Determination of furfurals in baby food samples after extraction by a novel functionalized magnetic porous carbon.

Herein, a novel polypyrrole-polyaniline functionalized magnetic porous carbon (MPC@PPy-PANI) composite material was fabricated and utilized for the separation/extraction of furfurals from baby food and dry milk samples. In this way, magnetite@silica nanoparticles were first synthesized, and then a magnetic metal-organic framework (MMIL-101(Fe)) was prepared. After that, the MMIL-101(Fe) was pyrolyzed in a neutral atmosphere to obtain MPC. Ultimately, the MPC was functionalized with a co-polymer of aniline-pyrrole via oxidation polymerization. The synthesis of MPC@PPy-PANI was confirmed with FT-IR spectroscopy, SEM, TEM, VSM, and XRD techniques. Furfural and hydroxymethyl furfural were selected as the model analytes, which were separated/quantified on an HPLC-UV instrument. The LODs, LOQs, and linear dynamic ranges (LDRs) were in the range of 0.3-0.7 µg kg-1, 1.0-2.5 µg kg-1, and 1.0-600 µg kg-1, respectively. Repeatability of the method was studied as an RSD parameter, and was located in the range of 5.5-6.8% (within-day, n = 5) and 8.2-9.4% (between-day, n = 3 days). The applicability of the proposed method was established by analyzing several baby food and dry milk samples. The relative recovery (RR%) and repeatability were located in the range of 86-111% and 3.3-10.1%, respectively, showing excellent accuracy and precision of the method.

Development of high-resolution melting (HRM) assay to differentiate the species of Shigella isolates from stool and food samples.

Shigella species, a group of intracellular foodborne pathogens, are the main causes of bacillary dysentery and shigellosis in humans worldwide. It is essential to determine the species of Shigella in outbreaks and food safety surveillance systems. The available immunological and molecular methods for identifying Shigella species are relatively complicated, expensive and time-consuming. High resolution melting (HRM) assay is a rapid, cost-effective, and easy to perform PCR-based method that has recently been used for the differentiation of bacterial species. In this study, we designed and developed a PCR-HRM assay targeting rrsA gene to distinguish four species of 49 Shigella isolates from clinical and food samples and evaluated the sensitivity and specificity of the assay. The assay demonstrated a good analytical sensitivity with 0.01-0.1 ng of input DNA template and an analytical specificity of 100% to differentiate the Shigella species. The PCR-HRM assay also was able to identify the species of all 49 Shigella isolates from clinical and food samples correctly. Consequently, this rapid and user-friendly method demonstrated good sensitivity and specificity to differentiate species of the Shigella isolates from naturally contaminated samples and has the potential to be implemented in public health and food safety surveillance systems.

RAPD and ERIC-PCR coupled with HRM for species identification of non-dysenteriae Shigella species; as a potential alternative method.

OBJECTIVE: Species identification of Shigella isolates are so prominent for epidemiological studies and infection prevention strategies. We developed and evaluated RAPD and ERIC-PCR coupled with HRM for differentiation of non-dysenteriae Shigella species as potential alternative methods. After isolation of eighteen Shigella strains from faecal specimens collected from children under 2 years of age with diarrhea (n = 143), the species of the isolates were identified by slide agglutination assay. Also, species were identified using developed RAPD-PCR-HRM and ERIC-PCR-HRM techniques. Differentiation of the data sets was measured by principal component analysis as a dimension reduction method. Then, sensitivity and specificity of the methods were evaluated. RESULTS: We found RAPD-PCR-HRM method with high sensitivity and specificity (100 and 85% respectively) to identify non-dysenteriae Shigella species in clinical specimens. However, sensitivity and specificity of ERIC-PCR-HRM were evaluated 33 and 46% respectively and significantly lower than that of RAPD-PCR-HRM assay. Regardless of inherent poor reproducibility of DNA fingerprinting-based methods, RAPD-PCR-HRM assay can be considered as a potential alternative method to identify non-dysenteriae species of Shigella in clinical specimens. As we observed in the current study, HRM technique is more rapid, inexpensive, and sensitive than gel electrophoresis method to characterize PCR amplicons.

Synthesis and characterization of a novel magnetic porous carbon coated with poly(p-phenylenediamine) and its application for furfural preconcentration and determination in baby food and dry milk powder samples.

The aim of the current research is to develop a MSPE method for the determination of furfural in baby food and dry milk samples. In this regard, a novel magnetic porous carbon composite coated with poly(p-phenylenediamine) was fabricated, characterized, and then applied to the preconcentration/extraction of furfurals from baby food and dry milk powder samples. Initially, magnetic nanoparticles (Fe3O4) were synthesized, and then coated with a metal-organic framework layer named MIL-101(Fe). Afterward, the magnetic MIL-101(Fe) was subjected to calcination under a nitrogen atmosphere and magnetic porous carbon was achieved. Finally, a layer of poly(p-phenylenediamine) was coated on the magnetic porous carbon. The structure of the nanocomposite was investigated with various methods, including FT-IR spectroscopy, electron microcopies (SEM and TEM), VSM, and XRD. The fabricated nanocomposite was applied in magnetic solid-phase extraction of furfural and hydroxymethyl furfural and their determination with liquid chromatography. The effect of experimental variables was explored by using an experimental design approach. The LODs and linear range for the target furfurals were 1.0-2.0 µg kg-1 and 3.0-500 µg kg-1, respectively. The method's repeatability was explored using RSD values and was found to be in the range of 5.2-6.4% (one-day, n = 5) and 9.1-10.8% (day to day, n = 3). Eventually, this new method was employed for the extraction/quantification of target compounds in baby food and dry milk powder samples.

Incorporation of nanoencapsulated garlic essential oil into edible films: A novel approach for extending shelf life of vacuum-packed sausages.

The efficacy of chitosan (CH) and whey protein (WP) films impregnated with garlic essential oil (GEO, 2% v/v) or nanoencapsulated GEO (NGEO, 2% v/v) to extend the shelf life of refrigerated vacuum-packed sausages were assessed and compared during 50 days. The primary evaluation of GEO and NGEO showed that GEO had a considerable amount of active compounds diallyl sulfide derivatives (~67%) and the mean size and zeta potential of NGEO were 101 nm and -7.27 mV, respectively. Based on the microbiological and lipid stability analysis of the sausages, all active films retarded lipid oxidation and the growth of main spoilage bacterial groups compared to the control, and CH film containing NGEO exhibited the best result with the peroxide value, thiobarbituric acid reactive substances and aerobic plate count of 0.37 (meq/kg lipid), 0.47 (mg malondialdehyde/kg) and 3.69 (log CFU/g), respectively, on day 50. The nanoencapsulation of GEO made no significant differences in the sensory properties comparing to free-GEO samples (P < .05).

Viability of microencapsulated and non-microencapsulated Lactobacilli in a commercial beverage.

The survival rate of free and encapsulated L. acidophilus and L. rhamnosus into Doogh beverage and simulated gastrointestinal conditions during 42-day were studied. Microencapsulation considerably protected both L. acidophilus and L. rhamnosus in Doogh beverage storage and in gastrointestinal conditions. Microencapsulation provided better protection to L. acidophilus than to L. rhamnosus during Doogh storage. In beverages containing the free form of bacteria, pH and acidity changes were greater than those of microencapsulated and control groups. More activity of the free probiotic bacteria (during a 42-day period especially after 21-day) produced more acid and metabolites inside the product, thereby reducing the organoleptic properties scores, However, acidity, pH and organoleptic characteristics of Doogh containing microencapsulated bacteria did not change considerably. In conclusion, this study suggests that the encapsulation and double coating of L. acidophilus and L. rhamnosus can increase the viability of them in Doogh beverage and in simulated GI conditions.

The effect of Zataria multiflora Boiss Essential oil on the growth and citrinin production of Penicillium citrinum in culture media and cheese.

The effect of Zataria multiflora Boiss Essential oil (EO) on the growth, spore production, and citrinin production of Penicellium citrinum PTCC 5304 in the culture media as well as Iranian ultra-filtered white cheese in brine was investigated. Radial growth and spore production on the potato dextrose agar (PDA) were effectively inhibited by EO in a dose-dependent manner. At 200 ppm, the radial growth and sporulation declined by 92% and 100%, respectively. The growth was completely prevented at 400 ppm of EO on PDA and the minimum fungicidal concentration (MFC) of the oil was estimated at 400 ppm. Furthermore, the Zataria multiflora also significantly suppressed the mycelial growth and citrinin production in broth medium at all investigated concentrations (P < 0.05). At 150 ppm of EO, the citrinin accumulation and mycelial growth reduced by 88.6% and 89.6%, respectively. The EO was tested at all concentrations and the findings show an inhibitory effect of P. citrinum against the radial fungal growth and citrinin production in cheese. However, no concentration of EO could completely inhibit the growth and production of citrinin in cheese. We therefore concluded that Zataria multiflora has the potential to substitute the antifungal chemicals as a natural inhibitor to control the growth of molds in foods such as cheese.

Phenotypic and genotypic characterization of antibiotic resistance of methicillin-resistant Staphylococcus aureus isolated from hospital food.

BACKGROUND: Pathogenic biotypes of the Methicillin-resistant Staphylococcus aureus (MRSA) strains are considered to be one of the major cause of food-borne diseases in hospitals. The present investigation was done to study the pattern of antibiotic resistance and prevalence of antibiotic resistance genes of different biotypes of the MRSA strains isolated from various types of hospital food samples. METHODS: Four-hundred and eighty-five raw and cooked hospital food samples were cultured and MRSA strains were identified using the oxacillin and cefoxitin disk diffusion tests and mecA-based PCR amplification. Isolated strains were subjected to biotyping and their antibiotic resistance patterns were analyzed using the disk diffusion and PCR methods. RESULTS: Prevalence of S. aureus and MRSA were 9.69 and 7.62%, respectively. Meat and chicken barbecues had the highest prevalence of MRSA. Prevalence of bovine, ovine, poultry and human-based biotypes in the MRSA strains were 8.10, 8.10, 32.43 and 48.64%, respectively. All of the MRSA strains recovered from soup, salad and rice samples were related to human-based biotypes. MRSA strains harbored the highest prevalence of resistance against penicillin (100%), ceftaroline (100%), tetracycline (100%), erythromycin (89.18%) and trimethoprim-sulfamethoxazole (83.78%). TetK (72.97%), ermA (72.97%), msrA (64.86%) and aacA-D (62.16%) were the most commonly detected antibiotic resistance genes. CONCLUSIONS: Pattern of antibiotic resistance and also distribution of antibiotic resistance genes were related to the biotype of MRSA strains. Presence of multi-drug resistance and also simultaneous presence of several antibiotic resistance genes in some MRSA isolates showed an important public health issue Further researches are required to found additional epidemiological aspects of the MRSA strains in hospital food samples.

Antimicrobial evaluation of novel poly-lactic acid based nanocomposites incorporated with bioactive compounds in-vitro and in refrigerated vacuum-packed cooked sausages.

Biodegradability and antimicrobial activity of food packaging materials are among the most attractive parameters in modern food industries. In order to develop biodegradable poly-lactic acid (PLA) film to antibacterial nanocomposites, different concentration of Zataria multiflora Bioss. essential oil (ZME), propolis ethanolic extract (PEE) and cellulose nanofiber (CNF) were incorporated to the polymer by solvent casting method. The resulting films were characterized by mechanical and physical tests and their antimicrobial application was evaluated in-vitro against four common foodborne pathogens and in vacuum-packed cooked sausages during refrigerated storage. Mechanical examination revealed that addition of ZME and PEE made films more flexible and incorporation of CNF improved almost all mechanical parameters tested. Moreover, according to physical analysis, incorporation of 0.5% v/v ZME to the composite primary solutions improved water vapor permeability of the resulting films. Almost all of the active films were effective against the tested bacteria except for PLA/PEE films, and maximum antibacterial effects recorded for the films containing both ZME and PEE. Based on the microbiological and sensory evaluation of the sausages, all of the PLA/1%ZME/PEE composites increased the shelf life to >40days. The results indicate that incorporation of natural antimicrobial substances such as ZME and PEE to packaging material could be an interesting approach in development of active packaging material without significant negative effect on polymer technical properties.

Effect of PLA films containing propolis ethanolic extract, cellulose nanoparticle and Ziziphora clinopodioides essential oil on chemical, microbial and sensory properties of minced beef.

This study was conducted to examine the effects of polylactic acid (PLA) film containing propolis ethanolic extract (PE), cellulose nanoparticle (CN) and Ziziphora clinopodioides essential oil (ZEO) on chemical, microbial and sensory properties of minced beef during storage at refrigerated temperature for 11days. The initial total volatile base nitrogen (TVB-N) was 8.2mg/100g and after 7days reached to 29.1mg/100g in control, while it was lower than 25mg/100g for treated samples. At the end of storage time in control samples peroxide value (PV) reached to 2.01meqperoxide/1000g lipid, while the values for the treated samples remained lower than 2meqperoxide/1000g lipid. Final microbial population decreased approximately 1-3logCFU/g in treated samples compared to control (P<0.05). Films containing 2% ZEO alone and in combination with different concentrations of PE and CN extended the shelf life of minced beef during storage in refrigerated condition for at least 11days without any unfavorable organoleptic properties.

Effects of Zataria multiflora Boiss. essential oil on growth and gene expression of enterotoxins A, C and E in Staphylococcus aureus ATCC 29213.

Staphylococcal food poisoning results from the consumption of food in which enterotoxigenic staphylococci have grown and produced toxins. The present study was conducted with three principal aims: i) to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Zataria multiflora Boiss. essential oil (EO) against Staphylococcus aureus ATCC 29213, ii) to evaluate the effect of subinhibitory concentrations (subMIC) of EO on the growth of bacteria over 72 h (at 25 and 35 °C), and iii) to investigate the expression of genes involved in the production of staphylococcal enterotoxins SEA, SEC and SEE over 72 h at 35 °C. The MIC and MBC of Z. multiflora Boiss. EO were 0.03 and 0.04%, respectively. Colony counting at 24, 48 and 72 h of 3 day cultures grown in the presence of 75%MIC of the EO showed that the growth rate was reduced 2.16, 2.78 and 2.91 log 10 cfu/ml at 25 °C, and 1.34, 2.35 and 2.57 log 10 cfu/ml at 35 °C, respectively, compared to control cultures. SubMIC levels of EO also significantly decreased the expression of staphylococcal enterotoxin (SE)-related genes and therefore the production of SEs in a dose dependent manner. For example, when cultured with 75% MIC, the transcriptional levels of sea, sec, see and agrA were decreased 11.7, 9.3, 10.45 and 10.3 fold after 18 h and 13.9, 11.21, 12.44 and 12.52 fold after 72 h in comparison to control, respectively.

Effects of Mentha longifolia L. essential oil and nisin alone and in combination on Bacillus cereus and Bacillus subtilis in a food model and bacterial ultrastructural changes.

In the face of emerging new pathogens and ever-growing health-conscious customers, food preservation technology remains on the top agenda of food industry. This study was aimed at determining the effects of the essential oil of Mentha longifolia L., alone and in combination with nisin, on Bacillus cereus and Bacillus subtilis at 8°C and 25°C in a food model (commercial barley soup) during 15 days. The essential oil alone at 8°C inhibited bacterial growth significantly compared with the control (p < 0.05). However, at 25°C, none of the concentrations of the essential oil alone showed inhibitory effect on bacterial growth. At 8°C, the combination effect of the essential oil and nisin on bacteria was noted at 0.25 µg mL(-1) for nisin and 0.05 µL mL(-1) for the essential oil (p < 0.05). The combination of nisin and the essential oil demonstrated significant inhibitory effects on the vegetative forms of bacteria at 25°C, although it was comparable to that of nisin alone at the same concentrations. Electron microscopy studies revealed a great deal of damage to B. cereus treated with a combination of nisin and the essential oil. However, the combination of nisin with the essential oil led to a complete destruction of cell wall and cytoplasm of vegetative cells of B. subtilis.

Effect of Zataria multiflora Boiss. essential oil on colony morphology and ultrastructure of Aspergillus flavus.

The mode of inhibitory action of Zataria multiflora Boiss. essential oil (EO) on the fungus, Aspergillus flavus, was studied by colony morphology examination, light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The EO at concentrations used in this study suppressed the size of the colony as well as sporulation. SEM of mycelia treated with given concentrations of EO showed morphological alterations ranging from loss of turgidity and uniformity of mycelia at low concentrations of EO to evident destruction of the hyphae at higher concentration of EO. Semi-thin sections of mycelia exposed to different concentrations of EO were analysed by light microscopy and revealed that the major change at level as low as 50 ppm of EO was limited to vacuolisation of cytoplasm resulting in cell swelling, while at higher concentrations, detachment of the cell membrane from the cell wall, deformation of mycelia and shedding the cytoplasm from the cell were the main alterations. These damages were well documented by TEM, which showed that the main sites of action of EO were the plasma membrane and cell wall. In conclusion, morphological and structural changes observed in this study may be one of the mechanisms involved in growth inhibition of the fungi and reducing aflatoxin production.

Effect of Zataria multiflora Boiss. essential oil, nisin, and their combination on the production of enterotoxin C and alpha-hemolysin by Staphylococcus aureus.

The effect of different concentrations of Zataria multiflora Boiss. essential oil (EO; 0%, 0.005%, and 0.015%), nisin (0, 0.125, and 0.25 microL/mL), and their combinations on the production of staphylococcal enterotoxin C (SEC) and alpha-hemolysin (alpha-toxin) by Staphylococcus aureus at different inoculation levels (10(3), 10(4), and 10(5) cfu/mL) in brain heart infusion broth during storage at 35 degrees C for up to 43 days was evaluated. The SEC production was significantly (p < 0.05) inhibited and the hemolysis due to alpha-toxin was significantly (p < 0.05) reduced by EO concentration at levels 0.015% and 0.005%, respectively. Significant (p < 0.05) inhibitory effect of EO on SEC production at level 0.005% was observed when it was used in combination with nisin = 0.125 microL/mL. The significant (p < 0.05) synergistic effect of EO = 0.005% and nisin = 0.125 microL/mL was also observed as more reduction of hemolysis due to alpha-toxin than EO = 0.005% alone. Further, EO significantly (p < 0.05) prevented SEC production by S. aureus during the manufacturing process of a traditional Iranian white brined cheese (as a food model) even at its lowest concentration (5 microL/100 mL), in this study.

Effect of Zataria multiflora Boiss. essential oil on growth and aflatoxin formation by Aspergillus flavus in culture media and cheese.

The effect of Zataria multiflora Boiss. essential oil (EO) against growth, spore production and aflatoxin formation by Aspergillus flavus ATCC 15546 was investigated in synthetic media as well as Iranian ultra-filtered white cheese in brine. EO effectively inhibited radial growth and spore production on potato dextrose agar (PDA) in a dose-dependent manner. At 200 ppm, the radial growth and sporulation reduced by 79.4% and 92.5%, respectively. The growth was completely prevented at EO400 ppm on PDA, and minimum fungicidal concentration (MFC) of the oil was estimated at 1000 ppm. The oil also significantly suppressed mycelial growth and aflatoxin synthesis in broth medium at all concentrations tested (P<0.05). At 150 ppm of EO, the mycelial growth and aflatoxin accumulation reduced by 90% and 99.4%, respectively. The EO at all concentrations tested, had an inhibitory effect against radial fungal growth and aflatoxin production by A. flavus in cheese. However, no concentration of EO examined was able to completely inhibit the growth and aflatoxin production in cheese. The results suggested the potential substitution of the antifungal chemicals by this EO as a natural inhibitor to control the growth of molds in foods such as cheese.

Crocus sativus L. (petal) in the treatment of mild-to-moderate depression: a double-blind, randomized and placebo-controlled trial.

Depression is a major worldwide health problem. Indeed, by 2020, depressive disorders are estimated to represent the second largest disease burden worldwide. Although a variety of pharmaceutical agents are available for the treatment of depression, psychiatrists find that many patients cannot tolerate the side effects, do not respond adequately, or finally lose their response. Our objective was to assess the efficacy of petal of Crocus sativus in the treatment of mild-to-moderate depression in a 6-week double-blind, placebo-controlled and randomized trial. Forty adult outpatients who met the Diagnostic and Statistical Manual of Mental Disorders, fourth edition for major depression based on the structured clinical interview for DSM IV participated in the trial. In this double-blind, placebo-controlled and randomized trial, patients were randomly assigned to receive capsule of petal of C. sativus 30 mg/day (BD) (Group 1) and capsule of placebo (BD) (Group 2) for a 6-week study. At 6 weeks, petal of C. sativus produced a significantly better outcome on Hamilton Depression Rating Scale than placebo (d.f.=1, F=16.87, p<0.001). There were no significant differences in the two groups in terms of observed side effects. The results of this study indicate the efficacy of petal of C. sativus in the treatment of mild-to-moderate depression. A large-scale trial is justified.