Introduction & Objective: Non-Alcoholic fatty liver (NAFLD) is defined as a spectrum of clinical scenarios which is pathological deposition of fat droplets in the liver of patients who have no history of Alcohol use. This study compared the effect of low calorie diet with and without sibutramine on body weight and liver function in patients with NAFLD.Materials & Methods: This clinical trial study was conducted in 2010 at Tabriz University of Medical Sciences, on 40 obese patients with non-Alcoholic fatty liver. Patients were randomly divided into two equal groups of intervention and control groups. Group one received 15 mg daily sibutramine capsules half an hour before lunch and a weight loss diet based on ideal body weight. The other group only had diet control for weight reduction. Before and after 3 months of intervention, weight changes, fasting glucose, glycosylated hemoglobin HbA1c, levels of liver enzymes and ultrasound evaluation was repeated. Data were analyzed using the SPSS software and the paired T test, Mann-Whitney and McNemar test.Results: The mean age of the subjects was 38.90±7.00 in the sibutramine group and 36.55±7.87 for the control group. After three months, the average weight loss in sibutramine group was significantly more than the control group (sibutramine group13 kg and control group 4 kg (p<0.05). Improvement in liver echogenicity in sibutramine patients was 90% and 50% of diet group patients. ALT changes in the sibutramine group and control group was 7.50±15.11 and 6.15±28.23 respectively, which was statistically significant in the sibutramine group. AST changes were 4.38±13.37 and 1.70±18.37 in sibutramine and control group respectively. The changes were not statistically significant.Conclusion: Overall, findings of this study suggest that sibutramine is effective in liver function improvement and treatment of NAFLD patients.

Introduction: Wetland ecosystems, especially marine coastal wetlands of the most important and also the most vulnerable are the worldchr('39')s environmental resources. Which has always been sensitive to the fragility of coastal areas, high population density and intensive human activities are faced with the threat of destruction. Based on this, monitoring the trend of the changes in wetlands and their surrounding lands can be effective in the management of these valuable ecosystems. Investigating the environmental risk is a suitable instrument for evaluating and ensuring understanding of the relationships between stressor factors and environmental effects especially in wetland ecosystems. In general, application of methods of evaluating environmental risk is one of the important tools in studying environmental management along with identifying and mitigating potential environmental damaging factors in wetland regions in order to achieve sustainable development. Today, multi-criteria decision-making methods are employed in evaluating the risk in many studies. This study is based on multi-criteria decision-making methods to identify and analyze the risks threatening Tyab-Minab International wetland located in Hormozgan province was conducted. Materials and methods: Based on the methodology to identify and prioritize risks Delphi, AHP and TOPSIS techniques were used to determine the risk priority number. In the first phase of this study, to identify and screen the main criteria of project selection, Delphi method was used. In this study, the panel of interest was determined based on a combination of experts with different expertise and out of a sample of 20 individuals, in which experts with various expertise gave a score from 1 to 5 (Likert scale) to each criterion. In this way, 32 criteria were identified as the most important and considerable risk for Minab Wetland and further proceeded to the second phase for prioritization and analysis. In this stage, multi-criteria decision-making methods were used, in which hierarchical analysis process was employed for prioritizing the criteria using Expert Choice 11 software. The indices of risk evaluation including the impact intensity, incidence probability, and the sensitivity of the receptive environment in environmental risk evaluation of wetlands do not have an equal value and significance. For this purpose, to weight the factors effective in estimating risk level and for prioritization of risk options, the technique for order of preference by similarly to ideal solution (TOPSIS) and Excel software were benefited from for calculations. The spectrum of scoring to each of the indices of incidence probability, impact intensity, and the sensitivity of the receiving environment was chosen from very low (1) to very high (9) based on hour spectrum. Following investigation of the types and frequency of indices along with the method of score determination of these indices, three indices of risk intensity (C1), risk incidence probability (C2), and the sensitivity of the receiving environment (C3) were chosen for risk ranking using TOPSIS model. Next, after determination of risk priority number using TOPSIS, the risk levels were calculated and evaluated using normal distribution method for each risk. To determine the degree of risk-taking, risks are organized in a descending order, where the elements of the number of the class and the length of the class are determined based on Relations 1 and 2 (n is the number of risks). Next, the risks are categorized based on these classes. Considering the concept of ALARP, the risks under investigation are divided into high risks, medium risks, and low risks. In this study, considering the number and length of classes, the studied risks were categorized in six levels (critical, intolerable, considerable, medium, tolerable, and trivial risks). (2) (1) the number of classes=1+3. 3 log (n) the length of the classes= the greatest risk value-the smallest risk value/the number of classes Results and discussion: In the first step, the final indices of the wetlandchr('39')s environmental risk were identified and the development of hierarchical tree and classification of the risks threatening wetlands along with their incidence probability in two groups of natural and environmental criteria was performed. Eventually, the final weight of criteria resulting from paired comparisons was obtained in Expert Choice 11 to achieve the score of incidence probability of each risk. Based on the results, among the natural, social, economic, physiochemical, biological, and cultural criteria, drought and climate change, increase urban and rural development, Smugling of fuel, oil pollution, reduce the density of vegetation, indiscriminate exploitation of groundwater were of high priority. The results obtained from ranking the the risks threatening Minab Wetland using TOPSIS suggest that oil pollution, dam construction upstream, persistent drought and climate change, and sometimes Alcohol and fuel smuggling and illegal overfishing the priorities are first to fifth. Also Results showed that the respectively based on (Cj+) oil pollution (0/9109), dam construction (0/8121), the drought and climate changes (0/8063) and the smuggling of fuel (0/7520) are in Unbearable level. Overall, the results indicated that same as this research, wetland ecosystems are subject to many threatening factors, resulting in ecological imbalance and abnormal appearance of the wetland, putting the wetland entity into danger of extinction in terms of fauna and flora. Conclusion: Nowadays, for assessment of environmental risk, various methods are used, each of which has positive and negative points given the studied environment and the conditions governing it. Therefore, one cannot reject or approve one method with total confidence. By employing novel methods in risk evaluation, the intensity of risk incidences and, in turn, the damages and losses incurred to the environment can be prevented or at least mitigated. Further, it is also possible to move in line with proper and optimal management of environmental resources, especially wetlands and with sustainable development. Undoubtedly, understanding and recognition of the factors threatening wetlands, according to the importance and the impact of them, Prevent and cope with the threats and accurate project preparation and implementation of wetland conservation plans and environmental management.

Background & Aims: Breast cancer is one of the most common cancers today and how to deal with it is one of the challenges for oncologists today (1). One of the new treatment strategies is the use of nanotechnology in the field of cancer (1, 2). Nanoparticles are the most common elements in nanoscience and technology, whose interesting properties have led to a wide variety of applications (5). In this regard, graphene quantum dots (GQDs) can be used as an excellent option for nanomedical fields such as the release of anti-cancer drugs, cell culture, and tissue engineering (7, 8). Considering the role of nanoparticles as well as the antitumor effects of graphene compounds, it seems the use of these nanoparticles in 3D culture can be one of the useful strategies in suppressing cancer cells with fewer side effects (9). Therefore, the aim of this project was to compare the apoptotic effects of graphene quantum dots on the MCF-7 breast cancer cell line cultured in hydrogel scaffolds. Methods: In this study, the MCF-7 breast cancer cell line was used as cancer cells. For cell culture in fibrin hydrogel scaffold, M199 medium containing 10% FBS serum and 1% penicillin/streptomycin and fibrinogen powder with a concentration of 3 mg/ml was used and it was added into each plate well of 24 wells and then cell suspension was added. Then, for 500 μ, l of scaffolding and cells, 15 μ, l of thrombin with a concentration of 120 u/ml was added in each well and it was immediately gently aspirated until the culture medium became gelatinous and the cell culture plate was transferred to the incubator. After 2 hours of cell incubation in the incubator (Sina, Iran), On the gelatinous scaffolding of each 24 well plates, 500 μ, l of M199 medium containing 10% FBS serum and 1% antibiotic was added and transferred to the incubator again. Cell treatment was performed by first removing the culture medium from each well and concentrations of 1, 5, 7, and 10 mg/ml of graphene quantum dots were added. But only the culture medium was added to the control group. The plate was transferred to the incubator and treated for 1, 3, and 5 days. In this study, MCF-7 cells were cultured on a fibrin hydrogel scaffold for SEM imaging and after 24 hours of incubation, the medium was gently removed on each well. 300 μ, l of PBS was added to each 24-well plate and wash three times and 300 μ, l of 2. 5% glutaraldehyde solution was added to each well and maintained at room temperature for 2 hours. Dehydration was done with increasing concentration of Alcohol (30, 50, 70, 80, 90, 100, 100%) and then close the door of the plate and maintained in the freezer for 2 hours and freeze it to dry and dehydrate the fibrin hydrogel. MTT method and acridine orange and DAPI stains were used to measure cell survival and morphology. One-way ANOVA and t-test were used for statistical analysis. The chart was drawn in Excel 2016. P<0. 05 was considered a significant difference for samples. Results: The results of scaffold morphology study using SEM electron microscope, showing the desired mechanical properties and suitable porosity for the scaffold. Therefore, the presence, growth, proliferation, and proper adhesion of cells to the scaffold and the natural morphology of the cells were confirmed. The results of the MTT test showed that the viability and survival of MCF-7 cells after exposure to the mentioned doses of graphene quantum dots were recorded (98, 81, 50, and 31%) respectively. This indicates that the viability of MCF-7 cells is significantly dependent on the concentration of graphene quantum dots. IC50 concentration of GQDs affecting MCF-7 cells was determined to be 7 mg/ml and after 1, 3, and 5 days decreased in a time-dependent manner so that after 24 hours, the average cell survival decreased to 50%, after 3 days to 35% and after 5 days to 22%. These data also showed the cytotoxic effect of IC50 concentration of graphene quantum dots on cells in a time-dependent manner. Also, the results of acridine and DAPI staining in GQDs-treated groups showed compact, fragmented, single, and orange nuclei, which indicated the onset of apoptosis in MCF-7 cells being treated. While in the control group cells that did not receive GQDs, the nuclei were clear, round, large, and in the form of light green clusters, which was a sign that the control group cells were alive. Conclusion: In this study, the effects of graphene quantum dots on MCF-7 cancer cells cultured on fibrin hydrogel were investigated. The results showed that GODs had a cytotoxic effect on MCF-7 cancer cells, so these effects were dependent on the time and dose in the experiment. In fact, with increasing concentration and time, the toxicity of graphene quantum dots increased. In this study, the IC50 for GODs was calculated to be 7 mg/ml by MTT test. Examination of morphological changes with an inverted microscope and also, qualitative studies of the nucleus characteristics of control and treatment cells confirmed the induction of apoptosis in the treated cells. In addition, the results of acridine orange staining were another confirmation of these findings. As mentioned, graphene and its many derivatives, as well as graphene quantum dots, which is one of the newest products of graphene, have been used for valuable research. According to the results of this study, GQDs have a toxicity effect and induce dose-dependent and time-dependent apoptosis in cultured breast cancer cells on fibrin hydrogel. However, more studies are needed to determine the molecular mechanism involved. In addition, it is suggested that the effects of GQDs on other cancer cells and normal cell lines be investigated and compared.

Introduction: Edible coatings and films are thin layers of edible compounds and biopolymers that are placed on the surface of food or among its compounds and their use is one of the methods. They are important for controlling physicochemical, microbial and physiological changes in food. One of the main functions of food packaging is to prevent or minimize the transfer of moisture between the food and the surrounding air. Therefore, in order to optimize the food environment and increase the shelf life of the food, water vapor permeability should be as low as possible. On the other hand, about 125 million tons of plastic are produced in the world, of which about 30 million tons are consumed in the packaging sector. Pollution from packaging materials produced from petroleum derivatives and problems caused by various methods of decontamination (such as burial, incineration and recycling) have attracted the attention of researchers in recent years to find suitable alternatives. For this type of packaging, it has been pointed out that films and edible coatings can be the best alternatives in this regard. A new generation of food packaging includes substances with antimicrobial properties. This packaging technology can play an important role in increasing the durability of food and safety. Metal nanocomposites with a combination of metal nanoparticles and polymer film have recently received a lot of attention. Among polymers, natural biodegradable polymers have been widely used that have advantages such as: edible, biocompatibility, attractive appearance, non-toxic, non-polluting and low cost. Among natural polymers, chitosan, as a cationic biopolymer, has been widely studied for various applications due to its excellent bioactivity and solubility in the aquatic environment and its resistance to oxygen penetration. As a result, it now has a variety of potential applications in medical products, food packaging film, bone replacement and artificial skin. Among metallic nanoparticles, silver nanoparticles have long been known as antimicrobial substances. The antimicrobial activity of these nanoparticles may be related to several mechanisms that destroy the cell membrane structure. The aim of this study is to prepare a suitable packaging using Chitosan-Silver film and to strengthen its antimicrobial and physicochemical properties by adding cumin extract. Cumin cyminum is a plant of the Apiaceae family, annual, fragrant, hairless, herbaceous stem with bifurcated and sometimes trifoliate branches. The stem of the plant is grooved and has a peripheral metropolitan texture. Green cumin fruit contains 2-5% essential oil, most of which consists of paracetamol, alpha and beta-p-nen, comic Alcohol, comic aldehyde, alpha and beta-flandren, eugenol, prialaldehyde, alpha-terpineol and myrison. Material and methods: In this study, the physicochemical and microbial properties of silver nanocomposites (Ag) and chitosan (Ch) containing cumin extract (E) were investigated. Initially, cumin extract was extracted by extraction and extraction and water / ethanol extraction methods. The extract was tested for total phenol, tocopherol. The film is made of chitosan / silver nanocomposite (0. 15 mol / liter) with three concentrations of 0. 2, 0. 4 and 1% cumin extract and is subjected to physicochemical tests (thickness measurement, water vapor passage rate, film solubility, resistance Stretching, scanning electron microscopy (SEM) and increasing the length of the film at the moment of rupture) and microbial (diameter of non-growth aura) were included. All experiments were performed in a completely randomized trial in three replications. The averages were compared with SPSS 23 software based on Duncan's tests at five percent. The resulting graphs were plotted in 2013 excel software and compared. Results: The results showed that the extract of green cumin extracted by masonry method had high phenolic compounds (74. 17 mg / g galicacid / g dry weight) and tocopherol (24. 05 mg alphatochofrol / g dry weight). The results of the experiments showed that with increasing cumin extract up to one percent of film thickness (0. 93 mm), water vapor permeability rate (3. 81 g / s. m2, film solubility (26. 45%) and increasing film length in The moment of rupture (30. 21%) showed a significant increase (P <0. 05), but the tensile strength of the film (16. 10 MPa) decreased with increasing concentration. As can be seen, in general, the addition of cumin extract causes Decreased tensile strength of bituminous / silver biocomposite films has been reduced. The results at 95% probability showed that the addition of the extract to the film formulation significantly reduced the tensile strength and continued to increase with increasing the concentration of the extract. The lowest tensile strength of the film was observed in 1% cumin extract. The images of the electron microscope show that the surface of the control films is perfectly smooth and smooth and free of any particles, while at the level of films containing nanoparticles Nano-silver particles are clearly visible. Also, no accumulation and accumulation of nanoparticles is observed at the film surface and they have a uniform distribution. The results of the microbial test are not clear. As the concentration of cumin extract increased by up to one percent, the antimicrobial properties of chitosan / silver nanocomposite increased. > Ashrashia has been general. Conclusion: Chitosan / Silver Biocomposite as a biodegradable film with high physical properties has a high capability as a coating. Cumin extract is also rich in antioxidants. The composition of cumin extract has high antioxidant properties as well as high antimicrobial activity. Cumin extract in combination with cumin extract, it can be used as a coating to cover food. The effect of oral coating of chitosan / silver biocomposite with cumin extract was studied in terms of physicochemical and microbial properties. At the end of the results, it was shown that the film has a high performance for covering and storing food. Therefore, the results of this study can be used to cover food. The results showed that with increasing the percentage of extract, the antimicrobial properties of the prepared films increased. In all antimicrobial tests, chitosan / silver biocomposite film containing 1% extract had the highest inhibitory or non-growth diameter.

Introduction: Sugar beet is adaptable to be cultivated in different weather conditions and regions. In 2015, the overall area for sugar beet cultivation was estimated about 105, 000 hectares which 19000 of this was in Khorasan Razavi province. Sugar beet is considered as a valuable agricultural crop both for economy and employment. Nevertheless, producing this crop faces many challenges including the high number of pests. One of these pathogenic factors is the nematodes. Among plant-parasitic nematodes cyst nematodes are a large group with economic importance in different countries. These nematodes cause much damage to agricultural crops. Among the different cyst nematode genera Globodera and Heterodera have species which are important due to economic damage. Materials and Methods: During the year of 2016, 22 samples of soil and roots of sugar beet in cyst nematodes contaminated field in Khorasan Razavi were gathered. Cyst nematodes were extracted by the use of a small clip and a binocular and put in petri dish with some water. White females were also taken from the root by a delicate needle and put in sterile distilled water after washing. Separating and purifying fungi were done in 3 parts: separating fungi from cysts and females, separating fungi from eggs and larvae nematode, and making single spore fungi and pure culture. The cysts and separated females were washed by distilled water for several times and antisepticised for 1, 2 or 3 minutes in 10% Sodium Hypochlorite, and 10% and 20% Ethyl Alcohol. Cysts and females were washed again with sterile distilled water and sterile sifter was used. The cysts and females in petri dish containing PCA, PDA, CMA and MEA culture mediums were separately taken and cultured by a needle under laminar. Four cysts were placed at the 4 sides of each 8cm petri dish containing the above mentioned culture mediums. 16 brown cysts from each soil sample were cultured. Two petri dishes from each sample were kept and checked consistently in two hot and cold temperatures in incubator with 20-25 and incubator with 8-10. After 7-14 days the grown fungi were taken to a new culture medium of PDA for a better development and in later stages they were purified on WA culture medium by single spore or hyphal tip methods. Purified fungi were kept in test tubes containing PCA food environment and 4 temperature and also on sand for later studies. Cyst shells were destructed by cyst crusher (homogenizer) and their eggs and mash were released. Released eggs from cysts were formed into suspension in sterile water. 0. 5-1 ml of suspension of eggs and larvae were taken by sterile micropipette and diffused on petri dish containing water agar culture medium. These culture mediums were kept and checked regularly in dark in the incubator with 20-25. Grown fungi from these eggs and larvae were taken to a new culture medium and purified by single spore and hyphal tip methods. WA culture medium was used for single spurring and purifying fungi. PCR based methods: morphological and molecular identification, were used to identify the fungi isolates. Mycelium fungi growth phases, fungi DNA extraction, PCR reaction and electrophoresis were done to identify molecular fungi. Lipase and chitinase assays were performed on the isolates. Results and Discussion: For Lipase test, each fungal isolate was cultured on peptone agar media and after 7 days isolates were examined. Around the colony of the isolates produced by the lipase enzyme formed a precipitate or a colorless aura, which is due to the formation of calcium salts from free lauric acid by the lipase enzyme, which indicates the positive activity of lipase. Among the isolates, Colletotrichum gloeosporioides had the highest sediment content, indicating the activity of lipase enzyme and the highest isolate in this test. After that, Neonectria macrodidyma and Penicillium chrysogenum showed the highest activity of lipase enzyme activity. Fusarium oxysporum showed no sediment and the lowest level of lipase enzyme activity was observed in this fungus. For chitinase test, isolates were cultured on colitic kitein medium. Around the colony and also the color of the culture medium, the chitinase-producing isolates changed the violet color, which indicates the positive activity of the chitinase enzyme in these isolates. Relative analysis of chitinase activity showed that the isolate of P. chrysogenum had the largest and fastest change in color to violet, indicating the highest production of chitinase enzyme by this fungus. The lowest chitinase production was by C. gloeosporioides, which showed the slightest and slowest changes in color compared with other isolates and the weakest isolate was introduced for the production of chitinase enzyme. Conclusion: In this research, different fungi were isolated from sugar beet cysts. Most fungal isolates belonged to Torbat-e-Haidiriyah and Fariman, and the least isolates belonged to Khaf. The highest frequency of Fusarium isolates was found to be 37. 35%. Isolation of Simplicillium lanosoniveum fungi, P. chrysogenum, C. gloeosporioides was the first reported cyst nematode in sugar beet. In the relative analysis of lipase activity, it was found that P. chrysogenum and C. gloeosporioides fungi exhibited the highest amount of lipase production, which was the highest marker in this test. Relative analysis of chitinase activity showed that P. chrysogenum had the largest and fastest color change to purple, indicating the highest production of chitinase enzyme by this fungus. The lowest chitinase production was obtained by isolate C. gloeosporioides, which showed the slightest and slowest changes in color compared with other isolates. The fungus P. chrysogenum showed the best results in both tests, this fungus is, therefore, recommended for further research with the observation of the necessary points.

12Introduction: Today, the problem that the beverage industry faces and is largely unchanged, and may be added to its complexity day by day is to provide a healthy, durable, and acceptable product. One of the main steps in this regard can be replacing the preservatives and chemical additives with their natural varieties. Since Iran is very diverse and rich in vegetation, especially in medicinal plants, due to its special geographical conditions, and on the other hand, since the medicinal plants have the known antimicrobial and antioxidant properties, they can be used as a substitute for the chemical preservatives in the beverage industry. In this regard, the possibility of production of saffron gaseous beverage has been studied in this research. Materials and methods: Initially, in order to produce the saffron gaseous beverages, the raw materials including saffron extract (Zardband Company), Sugar (Hedieh Company), Orang Serum Agar, Kant Agar Plate, Dichloran Rose-Bengal, Sodium Benzoate, Dipotassium Oxalate, Lead Acetate, Normal Sodium Hydroxide, activated carbon, citric acid, CO2 gas, ethyl Alcohol, 70% ethanol and sodium hydroxide as well as materials used in the microbial tests including Lactobacillus agar medium (MRS Agar), Orange-Serum Agar, Dichloran Rose-Bengal (all from Merck, Germany) were prepared. Next, the treatments of research including T1 (65% sugar and 0. 6% saffron), T2 (65% sugar and 4% saffron), T3 (65% sugar and 2% saffron), T4 (70% sugar and0. 6% saffron), T5 (70% sugar and 4% saffron), T6 (70% sugar and 2% saffron), T7 (75% sugar and 0. 6% saffron), T8 (75% Sugar and 4% saffron) and T9 (75% sugar and 2% saffron) were considered. In order to prepare the treatments, the syrup tanks were prepared. Usually, for each 7-unit syrup tank which is equal to 11. 659 liters, 1, 750 gallons of purified water were poured into the tank. The tank mixer was then turned on and the sugar was added according to the formulation of making the desired beverage to dissolve all the crystals of sugar in the water. Since the consumed sugar had foreign objects, the prepared syrup was not clear and clean, so it was completely transparent and clear by passing the material from special filters. In order to eliminate the pathogenic microorganisms, the syrup was pasteurized. After pasteurization, the syrup was directed to the steel tanks of the extract. The capacity of the extract tank was 10 units (17032. 5 liters), 6 units (10219. 5) and 4 units (6813 liters). Next, the extract was added at the same time as the syrup was added to the tanks. After mixing the concentrated extract and the syrup, the mixer was turned off and the mixture was placed in the same state for 15 minutes to remove its bubbles. It is recommended that the made extract will be kept in the tank for 24-12 hours in order to achieve better maturity. The prepared extract was directed by a transfer pump to a water and extract mixer (Intermix, Flumix or Perry Mix), to mix the water entered from the refinery with the ratio of 1 to 5 for the products with the brix less than 11 or with the ratio of 1 to 5. 5 for the products with the brix below 10 and form the beverage drink. To improve the work efficiency and increase the quality of extract made, the solid materials such as citric acid and sodium benzoate were added to the syrup tanks by the additive tanks to allow the filtration. After the completion of each treatment, the samples were subjected to physicochemical, microbial and sensory tests. In the same regard, in order to analyze the data of research, a factorial experiment in a completely randomized block design was used. The mean comparison was performed by Duncan's multiple range test at the probability level of 1%=α and analyzed by SPSS software, version 16. Results and discussion: According to the results, by adding sugar and saffron extract, the amount of brix was significantly increased and it seems the sucrose to be the main reason for the increase of brix because there is a direct relationship between the concentration of sucrose and brix. Also, by adding sugar and saffron extract to the beverages produced, the pH and acidity levels decreased and increased, respectively, but they were within the standard range. On the other hand, by adding sugar and saffron extract, the density of samples did not change significantly, but the amount of dry matter increased significantly, among which the increase in the amount of dry matter can be attributed to an increase in the sugar and saffron extract in the beverage. In this regard, the ash content of treatments and total sugar content of the samples were significantly increased due to increasing the amount of saffron extract and increasing the sugar content. According to the results obtained, the amount of mesophilic bacteria increased with the addition of sugars, but it remained within the standard range. Most of the mesophilic bacteria belonged to the sample containing 70% sugar. According to the results of sensory tests, the sweetness of beverage increased by increasing the sugar content, but in the samples in which the amount of saffron increased, the amount of sweetness showed no significant difference with the first sample. Since the saffron had a bitter and astringent taste, increasing the amount of sugar made the taste desired. On the other hand, as the amount of saffron increased, the color of samples was more attractive and their flavor was more favorable. In a general conclusion and based on the results of research, it was determined that it is possible to produce saffron gaseous beverage based on its chemical and medicinal properties, which could be an appropriate substitute compared to other beverages among which the treatment containing 75 % Sugar and 2% saffron extract was introduced as the most desired treatment.

Introduction: Acorn kernel contains a high percentage of starch that can be used as a source of energy in animal’ s diet, but it has a high percentage of tannin, which can have deleterious effects on the animals. Tannins are bitter phenolic compounds with a diverse structure that are divided into two groups of condensed and hydrolyzable. Condensed tannins are flavonoid polymers of the proanthocyanidins compounds. Hydrolyzable tannins have a polyhydric Alcohol at their core, the hydroxyl groups of which are esterifies with gallic acid. They may have long chains of gallic acid coming from the central glucose core (Reed 1995). Tannins have a high tendency to bind to proteins such as enzymes (Osawa et al. 2000). The harmful consequences of the high percentage of tannin in the diet of ruminants can reduce feed intake, have a negative effect on the activity of rumen bacteria, reduce the efficiency of digestive enzymes in the livestock and also reduce the availability of nutrients, which ultimately reduce growth and production of these animals (Salinger et al. 1996). Also, the high levels of tannin compounds in the poultry diet can reduce feed intake, growth rate, digestion of protein and amino acids, intestinal and pancreatic activity (Armstrong et al. 1974; Brand et al. 1989; Mahmood et al. 1997, 2008). Various chemical and physical methods such as sodium bicarbonate, polyethylene glycol, soaking in water, cooking or steaming, have been reported to reduce the amount of tannin and its harmful side effects (Frutos et al. 2004). Now a day, bioprocessing has vast applies in livestock and poultry feed, using the enzymatic capabilities of microorganisms. Furthermore, Lactobacillus plantarum is a safe microorganism that has the potential for high enzymes to break down and eliminate tannin compounds (Curiel et al. 2009). This bacterium has vast applications for food processing in the industries (Siezen et al. 2011). Material and methods: The Experiment was done in a completely randomized design with 7 treatments and 3 replicates. Experimental groups were treatments with or without bacteria, aerobic or anaerobic condition and 5 or 10 days processing periods. The moisture content of all samples was adjusted to 50% by sterile water. For each gram of experimental sample, 10 7 CFU of Lactobacillus plantarum bacteria were added and then, placed into the incubator at 37 ° C. In this experiment, phenolic and tannin compounds, pH, soluble carbohydrate and volatile fatty acids (acetic acid, propionic acid and butyric acid) were measured. Phenolic and tannin compounds were measured based on Makkar (2000) method. Soluble carbohydrate was measured by spectrophotometric method with standard raffinose solution (Dubois 1956) and volatile fatty acids (acetic, propionic and butyric acid) were measured by gas chromatography. Soluble carbohydrate in four treatments (zero day and ten days without bacteria, ten days with bacteria in aerobic and anaerobic conditions) and volatile fatty acids in three treatments (10 days without bacteria and 10 days with bacteria in aerobic and anaerobic conditions) measured. These treatments were selected to interpret and justify the main events of fermentation processes. Data were analyzed by SAS software (2011). The comparison of the means was done using Duncan's multiple range test at P<0. 01. Results and discussion: The findings of the experiment indicate that the highest reductions in the content of total phenol, non-tannin phenol, total tannin, condensed tannin and hydrolyzable tannin (39, 24, 49, 49. 5 and 59 %, respectively, see table 1 and 2) were observed after 5 days of processing and there was no significant difference between 5 and 10 days of processing. It is assumed that significant reduction in phenol and tannin compounds was the consequence of Lactobacillus plantarum enzymatic activity. A research showed that Lactobacillus plantarum were reduced phenolic compounds of pomegranate juice (Mousavi et al., 2013). Stopping the decline in phenolic compounds after the 5 days could result in lowering the pH to below 4 (Table 3) and cessation the biological activity of the Lactobacillus plantarum (Hunget 1957), or decreasing the efficiency of the enzyme tanase (tannin aceyl hydrolase). Also, there was no significant difference between the total phenol, non-tannin phenol (Table 1), total tannin, condensed tannin and hydrolyzable tannin percentage (Table 2) of aerobic and anaerobic treatments, which showed that the biological processes reduced phenolic compounds in aerobic or anaerobic conditions with the same efficiency. It has been reported that the enzymatic processes of breaking tannins compounds by bacteria in aerobic conditions are slightly different from the anaerobic conditions, resulting in different end products (Bhat et al. 1998). Our research findings show that the acidity of experimental samples in aerobic treatments was more than anaerobic in the 5 or 10 days of bioprocessing period (P<0. 01). Smetankova et al. (2012) showed that Lactobacillus plantarum grows faster in aerobic conditions and therefore, the pH decreases more quickly. After the processing, the amount of fatty acids (acetate, propionate and butyrate) increased, but the percentage of soluble carbohydrate decreased (P<0. 01). Table 4 shows that acorn kernel contains high percentage of soluble carbohydrate (on day zero), which is required for the growth and biological activity of Lactobacillus plantarum bacteria. Table 5 presents the results of statistical analysis of the treatments for the percentage of volatile fatty acids. The results showed that the percentage of acetic acid, propionic acid and butyric acid after 10 days of aerobic or anaerobic bacterial processing increased significantly, in comparison with non-bacterial treatment, but no significant difference was observed between aerobic and aerobic conditions, which indicates that in this study the activity of bacteria for the production of volatile fatty acids was not different in both aerobic and anaerobic conditions after ten days. Conclusion: For the first time, this study has shown that in both aerobic and anaerobic conditions, the tannins compounds were broken down and reduced in equal proportion. The results of the experiment have shown that Lactobacillus plantarum bacteria can be used for bioprocessing of acorn kernel to reduce its tannins.

Introduction: Aflatoxins are a large group of mycotoxins that are produced through Polyketide pathway by specific species of Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius. These pesticides are known to be the most dangerous mycotoxins affecting human and livestock health (Pleadin et al. 2014). Several hundred mycotoxins have been identified, and more than 25% of the world annual grain production is contaminated with mycotoxin (Smith et al. 2016). Among the 400 known mycotoxins, Aflatoxin B1, B2, G1, and G2 are the most important food and feed mycotoxins (Costanzo et al. 2015). Aflatoxin M1 and M2 are the hydroxyl metabolite of aflatoxin B1 and B2 that can be found in milk or other animal products (Hussein and Brasel. 2001). At the first level, the main manifestations of mycotoxins exposure in animals are reductions in feed intake and weight gain. At the second level, mycotoxins affect the quantity of animal products. The third level of influence is the safety and quality of the products from exposed animals (Wang et al. 2019). The present study was designed to detect contamination of aflatoxin M1 in milkand, aflatoxin B1 in feed and subsequent molecular isolation and identification of Aspergillus flavus species. Material and methods: In this study, 10 milk samples from milk reservoirs and 10 feed samples from a total mixed ration of livestock from dairy farms of East Azarbaijan province (Tabriz and Marand) were collected. After preparation of samples, the experiment was conducted using competitive ELISA method. The principles were as follows: after adding standard solutions or samples to the wells, aflatoxin M1 was bonded from specimens or standards to specific antibody binding sites. After 30 minutes of incubation step, unbound reagents were removed during a single wash step. Horseradish peroxidase (HRP) aflatoxin M1 was added to the wells and after one 15 minutes of incubation, the unlinked conjugate was removed during the washing step. Then, some aflatoxin M1-HRP was coherent by adding a substrate/chromogen (H2O2/TMB) solution. In the presence of colorless chromogen, mixed conjugated aflatoxin and M1-HRP agent was converted to colored product. The addition of sulfuric acid caused the suspension of the substrate reaction and finally, the light intensity was measured by a photometric method at 450 nm. Optical density had an inverse relationship with the concentration of aflatoxin in the sample. To isolate the fungus, first 2 g of the standardized feed were weighed and milled in a falcon containing 18 ml of physiological serum and then, mixed well with a vortex for five minutes. A portion of the diluted feed was removed and cultured on plots containing a Potato Dextrose Agar medium at several locations. Plates were incubated for 7 days at 25 ℃ . DNA was extracted from Potato Dextrose Broth (PDB) medium. The resulting mycelium mass was frozen and converted to a uniform powder by liquid nitrogen. DNA extraction was carried out by placing the samples in a buffer and purification with organic solvents such as chloroform/isoamyl Alcohol and finally, curing with cold isopropanol. The resulting DNA was stored at-20 ° C. In order to evaluate the actuary of identification for Aspergillus flavus, the primer sequence of AFLA-F and AFLA-R gene was aligned using the BLAST software (GenBank) to find similarity rate within resisted reference sequences. Each PCR reaction consists of: 6 μ l of PCR Master Mix, 2 μ l extracted DNA, 0. 2 μ l of each recipe primer, 1. 6 μ l of distilled water. Then, 10 μ l of the final volume of reaction was placed on thermosecler device. A PCR program for amplification of the targeted PCR fragment was fixed based on following temperature: Initial denaturant at 95℃ for 10 min, {denaturant at 95℃ for 1 min, annealing at 66℃ for 2 min, extension at 72℃ for 2 min (total 34 cycle)} and the final amplification at 72℃ for 5 minutes (Zachová et al. 2003). Isolated strains of Aspergillus strains were verified using the PCR method; its reaction products were detected in 1% agarose gel by electrophoresis. Results and discussion: The results showed that all milk and feed samples were contaminated with aflatoxin, but the contamination rate of milk samples was lower than the standard values of Iran, America, and the European Union (0. 1, 0. 5 and 0. 05 μ g / Kg). Among the 10 collected samples, only two edible samples with aflatoxin B1 contamination were higher than the Iranian and European standards (5 μ g / kg). The contamination level of milk and feed samples were observed in the range of 0. 021-0. 05 and 1. 1-6. 9 μ g / kg, respectively. Statistically, there was a significant difference between the mean of contamination of aflatoxin M1 in milk and B1 in feed in the region with national and international standards and the mean of M1 and B1 contamination was lower than these standards. The level of aflatoxin M1 in milk was detected by HPLC method, indicating that the infection rate of 10 samples was 0. 02-0. 31 μ g / l (Besufekad et al. 2018). In another study, 178 wheat samples were collected in China and reported 18. 8% of the samples contaminated with aflatoxin B1 (Liu et al. 2016). The results of the fungal species also showed that the analyzed samples did not show any bands in the 413bp range. As a result, it can be said that the dominant species and the main cause of contamination were not Flavus species. Wang et al. (2016) reported that aflatoxins are mainly produced by the genus Aspergillus, and are commonly found in food and feed in humid and warm environments. Research results in India show that among the 15 collected samples, only 9 samples (60%) were infected with Aspergillus. Seven samples were detected as Aspergillus flavus and two samples as Aspergillus niger (Khare et al. 2018). Conclusion: Milk composition, body mass gain, immunity, and reproductive performance are affected in dairy ruminants by feeds contaminated with aflatoxins. It is expected that by controlling animal feed agaisnt aflatoxin and reducing its levels in feed by improving production and storage conditions, a suitable method for preventing contamination of milk and its products will be adopted to help improve the health of the community.

Background and the purpose of the study: Several plant essential oils, as well as terpenes present in essential oils, have shown gastroprotective activity. The aim of the present work was to evaluate the gastroprotective activity of a-terpineol, a monoterpene Alcohol which is present in essential oils of various plants.Methods: The gastroprotective activity of a-terpineol was evaluated in rats by assessing the changes in ethanol and indomethacin-induced gastric ulcer scores and on gastric secretory volume and total acidity in pylorus-ligated rats. Alpha-terpineol was administrated orally at the doses of 10, 30, and 50 mg/kg one hour before administration of the ulcer inducing agents by the pylorus ligation procedure. The involvement of endogenous prostaglandins in the protective effect of a-terpineol in ethanol-induced gastric lesions test was assessed by administration of indomethacin (10 mg/kg, s.c.) 30 min before oral administration of a-terpineol at the dose of 50 mg/kg.Results:  a-terpineol presented gastroprotective activity against ethanol-induced ulcers at the doses of 10, 30, and 50 mg/kg. Epoxy-carvone at the dose of 10 mg/kg did not present gastroprotective activity against ulcer induced by indomethacin, but at the doses of 30 and 50 mg/kg it attenuated the gastric damages induced by this agent significantly. Pretreatment with indomethacin did not prevent the gastroprotective effect of a-terpineol on ethanol-induced ulcers. Alpha-terpineol also did not affect the gastric secretion in pylorus-ligated rats.Major conclusion: The results suggest that a-terpineol presents gastroprotective action which does not involve either an increase in the synthesis of endogenous prostaglandin or a decrease in the gastric acid secretion.

Introduction: Pistachio nut is one of the popular tree nuts. Among the different species of the genus Pistacia, only the fruits of Pistacia vera attain optimal size to be acceptable to consumers as edible nuts. Contamination of pistachio by Aspergillus species and their mycotoxins is the most important problem for consumption and export of this product. Aflatoxins are potent toxic, carcinogenic and mutagenic secondary metabolites primarily produced by two fungal species, Aspergillus flavus and Aspergillus parasiticus. Aspergillus flavus produces AFB1 and AFB2, while Aspergillus parasiticus produces AFB1, AFB2, AFG1 and AFG2. Among four main groups of af latoxins, AFB1 is the most potent carcinogenic compound. Therefore, identification of toxigenic fungi is necessary for evaluating the foods quality and the presence of mycotoxins. The current methods being used for assessing fungi presence in foods based on cultivation methods and microscopic characteristics are time-consuming and labor-intensive. Recently, molecular techniques such as polymerase chain reaction (PCR) due to high sensitivity, specificity and rapidity has been introduced as powerful tools for detecting toxigenic fungi. Many genes involved in the biosynthesis of these mycotoxins have been identified and their DNA sequences have been published. PCR methods can be used to detect of aflatoxigenic Aspergilli based on structural genes (nor1, ver1 and omtA) encoding key enzymes in aflatoxin biosynthesis pathway and the regulatory geneaflR.Materials and method: Pistachio samples were collected from different cultivation regions of two towns including Gonabad and Feyzabad. Samples were packed in sterile plastic bags and immediately transferred to the laboratory. The moisture content of samples was determined using thermal method and drying in at 95-100°C. Among fungal isolates 30Aspergillus genus were detected and purified by cultural-based methods using PDA (potato dextrose agar) medium. Colonies of the fungus were transferred to PDB (potato dextrose broth) medium and incubated for 5 days at 28°C with shaking at 150 rpm. The mycelium was frozen in liquid nitrogen and ground to a powder for later DNA isolation. DNA was extracted with CTAB (cetyl trimethyl ammonium bromide) extraction buffer, then was purified with organic solvents such as chloroform/isoamyl Alcohol and finaly was precipitated by isopropanol. Aspergillus genus were detected using polymerase chain reaction by specific primer pair Asp1/Asp2 for amplification of 18S rRNA region. Furthermore, aflatoxigenic genes were detected by three sets of primers (APA-450/APA-1482, ver1/ver2 and OMT-208/OMT-1232). PCR was performed in a volume of 25 μl containing 0.5 ml of each primer, 12.5 ml of Taq DNA polymerase master mix red, 10.5 ml of sterile distilled water and 1 ml of genomic DNA as template. A PCR consisted of an initial denaturing step of 5 min at 94°C followed by 35 cycles (30 s at 94°C, 35 s at 65°C and 40 s at 72°C) finished by a final extension step at 72°C for 10 min. The PCR products were analyzed by electrophoresis on a 1% agarose gel in TBE.Results and Discussion: Among fungal isolates 30 Aspergillus genus were detected using microscopic characterstics and colony color. Under the microscope, conidia were one-celled, spherical, hyaline or pigmented and they formed long chains.12 and 4 out of 30 samples hadomtA and ver1 genes respectively. No observation was found foraflR regulatory gene in the fungal isolates. The results showed that although some isolates had one or two structural genes in the aflatoxin biosynthetic pathway, they could not produce aflatoxin due to not having anyaflR gene. Coefficient of correlation was calculated to find the relationship between the existence of Aspergillusmolds and aflatoxigenic genes in pistachio. The statistical results indicated that there is a significant correlation between the enumeration of Aspergillus molds and the existence of genes (omtA and ver1) in different moisture domains (p<0.05) while no significant correlation was identified between the enumeration of Aspergillus molds and the existence of genes in different domains of enumeration of mesophilic bacteria, yeasts and molds. Contamination of nut seeds by fungi occurs during growth, harvesting, transport and storage. The production of aflatoxin is affected by different factors, such as genetic properties of the producing fungi, temperature, moisture content, the chemical composition of food and antimicrobial agents produced by other microorganisms. Water stress and temperature are the most relevant environmental factors which influence fungal growth and mycotoxin production. Other studies showed that there was a good correlation between the expression of an early structural gene (aflD) and aflatoxin B1 production in peanut seeds. Also previous studies have shown that there was a significant relationship between A.flavus contamination in the peanuts and pistachio with high humidity (p<0.05). Since other factors such as temperature, pH and chemical composition of pistachio can affect the existence of Aspergillus molds and expression of aflatoxigenic genes, the influence of these factors on existence of Aspergillus molds and genes involved in aflatoxin biosynthesis pathway need to be investigated.