Organoleptic and microbiological quality changes of catla (Catla catla) in immediate and delayed ice storage and at ambient temperature

An investigation was carried out on the quality changes of Catla (Catla calla) stored immediately (0 h) in ice, after 6 hours in ice and at ambient temperature. The samples were examined for organoleptic and microbiological parameters in summer. Organoleptically, the acceptability of fish varied between 16-20 days in both the iced storage conditions and 12-13.5 hours at ambient temperature (28°C). When fish were organoleptically just acceptable on the 16th day of storage, bacterial load were 6.23 and 6.17 log10 cfu/g, respectively for 0 hour and after 6 hours iced fish. But on the 20th day of storage, when fish were just unacceptable SPC were 6.51 and 6.62 log10 cfu/g. In case of ambient temperature storage condition standard plate count was 8.36 log10 cfu/g on 13.5 hours, when fish were organoleptically just unacceptable. At the time of rejection for fish stored in ice (0 hour and after 6 hours) on 20th day, gram negative and gram positive values were 55.45%, 44.55% and 44.52%, 55.48% respectively. While fish were rejected after 13.5 hours at ambient temperature gram negative and gram positive bacteria were found as 43.02% and 56.98%. The differences in SPC, gram positive and gram negative bacteria between the storage times were statistically significant (p<0.05).

Community-based tarball monitoring in the Maldives

Monitoring oil pollution by using students to count tarballs on beaches. Samples were taken between 2006 and 2010; a significant relationship was demonstrated between the abundance of tarballs and exposure to shipping lanes. The contribution of local communities to monitoring marine pollution was also demonstrated.

Effects of gamma radiation and -20°C temperatures on the shelf life of Hilsa, Tenualosa ilisha (ham.-Buch. 1822)

The combined effect of radiation and refrigeration on the shelf life of hilsa, Tanualosa ilisha was studied by monitoring the microbiological, chemical and sensory changes of unirradiated and irradiated fish samples using low dose irradiation, doses of 300 krad, 600 krad and 900 krad. Irradiation (900 krad) dramatically reduced population of bacteria, namely total viable counts 48.850cfu per gm for unirradiated, 31.850cfu per gm and 19.600cfu per gm of 300 krad and 600 krad, respectively. The effect was more pronounced at the higher dose (900 krad), total viable count were 14.100cfu per gm. Another microbial indicator total mould counts (TMC) was 8.750cfu per gm, 6.350cfu per gm, and 19.600cfu per gm for 300 krad and 600 krad, respectively. The effect was more pronounced at the higher dose (900 krad) where total viable counts were 14,100cfu per gm. Total volatile nitrogen values increased slowly attaining a value of 101.02mgN per 100gm for unirradiated T. ilisha during refrigerated storage, whereas for irradiated fish, lower values of 71.13, 59.33 and 47.03mgN per 100gm muscle were recorded. Sensory evaluation showed a good correlation with bacterial populations on the basis of overall acceptability scores.

Microbiological quality study of Macrobrachium rosenbergii (de Man 1879) during storage at -20°C temperature

The shelf life of fresh water prawn Macrobrachium rosenbergii by applying low temperature was investigated. M. rosenbergii preserved at -20°C was subjected for quality assessment before storage and at 15, 30, 45, and 90 days of storage period. The quality assessments as done microbiological viz. total bacterial count (TBC), total mould count (TMC), total yeast count (TYC), total coliform count (TCC) and salmonella count. All the samples were acceptable during 90 days because the upper limit of all spoilage indicator was not exceeding within the experimental time period.

Effect of gamma radiation in combination with freezing on the microbiological changes in frozen shrimp Penaeus monodon

In this study gamma radiation (3, 6 and 9 kGy) in combination with low temperature (-20°C) were applied to retain the quality and shelf-life of shrimp, Penaeus monodon for a longer period. The quality was assessed by monitoring microbiological changes (TBC, TMC, TYC, TCC and Salmonella count) in irradiated and non-irradiated (control) samples. Among microbiological indicators of spoilage, total bacterial count (TBC) values for irradiated shrimps were found to be 1875, 1625 and 1525 cfugˉ¹ of sample at 3, 6 and 9 kGy respectively after 90 days whereas for non-irradiated samples it was found 2475 cfugˉ¹ of sample. Total moulds count (TMC) value for non-irradiated samples after 90 days were found 425 cfugˉ¹ sample whereas that for irradiated shrimps at 3, 6 and 9 kGy were found to be 275, 250 and 200 cfugˉ¹ sample respectively. Total yeast count (TYC) value for non-irradiated samples after 90 days were found 4125 cfugˉ¹ sample whereas that for irradiated shrimps at 3, 6 and 9 kGy were found to be 2850, 2150 and 1725 cfugˉ¹ sample respectively. Total coliform count and Salmonella count showed that those were absent during 90 days storage period. From this study, it was clear that gamma radiation in combination with low temperature showed shelf-life extension (90 days) in each dose of radiation used but during the use of 9 kGy radiation, Penaeus monodon showed best quality.

Comparative effect of Pediococcus acidilactici and Lactococcus lactis on growth performance, survival and enzyme activity of western white leg shrimp (Litopenaeus vannamei)

This study was done in Shahid Kiani Marine Aquaculture Development Center, Choebde, Abadan in order to evaluate the effects of Pediiococcus acidilactici, Lactococcus lactis and vitamin C on growth performance, survival, enzymatic activities and immune responses of L. vannamei during three months. Treatments were included control group, Pediiococcus and Lactococcus treatments which fed with diet containing 1×10P9P cfu gP_1P bacteria and vitamin C. At the end of the experiment, the growth factors, immune parameters, digestive enzymes, intestinal, histology of intestine, carcasses and microbial flora (bacterial total count and lactic acid count) were evaluated. The results indicated that administration of lactobacillus had significant effects on the growth factors as the highest weight, increase specific growth rate, relative growth rate, feed conversion ratio and protein efficiency in the shrimps received pediococcus and then Lactococcus (P<0.05). The best immune function was also observed in the shrimps fed by probiotics, so that proteins and hemoglobin̛ hemolymph, phenoloxidase activity and challenged with V. parahaemolyticus showed a statistical difference comparing to the control group and the group received vitamin C (P<0.05). Some digestive enzymes, in pediococcus treatment showed a significant increase when compared to other treatments (P<0.05). Significant changes in bacterial intestinal flora were observed in probiotic groups compared with control and vitamin C groups (P < 0.05). Histological results showed the positive effects of probiotics in the gut (P < 0.05). While these supplements cannot caused to significant impacts on the shrimp carcass composition (P ˃ 0.05). As a result pediococcus group had the best performance among treatments.

Application of whey protein incorporation with Cinnamon oil as a preservative in Huse huso fillet under chilled storage condition

Biodegradable protein-based film was developed by incorporating cinnamon essential oil (CEO) into whey protein concentrate (WPC) at level of 0.8% and 1.5% v/v. Then physical and mechanical properties of the films were evaluated. Adding CEO to the WPC matrix decreased the water vapour permeability of the films and water solubility. Films containing CEO showed significant antibacterial activity both gram-positive and gram-negative strains and exhibited significant inhibitory effect on the studied fungi. In continue, the effect of whey coating and whey coating incorporated with 1.5% CEO on quality and shelf life of Huso huso fillet during refregrated (4±1°C) storage period were also investigated. The control and treated fish samples were analyzed for microbiological (total viable count, psychrophilic counts), chemical (PV, TBA, FFA, pH, TVB-N), and sensory characteristics in 4-day intervals up of microbial, chmical and sensoy analyses indicated lower levels of PV, TBA, FFA, pH, TVB-N in coasted sampels and specially, those with CEO while were kept in refrigerator. Based on results, whey protein edible coating contain 1.5% cinnamon essential oil could enhance preserving ability Huso huso during storage cold.

Application of encapsulated nisin and sodium citrate for shelf life increasing of Rainbow trout (Onchorhynchus mykiss) fillet packaged with MAP and their effect on Staphylococcus aureus

Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. It has been proposed that reduced efficacy of nisin in foods can be improved by technologies such as encapsulation to protect it from interferences by food matrix components. The aim of this study was using of spray dried encapsulated nisin with zein in concentration of (0.15 and 0.25 g/kg) and sodium citrate (1.5 and 2.5%) and treatments with both of them to extent the shelf life of filleted trouts packaged by Modified Atmosphere Packaging (45% CO2, 50% N2 ,5% O2) and stored at 4±1 °C for 20 days. Furthermore, to evaluate the antimicrobial efficiency of encapsulated nisin and soudium citrate the trouts fillets was inoculated with Staphylococcus aureus as an index pathogenic bacteria. Assessment of chemical spoilage indexes such as (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) , microbial parameters (Total Plate Count, Psychrotrophic count, Lactic acid bacteria count), Staphylococcus aureus cont in treatments which were inoculated with 5 logcfu/g of this bacteria and sensory evaluation of fillets including (smell, color, texture and total acceptability) was carried out in days of 0, 4, 8, 12, 16 and 20. The results revealed that treatment with both exposure of nisin and sodium citrate showed significantly lower chemical spoilage indexes in comparison with controls (vaccum packed and MAP) (P<0.05). Furthermore, (nisin 0.25 g/kg sodium citrate 2.5%) treatment which was exposed to the maximal level used of both materials was significantly the lowest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 9.95 (meq O2/kg) , 1.55 (mgMA/kg), 29.65 (mgN/100g) and 6.65 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was esstimated 20 days.The control (vaccum packed) treatment was significantly the highest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 15.17 (meq O2/kg), 3.03 (mgMA/kg), 38.4 (mgN/100g) and 6.95 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was estimated 11 days. Also, in microbial point of view (nisin 0.25 g/kg- sodium citrate 2.5%) treatment was the lowest treatment with Total Plate Count, Psychrotrophic count, Lactic acid bacteria count and Staphylococcus aureus count of 6.7, 6.83, 5.25 and 6.04 logcfu/g respectively, and conrol (vaccum packed) treatment was the highest treatment with 9.15, 9.41, 7.7 and 9.01 logcfu/g respectively. According to the lower results of chemical and microbial indices and higher sensory evaluated scores assessed in this research for encapsulated nisin in comparison with free nisin , it was concluded that encapsulation of nisin with zein capsules may improve the efficiency of nisin. The measuremented values of Mass yield, Total solids content of capsules, Encapsulation efficiency, In vitro release kinetics in 200 hour for encapsulated nisin in this study was 49.89, 62, 98.31 and 69% respectively and Encapsulated particle size was lower than 674.21 μm for 90% of particles. As a consequence, nisin , in particular encapsulated nisin, and sodium citrate alone or together with and Modified Atmosphere packaging might be considered as effective tools in preventing the quality degradation of the fillets, resulting in an extension of their shelf life.

Determination of proximate composition, shelf life and fatty acid profiles of breaded kilka (Clupeonella cultriventris) during frozen storage

The present study aimed production of a new product with various texture and sensory properties in chase of the impetus for increasing human consumption considering suitable resources of Kilka fish in Caspian Sea. Following deheading, gutting, and brining, common Kilka were battered in two different formulations, i.e. simple batter and tempura batter, via automated predusting machinery and then, they were fried through flash frying for 30 seconds at 170°C in sunflower oil after they were breaded with bread crumbs flour. The products were subjected to continuous freezing at -40°C and were kept at -18°C in cold storage for four months once they were packed. Chemical composition (protein, fat, moisture, and ash), fatty acid profiles (29 fatty acids), chemical indices of spoilage (peroxide value, thiobarbituric acid, free fatty acids, and volatile nitrogen), and microbial properties (total bacteria count and coliform count) were compared in fresh and breaded Kilka at various times before frying (raw breaded Kilka), after frying (zero-phase), and in various months of frozen storage (phases 1, 2, 3, and 4). Organoleptic properties of breaded Kilka (i.e. odor, taste, texture, crispiness, cohesiveness of batter) and general acceptability in the phases 0, 1, 2, 3, and 4 were evaluated. The results obtained from chemical composition and fatty acid profiles in common Kilka denoted that MUFA, PUFA, and SFA were estimated to be 36.96, 32.85, and 29.12 g / 100g lipid, respectively. Levels of ù-3 and ù-6 were 7.6 and 1.12 g / 100 gr lipid, respectively. Docosahexaonoic acid (20.79%) was the highest fatty acid in PUFA group. ù-3/ù-6 and PUFA/SFA ratios were 7.6 and 1.12, respectively. The high rates of the indices and high percentage of ù-3 fatty acid in common Kilka showed that the fish can be considered as invaluable nutritional and fishery resources and commonsensical consumption of the species may reduce the risk of cardiovascular diseases. Frying breaded Kilka affected overall fat and moisture contents so that moisture content in fried breaded Kilka decreased significantly compared to raw breaded Kilka, while it was absolutely reverse for fat content. Overall fat content in tempura batter treatment was significantly lower than that of simple batter treatment (P≤0.05). Presence of hydrocolloids, namely proteins, starch, gum, and other polysaccharides, in tempura batter may prohibit moisture evaporation and placement with oil during frying process in addition to boosting water holding capacity through confining water molecules. During frying process, fatty acids composition of breaded Kilka with various batters changed so that rates of some fatty acids such as Palmitic acid (C16:0), Stearic acid (C18:0), Oleic acid (C18:1 ù-9cis), and linoleic acid (C18:3 ù-3) increased considerably following frying; however, ù-3/ù-6, PUFA/SFA, and EPA+DHA/C16:0 ratios (Polyan index) decreased significantly after frying. ù-3/ù-6, PUFA/SFA, and EPA+DHA/C16:0 ratios in tempura batter treatment were higher than those of simple batter treatment which is an indicator of higher nutritional value of breaded Kilka with tempura batter. Significant elevations were found in peroxide, thiobarbituric acid, and free fatty acids in fried breaded Kilka samples compared to raw samples which points to fat oxidation during cooking process. Overall microorganism count and coliform count decreased following heating process. Both breaded Kilka samples were of high sanitation quality at zero-phase according to ICMSF Standard. The results acquired from organoleptic evaluation declared that odor, cohesiveness, and general acceptability indices, among others, had significant differences between the treatments (P≤0.05). In all evaluated properties, breaded Kilka with tempura batter in different phases gained higher scores than breaded Kilka with simple batter. During cold storage of various treatments of breaded Kilka, total lipid content, PUFA, MUFA, ù-3, ù- 3/ù-6, PUFA/SFA, Polyen index decreased significantly. The mentioned reductions in addition to significant elevation of spoilage indices, namely peroxide, thiobarbituric acid, and free fatty acids, during frozen storage, indicate to oxidation and enzymatic mechanism activity during frozen storage of breaded Kilka. Considering sensory evaluation at the end of the fourth month and TVB-N contents exceeded eligible rate in the fourth month, shelf life of the products during frozen storage was set to be three months at -18°C. The results obtained from statistical tests indicate to better quality of breaded Kilka processed with tempura batter compared to simple batter in terms of organoleptic evaluation, spoilage indices, and high quality of fat in various sampling phases.

Study of effect Rosmarinus officinalis oil and nisin on the growth of Streptococcus iniae in lab conditions and fillets of rainbow trout (Oncorhynchus mykiss)

Fish are an important part of a healthy diet since they contain high quality protein, but typically present a low fat percent when compared to other meats. Fish is an extremely perishable food commodity. On the other hand, food borne diseases are still a major problem in the world, even in well-developed countries. The increasing incidence of food borne diseases coupled with the resultant social and economic implications means there is a constant striving to produce safer food and to develop new antimicrobial agents concerns over the safety of some chemical preservatives and negative consumer reactions to preservatives they perceive as chemical and artificial, have prompted on increased interest in more ‘‘naturalgreen’’ alternatives for the maintenance or extension of product shelf-life. Particular interest has focused on the potential applications of plant essential oils. However, to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects and multiple hurdles can be devised. In this study, were evaluated the effects of different concentrations of Rosmarinus officinalis and nisin and storage time (15 days) on growth of Streptococcus iniae GQ850377 in a lab conditions and a food model system (fillets of rainbow trout) in 4 and 8 °C. In addition, we also studied multi factorial effects of four different concentration of rosemary, three different concentrations of nisin, two different levels of pH in 3 temperature 4,15 and 37 °C on log% of S.iniae during 43 days in BHI broth. The results on growth of S. iniae were evaluated using SPSS 20.0 statistical software and analyzed the logarithm of total count of the bacterial by Tukey Test. Results were considered statistically significant when P<0.05. MIC and MBC values of rosemary and nisin were 0.03, 0.075 % and 5, 40 μg/mL, respectively. The growth of S. iniae was effected significantly (P<0.05) by rosemary and nisin and also combination of rosemary and nisin in 4 and 8 °C. Samples treated with 0.135 and 0.405 % of rosemary showed a significant decrease on the growth of the bacteria compared with control sample(P<0.05). The most ١٤٦ inhibitory effects were seen in samples treated with 0.135 and 0.405% of rosemary until 9 days after storage. Also, the synergism effects of rosemary and nisin on the growth rate of bacteria was significant (P<0.05) compared with untreated samples and samples treated with the rosemary or nisin, only. Synergistic effects was observed at concentration of 0.405% rosemary and 0.75 μg/mL nisin in both temprature. Results of this study showed that different concentration of rosemary a significant inhibitory effect (P<0.05) on log% of S. iniae, in BHI broth in pH 5.5 and 7 in 4,15 and 37 °C during 43 days. In concentration of 0% rosemary (control) in pH 5.5 and 7 and 37°C, log% were 1.099 and 3.15, whereas in concentration of 0.015% rosemary were -4/241 and 1.454, respectively. The use of essential oils may improve food safety and overall microbial quality. If essential oils were to be more widely applied as antibacterials in foods, the organoleptic impact would be important. In addition, it is recommended to apply essential oils or their compounds as part of a hurdle system and to use it as an antimicrobial component along with other preservation techniques. Thus essential of R. officinalis with high antibacterial activity selected in this study could be a potential source for inhibitory substances against some food-borne pathogens and they may be candidates for using in foods or food-processing systems.

Effect of thawing methods on the chemical, biochemical, microbiological and sensory indices of Caspian Sea kutum (Rutilus frisii kutum) and rainbow trout (Oncorhynchus mykiss)

Effects of different thawing method i.e. in a refrigerator, in water, at air ambient temperature and in a microwave oven on proximate, chemical (PV, TBA, FFA, TVB-N, SSP, FA), biochemical (pH, WHC,ThL), microbial (total viable, psychrotrophic, coliform, Shewanella and yeast-mould count) and sensory analysis were carried out on frozen whole Caspian sea Kutum (Rutilus frisii kutum) and Rainbow trout (Oncorhynchus mykiss) carcasses. The values of ash, protein, SSP, WHC, PUFA, PUFA/SFA. EPA+DHA/C16:0, pH, and microbial count of thawed samples decreased significantly while fat, PV, TBA, FFA, TVB-N, SFA and MUFA increased compared to the fresh fish (unfrozen) as control samples. Also, sensory evaluation all of thawed samples showed a significant (p<0.05) quality loss compared to the fresh fish as control samples. The lowest chemical and biochemical values as well as microbial growth were determined in water thawed samples. Therefore, based on this study thawing in water is most suitable for frozen whole rainbow trout.

Application of combined effect of sodium acetate and nisin on reduction of contamination by Listeria monocytogenes in vacuum packaged grass carp (Ctenopharyngodon idella) fillets kept at 4°C

In this study microbiological , chemical quality and fatty acid composition of grass carp (Ctenopharyngodon idella) fillets treated by dipping in sodium acetate (%1 and %3), nisin (% 0.1 and % 0.2) and combination of sodium acetate and nisin was evaluated during 16 days of refrigerated of 4°C Antilisterial effect of nisin was enhanced with the increased concentration of sodium acetate. At day 12 post storage, Listeria monocytogenese count was higher in the control group than the recommended value, however in sodium acetate and nisin treated samples, the count was lower (5.17-5.91 log cfu/g). With increasing the concentrations of sodium acetate, mesophilic counts were lower. Regarding nisin, better results was obtained by applying %0.1 nisin. Greater inhibition of mesophile bacteria was observed when combination treatment was used. The number of lactobacillus was lower when higher concentrations of sodium acetate and nisin were used. Total Volatile Nitrogen values at the end of the experiment were lower in the samples treated with both nisin and sodium acetate and the better results were obtained in combination treatments. Peroxide (PV) at the end of the experiment was 1.9 meq/kg in control, and the lowest values were observed for the treatments 3(%0 sodium acetate +% 0.2 nisin) and 9(%3 sodium acetate +% 0.2 nisin) between 1.08 and 1.62 meq/kg without significant difference. Thiobarbituric acid (TBA) levels at the end of experiment have been shown to be 0.46 mg malonaldehyde per kg in the control. On the other hand treatments 9 had the TBA values of 0.19 mg malonaldehyde per kg which was significantly lower than that of control. Polyunsaturated fatty acids increased by increasing the sodium acetate doses and instead saturated fatty acids and n-6/n-3 ratio decreased. The ratio of UFA/SFA and also C22:6/C16:0 increased when a higher concentration of sodium acetate has been used. The best result obtained by using 3% of sodium acetate but no such relation with nisin was observed.

Effect of organophosphate, diazinon on some immunophysiological, haematological and histological variable of Rutilus frisii kutum (Kamensky,1901)

The acute toxicity and effects of diazinon on some haematological parameters of kutum (Rutilus frisii kutum, Kamensky, 1901) weighing 613.33 g±157.06 g were studied under static water quality conditions at 15°C ± 2ºC in winter and spring 2009. The effective physical and chemical parameters of water were pH= 7-8.2, dh= 300mg/L (caco3), DO= 7 ppm and T= 15°C±2ºC. The first test was primarily to determine the effects of acute toxicity (LC5096 h) of the agricultural toxicant diazinon (emulsion 60%) on kutum male brood stocks. For this purpose, 4 treatments were used to test toxicity; each treatment was repeated in 3 tanks with 9 fish per treatment and with 180 litres water capacity. After obtaining the final results, the information was analysed statistically with Probit version 1.5 (USEPA, 1985), and we determined the LC10, LC50 and LC90 values at 24 hours, 48 hours, 72 hours and 96 hours; the maximum allowable concentration value (LC5096 h divided by 10) (TRC, 1984); and the degree of toxicity. The second stage of testing consists of four treatments: LC0= 0 as experimental treatment, treatment A with a concentration of LC1= 0.107 mg/L, treatment B with concentration of LC5= 0.157 mg/L, treatment C with concentration of MAC value= 0.04 mg/L. Male brood stocks of kutum were treated with these concentrations for 45 days. Experiments were carried out under static conditions based on the standard TRC, 1984 method over 45 days. Our results show that long-term exposure to diazinon causes a decrease in the erythrocyte count (RBC), haemoglobin (Hb), haematocrit (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), leucocyte count (WBC), lymphocyte, testosterone, iron (Fe), sodium (Na), lactate dehydrogenase (LDH), and cholinesterase (CHeS). In addition, diazinon also causes an increase in prolymphocyte, aspartate aminotransferase (AST), cholesterol, alkaline phosphatase (ALP) and adrenaline (P<0.05). There are no significant effects on monocyte, eosinophil, magnesium (Mg), chloride (Cl), glucose (BS), urea (BUN), uric acid (U.A), triglyceride (TG), calcium (Ca), albumin (Alb), total protein (TP), cortisol, noradrenaline and high density lipoprotein (HDL) levels in kutum male brood stocks (P>0.05). Pathology results showed toxin diazinon no effect on average weight and fish body length, the average weight of heart, brain, spleen, liver, kidney and liver index but caueses decrease of gonad weigth and gonad index and also, cause complications of tissue necrosis, vascular congestion, inflammation in the liver, a sharp reduction in the number of glomeruli, necrosis, vascular congestion and haemorage in the kidney, capsule thickening and fibrosis, atrophy, vascular congestion, macrophages release increased, increasing sediment Hemosiderine and thickening of artery walls in the spleen, atrophy, fibrosis and necrosis in testis , vascular congestion, increased distance between the myocardium and fibrous string in heart and neuronal loss, vascular congestion and edema in the brain of kutum male brood stocks.

Comparative study of seasonal changes in some blood parameters and genetic polymorphisms of serum antibodies and antigens on red blood cells surface in immature Caspian salmon

Seasonal sampling from 40 immature Caspian salmon were performed in summer, autumn, winter and spring. The maximum ranges of RBC counts, Hct, Hb, WBC count and clotting times were observed in spring, summer, spring, spring and winter, respectively. The minimum amounts of these factors were counted in summer, winter, winter, winter and winter, respectively. Blood Samples were taken from healthy smolt, immature and adult Caspian salmon in spawning time. Hematological determinations and biochemical serum analysis were performed in 101 fish in the three samples. The ranges of hematological values for sample mean were counted. Red blood cell counts were 866600 mm3 and 1259400 mm3 in smolt and adult respectively. Hematocrit was 48.39% in smolt and 44.29% in adult. Hemoglobin was 8.85 gr/dl in smolt and 10.91 gr/dl in adult. White blood cell count was 8781.58 mm3 in smolt and 5217.55 mm3 in adult and mean were differential of WBC, Lymphocyte 90.57%in smolt and73.22% in adult. Neutrophil was 5.12% in smolt and 16.92% in adult, Monocyte were 1.27% in smolt and 4.24% in adult, Clotting time was 282.34 Seconds in smolt and 291.47 seconds in adult MCV, MCH and MCHC also meagered in smolt and adult. Biochemical parameter in immature and mature Caspian salmon meagered .Glucose concentration was 2.97 mmol.l- in immature and 1.99 mmol.l- in mature .Cholesterol concentration was 4.26 mmol.l- in immature and 7.06 mmol.l- in mature. Triglyceride amount was 2.35 mmol.l- in immature and 2.47 mmol.l- in mature and Calcium was 2.47 in immature and 2.61 mmol.l- in mature. An in situ study was made on erythrocytic isoantigens and hetero-antigen and their corresponding iso-and hetero-antibodies of sera by means of hemoagglutination tests on the blood sample, of 450 immature and 50 mature Caspian salmon. The absence of erythrocyte iso-antigens and hetero-antigen and their corresponding iso-and hetero-antibodies were shown by the experimental. It could be indicated an intra-specific variation and differences in species for kelardasht hatchery.

The genetic diversity of Acipenser stellatus populations in the Caspian Sea was studied using microsatellite technique

A total of 361 caudal fin samples were collected from adult A. stellatus specimens caught in the north Caspian Sea, including specimens from Kazakhstan (Ural River), Russia (Volga River), Azerbaijan (Kura River), specimens caught in the south Caspian Sea including specimens from Fishery Zone 1 (from Astara to Anzali), Fishery Zone 2 (from Anzali to Ramsar), Fishery Zone 3 (from Nowshahr to Babolsar), Fishery Zone 4 (from Miyankaleh to Gomishan) as well as from specimens caught in Turkmenistan (all specimens were collected during the sturgeon stock assessment survey). About 2 g of fin tissue was removed from each caudal fin sample, stored in 96% ethyl alcohol and transferred to the genetic laboratory of the International Sturgeon Research Institute. Genomic DNA was extracted using phenol-chloroform method. The quality and quantity of DNA was assessed using 1% Agarose gel electrophoresis and Polymerase Chain Reaction (PCR) was conducted on the target DNA using 15 paired microsatellite primer. PCR products were electrophoresed on polyacrylamide gels (6%) that were stained using silver nitrate. Electrophoretic patterns and DNA bands were analyzed with BioCapt software. Allele count and frequency, genetic diversity, expected heterozygosity and observed heterozygosity allele number, and the effective allele number, genetic similarity and genetic distance, FST and RST were calculated. The Hardy Wienberg Equilibrium based on X2 and Analysis of Molecular Variance (AMOVA) at 10% confidence level was calculated using the Gene Alex software. Dendrogram for genetic distances and identities were calculated using TFPGA program for any level of the hierarchy. It is evident from the results obtained that the 15 paired primers studied, polymorphism was observed in 10 pairs in 12 loci, while one locus did not produce DNA bands. Mean allele number was 13.6. Mean observed and expected heterozygosity was 0.86 and 0.642, respectively. It was also seen that specimens from all regions were not in Hardy Wienberg Equilibrium in most of the loci (P≤0.001). Highest Fst (0.063) was observed when comparing specimens from Fishery Zone 2 and Fishery Zone 4 (Nm=3.7) and lowest FST (0.028) was observed when comparing specimens from the Volga River and those from the Ural River (8.7). Significant differences (P<0.01) were observed between RST recorded in the specimens studied. Highest genetic distance (0.604) and lowest genetic resemblance (0.547) were observed between specimens from Fishery zones 2 and 4. Lowest genetic distance (0.311) and highest genetic resemblance (0.733) was observed between specimens from Turkmenistan and specimens from Fishery zone 1. Based on the genetic dendrogeram tree derived by applying UPGMA algorithm, A. stellatus specimens from Fishery zone 2 or in other words specimens from the Sepidrud River belong to one cluster which divides into two clusters, one of which includes specimens from Fishery zones 1, 3 and 4 and specimens from Turkmenistan while the other cluster includes specimens from Ural, Volga and Kura Rivers. It is thus evident that the main population of this species belongs to the Sepidrud River. Results obtained from the present study show that at least eight different populations of A. stellatus are found in the north and south Caspian Sea, four of which are known populations including the Ural River population, the Volga River population, the Kura River population and the Sepidrud River populations. The four other populations identified belonging to Fishery zones 1, 3, and 4 and to Turkmenistan are most probably late or early spawners of the spring run and autumn run of each of the major rivers mentioned. Specific markers were also identified for each of the populations identified. The Ural River population can be identified using primers Spl-68, 54b and Spl-104, 163 170, 173, the Volga River population can be identified using primers LS-54b and Spl-104, 170, 173 113a and similarly the population from the Kura River can be identified using primers LS-34, 54b and Spl-163, 173 and that from the Sepidrud River can be identified using primers LS-19, 34, 54b and Spl-105, 113b. This study gives evidence of the presence of different populations of this species and calls for serious measures to be taken to protect the genetic stocks of these populations. Considering that the population of A. stellatus in Fishery zone 2 is an independent population of the Sepidrud River in the Gilan Province, the catch of these fishes in the region needs to be controlled and regulated in order to restore the declining stocks of this species.