Sonoprocessing improves phenolics profile, antioxidant capacity, structure, and product qualities of purple corn pericarp extract.

For the first time, purple corn pericarp (PCP) was converted to polyphenol-rich extract using two-pot ultrasound extraction technique. According to Plackett-Burman design (PBD), the significant extraction factors were ethanol concentration, extraction time, temperature, and ultrasonic amplitude that affected total anthocyanins (TAC), total phenolic content (TPC), and condensed tannins (CT). These parameters were further optimized using the Box-Behnken design (BBD) method for response surface methodology (RSM). The RSM showed a linear curvature for TAC and a quadratic curvature for TPC and CT with a lack of fit > 0.05. Under the optimum conditions (ethanol (50%, v/v), time (21 min), temperature (28 °C), and ultrasonic amplitude (50%)), a maximum TAC, TPC, and CT of 34.99 g cyanidin/kg, 121.26 g GAE/kg, and 260.59 of EE/kg, respectively were obtained with a desirability value 0.952. Comparing UAE to microwave extraction (MAE), it was found that although UAE had a lower extraction yield, TAC, TPC, and CT, the UAE gave a higher individual anthocyanin, flavonoid, phenolic acid profile, and antioxidant activity. The UAE took 21 min, whereas MAE took 30 min for maximum extraction. Regarding product qualities, UAE extract was superior, with a lower total color change (ΔE) and a higher chromaticity. Structural characterization using SEM showed that MAE extract had severe creases and ruptures, whereas UAE extract had less noticeable alterations and was attested by an optical profilometer. This shows that ultrasound, might be used to extract phenolics from PCP as it requires lesser time and improves phenolics, structure, and product qualities.

CAM-21, a novel lytic phage with high specificity towards Escherichia coli O157:H7 in food products.

Escherichia coli O157:H7 is a foodborne pathogen that has become a serious global concern for food safety. Despite the application of different traditional biocontrol methods in the food industry, food borne disease outbreaks linked to this organism remain. Due to their high specificity, lytic bacteriophages are promising antimicrobial agents that could be utilized to control pathogens in foods. In this study, a novel Escherichia phage, CAM-21, was isolated from a dairy farm environment. CAM-21 showed targeted host specificity towards various serotypes of Shiga toxin-producing E. coli, including O157:H7, O26, O103, and O145. Morphological analyses revealed that CAM-21 has a polyhedron capsid and a contractile tail with a diameter of about 92.83 nm, and length of about 129.75 nm, respectively. CAM-21 showed a strong inhibitory effect on the growth of E. coli O157:H7, even at a multiplicity of infection (MOI) of as low as 0.001. Phage adsorption and one-step growth analysis indicated that the target pathogen was rapidly lysed by CAM-21 that exhibited a short latent time (20 min). Electron microscopic and genomic DNA analyses suggested that CAM-21 is a lytic phage, classified as a new species in the Tequatrovirus genus of the Myoviridae Family. Based on whole genome sequencing, CAM-21 has a double-stranded DNA with 166,962 bp, 265 open reading frames and 11 tRNA. The genome of CAM-21 did not encode toxins, virulence factors, antibiotic resistance, lysogeny or allergens. Phylogenetic and genomic comparative analyses suggested that CAM-21 is a T4-like phage species. The growth of E. coli O157:H7 was effectively controlled in milk, ground beef and baby spinach at MOIs of 1000 and 10,000. CAM-21 significantly (P ≤ 0.05) reduced the bacterial counts of the treated foods, ranging from 1.4-2.0 log CFU/mL in milk to 1.3-1.4 log CFU/g in ground beef and baby spinach. These findings suggest that the lytic phage, CAM-21, is a potential candidate for controlling E. coli O157:H7 contamination in foods.

Duplex detection of foodborne pathogens using a SERS optofluidic sensor coupled with immunoassay.

Herein, we developed a surface-enhanced Raman spectroscopy (SERS) optofluidic sensor coupled with immunoprobes to simultaneously separate and detect the foodborne pathogens, Escherichia coli O157:H7, and Salmonella in lettuce and packed salad. The method consists of three steps of (i) enrichment to enhance detection sensitivity, (ii) selective separation and labelling of target bacteria by their specific antibody-bearing SERS-nanotags and (iii) detection of tagged bacterial cells using SERS within a hydrodynamic flow-focusing SERS optofluidic device, where even low counts of bacterial cells were detectable in the very thin-film-like sample stream. SERS-nanotags consisted of different Raman reporter molecules, representing each species, i.e., the detection of Raman reporter confirms the presence of the target pathogen. The anti-E. coli antibody used in this study functions against all strains of E. coli O157:H7 and the anti-Salmonella antibody used in this work acts on a wide range of Salmonella enterica strains. Bacterial counts of 1000, 100, and 10 CFU/ 200 g sample were successfully detected after only 15 min enrichment. Our method showed a very low detection limit value of 10 CFU/ 200 g sample for the bacterial mixture in both lettuce and packed salad, proving the efficiency and high sensitivity of our method to detect multiple pathogens in the food samples. The total analysis time, including sample preparation for simultaneous detection of multiple bacteria, was estimated to be 2 h, which is much less than the time required in conventional methods. Hence, our proposed protocol is considered a promising rapid and efficient approach for pathogen screening of food samples.

Detection of virulence and extended spectrum ß-lactamase genes in Salmonella by multiplex high-resolution melt curve real-time PCR assay.

AIMS: Develop and standardize multiplex high-resolution melt curve (HRM) real-time PCR assays for simultaneous detection of Salmonella virulence and extended spectrum ß-lactamase (ESBL) genes in food. METHODS AND RESULTS: Two sets of multiplex real-time PCR assays targeting six virulence and three ESBL genes with internal amplification control were standardized. The first assay detected hilA, fimH, sipA, blaTEM and blaSHV, and the second detected invA, fimA, stn and blaCMY . The PCR assays were validated with DNA samples from 77 different Salmonella strains. The assay specificity was tested with DNA from 47 non-Salmonella strains. Melt curve analyses showed distinct, well-separated melting peaks of each target gene detected by their respective melting temperatures (Tm ). Different food samples were spiked with 10, 102 and 103  CFU/ml of Salmonella. The optimized assays were able to detect all target genes in concentrations of as low as 10 CFU/ml in 25 g foods within 10 h of enrichment. CONCLUSIONS: Multiplex HRM real-time PCR assays can be used as rapid detection methods for detecting Salmonella in foods. SIGNIFICANCE AND IMPACT OF STUDY: The assays developed in this study will allow for accurate detection of virulence and ESBL genes in Salmonella that are present in low concentrations in food samples.

Hazard Analysis and Risk-Based Preventive Controls (HARPC): Current Food Safety and Quality Standards for Complementary Foods.

Food safety is imperative, especially for infants and young children because of their underdeveloped immune systems. This requires adequate nutritious food with appropriate amounts of macro- and micronutrients. Currently, a well-established system for infant food is enforced by the regulatory bodies, but no clear system exists for complementary food, which is consumed by children from the age of 6 month to 24 months. As the child grows beyond 6 months, the need for nutrients increases, and if the nutritional needs are not fulfilled, it can lead to health problems, such as stunted growth, weak immune system, and cardiovascular diseases. Hence, it is important to have regulatory bodies monitoring complementary food in a similar capacity as is required for infant formula. The objective of this review is to provide an overview of the existing regulatory bodies, such as the Codex Alimentarius, International Standard Organization (ISO), Food and Drug Administration (FDA), etc., and their regulations specifically for infant formula that can be adopted for complementary foods. This study focuses on the development of a hazard analysis and risk-based preventive controls (HARPC)-based food safety plan to ensure safe food processing and prevent any possible outbreaks.

Zinc Oxide and Silver Nanoparticle Effects on Intestinal Bacteria.

The application of nanoparticles (NPs) for food safety is increasingly being explored. Zinc oxide (ZnO) and silver (Ag) NPs are inorganic chemicals with antimicrobial and bioactive characteristics and have been widely used in the food industry. However, not much is known about the behavior of these NPs upon ingestion and whether they inhibit natural gut microflora. The objective of this study was to investigate the effects of ZnO and Ag NPs on the intestinal bacteria, namely Escherichia coli, Lactobacillus acidophilus, and Bifidobacterium animalis. Cells were inoculated into tryptic soy broth or Lactobacilli MRS broth containing 1% of NP-free solution, 0, 12, 16, 20 mM of ZnO NPs or 0, 1.8, 2.7, 4.6 mM Ag NPs, and incubated at 37 °C for 24 h. The presence and characterization of the NPs on bacterial cells were investigated by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDS). Membrane leakage and cell viability were assessed using a UV-visible spectrophotometer and confocal electron microscope, respectively. Numbers of treated cells were within 1 log CFU/mL less than those of the controls for up to 12 h of incubation. Cellular morphological changes were observed, but many cells remained in normal shapes. Only a small amount of internal cellular contents was leaked due to the NP treatments, and more live than dead cells were observed after exposure to the NPs. Based on these results, we conclude that ZnO and Ag NPs have mild inhibitory effects on intestinal bacteria.

Preparation of cellulose nanofibril/titanium dioxide nanoparticle nanocomposites as fillers for PVA-based packaging and investigation into their intestinal toxicity.

The aims of this work were to synthesize cellulose nanofibril/titanium dioxide nanoparticle (CNF/TiO2NP) nanocomposites, evaluate the use of CNF/TiO2NP nanocomposites in PVA-based films, and investigate the intestinal toxicity of the nanocomposites. Via a mixing method, CNF/TiO2NP nanocomposites were synthesized. The addition of the nanocomposites significantly enhanced the tensile strength, Young's modulus, and light barrier capacity of PVA films. Moreover, a high concentration of CNF/TiO2NP nanocomposites (10 mg/mL) had no appreciable effect on the growth of Escherichia coli P-24, Lactobacillus acidophilus ADH, and Bifidobacterium animalis Bif-6 cells. The nanocomposites did not exhibit significant toxicity to cancerous and normal colon cells even when their concentrations increased to a high level of 1000 µg/mL. The results indicate that CNF/TiO2NP nanocomposites can potentially be used as functional fillers for a PVA-based packaging system.

Antimicrobial effect and toxicity of cellulose nanofibril/silver nanoparticle nanocomposites prepared by an ultraviolet irradiation method.

The objective of this study was to synthesize a novel antimicrobial cellulose nanofibril/silver nanoparticle (CNF/AgNP) nanocomposite by an ultraviolet (UV) irradiation method and evaluate the toxicity of the nanocomposite to human colon cells. AgNPs coated on CNFs have an average size of ˜28 nm and exhibited a surface plasma resonance absorption peak at 402 nm. Coating AgNPs on CNFs interfered with the formation of intra-chain and inter-chain hydrogen bonds of cellulose. Moreover, the CNF/AgNP nanocomposite exhibited significant antimicrobial activities against two important food-borne pathogens, including Escherichia coli O157:H7 and Staphylococcus aureus. No apparent toxicity of the CNF/AgNP nanocomposite to Caco-2 and FHC human colon cells was observed, except when a high content of (≥500 µg/m L) of the nanocomposite was used for which a significant decrease of cell viability was observed. The nanocomposite's toxicity was related to the content, size, and surface charge of UV-synthesized AgNPs on CNFs. These results indicate that the antimicrobial CNF/AgNP nanocomposite prepared by UV irradiation method can be potentially used as an active filler applied in food packaging materials.

Cellulose nanofibril/silver nanoparticle composite as an active food packaging system and its toxicity to human colon cells.

Cellulose nanofibrils (CNFs) and silver nanoparticles (AgNPs) are nano-sized fillers widely used for enhancing the active functions of food packaging materials. However, nanomaterials may pose unexpected toxic effects on humans. Thus, the objective of this work was to develop a novel CNF/AgNP composite and investigate its properties and cytotoxicity. The CNF/AgNP composite was prepared via a reduction method using NaBH4. The AgNPs embedded in the composite showed an average size of 10.72 ±â€¯4.96 nm and a surface plasmon resonance (SPR) absorption peak at 397 nm. AgNPs were distributed individually in the composite after synthesis, but aggregated during film preparation. The formation of AgNPs disrupted hydrogen bonds between the hydroxyl groups of cellulose, weakening the hydrogen-bond intensity, as shown by FTIR. Silver ions were efficiently released from the composite film in the first 24 h. The CNF/AgNP composite exhibited inhibitory effects on Escherichia coli O157:H7 and Listeria monocytogenes. The composite (50-1000 µg/mL) did not significantly reduce the viability of Caco-2 and FHC colon cells, although the uptake of AgNPs through an endosomal mechanism was observed. These results suggest that the as-prepared CNF/AgNP composite could potentially be used as an antimicrobial material in active food packaging systems.

Antimicrobial Coatings for Food Contact Surfaces: Legal Framework, Mechanical Properties, and Potential Applications.

Food contact surfaces (FCS) in food processing facilities may become contaminated with a number of unwanted microorganisms, such as Listeria monocytogenes, Escherichia coli O157:H7, and Staphylococcus aureus. To reduce contamination and the spread of disease, these surfaces may be treated with sanitizers or have active antimicrobial components adhered to them. Although significant efforts have been devoted to the development of coatings that improve the antimicrobial effectiveness of FCS, other important coating considerations, such as hardness, adhesion to a substrate, and migration of the antimicrobial substance into the food matrix, have largely been disregarded to the detriment of their translation into practical application. To address this gap, this review examines the mechanical properties of antimicrobial coatings (AMC) applied to FCS and their interplay with their antimicrobial properties within the framework of relevant regulatory constraints that would apply if these were used in real-world applications. This review also explores the various assessment techniques for examining these properties, the effects of the deposition methods on coating properties, and the potential applications of such coatings for FCS. Overall, this review attempts to provide a holistic perspective. Evaluation of the current literature urges a compromise between antimicrobial effectiveness and mechanical stability in order to adhere to various regulatory frameworks as the next step toward improving the industrial feasibility of AMC for FCS applications.

Single Locked Nucleic Acid-enhanced nanopore genetic discrimination of pathogenic serotypes and cancer driver mutations.

Rapid and accurate detection of single-nucleotide polymorphism (SNP) in pathogenic mutants is crucial for broad fields from food safety monitoring to disease diagnostics and prognosis. Here, we developed a nanopore single-molecule sensor, coupled with the locked nucleic acid (LNA) technique, to accurately discriminate SNPs for detection of Shiga toxin producing Escherichia coli (STEC) O157:H7 pathogen serotype, and cancer-derived driver mutations EGFR L858R and KRAS G12D. This sensitive method, with a simplified, low cost, easy-to-operate LNA design, can be applied in food science and medical detection that need rapid and accurate determination of genetic variations.

Detection of viable Escherichia coli in environmental water using combined propidium monoazide staining and quantitative PCR.

The objectives of this study were to specifically detect viable Escherichia coli in environmental waters by targeting the ycjM gene in a propidium monoazide (PMA)-qPCR assay. PMA is a viability dye that can inhibit the amplification of DNA from dead cells, thus allowing for the detection and quantification of only viable cells. The ycjM primers were used to target E. coli that directly originated from the feces of warm blooded animals, and avoid false positive detection caused by "naturalized" E. coli that can exist in the environment. In this study, tap water and environmental waters were inoculated with E. coli isolated from animal feces. Following cell collection, samples were treated with PMA, followed by DNA isolation and qPCR detection. For pure cultures, 5 µM PMA with a 10-min light exposure was efficient at inhibiting the amplification of DNA from 105 CFU/mL dead E. coli cells, with a detection limit of 102 CFU/100 mL viable cells. For tap and environmental waters collected in the winter, a 10 µM PMA was required and as low as 103 CFU/100 mL viable cells could be detected in the presence of 105 CFU/100 mL dead cells. For water samples collected during the summer, 102 CFU/10 mL viable cells could be detected in the presence of 104 CFU/10 mL dead cells, after a 20 µM PMA treatment. No significant differences were found among the PMA-qPCR assay and two other standard culture-based methods for detection of viable E. coli in environmental water. In conclusion, with proper pretreatment of environmental water samples, this PMA-qPCR assay that targets the ycjM gene could quantify viable E. coli cells that directly come from the feces of warm-blooded animals, and therefore effectively and accurately indicate the quality of environmental water.

Green synthesis of silver nanoparticles using turmeric extracts and investigation of their antibacterial activities.

This study aimed to develop an environmentally friendly and cost-effective approach to synthesize green silver nanoparticles (AgNPs) from silver precursors. Green synthesis of AgNPs was accomplished using the aqueous extract of turmeric powder, in which plant biomaterials were used as a reducing as well as a capping agent. After 24 h of reaction, the yellow color of the extract was changed to dark brown-reddish due to the reduction of silver ions to AgNPs. AgNPs were characterized using UV-vis spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM), and energy dispersive X-ray spectroscopy (EDS). The maximum absorbance of the UV-vis spectra was at 432 nm. TEM analysis reveals that the shape of most of the biosynthesized AgNPs was spherical forms and the average particle size was 18 ± 0.5 nm. EDS analysis exhibits strong signals of silver element. In addition, green synthesized AgNPs show high and efficient antimicrobial activities against two food-borne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes). TEM and scanning electron microscopic images reveal that there were significant shrinkage and damage of bacterial cell wall, and leakage or loss of bacterial intracellular contents. A significant reduction (P ≤ 0.05) of bacterial counts just after 4 h of exposure was observed. These results indicate that green synthesized AgNPs can be utilized as an antimicrobial means to inhibit the growth of pathogenic bacteria for applications in agricultural and food industries.

Rapid determination of thiabendazole in juice by SERS coupled with novel gold nanosubstrates.

The aim of this study was to develop surface-enhanced Raman spectroscopy (SERS) methods in combination with novel gold nanomaterial-based substrates for rapid measurement and quantification of pesticides extracted from lemon, carrot, and mango juice. Facile synthesis of a sensitive and robust SERS substrate was achieved by assembling gold nanorods (AuNRs) into vertically aligned arrays on silicon slides. The nanorod arrays were orderly aligned and can induce vigorous electromagnetic field for SERS measurement. The synthesized SERS substrate was utilized for detection and quantification of thiabendazole in juice samples using partial least squares analysis with R values of 0.99, 0.98, and 0.99 for lemon, carrot, and mango juice, respectively. The detection limits of thiabendazole were 149, 216, and 179 µg/L in lemon, carrot, and mango juice, respectively. These results demonstrate that SERS combined with AuNR substrates is a quick, convenient, and highly sensitive technique for detection of thiabendazole residues in fruits juice.

Single Locked Nucleic Acid-Enhanced Nanopore Genetic Discrimination of Pathogenic Serotypes and Cancer Driver Mutations.

Accurate and rapid detection of single-nucleotide polymorphism (SNP) in pathogenic mutants is crucial for many fields such as food safety regulation and disease diagnostics. Current detection methods involve laborious sample preparations and expensive characterizations. Here, we investigated a single locked nucleic acid (LNA) approach, facilitated by a nanopore single-molecule sensor, to accurately determine SNPs for detection of Shiga toxin producing Escherichia coli (STEC) serotype O157:H7, and cancer-derived EGFR L858R and KRAS G12D driver mutations. Current LNA applications that require incorporation and optimization of multiple LNA nucleotides. But we found that in the nanopore system, a single LNA introduced in the probe is sufficient to enhance the SNP discrimination capability by over 10-fold, allowing accurate detection of the pathogenic mutant DNA mixed in a large amount of the wild-type DNA. Importantly, the molecular mechanistic study suggests that such a significant improvement is due to the effect of the single-LNA that both stabilizes the fully matched base-pair and destabilizes the mismatched base-pair. This sensitive method, with a simplified, low cost, easy-to-operate LNA design, could be generalized for various applications that need rapid and accurate identification of single-nucleotide variations.

Effect and mechanism of cellulose nanofibrils on the active functions of biopolymer-based nanocomposite films.

Cellulose nanofibrils (CNFs) are superfine cellulose fibrils with a nanoscale diameter and have gained increasing attention due to their great potential in the food industry. However, the applications of CNFs in active food packaging are still limited. The objectives of this study were to develop biopolymer-based edible nanocomposite films using CNFs, corn starch, and chitosan, and to investigate the effect and mechanisms of CNFs on the active functions and properties of the nanocomposite films. Important functional properties of the films were measured and the films were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and Zetasizer. The results demonstrate that CNFs increased the rigidity of the films due to more hydrogen bonds being induced by CNFs (≥60%). Incorporating a high content of CNFs (≥60%) in the film resulted in enhanced filling effect on the structure of the biopolymer films, which significantly improved the light barrier, oxygen barrier and water vapor barrier capacities. As CNF content increased to 100%, the film opacity increased by 59%, while the peroxide value of corn oil protected with edible films was reduced by 23%. Furthermore, the antimicrobial properties of the edible films with 80% and 100% CNFs were increased by up to 2logCFU/g on day 8 in a beef model, due to more positive charges in the films and improved blocking effects on oxygen. These results demonstrate that CNFs can effectively enhance the antimicrobial effect and barrier properties of biopolymer-based nanocomposite films and have great potential in applications of active packaging for food products.

Biocontrol of the internalization of Salmonella enterica and Enterohaemorrhagic Escherichia coli in mung bean sprouts with an endophytic Bacillus subtilis.

Internalization of Salmonella enterica and enterohaemorrhagic Escherichia coli (EHEC) in seed sprouts poses a health risk to consumers, and the conventional sanitization methods are not always effective to reduce this risk. This study initiated a biocontrol approach to limit the internalization using endophytic Bacillus subtilis strains, which were isolated from the inner tissue of mung bean seeds or lettuce stems. By using the deferred agar method, 12 strains of B. subtilis out of 94 putative Bacillus isolates displayed inhibitory activity against at least one of the pathogenic indicators, S. enterica Typhimurium ATCC 14028 and E. coli O157:H7 505B. Two B. subtilis isolates (LCA1 and M24) showed a broad inhibitory spectrum against multiple strains of S. enterica and EHEC, Staphylococcus aureus sp., Klebsiella pneumoniae ATCC 700603, and Listeria monocytogenes Scott A, while the laboratory B. subtilis strain 168 was only moderately inhibitory against L. monocytogenes. To facilitate the tracking of the three B. subtilis strains (LCA1, M24, and 168) in the mung bean sprouts, the three strains were genetically engineered to carry the chloramphenicol acetyltransferase (cat), generating the strains LCA1-cat, M24-cat, and 168-cat, respectively. Data of the study using the cat-tagged strains demonstrated that both the two vegetable-associated and the laboratory B. subtilis strains could internalize in mung bean sprouts during the sprouting, but the latter displayed about 1.2 lg CFU/g of seeds lower in internalization. Overall, the presence of the three B. subtilis strains could significantly reduce the internalization of S. enterica or EHEC cocktail in mung bean sprouts during the sprouting. Among them, LCA1 showed the greatest inhibition against the EHEC cocktails with a reduction of about 2.0lg CFU/g of seeds by the end of sprouting (day 5), while 168 had the smallest reduction at about 0.6lg CFU/g of seeds. In addition, the three strains demonstrated a similar inhibition against the S. enterica cocktails by a reduction of about 1.1-1.4lg CFU/g of seeds by day 5. Results of this study suggest that the source (native vs. alien) of B. subtilis isolates may not affect the efficacy of the inhibition, but it might be affected by the production of antimicrobial substance and/or nutrition/space competition. The results also indicate that strain LCA1 may be useful as a biocontrol agent to reduce Salmonella and EHEC contamination in seed sprouts.

Cellulose nanofibers coated with silver nanoparticles as a SERS platform for detection of pesticides in apples.

A nanocomposite based on cellulose nanofibers (CNFs) coated with silver nanoparticles (AgNPs) was developed in this study as a flexible and effective substrate for use in surface-enhanced Raman spectroscopy (SERS) analysis. An effective Raman indicator molecule, 4-aminothiophenol (pATP), was used to characterize AgNPs impregnated on CNFs. The CNF-AgNP films were used in SERS analysis to detect thiabendazole (TBZ) pesticides in apples. The influence of pH on the SERS spectra of TBZ was investigated because TBZ is a neutral molecule that has a low affinity for AgNPs. The pH of TBZ solution was decreased to below the TBZ's pKa, thus enabling the electrostatic attraction between TBZ and AgNPs. CNFs can prevent the uncontrolled aggregation of AgNPs in low pH environment and serve as an effective AgNP/nanocellulose platform for SERS analysis. Results of this study demonstrate that CNF-AgNP nanocomposites can be used to rapidly detect TBZ pesticides in various food products.

Engineered Nanoparticles as Potential Food Contaminants and Their Toxicity to Caco-2 Cells.

Engineered nanoparticles (ENPs), such as metallic or metallic oxide nanoparticles (NPs), have gained much attention in recent years. Increasing use of ENPs in various areas may lead to the release of ENPs into the environment and cause the contamination of agricultural and food products by ENPs. In this study, we selected two important ENPs (zinc oxide [ZnO] and silver [Ag] NPs) as potential food contaminants and investigated their toxicity via an in vitro model using Caco-2 cells. The physical properties of ENPs and their effects on Caco-2 cells were characterized by electron microscopy and energy dispersive X-ray spectroscopic (EDS) techniques. Results demonstrate that a significant inhibition of cell viability was observed after a 24-h of exposure of Caco-2 cells to 3-, 6-, and 12-mM ZnO NPs or 0.5-, 1.5-, and 3-mM Ag NPs. The noticeable changes of cells include the alteration in cell shape, abnormal nuclear structure, membrane blebbing, and cytoplasmic deterioration. The toxicity of ZnO NPs, but not that of Ag NPs after exposure to simulated gastric fluid, significantly decreased. Scanning transmission electron microscopy shows that ZnO and Ag NPs penetrated the membrane of Caco-2 cells. EDS results also confirm the presence of NPs in the cytoplasm of the cells. This study demonstrates that ZnO and Ag NPs have cytotoxic effects and can inhibit the growth of Caco-2 cells.