prevalence of foodborne pathogens, and to identify sources of dominating microorganisms, which could make a contribution to the field of food safety

Selective removal and inactivation of bacteria by nanoparticle composites prepared by surface modification of montmorillonite with quaternary ammonium compounds.

The purpose of the present study was to prepare new nanocomposites with antibacterial activities by surface modification of montmorillonite using quaternary ammonium compounds that are widely applied as disinfectants and antiseptics in food-processing environments. The intercalation of four quaternary ammonium compounds namely benzalkonium chloride, cetylpyridinium chloride monohydrate, hexadecyltrimethylammonium bromide, tetraethylammonium chloride hydrate into montmorillonite layers was confirmed by X-ray diffraction. The antibacterial influences of the modified clay variants against important foodborne pathogens differed based on modifiers quantities, microbial cell densities, and length of contact. Elution experiments through 0.1 g of the studied montmorillonite variants indicated that Staphylococcus aureus, Pseudomonas aeroginosa, and Listeria monocytogenes were the most sensitive strains. 1 g of hexadecyltrimethylammonium bromide intercalated montmorillonites demonstrated maximum inactivation of L. monocytogenes populations, with 4.5 log c.f.u./ml units of reduction. In adsorption experiments, 0.1 g of tetraethylammonium chloride hydrate montmorillonite variants significantly reduced the growth of Escherichia coli O157:H7, L. monocytogenes, and S. aureus populations by 5.77, 6.33, and 7.38 log units respectively. Growth of wide variety of microorganisms was strongly inhibited to undetectable levels (

Behavior of Escherichia coli O157:H7 on damaged leaves of spinach, lettuce, cilantro, and parsley stored at abusive temperatures.

Recent foodborne illness outbreaks associated with the consumption of leafy green produce indicates a need for additional information on the behavior of pathogenic bacteria on these products. Previous research indicates that pathogen growth and survival is enhanced by leaf damage. The objective of this study was to compare the behavior of Escherichia coli O157:H7 on damaged leaves of baby Romaine lettuce, spinach, cilantro, and parsley stored at three abusive temperatures (8, 12, and 15 degrees C). The damaged portions of leaves were inoculated with approximately 10(5) CFU E. coli O157:H7 per leaf. The pathogen grew on damaged spinach leaves held for 3 days at 8 and 12 degrees C (P < 0.05), with the population increasing by 1.18 and 2.08 log CFU per leaf, respectively. E. coli O157:H7 did not grow on damaged Romaine leaves at 8 or 12 degrees C, but growth was observed after 8 h of storage at 15 degrees C, with an increase of less than 1.0 log. Growth of E. coli O157:H7 on Romaine lettuce held at 8 or 12 degrees C was enhanced when inocula were suspended in 0.05% ascorbic acid, indicating the possibility of inhibition by oxidation reactions associated with tissue damage. Damaged cilantro and Italian parsley leaves held at 8 degrees C for 4 days did not support the growth of E. coli O157:H7. Behavior of the pathogen in leaf extracts differed from behavior on the damaged tissue. This study provides evidence that the damaged portion of a leafy green is a distinct growth niche that elicits different microbial responses in the various types of leafy greens.

Evidence for probiotic potential of a capsular-producing Streptococcus thermophilus CHCC 3534 strain.

The purpose of this research was to evaluate the probiotic potential of an capsulated Streptococcus thermophilus CHCC 3534 strain. The strain tolerates 0.4% oxgall (bile) and was sufficiently resistant to pH as low as 2.5 for 3 hours of exposure. The strain demonstrated high adherence to human intestinal mucus, and showed unique resistance to different antibiotics. Crude extracts ofS. thermophilus CHCC 3534 contained a diffusible antimicrobial compound "bacteriocin" with a broad spectrum that inhibited the growth of closely related lactic acid bacteria and a number of food spoilage bacteria including Salmonella typhimurium and Staphylococcus aureus. The bacteriocin was heat stable, resistant to pH, inactivated by proteolytic enzymes, and resistant to a-amylase and lipase. A SDS-PAGE analysis of the partially purified bacteriocin revealed one component with a molecular weight ranging from 14.4 to 18.4 kDa. The strain may have industrial significance and represents an interesting candidate for use in biopreservation, probiotic food formulations and in the control of spoilage caused by food borne pathogens.

Are senior UAE medical and nursing students ready for interprofessional learning? Validating the RIPL scale in a Middle Eastern context.

The success of interprofessional education initiatives depends substantially on attitudes and readiness of health care students to this type of learning. To our knowledge, nothing is known about this subject in a non-western educational context. Using a self-administered extended 29 item version of the Readiness for Interprofessional Learning Scale (RIPLS), this study examined attitudes and readiness for interprofessional education of senior medical (n = 90) and nursing (n = 88) students' from the United Arab Emirates University and Institute of Nursing. The researchers also tested the validity of the RIPLS in this Middle Eastern context. Three main factors, comprising 20 statements emerged from statistical analysis of the data. Sub-scales were labelled "teamwork and collaboration", "professional identity" and "patient-centredness" and each had a strong internal consistency (0.86, 0.80, 0.80 respectively). Both groups of students believed that there are potential academic and clinical benefits of interprofessional learning. Nevertheless, analysis of variance indicated significant differences between the two groups of students in respect of key statements on each factor. The extended RIPLS was validated for use in an undergraduate Middle Eastern community and thus provides programme developers and evaluators with a useful tool to assess medical and nursing students' readiness for interprofessional learning in this context.