Food Biotechnology
Soheyl Reyhani Poul; sakineh Yeganeh; Reza Safari
Abstract
Introduction: One of the synthetic and harmful preservatives used in sausage is sodium nitrite. Sodium nitrite in meat products helps to increase the storage period and marketability of the product by preventing the growth of anaerobic bacteria, especially clostridiums, exerting an antioxidant effect, ...
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Introduction: One of the synthetic and harmful preservatives used in sausage is sodium nitrite. Sodium nitrite in meat products helps to increase the storage period and marketability of the product by preventing the growth of anaerobic bacteria, especially clostridiums, exerting an antioxidant effect, stabilizing the red color of the meat and improving the taste. Besides these benefits, sodium nitrite is very dangerous for health and it can cause malignant diseases. For this reason, it is necessary to replace this substance with a natural preservative. Pigments extracted from aquatics such as astaxanthin with antioxidant activity, antimicrobial properties and pink color may be a good substitute for sodium nitrite. But these pigments must be nanoencapsulated first due to their sensitivity to food production conditions, including high temperature. The aim of the current research at the first is to extract astaxanthin from Haematococcus microalgae by acid-acetone method and pigment nanoencapsulation with maltodextrin-caseinate combination coating. Then, the nanocapsules produced with different proportions will replace sodium nitrite in the sausage formulation, and oxidative and microbial spoilage tests, color and sensory evaluation will be performed for different treatments.Materials and Methods: First, astaxanthin pigment was extracted from Haematococcus microalgae (Haematococcus pluvialis) using acid-acetone technique. Then, the extracted pigment was nanoencapsulated with maltodextrin-sodium caseinate combined coating and the resulting nanocapsules in the form of treatments A (120 mg/kg sodium nitrite), B (120 mg/kg nanocapsules carrying astaxanthin), C (90 mg/kg sodium nitrite+30 mg/kg nanocapsules carrying astaxanthin), D (60 mg/kg sodium nitrite+60 mg/kg nanocapsules carrying astaxanthin) and E (30 mg/kg sodium nitrite+90 mg/kg nanocapsules carrying astaxanthin) replaced sodium nitrite in the sausage formulation. These treatments were evaluated in terms of oxidative spoilage, microbial spoilage, color indices and sensory properties during 28 days of storage at refrigerator temperature along with the control (without sodium nitrite and nanocapsules). This research was conducted in a completely randomized design. Data were analyzed by one-way analysis of variance and the difference between the means was evaluated by Duncan's test at 95% confidence level. Results and Discussion: According to the results, the lowest levels of thiobarbituric acid and peroxide value during the storage period were related to B, E and D (p>0.05) treatments (p<0.05). A and C treatments had no significant difference in terms of thiobarbituric acid and peroxide until day 14 (p>0.05), but with the passage of time this difference became significant and treatment A showed higher values (p<0.05). The results of this section showed that the power of astaxanthin in controling with oxidative spoilage is significantly greater than that of sodium nitrite, and if the purpose is only to control this type of spoilage, there is no need to replace or use sodium nitrite. The results showed that in the field of controlling microbial spoilage, sodium nitrite has more power than nanocapsules carrying astaxanthin. So that, the lowest amount of total volatile basic nitrogen (TVB-N) and the most standardized pH were related to treatments A, C and D (p>0.05) during the storage period (p<0.05). B and E Treatments (p>0.05) were ranked next (p<0.05). The results of this section showed that if from 120 mg/kg sodium nitrite to 60 mg/kg is replaced with astaxanthin carrying nanocapsules in the sausage formulation, the resulting product has the same antimicrobial power as product containing 120 mg/kg sodium nitrite. Evaluation of the color and sensory properties of treatments showed that A, C and D treatments are at a higher level than treatments B and E and the control in terms of color indices and general acceptance (p<0.05). Comparison of the color and sensory properties of the treatments on days 0 and 28 of storage at refrigerator temperature showed that the color and sensory indices remained constant in the formulated treatments, unlike the control.Conclusions: Nanocapsules carrying astaxanthin with combined maltodextrin-sodium caseinate coating as a natural product with many properties in health, control and prevention of various diseases, have a high efficiency to replace the harmful sodium nitrite preservative in sausage formulation. So that, if 30 to 60 mg/kg of the permissible limit of 120 mg/kg of sodium nitrite in the sausage formulation is replaced with nanocapsules carrying astaxanthin, the resulting product will be similar to the product containing 120 mg/kg of sodium nitrite in terms of shelf life, resistance to oxidative and microbial spoilage, color indices and sensory evaluation.
Food Biotechnology
Shohreh Nikkhah; Fakhri Shahidi; Mohebbat Mohebbi; Farideh Tabatabaei Yazdi
Abstract
IntroductionCucumber is an economically important crop, containing vitamins, minerals, antioxidants, and flavonoids. However, due to loss of weight and firmness, microbial contamination, mechanical damage, and yellowing, the storage duration of cucumber is limited to 3–5 days at room temperature. ...
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IntroductionCucumber is an economically important crop, containing vitamins, minerals, antioxidants, and flavonoids. However, due to loss of weight and firmness, microbial contamination, mechanical damage, and yellowing, the storage duration of cucumber is limited to 3–5 days at room temperature. Therefore, pretreatments are crucial for prolonging its shelf life. Chitosan is a cationic polysaccharide and can interact electrostatically with anionic, partially demethylated pectin. Besides, chitosan has inhibitory effects on fungal rot and prevents weight loss in fruits. Pectin can form excellent films. Because of increasing demand to reduce synthetic chemicals as antimicrobial agents, substances derived from plants, such as essential oils, can play a significant role in the future. Several essential oils and essential oil components have shown antimicrobial activity against spoilage and pathogenic microorganisms during fruit and vegetable storage. Essential oils of thyme and cinnamon contained phenolic groups have been found to be most consistently effective against microorganisms, however, essential oils are volatile and irritant. Therefore, forming an inclusion complex using b-cyclodextrin can improve solubility, control volatile, and induce off-flavors and unpleasant odor of the essential oils. The objectives of this study were to develop the microencapsulated thymol (thyme) and trans-cinnamaldehyde (cinnamon) essential oils to produce antimicrobial agents and subsequently evaluate the effectiveness of edible coating made of chitosan and pectin containing microencapsulated trans-cinnamaldehyde or thymol essential oils to improve qualitative and quantitative characteristics and shelf life of cucumber.Materials and MethodsThe inclusion complexes of trans-cinnamaldehyde and thymol in beta-cyclodextrin (CD) were prepared separately by freeze-drying. Each essential oil was dispersed in 1000 ml of beta-cyclodextrin aqueous solution (16 mmol/L, 18.15 g) in molecular ratio 1:1 (2.4 gr thymol, 2.11 gr trans-cinnamaldehyde) and mixed in a laboratory stirrer for 24 hour at room temperature , then frozen (-70 ºc) and freeze-dried (<20Pa, 48 h). Lyophilized samples were stored inside a freezer (-20 ºc) until further use. Cucumbers cv. Nagene with uniform size, appearance, ripeness and without mechanical damage or fungal contamination were selected. Then They were then sanitized by immersion in chlorine solution (150 mg/kg) for 1 min and air dried. Edible coatings were prepared as three immersion solutions of chitosan, pectin, and calcium chloride (CaCl2). The fruits were coated with pectin (1%) and chitosan (0-0.5%-1%) containing beta-cyclodextrin microencapsulated trans-Cinnamaldehyde or thymol each (0-0.25%-0.5%). After coating by chitosan, the fruits were immersed in 1% Calcium chloride solution to induce crosslinking reaction. After dipping step, fruits dried for 8 minutes at room temperature to remove the excess solution attached to the surface .Uncoated fruits served as control. Then fruits were preserved in cold storage (temperature: 10ºc; relative humidity: 90-95%) for 15 days. chemical (total soluble solids, titratable acidity) and physical (total color difference, Hardness, and weight loss) Characterization of fruits were measured immediately after harvest and after 5, 10 and15 days. Microbial tests (total count, mold, and yeast) were done at the end of preservation time. Analytical data were subjected to analysis of variance and factorial adopted completely randomized design and a Duncan comparison test was used. Results and DiscussionThe results showed that weight loss, total soluble solids, and the total color difference increased and hardness and titratable acidity decreased gradually in all samples during cold storage (<0.05). Chitosan and essential oils slowed down this rising or decreasing trends. Interactive effects of chitosan, essential oil type, essential oil concentration, and storage time had positive effects on these quality attributes. The fruits coated with the highest concentration of chitosan (1%) and thymol (0.5%) essential oils showed the least weight loss, loss of hardness, and color change throughout 15 days of storage. Besides thymol in comparison with trans-Cinnamaldehyde was more efficient to prevent yeasts and molds on the surface of cucumber. By increasing chitosan and essential oil amounts, the ability of inhibiting microbial growth by coating is enhanced. ConclusionThe results of chemical, physical and microbial tests, showed that multi-layer coating solution containing chitosan 1% with thymol 0.5% was effective in extending the shelf life of cucumber. The combined usage of microencapsulated thymol essential oil and chitosan-based coating on cucumber could be considered a healthy and effective treatment that reduces microbial spoilage and preserves quality and color characteristics in cucumber and represents an innovative method for commercial application. Therefore, this coating can be used as an alternative to chemical fungicides to prevent fungal rot of cucumber and other fruits, however, it is suggested that more studies should be done in this field.
Food Biotechnology
Behrooz Alizadeh Behbahani; Mostafa Rahmati-Joneidabad; Mohammad Noshad
Abstract
IntroductionThe use of safe ingredients to preserve food is steadily increasing. The high time and cost of production and approval of synthetic food additives and the reduction of public acceptance of these compounds have caused serious problems in their utilization. Excessive use of synthetic preservatives, ...
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IntroductionThe use of safe ingredients to preserve food is steadily increasing. The high time and cost of production and approval of synthetic food additives and the reduction of public acceptance of these compounds have caused serious problems in their utilization. Excessive use of synthetic preservatives, which some of them are suspected to be toxic, has completely eliminated these additives and led to the use of natural alternatives to preserve or extend the shelf life of food products. Many plant-based bioactive compounds are good alternatives to synthetic antimicrobial and antioxidant supplements. Plant extracts have significant biological activity including antioxidant, antibacterial, and antifungal properties, which has increased their use in food products. In addition, plant-derived antimicrobial compounds have been considered in the pharmaceutical industry to control microbial pathogens. Natural antioxidant and antimicrobial compounds are receiving a lot of research and industrial attention in food preservation technologies. In the last 2 decades, the use of herbal medicines rich in bioactive molecules (including polyphenols, carotenoids and flavonoids) with medicinal and health effects such as delaying the onset of some diseases such as cardiovascular disorders, diabetes, and cancer have increased.The plant Prosopis farcta grown in arid and semi-arid regions. In Iran, it is found in the southern regions of the country. In traditional medicine, this plant is used to prevent hyperlipidemia and hyperglycemia, to treat hemorrhoids, intestinal diseases and diarrhea, and leprosy, and to reduce abortion. In addition, antimicrobial and antioxidant properties of various species of Prosopis have been reported. Accordingly, in this study, after examining the of total phenols and flavonoids concentrations, the antioxidant and antimicrobial properties of ethanolic extract of Prosopis farcta were determined. Materials and MethodsThe ethanolic extract of P. farcta was obtained maceration method. Total phenol content (by Folin-Ciocalteu reagent method), total flavonoid content (by aluminum chloride method), antioxidant activity (by DPPH and ABTS free radical scavenging and beta-carotene bleaching methods), and antimicrobial effect against Escherichia coli, Shigella dysentery, Staphylococcus aureus, and Bacillus subtilis (by disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum fungicidal concentration) of the extract were evaluated. Results and Discussion farcta ethanolic extract showed high phenol content (145.58 ± 1.30 mg GAE/g), while its total flavonoid content was 72.37 ± 1.48 mg QE/g. Antioxidant activity of ethanolic extract of melon root using different methods of DPPH and ABTS free radical scavenging and beta-carotene bleaching inhibition were 62.60, 71.82 and 54.50%, respectively. Antibacterial activity of P. farcta ethanolic extract against Escherichia coli, Shigella dysentery, Staphylococcus aureus, and Bacillus subtilis according to disk diffusion agar and well diffusion agar methods showed that the antimicrobial activity of the extract was concentration dependent and Shigella dysentery and Staphylococcus aureus were the most resistant and sensitive bacterial strains to the extract respectively. The minimum inhibitory concentrations of ethanolic extract of P. farcta root for Escherichia coli, Shigella dysentery, Staphylococcus aureus, and Bacillus subtilis were 8, 8, 4 and 4 mg/ml, respectively; while the minimum bactericidal concentrations for these bacteria were 128, 256, 32 and 64 mg/ml, respectively. ConclusionIn the present study, ethanolic extract obtained from the roots of P. farcta was identified as a rich source of phenolic and flavonoid compounds. The ethanolic extract showed effective antimicrobial and antioxidant properties. The results greatly indicated the promising effect of P. farcta root extract against Gram-positive and Gram-negative bacterial species. As the microbial resistance is constantly increasing, ethanolic extract of P. farcta root can be considered as a suitable complementary option to tackle this problem. In addition, the identification of individual components of P. farcta ethanolic extract and their biological functions or their combination with common antioxidant and antimicrobial agents could be the subject of future research.
Food Biotechnology
Soheyl Reyhani Poul; Sakineh Yeganeh; Zeynab Raftani Amiri
Abstract
Introduction Since heat treatments and special standards are not used in the production of traditional (homemade) tomato paste, fungal and bacterial spoilage in the product occurs extensively during storage in the refrigerator (4°C). Astaxanthin extracted from aquatics has antimicrobial activity ...
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Introduction Since heat treatments and special standards are not used in the production of traditional (homemade) tomato paste, fungal and bacterial spoilage in the product occurs extensively during storage in the refrigerator (4°C). Astaxanthin extracted from aquatics has antimicrobial activity and color similar to tomato and can probably be effective in preventing spoilage of tomato paste. In addition, astaxanthin has other properties in the field of preventing and controlling diseases and maintaining human health, which justifies its use in food formulations as an enrichment. Since heat, enzyme, acid, etc. treatments are practiced during the production of tomato paste, these factors may change the structure and thus the function of astaxanthin. For this reason, astaxanthin nanoencapsulation is necessary for its use in tomato paste formulation. Materials and Methods In this research, first, astaxanthin was extracted from Haematococcus pluvialis microalgae using the acid-acetone combined method. Then, this pigment was nanoencapsulated using maltodextrin-sodium caseinate coating and the resulting nanocapsules were used together with the pure form of astaxanthin in the formulation of tomato paste. The research treatments were control, tomato pastes containing 3 and 6% astaxanthin (A and B, respectively) and also 3, 6 and 9% nanocapsules carrying the pigment (C, D and E, respectively). These treatments were kept at refrigerator for 28 days and were evaluated (on days 0, 7, 14, 21 and 28) in terms of the total number of fungi, Howard's number (HMC), pH, fungal flora, total bacteria count, amount of lactic acid bacteria and sensory properties. This research was conducted in a completely randomized design. Data were analyzed by One-way Anova and the difference between the means was evaluated by Duncan's test at 95% confidence level. Results and Discussion The results showed that the fungi proliferation, total count and lactic acid bacteria were slower than the control during the storage period in the treatments containing astaxanthin and its carrying nanocapsules, and the minimum number of the mentioned microorganisms and Howard's number were related to treatments D and E (p>0.05). Treatments C, B and A were ranked next in this respect (p<0.05). The number of fungi in two treatments D and E from day 0 to 28 varied from 128 to 332 cfu/gr. Also, the Howard number of these treatments was recorded from 18 to 34% in the mentioned time period. However, these two indices in the control ranged from 121 to 792 cfu/gr and 18 to 91%, respectively, during the storage period. The count of total bacteria and the amount of lactic acid bacteria in the control on day 28 were equal to 8.9 cfu/gr and 311 mg/kg, respectively, but these two values were recorded in the E and D treatments on the same day, about 4.8 cfu/gr and 110 mg/kg, respectively. Counting the total number of fungi, bacterias and also Howard's number in control and other treatments showed that the effect of nanocapsules carrying astaxanthin on microbial growth and proliferation is significantly greater than pure astaxanthin (p<0.05). The pH of the treatments varied from 3.9 to 5.8 during the storage period and the most standardized pH (3.9-4.4) was recorded in C, D and E (p>0.05) treatments (p<0.05). The pH of two treatments A and B (p>0.05) was higher than the three mentioned treatments and lower than the control (p<0.05). This finding showed that nanocapsules carrying astaxanthin have a greater effect on controlling the pH of tomato paste than pure astaxanthin during storage at refrigerator (p<0.05). The identification of the fungal flora of the treatments on the 28th day confirmed that two genus of Penicillium and Aspergillus form the main flora of the product. The results of the sensory evaluation of the treatments on day 0 showed that adding astaxanthin and its carrier nanocapsules does not change the color, aroma, taste and texture indicators (subsequently the general acceptance) of tomato paste (p>0.05). On the 28th day, the mentioned sensory indices only in the two treatments D and E were not significantly different from the 0 day, but they changed negatively in the other treatments (p<0.05). Conclusion According to the findings of the present research, astaxanthin extracted from Haematococcus pluvialis microalgae has the ability to inhibit fungal and bacterial spoilage and stabilize the sensory properties of tomato paste stored at refrigerator. This properties were improved by adding nanoencapsulated pigment using maltodextrin-sodium caseinate combined coating. Since there were no significant differences between the two treatments containing 6% and 9% of nanocapsules carrying astaxanthin (D and E) in terms of quality indices and microbial spoilage, therefore, the treatment containing 6% nanocapsules is introduced as the optimal treatment.
Food Biotechnology
Mahdieh Mostafidi; Mohammad Reza Sanjabi; Naheed Mojgani; Sohyel Eskandari; Sepideh Arbabi Bidgoli
Abstract
Introduction The food and water contamination with heavy metals is increasing due to the environmental pollutions. Heavy metals are the elements with the density of more than 5 g/cm3 and have become a serious problem as a result of the urbanization and industrialization. These toxic metals pollute ...
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Introduction The food and water contamination with heavy metals is increasing due to the environmental pollutions. Heavy metals are the elements with the density of more than 5 g/cm3 and have become a serious problem as a result of the urbanization and industrialization. These toxic metals pollute water, soil, plants, and eventually foodstuffs and our bodies. Several methods exist to remediate heavy metal pollution in waters such as membrane filtration, ion exchange mechanisms, or by precipitation. Yet, these techniques are not cost effective, in some cases, and do produce wastes that need to be properly disposed of. Microbial bioremediation could be an alternative. The use of microbes for remediation of heavy metals has been well studied. Some microorganisms, especially soil bacteria, have the ability to tolerate these contaminants. In addition, certain bacterial strains are capable of binding to heavy metals or transforming them into less toxic forms. Low operating costs, usable in foodstuffs, selective removal for specific toxic metals, minimal use of chemicals (resulting in low sludge production) and high efficiencies at very low levels of heavy metals are some of the advantages of biosorption methods. In this regard, the purpose of this study was to investigate the ability of active and passive absorption of heavy metals by a number of Lactic Acid Bacteria (LAB) strains in laboratory environment and food. Materials and Methods Seven LAB isolates including Lacticaseibacillus casei (RTCC 1296-3), Lacticaseibacillus rhamnosus (RTCC 1293-2), Lactiplantibacillus plantarum (RTCC 1290), Limosilactobacillus fermentum (RTCC 1303), Enterococcus faecium (RTCC 2347), Lactobacillus helveticus (RTCC 1304) and Lactobacillus acidophilus (RTCC 1299) were obtained from Razi type culture collection (RTCC), located at Razi vaccine and Serum Research Institute, Iran. All isolates were cultured in MRS (Scharlau, Spain) broth medium, at 37 °C for 24 hours, under anaerobic conditions. Pure cultures were preserved for long term by freezing at -70°C with 20% Glycerol. Heavy metals including Nitrate of Pb (II), Cd (II) and Ni (II) were purchased from Merck (Darmstadt, Germany). All standard solutions were prepared from the stock solutions containing 1000 mgl-1 in distilled water. Other chemicals used in study including Nitric acid (65%) and Hydrogen peroxide (37%), were also purchased from Merck, Germany. This study was conducted in two in- vitro and in-vivo phases; in the in- vitro phase, seven strains of bacteria with probiotic properties (L. casei, L. rhamnosus, L. plantarum, L. fermentum, Ent. facium, L. helveticus and L. acidofilous) were screened and then their ability to bind to cadmium (Cd), Lead (Pb) and nickel (Ni) in aqueous solution was investigated. Then, in the in-vivo stage, three probiotic strains that had the highest biosorption efficiency in the previously stage were selected and their effect with a ratio of 1:1:1 and contact time of 15 and 30 minutes on the removal of these toxic metals in coriander, leek and parsley fresh vegetables was evaluated. The residual concentrations of heavy metals in solution were measured by Inductively Coupled Plasma Mass Spectrometer (ICP-MS; ELAN DRC-e, PerkinElmer SCIEX, Canada) and Morphology of bacteria cell surfaces incubated with metals were monitored by scanning electron microscopy (JEOL JSM 5400 LV, Japan). Results and DiscussionThe results of the in vitro stage showed that the most ability to heavy metals adsorption was related to the Ent. Facium bacterium which were equal to 79.75±0.11, 75.28±0.05 and 83.99±0.10% for Pb, Cd and Ni, respectively. In general, the removal efficiency of heavy metals by LAB bacteria in the inactive and killed state was significantly higher than the active removal efficiency of these bacteria, so that the highest percentage of passive absorption of lead, cadmium and nickel metals by inactive strains of L. casei, L. plantarum and Ent. Facium were 90.01, 81.98 and 86.56%, respectively. Electron microscopy observations and energy dispersive X-ray (EDX) analysis confirmed that the majority of these toxic metals significantly damage the surface of living cells by accumulating and binding on the surface of bacterial cells. A combination of three bacterial strains had a synergistic effect on the binding properties of toxic metals compared to the single state of these bacteria, so that in both active and inactive states, 90-99% of heavy metals from edible leafy vegetables were removed in less than 15 minutes. The results of this research generally showed that the binding capacity of dead biomass is significantly high and it is possible to dispose and reuse biomass in case of biological absorption.
Food Biotechnology
Asad Abbaspour Anbi; Masoud Seidgar; Masoud Neyriz Nagadehi
Abstract
The present investigation was done to study the effects of Lactococcus lactis (L. lactis) subsp. lactis on the shelf life of the vacuum-packaged Oncorhynchus mykiss. Fish fillets were prepared and divided into 5 different treatment groups including control (distilled water), 2% and 4% supernatant, and ...
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The present investigation was done to study the effects of Lactococcus lactis (L. lactis) subsp. lactis on the shelf life of the vacuum-packaged Oncorhynchus mykiss. Fish fillets were prepared and divided into 5 different treatment groups including control (distilled water), 2% and 4% supernatant, and 106 CFU/g L. lactis subspecies lactis. The pH, Thiobarbituric Acid Reactive Substances (TBARS), Total volatile Nitrogen (TVN), and Peroxide Value (PV) of the fillets were determined on days 0, 5, 10, and 15 while maintained at 4˚C. Protein expression and destruction were analyzed using the SDS-PAGE. The organoleptic assessment was done using five expert sensory panelists. Contents of TBARS, TVN, pH, and PV were increased throughout the storage period (P <0.05). An increase in the concentration of supernatant caused a significant decrease in the content of TBARS, TVN, pH, and PV (P <0.05). The highest and lowest contents of TBARS, TVN, pH and PV on 15th day were belonged to the control (3.367±0.04 mg MDA/kg) and pure bacteria (0.70±0.02 mg MDA/kg), control (87.20±6.40 mg/100g) and 4% supernatant (40.79±0.61 mg/100g), pure bacteria (6.23±0.04) and 4% supernatant (5.44±0.07) and control (12.22±0.01 meq/kg) and 4% supernatant (3.08±0.06 meq/kg) groups, respectively. Protein destruction was lower in the fillet samples treated with pure bacteria and 4% supernatant. The highest scores of the odor, flavor, texture, and color were obtained for fillets treated with 4% supernatant, pure bacteria, pure bacteria, and 4% supernatant and pure bacteria, respectively. The results revealed that treating O. mykiss fillets with 4% supernatant and 106 CFU/g of pure L. lactis subsp. lactis can extend the shelf life of O. mykiss fillets.
Food Biotechnology
Nasim Najafi; Hajar Abbasi
Abstract
Due to its health benefits, fresh sprouted cereals are considered popular food source. They are very sensitive and highly susceptible to microbial spoilage during transportation, processing, and storage. This phenomenon makes them potentially high-risk fresh products. This study aimed to assess the effect ...
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Due to its health benefits, fresh sprouted cereals are considered popular food source. They are very sensitive and highly susceptible to microbial spoilage during transportation, processing, and storage. This phenomenon makes them potentially high-risk fresh products. This study aimed to assess the effect of emulsion coating consisting of Dracocephalum kotschyi essential oil (0, 50, 150, 250, 300 ppm)-chitosan solution (0, 0.3, 0.38, 0.63, 0.75%) during the immersion time (10, 25, 55, 85, 100 s) on the microbial properties of fresh sprouted wheat stored at 4°C. The Response Surface Methodology (RSM) was adopted in modeling the independent variables’ effects. The results shown that increase in the essential oil and chitosan solution concentration reduced the microbial spoilage. High concentration of Dracocephalum kotschyi oil decreased the fungus population after 12 days. Coating of sprouted wheat at optimized level of independent variables (0.62% chitosan, 57 ppm Dracocephalum kotschyi oil and 29.49 s immersion time) reduced the microbial and fungal populations. This treatment can reduce weight loss, and maintain tissue firmness, total phenolic, and ascorbic acid content of the sprouted wheat during cold storage, with no effect on its sensory properties. Our findings indicate that nanoemulsion coating based on chitosan and Dracocephalum kotschyi oil at appropriate levels could be beneficial in maintaining sprouted wheat quality and increasing its shelf-life.
Food Biotechnology
Zohre Noruzi Motlagh; Mahmoud A. Mahdavi; Reza Gheshlaghi
Abstract
Introduction Carotenoids have many effects on human health. These compounds are produced by plants and microalgae. The extraction of carotenoids from microalgae such as Chlorella has received much attention, since microalgae grow all year round (regardless of the season) and at a much faster rate ...
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Introduction Carotenoids have many effects on human health. These compounds are produced by plants and microalgae. The extraction of carotenoids from microalgae such as Chlorella has received much attention, since microalgae grow all year round (regardless of the season) and at a much faster rate than plants in non-arable lands. The aim of this research was to optimize the concentrations of nutrients (nitrogen and phosphorous) in the growth medium of microalgae with the objective of maximizing carotenoids content. At the optimized nutrient conditions, the effect of phytohormones on production of carotenoids using Chlorella sorokiniana IG-W-96 was investigated. Materials and MethodsChlorella sorokiniana IG-W-96 was cultivated in BG11 growth medium with light intensity of 25000 lux and light: dark cycle of 16: 8 supplied with compressed air flow of 0.5 vvm containing 6% vol carbon dioxide. Under three concentrations of nitrate (0.04, 0.25, 1.5 ) and three concentrations of phophate (0.01, 0.04, 0.16 ) and carotenoid concentration was measured. Full factorial experimnetal design was performed and the resuts of the experiments were analyzed using Minitab (ver. 21.01.1). Finally, the best concentrations of nitrate and phosphate were chosen for pigments production, and at that concentration, naphthalene acetic acid (0, 2.5, 5, 7.5, 10 and 12 ppm) was added to the culture medium to check its effect on pigments production. By measuring the dry weight of C. sorokiniana, its growth rate was determined. After extracting the pigments with solvent, the concentration of the pigments was determined by measuring the amount of light absorption. Results and DiscussionDry weightThe results showed that the highest amount of dry weight was related to the treatment with nitrate amount of 0.25 , and nitrate more and less than this amount caused a decrease in growth. This result was not dependent on the amount of phosphate and was true for all phosphate concentrations. Nitrate reduction from 1.5 to 0.25 increased the growth of microalgae up to 81.8%, so that the dry weight of 0.88 reached 1.6 . However, reduction of nitrate from 0.25 to 0.04 decreased the dry weight by 65.6%. In order to reach the maximum growth rate, it is necessary to determine the appropriate concentration of each nutrient. CarotenoidsUnlike the dry weight, not only the pigment production did not decrease with the excessive of nitrate concentration, but also the maximum amount of pigment production was related to the treatment with the maximum amount of nitrate concentration. Based on the results obtained, the concentration of carotenoids was higher in the concentration of 1.5 of nitrate and 0.04 of phosphate (6.7 ).When the nitrate concentration was very low (0.04 ), changing the phosphate concentration had no significant effect on the production rate of any of the pigments. Only when the nitrate concentration was high (1.5 ), change in phosphate concentration caused a change in pigments concentration. The increase of phosphate concentration from 0.01 to 0.04 increased the carotenoids concentration to 1.65-fold. Of course, increasing phosphate concentration to 0.16 did not affect the pigments concentration. Based on the statistical analysis, the P-value<0.05 indicated that the effect of the factors and the model was significant. In this situation, in order to increase the production of carotenoids, naphthalene acetic acid was added to the phytohormone culture medium. At the optimal concentration of 2.5 ppm of naphthalene acetic acid, the concentration of carotenoids increased by 26.71% and reached 8.49 . However, phytohormone had no significant effect on dry weight. ConclusionCarotenoid production using microalgae could be maximized through optimization of nutrients concentrations (nitrate and phosphate) in the growth medium. Phytohormones could further increase the prodcution of carotenoids at optimum concnetrations.
Food Biotechnology
Masume Atharinia; Nasrin Zonourian; Sare Davarzani
Abstract
IntroductionTomato paste is one of the processed tomato products that has a long shelf life and is used as an important food ingredient all over the world. According to global statistics, Iran is among the top ten producers of tomato paste in the world, Iran ranks fourth to fifth in the world in the ...
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IntroductionTomato paste is one of the processed tomato products that has a long shelf life and is used as an important food ingredient all over the world. According to global statistics, Iran is among the top ten producers of tomato paste in the world, Iran ranks fourth to fifth in the world in the field of aseptic paste production. Alicyclobacillus bacteria are considered as a risk for pasteurized acidic food industries. These bacteria enter the product through soil-contaminated fruits, production equipment of the factories and finally produce metabolites such as guaiacol, causing an unpleasant taste in the product. Materials and Methods In order to investigate the microbial contamination of canned tomato paste in the country, 46 samples of canned tomato paste in the amount of 184 cans of 800 grams were purchased from the market. Regarding the purchase of samples from the market, we tried to buy a different production date and production series for each sample (approximately 4 cans for each brand from each production series). The purchased samples were sent to the Microbiology Department of the Standard Research Institute laboratory for microbiology tests. At the same time, the culture media of thermophilic bacteria (Orange Serum Agar, Thermoacidurans Agar from 4 available brands) were tested for performance control. The canned tomato paste samples were incubated at 30°C ± 1°C for 14 days and 55°C ± 1°C for 7 days. Results and Discussion The contents of both examined samples were tested separately for thermophilic bacteria, mesophilic bacteria, mold and yeast. Out of the 46 samples prepared with different production dates and production series, which were 46 cans of tomato paste, 28 samples were positive in terms of contamination with thermophilic bacteria. According to the number of contaminated samples, it was found that 60.86% of the samples were contaminated. Colonies grown on Thermoacidurans Agar medium were examined morphologically. For further investigations, gram staining was performed. All the stained colonies morphologically showed the form of gram-positive rod-shaped bacilli. Biochemical tests including catalase and oxidase were performed to identify Alicyclobacillus species. All the grown colonies were catalase positive and oxidase negative. The final identification of the species was done by performing molecular tests based on specific primers designed from Alicyclobacillus gene. These tests were performed in three stages: genomic DNA extraction, polymerase chain reaction and electrophoresis. Using the PCR method, the grown colonies were analyzed for two types of bacteria, Alicyclobacillus acidocaldarius and Bacillus coagulans. According to the results obtained from sequencing with designed primers in the NCBI database, it showed 100% similarity with the registered sequences, which are all different strains of the Alicyclobacillus acidocaldarius species. None of the colonies were detected as Bacillus coagulans species. Since Alicyclobacillus acidocaldarius was isolated from soil for the first time, the presence of these bacteria in the product indicates the contamination of raw materials with soil. ConclusionIn this research, the presence of Alicyclobacillus bacteria in canned tomato paste was confirmed. Due to the high heat resistance of this bacteria, there is a possibility of the presence of Alicyclobacillus in the all stages of tomato paste production, which have entered the product through the soil, and 95°C ± 3°C pasteurization temperature in 30 minutes is not effective in removing this bacteria completely. Most acidophilus thermophilic bacteria, such as Alicyclobacillus family, are not pathogenic bacteria. Their presence in food may make the food taste bad or smelly, but it does not pose a risk to the health of the consumer. Therefore, in order to reduce the risk of spoilage and to prevent the growth of bacterial spores in the product, it is essential not to expose the product to high temperatures for a long time. It is also necessary to perform rapid cooling after heat treatment and keep the product at a temperature below 30°C. AcknowledgementThis article is the result of a common research project of Microbiology and Biology Research Group of Standard Research Institute and Kermanshah Standard Regional Research Group. We hereby thank and appreciate the cooperation of the microbiology research group of the Standard Research Institute and the Kermanshah General Directorate of Standards. We are also very grateful to Rogin Talk Company as the employer of this project.
Food Biotechnology
َAlireza Hemmati; Ali Ganjloo; Kambiz Varmira; Mandana Bimakr
Abstract
Introduction It is believed that edible oils and fats with high levels of unsaturated fatty acids are susceptible to oxidation. Soybean oil as one of the four important edible oils has high content of polyunsaturated fatty acids and so prone to oxidation. Generally, lipid oxidation leads to deterioration ...
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Introduction It is believed that edible oils and fats with high levels of unsaturated fatty acids are susceptible to oxidation. Soybean oil as one of the four important edible oils has high content of polyunsaturated fatty acids and so prone to oxidation. Generally, lipid oxidation leads to deterioration of nutritional quality and organoleptic properties of edible oils and fats as well as accelerate the development or progression of cancer, mutagenesis, carcinogenesis, aging and cardiovascular diseases through the formation of free radicals. Therefore, edible oils and fats fortification with antioxidant compounds in order to protect them against oxidation is essential. In recent years, numerous studies were carried out on exploration of natural and safe antioxidant compounds due to the consumers concerns about potential health risk of synthetic antioxidants, such as butylatedhydroxyanisole (BHA), butylatedhydroxytolene (BHT), tert-butylhydroquinone (TBHQ) and propylgallate (PG). In this regard, TBHQ as the most powerful synthetic antioxidant is prohibited as food additive in Japan, Canada and Europe. Ferulago angulata Boiss which called chavir or chavil belongs to the family of Apiacea consisting of 35–40 species that 8 species grow in Iran. It was reported that Ferulago species are used in folk medicine for their tonic, digestive, sedative, aphrodisiac properties from ancient times. Therefore, in the current study, the oxidation development of soybean oil enriched with F. angulata essential oil (EO) during accelerated storage was investigated. Materials and Methods EO from freeze dried aerial parts of F. angulata was extracted through hydrodistillation using Clevenger type apparatus. Gas chromatography-mass spectrometry (GC-MS) was used to identify main components of the EO. Total phenolic and flavonoid content of the EO were assessed using Folin–Ciocalteu and aluminium chloride colorimetry methods, respectively. Antioxidant activity of EO was measured through 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) and reducing power (RP) tests. Then, the EO of F. angulata at three concentrations, i.e. 200 ppm (SO-200), 400 ppm (SO-400), and SO-Mixture (100 ppm TBHQ + 100 ppm EO) were added to soybean oil. The synthetic antioxidant of TBHQ at the concentration of 200 ppm was added as control. The effect of EO from freeze dried aerial parts of F. angulata on oxidative stability of soybean oil stored under accelerated conditions at 65 ºC for 24 days was evaluated through acidity, peroxide (PV), p-anisidine (p-An) and TOTOX values. Results and Discussion Extraction yield, total phenolic and flavonoid contents of EO of F. angulata were 2.5% v/w, 188 mg GAE/g and 70.90 mg QE/g respectively. Furthermore, DPPH free radical scavenging activity and RP were 55.45-13.21% and 3.61-2.72 in the concentration range of 1.6-4.6 mg/ml of EO, respectively. Based on GC-MS analysis, the EO contains 41 natural compounds, representing 96.97% of the total EO. F. angulata EO could effectively reduce the acidity, PV and p-An values. For control sample, the maximum values of acidity, PV peroxide, p-An and TOTOX were 1.52 mg KOH/g, 10.60 meq O2/kg, 12.48 and 33.68 respectively after 24 days under accelerated conditions. While these values were 0.085 mg KOH/g, 4.5 meq O2/kg, 9.16 and 18.16 respectively for the soybean oil containing the lowest concentration of EO of F. angulata. Conclusion The results confirmed the instability of soybean oil during storage as well as the ability of EO from F. angulata for soybean oil protection against oxidation. As a result, EO from aerial parts of F. angulata could be suggested as a natural and effective antioxidant to be used instead of TBHQ as a synthetic antioxidant for soybean oil stabilization.
Food Biotechnology
Hossein Zanganeh; Fakhri Shahidi; Seyed Ali Mortazavi; Behrooz Alizadeh Behbahani
Abstract
IntroductionOxidation reactions and microorganisms’ activity are considered as the most important factors affecting the quality of food products. Recently, in the light of the inefficiency of some chemical preservatives against microorganisms and the presence of toxic residues in food products, ...
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IntroductionOxidation reactions and microorganisms’ activity are considered as the most important factors affecting the quality of food products. Recently, in the light of the inefficiency of some chemical preservatives against microorganisms and the presence of toxic residues in food products, the use of natural antimicrobials and antioxidants has been increased. Natural antimicrobial compounds have the potential to control microbial contamination and reduce the use of antibiotics. Plant essential oils are natural compounds with the potential to be used as active ingredients in the food, pharmaceutical, and cosmetic industries. Various studies have shown that essential oils have antifungal, antibacterial, antiviral, and antioxidant activity. The essential oils are considered as superb preservatives with various biological functions. Essential oils are generally recognized as safe product (GRAS) which can be used as an alternative to synthetic additives.Grapefruit (Citrus paradisi) peel and fruit contain active ingredients such as acids, flavonoids, vitamin C, and potassium, and its essential oil is composed of terpenic hydrocarbons, such as citral, limonene, citronelal, and geraniol. Although plant essential oils have antimicrobial and antioxidant properties, one of the main problems of these natural compounds is their high volatility and instability. In this context, nanoemulsion formulations are frequently used to increase the stability and efficiency of these biologically active compounds. This study is therefore aimed to nanoemulsifying the grapefruit essential oil and evaluate its antioxidant and antimicrobial properties. Materials and Methodsβ-carotene, linoleic acid, ABTS (2,2’-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt), and DPPH (2,2-diphenly-1-picrylhydrazyl) were purchased from Sigma-Aldrich Co. (USA). Mueller Hinton Broth (MHB) and Mueller Hinton Agar (MHA) were supplied from Merck Co. (Darmstadt, Germany). Grapefruit peel was dried at ambient temperature and then powdered. The obtained powder was then transferred to a Clevenger device containing 750 ml of distilled water to perform the distillation extraction (3 h). The resulting grapefruit essential oil was stored at 4 °C until use. Grapefruit essential oil was prepared using the hydrodistillation method, and then nanoemulsified. The antioxidant activity of the nanoemulsified essential oil was investigated by DPPH and ABTS radical scavenging activity and beta-carotene/linoleic bleaching test. The nanoemulsified essential oil or methanolic (control) was mixed with DPPH solution and the mixture was then stored at ambient temperature for 30 min, in a dark place. The control sample was prepared by methanol. The absorbance of the samples was measured at 517 nm. To determine the ABTS-RS activity, the nanoemulsified essential oil was briefly charged with methanolic ABTS radical cation solution and the resulting mixture was left at room temperature for 30 min. Afterward, the absorbance was read at 734 nm. A spectrophotometric method was applied to monitor β-carotene/linoleate solution bleaching in the presence of the nanoemulsified essential oil. To do this, the absorbance of the solution was recorded at 490 nm after 120 min against the control sample at time zero and after 120 min. Antibacterial effect of the grapefruit essential oil nanoemulsion was also evaluated against Escherichia coli ATCC 25922, Salmonella typhi ATCC 6539, Pseudomonas aeruginosa ATCC 27853, Listeria innocua ATCC 33090, Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 14579, Bacillus subtilis ATCC 23857, Streptococcus pyogenes ATCC 19615, and Staphylococcus epidermidis ATCC 12228, based on disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum bactericidal concentration. Results and DiscussionsThe results showed that the nanoemulsion of grapefruit essential oil had a remarkable antioxidant effect of 42.27 mg/ml, 33.27 mg/ml and 54.54%, respectively, based on DPPH, ABTS, and beta-carotene-linoleic acid bleaching tests. According to disk diffusion agar and well diffusion agar results, the lowest inhibition zone was related to E. coli and the highest inhibition zone was observed in L. innocua. The minimum inhibitory concentration for L. innocua and S. aureus (the most sensitive bacteria) was 25 mg/ml, and E. coli, S. typhi, and P. aeruginosa had the highest inhibitory concentration. Also, the lowest bactericidal concentration was related to L. innocua and S. aureus bacteria and the highest concentration was observed for E. coli, S. typhi and P. aeruginosa. The nanoemulsified essential oil generally exhibited greater antibacterial activity against Gram-positive species. This could be mainly due to the difference in the cell wall composition of Gram-positive bacteria in comparison to Gram-negative; Gram-positive bacteria have a thicker mucopeptide layer in their cell wall, while Gram-negative bacteria have only a thin layer of mucopeptide and the wall structure is mainly composed of lipoprotein and lipopolysaccharide, thereby leading to a higher resistant to antibacterial agents According to the results, grapefruit essential oil nanoemulsion can be used as a natural antioxidant and antimicrobial agent to control oxidation reactions and the growth of spoilage and pathogenic microorganisms.
Food Biotechnology
Mostafa Rahmati-Joneidabad; Behrooz Alizadeh Behbahani; Mohammad Noshad
Abstract
IntroductionStrawberry and grapes are generally infected with pathogenic fungi (e.g., Aspergillus niger, Botrytis cinerea, Rhizopus stolonifera, etc.). Synthetic fungicides are commonly used as the first line of defense against post-harvest pathogens on packaging lines. However, disposal of toxic waste ...
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IntroductionStrawberry and grapes are generally infected with pathogenic fungi (e.g., Aspergillus niger, Botrytis cinerea, Rhizopus stolonifera, etc.). Synthetic fungicides are commonly used as the first line of defense against post-harvest pathogens on packaging lines. However, disposal of toxic waste is a costly process and the hazardous waste causes serious environmental problems. In addition, fungal pathogens have shown a worrying trend of resistance to these fungicides, thus shortening the shelf life of products. Compounds that can be equally effective in controlling pathogens, but preventing or minimizing the waste problems will be inevitable. The large volume of internationally processed agricultural products, as well as the increasing demand for organically produced fruits, emphasizes the need to replace synthetic fungicides with safer and biodegradable alternatives. Natural plant-derived products effectively meet this criterion and have great potential to influence modern agricultural research. Catechins and other polyphenols in green tea show strong antioxidant activity. Also, the antimicrobial activity of green tea extract against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans has been reported. Therefore, the present study was performed to prepare the ethanolic extract of green tea and to determine the content of total phenol, total flavonoids, antioxidant activity, and its antifungal effect against Aspergillus niger, Botrytis cinerea, and Rhizopus stolonifer (causing rot in strawberry and grapes). Materials and MethodsFresh green tea leaves were dried at room temperature and then powdered. Then, ethanol (70%) was added to the powdered leaves (solvent to powder ratio of 10:1 v/w) and the mixture was refluxed for 120 min. The resulting mixture was filtered through a filter paper and then concentrated under vacuum and finally dried in an oven.Total phenol content (by Folin-Ciocalteu reagent at 756 nm), total flavonoid content (spectrophotometrically at 510 nm), antioxidant activity (by DPPH and ABTS radical scavenging methods), and antifungal effect (by disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum fungicidal concentration) of the extract were evaluated. Results and DiscussionThe extract contained 175.60 mg GAE /g total phenol and 47.53 mg QE/g total flavonoids and its antioxidant activity using DPPH and ABTS free radical assays was 78.89% and 86.57%, respectively. The results of antifungal activity showed that the diameter of the growth inhibition zone increased significantly with increasing the concentration of the extract, and Botrytis cinerea and Rhizopus stolonifer were the most sensitive and resistant fungal strains to the extract, respectively. The minimum fungicidal concentrations for the strains of Botrytis cinerea and Rhizopus stolonifer were 64 and 512 mg/ml, respectively. ConclusionThe results of the present study showed that the ethanolic extract of green tea could be considered as potential source of natural antioxidant and antifungal agents. The presence of phenolic and flavonoid compounds may be responsible for the antifungal and antioxidant effects of the extract. However, due to the fact that this study was performed with the crude extract of green tea, it is difficult to identify compounds responsible for antifungal and antioxidant activity. On this point, only the separation of the components of the extract allows the detection of antifungal and antioxidant compounds. This study provides a basis for further researches, in particular the use of these antioxidants and antifungal compounds. Green tea extract is especially suitable for products with high sensitivity to lipid oxidation and infection with molds.
Food Biotechnology
Saeedeh Fatemizadeh; Mohammad Bagher Habibi Najafi; Dennis Sandris Nielsen
Abstract
IntroductionCronobacter sakazakii is an opportunistic pathogen, which has been linked to the contamination of powdered infant formula, and associated with outbreaks leading to fatalities in neonatal intensive care units. Few studies have explored the direct interaction between probiotics and C. sakazakii. ...
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IntroductionCronobacter sakazakii is an opportunistic pathogen, which has been linked to the contamination of powdered infant formula, and associated with outbreaks leading to fatalities in neonatal intensive care units. Few studies have explored the direct interaction between probiotics and C. sakazakii. In this study, the effect of a Lactiplantibacillus plantarum strain (M17) along with the standard strain Lactobacillus plantarum (ATCC 8014) and the well-characterized probiotic strain Lactobacillus rhamnosus GG on the adhesion of C. sakazakii to intestinal epithelial cells was analyzed. Materials and MethodsAcid and bile tolerance of M17 was evaluated in the presence of pepsin and pancreatin. L-arginine hydrolysis was investigated using an arginine-including medium. Auto-aggregation and co-aggregation assays were performed by absorbance measurement. Minimum inhibitory concentrations of the antimicrobials recommended by the European Food Safety Authority were established. Total lactic acid and the ratio of D/L lactate isomers were determined with a Megazyme enzymatic kit. The ability of the isolate to produce biogenic amines was tested by qualitative and quantitative monitoring. Hemolysis was assessed phenotypically on MRS agar enriched with sheep blood. The strain was tested for its capability to adhere to mucin and Caco-2 cells. The antagonistic effects of the strain against C. sakazakii were further evaluated in vitro on mucin and cultured Caco-2 cells. The LAB strain was added simultaneously with, before, and after C. sakazakii to Caco-2 cells for competition, exclusion and displacement assays, respectively. Data analysis was performed in R using one-way analysis of variance, and the experimental groups were compared with the controls using Tukey’s test. P values <0.05 were considered statistically significant. Results and DiscussionThere was no significant difference in the survival rate of M17 and L. plantarum ATCC 8014 at pH = 4. After 2 h of incubation at pH = 2.5, the survival rate of L. plantarum ATCC 8014 was estimated to be higher than strain M17, but this difference was not significant. After 4 hours of incubation at pH = 8, M17 showed a higher survival rate than L. plantarum ATCC 8014, and this difference was significant after transfer from pH = 4. These results confirm the appropriate viability of M17 in the gastrointestinal tract. Both M17 and L. plantarum ATCC 8014 developed the color yellow in the L-arginine hydrolysis assay, which confirms the safety of these strains. The percentage of auto-aggregation for M17, L. plantarum ATCC 8014, and Lactobacillus rhamnosus LGG was estimated at 24.38, 25.28, and 32 after 6 hours, respectively, and no statistically significant difference between the two isolates were noticed. Given the auto-aggregation and co-aggregation parameters of M17, this strain may constitute a defense mechanism against C. sakazakii. Strain M17 showed resistance to kanamycin and clindamycin antibiotics. With intrinsic resistance, the risk of transferring resistance genes is not only speculative, but practically impossible. Intrinsic resistance of lactic acid bacteria may be considered desirable because it ensures their survival when the host is treated with antibiotics. Both D and L isomers of lactic acid were produced by the studied strains. In humans, D(-)-lactic acidosis is a rare metabolic complication that has only been reported in individuals with short bowel syndrome). Clinical studies have shown that the consumption of probiotic bacteria producing D(-)-lactic acid is safe for children and does not cause a long-term increase in blood D(-)-lactic acid. The reference L. plantarum strain and M17 did not produce biogenic amine precursors, and had no ß-hemolytic activity. Mucin adhesion assay exhibited that M17 has less adhesion (12.10 ± 1.14 %) than L. plantarum ATCC 8014 (13.33 ± 2.30 %) and LGG (15.93 ± 2.06 %) although these differences were not statistically significant. However, the amount of adhesion for the positive control sample Escherichia coli K12 (25.19 ± 4.40 %) was significantly higher than those of the other strains. Compared to the positive control, M17 had a significantly lower adhesion rate (6.8 ± 1.41) to CaCo-2 cells. This value was estimated at 13.77 ± 3.53 % for the reference strain and 21.6 ± 7.54 % for Lactobacillus fermentum PCC (positive control). In antagonistic assays, M17 was able to reduce the adhesion of C. sakazakii to mucin and CaCo-2 cells in all three methods of exclusion/inhibition, competition and displacement. Statistical analysis of the results does not show a significant difference between M17 and LGG. Therefore, the performance of M17 is similar to that of the standard probiotic LGG. ConclusionLactic acid bacteria with acceptable ability to adhere to epithelial cells can be suitable for colonization in the intestine. They can act as a barrier to fight pathogens through various competitive mechanisms, such as co-aggregation with pathogens and adhesion. The M17 strain has an acceptable immune profile and probiotic properties because it shows an acceptable antagonistic activity against C. sakazakii invasion. AcknowledgementThis study was supported by Ferdowsi University of Mashhad (Research affairs) [project No.:46718] and the research infrastructure at the University of Copenhagen.
Food Biotechnology
Mina Seifzadeh
Abstract
Collagen has diverse general and biomedical applications and its important role in the future of society have made it a key biopolymer for human health and well-being. Therefore, the present study was conducted with the objectives of extracting collagen from the skin of farmed carp, determining the quality ...
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Collagen has diverse general and biomedical applications and its important role in the future of society have made it a key biopolymer for human health and well-being. Therefore, the present study was conducted with the objectives of extracting collagen from the skin of farmed carp, determining the quality characteristics of collagen, and comparing them. Collagen was extracted from the skin of carp fishes by acidic enzymatic method using 0.5 M acetic acid and pepsin in 48 hours. Collagen treatments (5 treatments) included collagen prepared from the skin of common species, grass carp, bighead, silver, and cow (control). Collagen treatments were not capable of hemolysis and did not show toxic effects on human fibroblast cells. Heavy metals (0.01-0.18 ppm) in collagen extracted from cultured carp species were within the standard range. The color (brightness) of experimental collagen (92.74-93.68) and control (92.38) showed no significant difference (p<0.05). Amino acids cysteine and tryptophan were not observed in collagen. Glycine and hydroxylysine amino acids (352 and 3 residues 1000g-1, respectively) had the highest and lowest amounts in collagen. Amino acids profile and collagen production efficiency (10.51-10.59%) did not show significant differences in carp fish species (p<0.05). Based on the results of the present study, production efficiency, safety and quality characteristics of collagen in cultured carp species did not show any significant difference (p<0.05), and no significant difference was observed between these characteristics and the control (p<0.05). Therefore, the skin of these species can be used to produce collagen and introduce it to the industry as a substitute for mammalian collagen.
Food Biotechnology
Shabnam Parichehreh; Gholamhossein Tahmasbi; Mohammad Eslampanah; Pejvak Khaki
Abstract
Iranian men are at risk of developing gastrointestinal cancer caused by H. pylori. It is very imperative to find effective methods to control this bacterium as there are currently no very effective treatments for it. Honey has been shown to have antimicrobial properties against various pathogens. ...
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Iranian men are at risk of developing gastrointestinal cancer caused by H. pylori. It is very imperative to find effective methods to control this bacterium as there are currently no very effective treatments for it. Honey has been shown to have antimicrobial properties against various pathogens. This study analyzed 15 honey samples from A. florea bees, collected from different floral and geographical origins, for their antimicrobial efficacy against H. pylori. Using atomic absorption measurements, the honey samples were also tested for their phenolic and flavonoid content, protein concentration, and mineral content. Antioxidant activity was determined using the FRAP, DPPH, and ABTS methods. The antibacterial activity of honey samples was investigated both in-vitro and in-vivo in the gastrointestinal tract of mice. Statistical analysis revealed a significant positive correlation between antioxidant activity and antibacterial activity. All honey samples showed antimicrobial activity in-vitro, among which jujube honey from Bushehr exhibiting the highest activity. Differences in antioxidant and antimicrobial activities were likely due to the flora of the plants and the geographic region from which the honey was harvested. Based on these results, A. florea honey may be used in the prevention and treatment of H. pylori-associated infections and inflammation of the gastrointestinal tract. This feature can be applied to the control of Helicobacter pylori along with other available measures.
Food Biotechnology
S.Javad Hosseini; Mostafa Shahidi Noghabi; Hossein Zamani; Gholamhossein Zohuri; Mahboubeh Sarabi
Abstract
The essential oils usually have a good effect against undesirable microorganisms; therefore, they can be utilized as natural antimicrobial agents in food or their packaging. In this research, the antimicrobial attributes of two essential oils (Oliveria decumbens and Pistacia atlantica gum), have been ...
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The essential oils usually have a good effect against undesirable microorganisms; therefore, they can be utilized as natural antimicrobial agents in food or their packaging. In this research, the antimicrobial attributes of two essential oils (Oliveria decumbens and Pistacia atlantica gum), have been investigated before and after thermal process (200°C - 10 minutes) against bacterial and mold spoilage in bread. Also, the compounds of essential oils were detected by gas chromatography-mass spectrometry. The main compounds of the essential oil of O. decumbens were carvacrol, thymol, and elemicin before and after thermal treatment. In the case of P. atlantica gum, only one prominent peak was observed in the chromatogram, which was related to the α-pinene. For both essential oils, the MIC and MFC against Aspergillus niger were 4000 and 8000 μL/ml, respectively. In comparison, the antimicrobial effect of both essential oils against Bacillus subtilis was higher than the mold. The amount of MIC and MBC were 125 and 250 μL/ml for Oliveria decumbens and 62.5 and 125 μL/ml for Pistacia atlantica gum, respectively. The results showed that these two essential oils have a promising effect against the main microorganisms of bread spoilage. The thermal process did not significantly affect the antimicrobial activity of Pistacia atlantica gum essential oil against A. niger but significantly decreased the antimicrobial activity against B. subtilis, while in the case of antimicrobial activity of Oliveria decumbens essential oil, the results were the opposite. Considering the fact that the most spoilage agents of the bread are molds so the use of Pistacia atlantica gum essential oil is recommended as natural preservatives in products that tolerate high heat treatment, such as bread and bakery products.
Food Biotechnology
Khadijeh Shirani Bidabadi; Shilla Safaeian; Rezvan Mousavi Nadushan; Nahid Rahimifard
Abstract
IntroductionSargassum and Padina are two genera of brown algae that are widely scattered in temperate regions. Sargassum species are categorized as tropical and sub-tropical brown seaweed which are valuable sources of bioactive compounds including dietary fibers, carotenoids, vitamins, and minerals. ...
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IntroductionSargassum and Padina are two genera of brown algae that are widely scattered in temperate regions. Sargassum species are categorized as tropical and sub-tropical brown seaweed which are valuable sources of bioactive compounds including dietary fibers, carotenoids, vitamins, and minerals. These brown algae demonstrate diverse biological activities such as antioxidant, antimicrobial, and anti‑Alzheimer, due to the presence of flavonoids, triterpenoids, flavonoids, sterols, polyphenols, and pheophytine. The genus Padina is scattered in many environmental conditions, mainly in the tropical marine waters, and belongs to the family Dictyotaceae. Some bioactive components isolated from Padina species have been demonstrated hypoglycemic, hypolipidemic, anti-obesity, hepatoprotective, cardioprotective, immunostimulatory, and antimicrobial activities., The aim of this study was to prepare an extract from two species of algae Padina and Sargsum by massaging and ultrasound assisted-methods as well as analyzing their compounds and investigating the antioxidant, antimicrobial and enzymatic properties of the extracts. According to the obtained results, ultrasound assisted method was a suitable method for extraction. This extract can be used as a combination of antioxidant, antimicrobial, anti-Alzheimer's and nitrate reducing agent in food additives. Materials and MethodsChemical materials were supplied by Sigma-Aldrich GmbH (Sternheim, Germany). The algal species utilized in the current investigation; namely, Padina distromatic and Sargassum angustifolium were collected from the coastal region of Chabahar bandar, Sistan and Baluchistan Province, Iran. To eliminate all the impurities and extraneous materials, they were washed by using distilled water and then dried at ambient temperature (24-48 h) until the constant weight. Extraction by maceration was compared with the extraction using ultrasonic assisted method. Determination of chemical compounds was parformed using GC-MS device. Investigation of antioxidant properties and total polyphenol and flavonoid content were also performed. The degree of free radical scavenging was done according to DPPH method. Evaluation of antimicrobial effect of algae extracts were the main challenges in our research. S. aureus ATCC25923, Listeria innocua ATCC 33090, E. coli ATCC 25922 and S. typhi ATCC 6539 were used for antimicrobial test. Determination of minimum growth inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were performed using wells in agar.The enzymatic activity was also determined. In this study, the activity of acetylcholinesterase was investigated using the method introduced by Ellman et al. (1961) and also the activity of nitric oxidase was determined using a kit protocol (Nvand-Iran). Factorial experiment in the form of a completely randomized design was used to analyze the data. Duncan's multiple range test was used to determine the difference between the means at the 95% confidence level, and SAS software version 2.9 will used for statistical analysis. Results and DiscussionThe current study investigated the antioxidant, antibacterial, anti-inflammatory, and anti-Alzheimer’s attributes of two brown algae namely Padina distromatic and Sargassum angustifolium which were collected from the coastal region of Chabahar Bandar, Iran. The results clearly indicated that the type of algae and extraction techniques used in this investigation highly affected phytochemical compositions, antioxidant, AChE inhibitory, scavenging, and antimicrobial activities. Considering both extraction yield and phytochemical components, extraction by ultrasound assisted method provided better results. Among all algal extracts, UPE presented the highest AChE inhibitory activity, and antibacterial activity and USE presented the highest antioxidant activities, total phenols and flavonoids, reflecting the presence of various bioactive components. The extracts of two various seaweeds utilized in the current study highlighted considerable inhibitory action against four pathogenic bacteria. According to the observations of the antibacterial assay, S. aureus was the most sensitive microorganism, while L. innocua was revealed as the most resistant bacteria to the extracts of P. distromatic and S. angustifolium. Further, the chemical components responsible for the antioxidant, AChE inhibitory, and antibacterial power were confirmed by GC-MS analysis. The findings of the current investigation confirmed the potential of the health benefits and therapeutic effect of brown marine algae. Thus, the extract of P. distromatic and S. latifolium could be an effective supplement to be incorporated into the products’ formulation in the food and pharmaceutical industries as well as in medication to alleviate several disorders such as Alzheimer.
Food Biotechnology
Mahsa Noktehsanj Avval; Marzieh Hosseininejad; Abolfazl Pahlavanlo; Hamid Bahador Ghoddusi
Abstract
Introduction
Lactic acid bacteria (LAB) and their bacteriocins are widely used as natural and safe preservatives in food products, to control both pathogenic and spoilage microorganisms. This study aimed to isolate and identify LAB from several traditionally produced fermented fruits and vegetables ...
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Introduction
Lactic acid bacteria (LAB) and their bacteriocins are widely used as natural and safe preservatives in food products, to control both pathogenic and spoilage microorganisms. This study aimed to isolate and identify LAB from several traditionally produced fermented fruits and vegetables from different parts of Iran, screening their potentials for producing bacteriocin-like substances production and evaluate their antimicrobial activities against various pathogens. The effect of heat treatment and different pH values on the stability of bacteriocins were also assessed and compared with commercial nisin for their possible application in the food industry as an alternative to chemical preservatives.
Materials and Methods
Lactic acid bacteria were isolated from several fermented products like hawthorn, mixed fruit pickles containing quince and apple, mango, and medlar. pickle, and tThe isolates were identified using phenotypic (physiological and biochemical) and genotypic (16S rDNA gene sequencing) methods followed by drawing phylogenetic tree based on the neighbor-joining method. The bacteriocins were prepared from the neutralized and cell-free supernatant (CFS). To precipitate the bacteriocins, ammonium sulfate (75%), potassium phosphate buffer, and methanol-chloroform were used, and extraction was completed with a high-speed centrifugecentrifugation. After freeze-drying, the precipitate was kept as crude bacteriocin. The bacteriocin activity was measured by the critical method, and the effect of heat, storage time and pH on the stability of bacteriocins was evaluated. The minimum inhibitory concentration (MIC) and the minimum inhibitory bactericidal concentration (MBC) of the examined bacteriocins were determined on against the pathogenic strains of Escherichia coli and Staphylococcus aureus and compared with commercial nisin.
Results and Discussion
In this research, from 162 isolated strains of LAB, four isolates (10A, S6, Sa, and Ab) were selected based on the highest amount of antimicrobial compounds and diameter of the inhibitory zone against pathogenic strains. then the isolates were identified as different strains of Lactiplantibacillus plantarum (previously classified as Lactobacillus plantarum). The phylogenetic position of the isolates was determined by drawing a phylogenetic tree. The drawn tree consists of two clusters and the first cluster consist of two sub-clusters, with two different strains of L. plantarum in each of them. In the next step, bacteriocin of the isolates was extracted using saturated ammonium sulfate and high-speed (23000g) centrifugingcentrifugation. Partially purified bacteriocins from different species showed high inhibitory effects on tested indicators, which were estimated, for L. plantarum 3360 (10A) and L. plantarum lb51 (Ab), 64000 AU/ml against Staphylococcus aureus and Escherichia coli. All selected bacteriocins indicated a stable effect at different temperatures of 60 and 121°C for 20 min and 4 and -20°C for 6 months, this effect was the examined bacteriocins were also stable against at acidic and alkaline pHs too. Also, the inhibitory property decreased under very acidic (pH < 3) and very alkaline (pH > 8) conditions, but this reduction was not significant at the 95% confidence level. Bacteriocins with 64000 AU/mL activity had higher antimicrobial properties against the pathogens compared to an equal amount of commercial NiseenNisin-S (680 AU/mL). The results of MIC and MBC showed that isolates 10A and Ab have the highest inhibitory properties compared to other extracted bacteriocins and/or nisin. Since heat and chemical preservatives are used in food preparation, the stability of bacteriocins against heat and different pH is important, therefore, after extraction and purification, the extracted bacteriocins can be used as a biological preservative in the production of various food products in the range of acidic and alkaline pH, including juices, meat products, and sauces. Encapsulation of these peptides and their application in food products needs further investigation.
Food Biotechnology
Elnaz Bayat asl; Marzieh Moosavi-Nasab; Mahbobe Fazaeli
Abstract
Introduction
Wheat germ is a valuable nutritional supplement and a by-product of the flour milling industry used for animal feed and oil extraction. Quinone compounds found in wheat germ have anti-cancer properties that are abundantly found in wheat germ. The aim of this study was to investigate the ...
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Introduction
Wheat germ is a valuable nutritional supplement and a by-product of the flour milling industry used for animal feed and oil extraction. Quinone compounds found in wheat germ have anti-cancer properties that are abundantly found in wheat germ. The aim of this study was to investigate the effect of fermentation conditions on the bioactive compounds in wheat germ with anti-cancer properties. For this purpose, the Saccharomyces cerevisiae 5022 and Lactobacillus plantarum strain 1058 were used for fermentation of wheat germ under different pH levels (4.5, 6, and 7.5) over different time (24, 48, and 72h). Response Surface Methodology was used to find the optimal fermentation conditions and to investigate the effects of above-mentioned conditions on DPPH radical scavenging activity, total phenolics and dimethoxy benzoquinone (DMBQ) content. Moreover, the amounts of bio-peptides and gamma aminobutric acid (GABA) were also determined under optimum conditions.
Materials and Methods
To accomplish the fermentation process, 10 g of wheat germ was suspended in 200 mL of sodium phosphate buffer solution. Bacterial and yeast cells were then separated from the culture medium by a centrifugation at 6,000×g for 5 min at room temperature. The harvested cells were then washed with sterile phosphate buffer multiple times, resuspended in water to achieve a cell population of 108 CFU/mL, and finally homogenized using a vortex unit. The yeast and bacterial cells were incubated at 28° C and 37° C, respectively, for 24, 48, and 72 h at pH levels of 4.5, 6.0, and 7.5. Upon the completion of each fermentation process, the obtained samples were lyophilized. Total phenolic content (TPC) was measured using the method adapted by Liu et al. (2017). Briefly, the Folin-Ciocalteu phenol reagent was diluted ten times using distilled water. Subsequently, 0.1 mL of the extract was mixed with 0.75 mL of the diluted reagent. After 10 min, 0.75 mL sodium carbonate solution (2% w/v) was added to the mixture and vortexed. The absorbance was measured at 765 nm by a spectrophotometer. The antioxidant activity of fermented wheat germs was assessed using the free radical scavenging activity of the samples evaluated through a DPPH radical assay. Briefly, 2 mL of wheat germ extract was diluted with 100 mL 90% methanol aqueous solution. The methanol extract was then mixed with 4 mL of DPPH stock solution. The tube was subsequently kept in the dark for 45 min. The absorbance of each sample was then read using a spectrophotometer at 517 nm (Adedoyin et al., 2013). Dimethoxy benzoquinone (DMBQ) content was measured by an HPLC system. Briefly, 10 g of lyophilized wheat germ sample was dissolved in 250 mL of distilled water and extracted three times by shaking with 200 mL of chloroform. The chloroform layers were collected, washed three times with distilled water, and exposed to sodium sulfate solution to induce drying of the sample. The filtrate was then evaporated using a vacuum evaporator at 30° C to achieve a stable dry material. The dried sample was thereafter dissolved in the mobile phase and injected into the HPLC column to determine the DMBQ content. The HPLC system was equipped with a C-18 column and a UV detector operating at 245 nm. The mobile phase consisted of 20% acetonitrile-80% water (v/v) mixture at a flow rate of 0.5 mL/min and a temperature of 25° C.
Results and Conclusion
The highest biological activity was found when fermentation proceeded by L. plantarum under pH 6 for 48 h. Under these optimal conditions, total phenol content (3.33 mg of GAE/g), free DPPH radical scavenging (86.49%), dimethoxy benzoquinone content (DMBQ) (0.56 mg/g), peptide content (607 μg/mL) and gamma aminobutyric acid (GABA) (19983.88 mg/kg) were significantly higher than those of raw non-fermented samples. During the fermentation process, increasing the pH levels led to enhancement of antioxidant activity, total phenolic and DMBQ contents up to 48 h followed by a decline. Also, the fermentation time had a positive effect in the amount of the antioxidant activity, while it allowed an increased followed by a decrease in the contents of total phenolic and DMBQ. These findings underscore the importance of fermentation conditions of wheat germ by L. plantarum and Saccharomyces cerevisiae and can potentially serve as a promising way for the development of valuable products with anti-cancer and antioxidant functions.
Food Biotechnology
Siamak Gheibi; Amir Pourfarzad; Ahad Rastkar Allahverdizadeh
Abstract
Introduction Marinated chicken meat is one of the traditional foods in Iran which is an indoor and outdoor popular food in different areas of Iran due to its convenience and pleasant taste. It is also one of the authentic foods which is categorized in Iranian traditional kebabs. This product is ...
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Introduction Marinated chicken meat is one of the traditional foods in Iran which is an indoor and outdoor popular food in different areas of Iran due to its convenience and pleasant taste. It is also one of the authentic foods which is categorized in Iranian traditional kebabs. This product is constituted of fresh pieces of chicken meat, diced in different pieces, marinated with salt, lemon juice, verjuice, vegetables, saffron, spices, fruits and oil and then supplied as fresh and/or frozen form. After the seasoning, it is grilled using a skewer. Therefore, it is necessary to investigate its microbial load, shelf life, and quality changes during storage. To the best of our knowledge, there are no reports on the chemical and microbial properties of processed chicken meat used as a factor to predict its shelf life. Thus, the present study was designed to examine the relationship between physicochemical, microbial and sensory properties of the marinated chicken meat during storage using symmetric multivariate (determination of correlation coefficients and principal components analysis) and asymmetric methods (partial least squares regression). Materials and Methods In this study, 30 samples of marinated chicken meat were collected from restaurants. Then, the asymmetric method (completely randomized factorial and partial least squares regression) and symmetric method (coefficients of determination and Principal components analysis) were used for the study of chemical (aw, TVN, pH, O/R) and microbial (total count, Staphylococcus aureusand Escherichia coli) and organoleptic characteristics of samples on the storage days of 0, 3, 6 and 9.An aw meter was used to measure the aqueous activity (aw), an automatic Kjeldahl device was used to measure total volatile nitrogen (TVN) and a pH meter was used to measure pH and redox potential (O / R). The dilution method was used to count the total bacteria. The cooked meat and bird parker media were used to identify Staphylococcus aureus. The MPN method was used to identify Escherichia coli. Sensory evaluations were carried out by 10 trained panelists. All assessors of the internal sensory panel assumed the basic odor test and color vision test. Each sample contained 50g marinated chicken meat, which was given to panelists in plastic containers at room temperature. The color, odor and overall acceptance of the marinated chicken meat were assessed. Each evaluator randomly evaluated the samples and served drinking water before each evaluation. Each parameter was scored in a 5-point scaling ranging from 1 (lowest) to 5 (highest). The data on the physicochemical, sensory and microbial properties of marinated chicken meat were statistically analyzed using Minitab 15 software (Minitab Inc., State College, PA, USA). The mean values of the treatments were compared by Duncan's multiple range test at a confidence level of 95%. Each experiment was performed in three replications. The PCA and PLSR modeling were also performed on the datasets. Results and Discussion Results from the statistical analysis suggest that the microbial and chemical parameters of marinated chicken meat were greatly capable of predicting its shelf life. Determination coefficients, principal components analysis (PCA), and partial least squares regression (PLSR) models are able to extract relevant information and offer an easy and promising approach for the interpretation microbial and chemical properties of samples and their correlation with shelf life. The obtained results indicated that three parameters including pH, redox potential and total count had the highest correlation with shelf life, recommended as the predictive components. The results of measuring water activity in chicken meat samples during storage at refrigerator temperature showed that the aw parameter did not change significantly during storage time in the samples while the pH, TVN and O / R parameters were significantly affected. Changes in the total count, Staphylococcus aureus and Escherichia coli were also significant during refrigeration. Sensory evaluation experiments showed that all samples had no signs of spoilage until the sixth day. ConclusionThe obtained results indicated that three parameters including pH, redox potential and total count had the highest correlation with shelf life, recommended as the predictive components. Overall, we can produce a product with higher quality and shelf-life if high quality raw materials are supplied with suitable pH values and also, hygienic production is considered.
Food Biotechnology
Ehsan Safari; Hassan Barzegar; Hossein Jooyandeh; Behrooz Alizadeh Behbahani; Mohammad Noshad
Abstract
Introduction
The addition of chemical preservatives increases the shelf life of food products, but prolonged and indiscriminate use of chemical preservatives increases the resistance of microorganisms and the health risks associated with theiruptake. Medicinal plants have a wide variety in the ...
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Introduction
The addition of chemical preservatives increases the shelf life of food products, but prolonged and indiscriminate use of chemical preservatives increases the resistance of microorganisms and the health risks associated with theiruptake. Medicinal plants have a wide variety in the world as well as in Iran. In recent years, the use of natural preservatives such as plant extracts and essential oils, due to their importance and role in controlling the growth of pathogenic microorganisms, has been proposed as an alternative to chemical preservatives. Black pepper is an aromatic medicinal plant. The specific properties of black pepper essential oil, such as its antimicrobial and antioxidant activity, have also been confirmed. Amphotericin B is one of the effective antibiotics for treating infections caused by pathogenic fungi. The mechanism of action of amphotericin B is to destroy fungal cells in such a way that by binding to ergosterol in the cell membrane of fungi, it creates pores and eventually destroys them. One of the most important and common antibiotics used in the treatment of infections caused by pathogenic bacteria is chloramphenicol. This antibiotic is effective against gram-positive and gram-negative bacteria due to its broad spectrum. The aim of this study was to identify bioactive functional groups, antioxidant potential, phenol and total flavonoid compounds and to evaluate the antimicrobial activity of black pepper extract against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus coagulans and Aspergillus niger.
Materials and Methods
In this study, the antimicrobial effect of black pepper aqueous extract was investigated against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus coagulans and Aspergillus niger by disc diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum bactericidal concentration methods. Total phenol and flavonoid contents of the species were determined by Folin-Ciocalteu and AlCl3 assays, respectively. Three biochemical assays, namely 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azinobis 3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) free radical scavenging and β- caroten/linoleic acid activity systems, were used to evaluate antioxidant activity. Identification of functional groups as well as the structure of organic compounds in black pepper extract was also performed by Fourier transform infrared spectroscopy (FTIR). To evaluate the synergistic effect of black pepper extract in combination with amphotericin B and chloramphenicol antibiotics, Sub-MIC was used.
Results and Discussion
The peaks observed in aqueous black pepper extract confirmed the presence of O-H (3000-3500 cm-1), C-H (2800-3000 cm-1), C=O (1613.62-1633.52 cm-1) and C-O (100.57-1038.82 cm-1) functional groups of bioactive compounds. The total phenol and flavonoids content of the extract were 45.12 mg GAE/g extract and its flavonoid content was 29.66 mg QUE/g extract which had an important role in its antioxidant activity. The aqueous black pepper extract had remarkable DPPH free radical scavenging activity (IC50=32.37 μg/ml), ABTS free radical scavenging activity (IC50=28.45 μg/ml) and beta-carotene bleaching inhibitory effect (46.45%), revealing the electron/hydrogen donating ability of the essential oil. The results of measuring the antimicrobial activity of extract by disk diffusion and agar well showed that black pepper extract showed more antimicrobial effect on gram-positive bacteria Staphylococcus aureus and Bacillus coagulans than gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa. This could be due to the difference in their cell wall structure. Aspergillus niger is the most sensitive species to aqueous black pepper extract. The minimum inhibitory concentrations of extract for Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus coagulans and Aspergillus niger were equal to 32, 16, 4, 8 and 4 mg/ml, respectively. The minimum bactericidal concentration of black pepper extract for two bacterial species, Escherichia coli and Pseudomonas aeruginosa was more than 512 mg/ml. Also, the minimum bactericidal concentration for Staphylococcus aureus and Bacillus coagulans was 128 and 256 mg/ml, respectively, and 128 mg/ml for Aspergillus niger. The results of interaction of black pepper extract with chloramphenicol antibiotic showed that the Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were synergistic, but antagonism was observed for the gram-positive Bacillus coagulans.
Food Biotechnology
Sara Shahryari; Alireza Sadeghi; Maryam Ebrahimi; Alireza Sadeghi Mahoonak; Ali Mayedi
Abstract
[1]Introduction: Evaluation of antimicrobial and probiotic properties of the microbiota isolated from fermented pseudo-cereals is important in order to prepare adjunct and starter cultures. Probiotics are live and active microorganisms that, if used in sufficient numbers, help the microbial balance of ...
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[1]Introduction: Evaluation of antimicrobial and probiotic properties of the microbiota isolated from fermented pseudo-cereals is important in order to prepare adjunct and starter cultures. Probiotics are live and active microorganisms that, if used in sufficient numbers, help the microbial balance of the gastrointestinal tract and improve its function. Probiotics are also used as a substitute for antibiotics and synthetic preservatives in the prevention and treatment of complications of many infectious and pathogenic gastrointestinal pathogens. Probiotic microorganisms include lactic acid bacteria (LAB) and yeasts, and despite numerous reports on the probiotic properties of LAB, the probiotic properties of yeasts are less reported. Probiotic yeasts are organisms resistant to antibacterial compounds and effective against pathogens, which can rapidly increase their population in the gastrointestinal tract. These yeasts have several properties such as antimicrobial effects, resistance to acid and bile, binding to mucosal surfaces, inhibitory activity against pathogens and also the inability to transmit antibiotic resistance genes. In the present study, after isolation of the predominant yeast from buckwheat sourdough, the probiotic and antifungal properties of the isolate were investigated. Based on the literature review, no study has been presented to evaluate the probiotic and antifungal capabilities of yeasts isolated from buckwheat sourdough. Materials and Methods: In the present study, predominant yeast was isolated from buckwheat sourdough, and then it was identified using PCR. Subsequently, probiotic properties of the isolate including survival in present of low pH and bile salt, antibacterial effect, antibiotic susceptibility assay, aggregation, hydrophobicity, haemolytic activity as well as its antifungal activity against Aspergillus niger and Aspergillus flavus were studied. After spontaneous fermentation of buckwheat, predominant yeast was isolated using ten-fold dilution of the sourdough sample and its spread plating. The predominant isolate was identified through PCR amplification of a 650 bp target sequence from its ribosomal gene and sequencing of the PCR product. Then, survival of the yeast was determined at pH=2 and 0.3% bile salt as simulated gastrointestinal conditions. Subsequently, simultaneous culture of the yeast with some food-borne indicator bacteria in chromogenic media was used to investigate the inhibitory activity of the isolate against the studied bacteria. After that, resistance of the yeast isolate against the common antibiotics and some antimycotic agents was evaluated using disc method. Co-aggregation ability and hydrophobicity capability of the isolate were also determined based on the absorbance tests. In vitro safety of the yeast isolate was checked through its cultivation on blood agar containing sheep blood. Next stage, overlay bioassay was conducted to investigate antifungal effect of the yeast on the selected fungi. Finally, one way analysis of variance (ANOVA) with the least significant differences (LSD) post hock at p<0.05 was used for statistical analysis of the data. Results and Discussions: Sequencing results of the PCR products led to the identification of Pichia kudriavzevii as predominant yeast isolated from buckwheat sourdough. Survival rate of the isolate after treatment in simulated gastrointestinal conditions was 79.26% in comparison with the control. Antibacterial effect of the isolate on Escherichia coli was also significantly (P<0.05) higher than the other studied food-borne agents. Meanwhile, the growth of Listeria monocytogenes was decreased 19.50% in the present of the isolate. Whereas, the yeast isolate had no inhibitory effect on Salmonella enterica. Hydrophobicity and auto-aggregation capabilities of the isolate were also 64.07 and 67.40%, respectively. Furthermore, the isolate showed resistance towards all of the antibiotics tested, while it was resistant against ketoconazole and fluconazole, and the yeast was semi-sensitive towards itraconazole as antimycotic agents. The yeast isolate had no hemolytic activity, and its antifungal activity against A. niger and A. flavus was also verified. Accordingly, it is concluded that P. kudriavzevii isolate exhibits proper potential to be used as probiotic or protective culture in fermentation industries.In accordance with the results, probiotic characteristics of P. kudriavzevii (as the predominant yeast isolated from buckwheat sourdough) were approved. Accordingly, the isolate can be used as a potential probiotic culture in food industries.
Food Biotechnology
Kobra Tajik Toughan; Mohammad Reza Edalatian Dovom; Seyed Ali Mortazavi; Ali Javadmanesh
Abstract
[1]Introduction: Poultry and meat products are the largest sources of non-typhoid salmonella infections in most countries. Studies have shown that raw foods of animal origin, especially poultry and its products, are the main source of contamination of kitchens and restaurants. In terms of growth conditions, ...
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[1]Introduction: Poultry and meat products are the largest sources of non-typhoid salmonella infections in most countries. Studies have shown that raw foods of animal origin, especially poultry and its products, are the main source of contamination of kitchens and restaurants. In terms of growth conditions, these microorganisms are resilient bacteria and easily adapt to their environmental conditions. Salmonella has been known to cause intestinal disease for many years and has been reported as the most important cause of food poisoning. According to Iranian and international standards, there should be no S. enteritidis or S. typhimurium in 25 grams of food. DNA-based methods for the identification and differentiation of Salmonella serovars have been designed and applied using specific primers at the genus and serovar levels. Therefore, they can be used as useful and rapid screening tests, as well as to supplement or replace conventional biochemical and serological tests. Real-time PCR, with the most accurate and reliable results using a fluorescence probe, which of course has a high cost. In this method, sequence specific fluorescence probes are used, and as a result, in the target molecule, screening and determination the presence or even the concentration of specific sequences is possible. Therefore, even in the presence of other types of nucleic acid molecules, the results are obtained quickly and have a high level of specificity. Under these conditions, if specific probes with different florescence dyes are used, even multiple targets can be detected in a single PCR reaction. The aim of this study was to identify S. enteritidis or S. typhimurium by PCR and Salmonella spp. by real time PCR method in poultry products. Material and Method: In total, 45 samples of poultry products, including chicken breast, liver and gizzard (15 samples each) were purchased from different regions of Mashhad and from various companies and transferred to the laboratory in accordance with hygienic standards. For each sample, 25 g of tissue was isolated and homogenized under sterile conditions and DNA extraction was then performed using a DNA extraction kit. The extracted DNA was evaluated by agarose gel electrophoresis. The purity and quantity of DNA extracted from each sample was examined by spectrophotometry method. In the next step, in order to identify the genus Salmonella, the samples were examined by real time PCR. In this method we used an internal control to ensure that negative results are not false negative due to inhibitors. The results of real time PCR showed that out of 45 samples, nine samples were infected with Salmonella. Then, these nine samples were evaluated for Salmonella typhimurium and Salmonella enteritidis infection by conventional PCR method. Result and Discussion: The results showed that out of nine samples that were positive in real time PCR test, seven samples were contaminated with Salmonella typhimurium, of which five samples were related to chicken breast and two to liver. Regarding Salmonella enteritidis infection, out of nine samples, only one sample was contaminated, which was related to chicken breast. Conventional methods have been traditionally used to enumerate target bacteria in food. However, these methods have some limitations and require considerable time and labor. Previous studies have already shown that real time PCR is more effective than conventional bacteriological methods for the detection of Salmonella spp. In a study by Whyte et al. (2002) The presence of Salmonella was assessed by traditional culture methods and by a Salmonella-specific polymerase chain reaction (PCR) test. Salmonella was recovered from 16% of samples using traditional culture methods. In contrast, the PCR assay proved to be more sensitive and detected Salmonella DNA in 19% of the examined samples (Whyte et al. 2002). Results of PCR with specific primers showed that reactions in real time PCR with general primers of Salmonella spp. were done correctly. Despite of accuracy and speed of real time PCR to detect DNA of microorganisms, further studies are developed to have more advantages. Loop-mediated isothermal amplification (LAMP) showed a higher sensitivity of Salmonella detection in compare to qPCR (Vichaibun & Kanchanaphum, 2020). Although LAMP could detect trace amount of Salmonella DNA but primer design for this reaction is very difficult. However, it is important to highlight that non-viable cells can be detected by real time PCR or other DNA-based methods, which does not occur in traditional methods of culture and isolation that require viable cells for quantification (Zeng et al., 2016).
Food Biotechnology
Behrooz Alizadeh Behbahani; Mohammad Noshad; Mostafa Rahmati-Joneidabad
Abstract
[1]Introduction: Oxidation and food pathogens are considered two important and influential factors affecting food quality and health. Recently, due to the increasing demand for natural products, the application of synthetic preservatives to control microbial growth and lipid oxidation have been decreased ...
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[1]Introduction: Oxidation and food pathogens are considered two important and influential factors affecting food quality and health. Recently, due to the increasing demand for natural products, the application of synthetic preservatives to control microbial growth and lipid oxidation have been decreased significantly. Therefore, natural antioxidant and antimicrobial compounds are receiving more attention in food preservation technologies. In the last 2 decades, the use of herbal medicines rich in bioactive molecules (including polyphenols, carotenoids and flavonoids) with medicinal and health effects such as delaying the onset of some diseases such as cardiovascular disorders, diabetes, and cancer have increased. Furthermore, secondary metabolites in plant extracts and essential oils are able to control and inhibit free radical-mediated reactions. The olive tree (Olea europaea) is an evergreen plant that grows in tropical and subtropical regions. Iran is one of the most important olive growers in the world due to its suitable conditions for olive cultivation. The leaves of the olive plant have a high potential for the production of various products such as tea and extracts. Olive leaf extract can be used as a raw material in the production of various products, due to exhibiting various biological activities such as antimicrobial and antiviral activity, lipid stabilizer, blood pressure regulator, antioxidant activity, and free radical scavenger. The leaves of the olive tree also contain various phenolic compounds, mainly Oleuropein and hydroxytyrosol, with antioxidant and antimicrobial activities. Therefore, in this study, the amount of phenolic and flavonoid compounds of olive leaf ethanolic extract and its antioxidant effect and antimicrobial properties on Escherichia coli, Enterobacter aerogenesis, Bacillus cereus and Listeria innocua were investigated. Materials and Methods: The olive leaf ethanolic extract was prepared through maceration method and its total phenolic content (Folin-Ciocalteu method), total flavonoids content (aluminum chloride colorimetric assay), antioxidant activity (ABTS and DPPH free radical scavenging methods), and antimicrobial effect on E. coli, E. aerogenesis, B. cereus and L. innocua (based on disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum bactericidal concentration) were determined according to standard methods. Data were analyzed by SPSS software through one-way ANOVA and Duncan test at p<0.05. Results and Discussion: The ethanolic extract of olive leaves contained 176.58 ± 0.72 mg GAE/g total phenol and 69.85 ± 0.26 mg QE/g total flavonoids. In addition, ethanolic extract of olive leaf was able to inhibit free radicals DPPH (70.62 ± 0.59%) and ABTS (76.15 ± 0.43%). The antimicrobial results showed that the antimicrobial effect of the extract depended on its concentration and type of bacteria. Antimicrobial effect was increased as a function of ethanolic extract, and Gram-positive bacteria (B. cereus and L. innocua) were more sensitive to ethanolic extract of olive leaf than Gram-negative bacteria (E. aerogenesis and E. coli). Generally, B. cereus and E. aerogenesis were the most sensitive and resistant microbial strains to ethanolic extract of olive leaf, respectively.The results of this study showed that the high antioxidant and antimicrobial activity of olive leaf ethanolic extract is mainly due to its phenolic and flavonoid compounds. Olive leaf ethanolic extract was able to neutralize DPPH and ABTS free radicals. Also, Gram-positive bacteria were more sensitive to ethanolic extract of olive leaf than Gram-negative bacteria. In general, the ethanolic extract of olive leaf can be used as a nutraceutical to control or prevent the growth of spoilage/infection-causing microorganisms and free radical reactions in food and the human body. However, more in-depth studies are needed to determine the mechanism of antimicrobial and antioxidant effects of olive ethanolic extract in vitro and in vivo.
Food Biotechnology
Mostafa Rahmati-Joneidabad; Behrooz Alizadeh Behbahani; Mohammad Noshad
Abstract
[1]Introduction: Economic losses can occur due to the growth of fungi on foods that lead to food spoilage and plant diseases. Fruits and vegetables are often exposed to microbial activity, caused by pathogenic fungi, during post-harvest storage. Diseases of food origin are a growing public health problem. ...
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[1]Introduction: Economic losses can occur due to the growth of fungi on foods that lead to food spoilage and plant diseases. Fruits and vegetables are often exposed to microbial activity, caused by pathogenic fungi, during post-harvest storage. Diseases of food origin are a growing public health problem. Thus, food safety has become a major public concern as microbial contamination increases the risk of foodborne illnesses and shortens the shelf life of foods. Infection with fungi such as Aspergillus, Rhizopus, and Penicillium species is considered as the primary cause of rapid spoilage of fresh produce, which reduces their quality and shelf life. Synthetic fungicides have been applied to solve this problem for many years. Nonetheless, the adverse effects of synthetic chemicals on human health and the emergence of fungicide-resistant strains have motivated the scientists and food industries to find out safe preservatives to control postharvest rot/diseases. On this point, natural antimicrobial agents such as plant extracts and essential oils are gaining more and more interest. In this study, we used Levisticum officinale Koch essential oil, which its antimicrobial and antioxidant activity has been reported in literatures. Materials and Methods: L. officinale Koch essential oil was obtained by hydrodistillation method and its total phenol content, total flavonoids, antioxidant activity (based on DPPH and ABTS free radical scavenging and β-carotene bleaching tests) and its antifungal effect against fungi causing apple and orange rotting (Alternaria alternata, Penicillium expansum, Penicillium digitatum, Penicillium italicum, and Botrytis cinerea) were examined according to antimicrobial tests of disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum fungicidal concentration. Results and Discussion:L. officinale Koch essential oil contained 61.27 ± 0.34 mg GAE/g and 20.14 ± 0.21 mg QE/g total phenol and flavonoids, respectively. Its antioxidant activity, based on DPPH free radical scavenging, ABTS free radical scavenging, and β-carotene bleaching inhibition were 69.72 ± 0.65%, 78.54 ± 0.3% and 57.50 ± 0.41%, respectively. L. officinale Koch essential oil was effective against all fungal species and the highest susceptibility was observed for Penicillium expansum. According to the results, L. officinale Koch essential oil can be used as a natural antifungal agent to prevent post-harvest diseases of fruits and vegetables.
