BACKGROUND: Numerous studies have investigated the positive effect of chitosan and nano-chitosan loaded clinoptilolite on growth performance, digestive enzyme activity, and intestinal histomorphology in different fish species. However, there are no data evaluating the potential effect of the composites on the fish stomach.OBJECTIVES: In the current study, the effects of chitosan and nano-chitosan loaded clinoptilolite on histological features and pepsin activity in the rainbow trout stomach were considered.METHODS: Chitosan and nano-chitosan loaded clinoptilolite were synthesized, and then two hundred and forty fish (~27.75 g) fish were distributed in eight groups each in three replicates. Ten days after adaptation, the fish were fed with eight diets, including control diet (CTR), clinpotilolite (T1), chitosan composites (T2, T3, T4), and nano-chitosan composites (T5, T6, T7) for 70 days. Afterward, all fish in each tank were anesthetized in clove extract (50 μl/l), and tissue samples were obtained for pepsin activity (n= 5) and histological assay (n = 5).RESULTS: The groups administrated with nanochitosan composites showed the highest pepsin activity (P˂0.05). Additionally, histological examinations exhibited a higher epithelial height, increased mucosal density, and oxynticopeptic cells hypertrophy in fish fed composites compared to the CTR group (P˂0.05). Meanwhile, nanochitosan composite administration could cause higher reaction of secreted granules to periodic acid–Schiff (PAS) staining.CONCLUSIONS: The findings demonstrated the potential application of chitosan and nano-chitosan loaded clinoptilolite composites for improvements in the histomorphology and pepsin activity of the rainbow trout stomach, resulting in higher growth performance.

BACKGROUND: Intensive aquaculture and commercialization production are believed to be needed for improving fish health status and probiotics are candidates to achieve this goal. OBJECTIVES: In this study, we utilized Lactobacillus plantarum and Lactobacillus pentosus as autochthonous food supplementation for biochemical investigation in rainbow trout. METHODS: In the current study, 240 juvenile rainbow trout with an average weight of 15 ± 2 g were randomly divided into four groups with three replications. Group 1, 2, and 3 were fed with 108 CFU/g of L. plantarum, L. pentosus, and the combination of these two probiotics, respectively. The control group was fed with the normal diet, for 60 days (2 % of body weight, 3times per day). Samples were taken at days 0 and 60 and biochemical parameters and hepatic enzymes activity were then investigated. RESULTS: Most biochemical factors had positive changes in the fish fed with probiotics, particularly in L. plantarum group (p < 0.05). Furthermore, the results exhibited a significant decrease in ALP level in the probiotic groups, particularly in L. pentosus group (p < 0.05). Meanwhile, AST enzyme activity in plantarum+pentosus group was higher than that in the other groups (p < 0.05). CONCLUSIONS: The present results indicated the effectiveness of food supplementation with probiotics in fish diet with the preference of probiotic to improve the biochemical factors and hepatic enzymes activity of rainbow trout, Oncorhynchus mykiss.

BACKGROUND: The use of natural immune stimulants is one of the most effective methods for strengthening immunity and preventing diseases in fish. OBJECTIVES: Due to the abundance of red algae (Laurencia caspica) in the Caspian Sea, the aim of this study was to investigate the effect of this algae on growth performance and blood indices of rainbow trout. METHODS: The present study was performed on 750 randomly selected rainbow trout (Oncorhynchus mykiss) in 5 experimental treatments including an algal extract-free diet (control), and diets supplemented with 0.5, 1, 1.5, and 2% of algal hydroalcoholic extract. During the experimental period, fish were sampled to measure growth performance and blood indices every two week for 8 weeks. RESULTS: Growth indices were not affected by the algal extract at the end of eight weeks of feeding. There were no significant differences in fish survival at different treatments. Total counts of red blood cells, white blood cells, hematocrit percentage, hemoglobin concentration and neutrophil and monocyte percentages were affected by algal extract with significant increases compared to the control group (p < /em><0.05). Results of serum biochemistry showed that significantly increased total protein, albumin and globulin and also significantly reduced ALT, AST and ALP compared to the control in rainbow trout. CONCLUSIONS: Based on the results of this study oral use of red algae (L. caspica) as an immune stimulant in rainbow trout (O. mykiss) was recommended.

BACKGROUND: The sources of fish oil for use in aquaculture is limited and different oil sources vary in fatty acid composition. OBJECTIVES: The main objective of this experiment was to determine the effects of plant oil sources such as soybean and canola oils with or without L-carnitine supplement on some blood biochemical parameters and growth performance of fingerlings rainbow trout. METHODS: In 3×2 factorial experimental design, 180 rainbow trout with 28±2 g initial body weight were randomly divided into six groups with three replicates and fed two times a day at ad libitum by dietary treatments during 10 weeks. The experimental diets contained three dietary oil sources such as fish, soybean and canola oil; and two supplemental dietaries L- carnitine levels (0 and 1 g.kg-1). Feed conversion ratio, specific growth rate, feed intake, body weight gain and blood biochemical parameters of fish such as albumin, total protein, triglyceride, cholesterol and high-density lipoprotein (HDL) were measured at the end of the experiment. RESULTS: Replacement of fish oil by soybean and canola oil had no significant effect on growth performance such as body weight gain, feed intake, feed conversion ratio, specific growth rate, hepatosomatic index, carcass weight and condition factor of fish. L-carnitine supplement significantly improved weight gain and feed conversion ratio and increased condition factor of fish (P<0.05). Soybean oil significantly increased cholesterol and triglyceride level, and canola oil decreased HDL and globulin level in blood serum of fish (P<0.05). CONCLUSIONS: Results of this study showed that fish oil could be wholly replaced by soybean or canola oil in the diet of rainbow trout and also physiological responses and growth performance of fish to carnitine supplement were affected by dietary oil sources.

BACKGROUND: Today the use of natural substances plays a major role in improving the growth and immunity of aquatic organisms. OBJECTIVES: The purpose of the present study was to evaluate the effects of Iron nanoparticles with Lactobacillus casei as a probiotic on growth parameters and probiotic counts in rainbow trout intestine. METHODS: Seven hundred and twenty fish with 12±0.49 g initial weight were prepared and after acclimatized to the laboratory conditions divided randomly into six groups. First group was selected as control, other groups were fed with a commercial diet supplemented with 108 CFU/g Lactobacillus casei (group 2), 50 µg/kg Iron nanoparticles (group 3), 100 µg/kg Iron nanoparticles (group 4), 50 µg/kg Iron nanoparticles with 108 CFU/g Lactobacillus casei (group 5) and 100 µg/kg Iron nanoparticles with 108 CFU/g Lactobacillus casei (group 6), respectively. Biometry and intestine bacteriologic examination were carried out at days 0 and 60. RESULTS: Results showed that growth parameters (weight gain, body weight gain, specific growth rate, daily growth rate, and condition factor and food conversion rate) were significantly higher in group 5 compared with the other groups. Also, intestine bacterial counts increased by using Iron nanoparticles in combination with Lactobacillus casei. The fish that received 100 µg/kg Iron nanoparticles and probiotic had significantly higher bacterial counts in their intestine compared with the other groups. CONCLUSSIONS: Based on the obtained results we conclude that diet supplementation with Iron nanoparticles and Lactobacillus casei as a probiotic could significantly improve growth parameters in rainbow trout. However, these results would warrant further study on the clinical application of these agents.

BACKGROUND: Many herbs such as fenugreek and orange have compounds with antioxidant properties, which can increase the shelf life of foods. Considering that fish are a popular food among consumers, they are susceptible to rapid corruption. OBJECTIVES: In this study, to improve rainbow trout (Oncorhynchus mykiss) fillet shelf life at refrigerated condition, orange juice concentrate and chitosan coating enriched with Fenugreek (Trigonella foenum-graecum) essential oil was used. METHODS: In the present study, 8 treatments were evaluated for 12 days at refrigerator temperature for Chemical indicators PH value, Total volatile nitrogen (TVN), Thiobarbituric Acid (TBA), Peroxide value (PV), Free Fatty Acid (FFA), and Sensory Properties. RESULTS: According to statistical results, fish fillets coated with chitosan enriched with 2% fenugreek essential oil and immersed in orange juice concentrate were lower than other groups for all chemical indicators and had a significant difference with the control group (P<0.05). In the DPPH test, the most inhibitory effect after BHT (butylated hydroxy toluene) was orange juice with 1.39 brix and then black pepper 2%, respectively. In the RP test, the absorbance of the coated sample with chitosan containing fenugreek essential oil and orange juice concentrate with BHT did not show any significant difference (P>0.05). Sensory evaluation also showed that the chitosan-coated sample containing fenugreek essential oil and orange juice concentrate improved the sensory index during storage compared to other groups, especially the control group. CONCLUSIONS: According to the results, it can be stated that the use of orange juice concentrate, chitosan coating and fenugreek essential oil have a significant effect on reducing the oxidation process of rainbow trout fillet at refrigerator temperature.

BACKGROUND: Nowadays, the tendency to use food packaging with antimicrobial activity of natural origin has increased. The use of edible and biodegradable films has been considered as an appropriate alternative to non-degradable plastic coatings. OBJECTIVES: In this study, the effect of coating with wheat starch and Pennyroyal (Mentha pulegium essential oil was investigated on microbial and physicochemical characteristics of rainbow trout fillet in cold conditions. METHODS: For this purpose, different treatments under the influence of starch, starch and Pennyroyal essential oil (1%), starch and Pennyroyal essential oil (2%) were prepared and physicochemical and microbial changes were evaluated at specified intervals (0, 4, 8, 12 and 16 days). RESULTS: In the microbial test, the count of total bacterial and other bacteria by starch and starch + 1% and 2%, the essential oil was decreased in different days, also the difference between the control group and other treatments was significant (p < /em>≥0.05). The highest effect of pH reduction belonged to starch + 1% and 2% essential oil in different studied days. The level of pH was not different among different treatments on day 0 of storage. However, there was a significant difference between the control group and treatments on the fourth, eighth, twelfth, and sixteenth days of storage (p < /em>≥0.05). The results of the oxidative test showed that on the 16th day of storage, the lowest level of TBA was related to the treatment with starch + 2% essential oil and the highest value was related to the control group. In fact, the application of the coating layer delayed the oxidation process. CONCLUSIONS: This study demonstrated antioxidant and antibacterial properties of the composition of edible starch and pennyroyal essential oil on the increased shelf-life of Rainbow trout fillet.

BACKGROUND: Antimicrobial agents can alter physiological status and immunity system of the host, and use of hematological indices are the appropriate marker for monitoring them. OBJECTIVES: The present study intended assessment of changed hematological indices of rainbow trout challenged with streptococcosis/lactococcosis agents following oral administration of florfenicol. METHODS: The purchased fish (55±7.5 g) were examined through a randomized blocks design in the replicated 6 treatments consisting of without/with the infectious challenge of each pathogen separately and without/with medicated feeding in 15 mg/kg-1 BW for 10 consecutive days. Doses of Streptococcus iniae (2.87×107 CFU/ml) and Lactococcus garvieae (6.8×105 CFU/ml) equal to 30% of LD50 values were applied in the main experiment. At the end of trial, blood was sampled via caudal vein. Measurement of hematocrit and hemoglobin has been accomplished according to standard methods, and the number of blood cells was counted by hematocytometer. RESULTS: Findings pointed out PVC%, Hb value and RBCs count of groups that received the drug were reduced significantly than control (p < /em><0.05). 10-day administration of flornfeicol in the mentioned dosage could cut down lymphocytes statistically (p < /em><0.05). On the other hand, monocytes, neutrophils, and other blood cells were enhanced following administration of therapeutical dosage (p < /em><0.05). CONCLUSIONS: It seems that oral consumption of florfenicol could improve innate immunity, especially through enhancement of hematocytes. However, due to reduced frequency of blood lymphocytes, and as the problem of anemia in fish following drug consumption is still observed, it is recommended that in case of reduction of losses and improvement of clinical symptoms, the lowest FDA-suggested levels of the antibiotic for treatment should be used.

BACKGROUND: Yersinia ruckeri is the etiological agent of enteric red mouth (ERM) or yersinioisis disease, one of the important bacterial diseases in the cultured salmonids. OBJECTIVES: The purpose of present study was detection of gyrA gene (quinolone resistance) in the Y. ruckeri bacterium. METHODS: In this study fish were evaluated in average size 8-12 cm from six rainbow trout farms in Chahar Mahal va Bakhtiyari province (Iran). In each farm 10 fish (totally 60) suspected to yersinioisis were randomly selected; sampling was done from lower part of intestine and cultured on Trpticase Soy Agar (TSA). The mediums were transferred to incubator and kept at 22 °C for 48 hours. Pure colonies which are grown on the mediums were tested by catalase, oxidase and gram staining, then those of gram-negative, catalase positive and  oxidase negative were diagnosed, and cultured on Waltman- Shots medium (as specific medium for Y. ruckeri). These mediums were incubated at 22 °C for 48 h. Colonies that were grown were tested by PCR method for Y.ruckeri detection. Then, in the identified strains of Y.ruckeri gyrA gene were detected by PCR test. RESULTS: The results of bacteriological, biochemical and molecular tests showed that three cases out of total isolates were identified as Y. ruckeri. In all isolates of Y. ruckeri, gyrA gene was identified by molecular test. CONCLUSIONS: Identification of quinolone resistance gene in Y. ruckeri isolates can be the reason of low efficacy of these classes of antibiotics in the aquaculture. ِTherefore, the policy of treatment should be changed specially in enteric red mouth disease.

BACKGROUND: Replenishing fish oil with vegetable oils and using dietary supplements plays an important role in the metabolism of fats in aquatic animals. Corn oil is one of the vegetable oils that can be a good alternative to fish oil. OBJECTIVES: The aim of this study was to investigate the effects of replacing fish oil with corn oil and the use of dietary supplements L-carnitine and betaine on the metabolism of fats and blood indices of rainbow trout. METHODS: 450 rainbow trout (9.12±0.26 g) were divided into four treatments (three replicates) and were fed with manual diet containing fish oil (1st treatment), corn oil (2nd treatment), fish oil with 500 mg/kg of food containing L-carnitine and betaine (3rd treatment) and corn oil with 500 mg/kg of food containing L-carnitine and betaine (4th treatment) for eight weeks. At the end of study, fatty acid profiles of the treatments were determined by gas chromatography (GC) and blood indices and their weights were also examined. RESULTS: At the end of the experiment, the weight index did not show a significant difference. In the replacement of fish oil with corn oil, the levels of PUFA n-3, C18:3n3, C20:3n3, C20:5n3 and C22:6n3 fatty acids significantly decreased in fish carcasses and the PUFA n-6, C18:2n6, C20:2n6 and C20:4n6 increased significantly, but no significant difference was observed in total SFA and MUFA fatty acids. L-carnitine and betaine increased the EPA accumulation in fish oil and corn oil significantly, and the number of white blood cells in corn oil, L-carnitine and betaine fed fish (fourth treatment) were increased compared to fish oil treatment (first treatment), significantly (p < /em><0.05). CONCLUSIONS: Considering the effect of corn oil, L-carnitine and betaine supplements on increasing the n-6 PUFA fatty acids, essential EPA and white blood cell counts, the use of corn oil and L-carnitine and betaine supplements in rainbow trout diet is recommended.