The survey on determination of malathion biomarker with genotoxicity and ecophysiological reactions in the Caspian roach (Rutilus rutilus caspicus)

In the present study possibility of Malathion biomarker with Genotoxicity and Ecophysiological reactions were determined in Caspian Roach (Rutilus rutilus caspicus). At fist LC50 value of Malathion, an organophosphate insecticide was determined. Then four groups of experimental fish (containing 30 fish in each group) were exposed to different concentrations of Malathion. e. 0, 0.01, 0.05 and 0/1 ppm respectively for 23 days and effects of Malathion on Hematological (RBC, WBC, Hb and Hct) and biochemical parameters (Glucose, triglyceride, urea, total protein and Albumin), some enzymes (SGPT, SGOT and ALP), Cortisol level, plasma cations (Na+ and K+) , histological changes (gill and liver) and finally DNA destruction were examined. Sampling was done in 3rd, 13th, 23rd days during exposure and also 30 days after recovery. Data analysis was done by SPSS (Ver.13) and graphs were drawn by Excel 2007. Results showed that WBC, RBC, Hb, Hct, some biochemical parameters and K+ of Mallation treatments were decreased significantly in compare to control group (P<0.05). Changes in enzyme were many different. No significant changes were observed in Na+ and cortisol levels (except in groups treated with 0.01 Mallation) (P>0.05). LC50 value of Malathion in Caspian Roach was 6.5 ppm. Histological examinations showed that Mallation cause tissue damages and there were more damages in longer times and in higher concentrations. Apoptotic cell and comet were observed as DNA destruction and they were more in treatments with higher Mallation concentrations for longer times.

Determination of bioaccumulation and microscopic survey of LAS pollutant effect on the liver, kidney and gill tissues of Rutilus frisii kutum in Caspian sea

Linear alkylbenzene sulfonate (LAS) are widely used in detergent industry. Due to contaminants entering the water, and the effects of their accumulation in fish, LAS, has a great importance in environmental pollution. In the present study, accumulation of LAS and its histological effects on gill tissue, liver and kidney of Caspian kutum (Rutilus frisii kutum) were studied. Caspian kutum is the most important and most valuable teleosts of the Caspian Sea. Due to releasing Caspian Kutum in rivers and Anzali Lagoon and unlimited entry of wastewater to the aquatic ecosystem, research on the impact of LAS on Caspian kutum is important. In the present study, fish exposed to sublethal concentrations of LAS (0.58, 1.16 and 2.32 mg/l) for 192 hours. Control treatments with three replicates at 0, 24, 48, 72, 96 and 192 hours were done. For assessments of the histological effects of LAS, tissue sections prepared and by using Hematoxylin - Eosin were stained, then the prepared sections, examined by light microscopy. For determination of the bio accumulation of LAS, the soxhlet extraction and solid phase extraction was performed to determine the amount of LAS using HPLC with fluorescence detector. According to results average of bioconcentration factor and LAS concentrations in fish had reached stable levels after approximately 72 h and thus represented steady state BCF values in this species. The value of steady-state bio-concentration factor of total LAS was 33.96 L.Kg- 1 and for each of the homologous C10-n-LAS, C11-n-LAS, C12-n-LAS and C13-n- LAS were 3.84, 6.15, 8.58 and 15.57 L.Kg-1 respectively. According to the results obtained in gills exposed to LAS, histopathological alteration include hypertrophy, lifting of lamella epithelium, edema, clubbing of lamellae hyperplasia, lamellar fusion and aneurysm were seen. In liver tissue exposed to three concentrations of LAS, congestion and dilation of sinusoids, irregular-shaped nuclei and degeneration in the hepatocyte, vacuolar degeneration and necrosis were observed. In kidney exposed to three concentrations of LAS, reduction of the interstitial haematopoietic tissue, degeneration in the epithelial cells of renal tubule, tubular degeneration, necrosis, shrinkage and luminal occlusion were observed. According to the results the most alteration due to exposure to LAS was seen in the gill tissue. None of the control samples showed histological effects of LAS.

Biological characteristics and determination of population dynamic parameters of Sphyraena jello in the coastal waters of Province Bushehr- Persian Gulf

Sphyraena jello, Pick handle Barracuda, is amongst the highly valuable and main commercial fisheries resources in the southern waters of Iran. Given such an economically significant position, this study, being conducted in 2006-2007, attempts to investigate its biological habit and characteristics in Iran’s water of the Persian Gulf. For the sampling purpose, three major landings namely Bushehr, Deylam and Genaveh were selected to obtain samples from commercial catches. The sampling is composed of 655 males and 515 females during a twelve month period. By studying the feeding through the counting method, it is revealed that, Liza subviridis characterized by %42.8 and Sepia pharaonis by %8.4 made the highest and lowest stomach content respectively. The findings showed that male fish in smaller size will mature sooner than females’ specimen but this gender ratio or proportion was not significantly different except during October and September. Such a difference between male and female in different months could be originated from longer residing of female group in spawning ground compared to male group. The previous spawning lasted during September –October, and there was a peak of spawning in feeding in August. The lowest fat proportion for both male and female genders was reported 0.10 and 0.11 respectively in October; but the highest level of condition factor was reported to be 0.59 and 0.63 during November and June.

Determination of some dominant genetic characteristics and morpho-meristic features for selection of rainbow trout (Oncorhynchus mykiss) broodstocks

In this study, Iranian and French male and female Oncorhynchus mykiss broodstocks were divided into two groups 50 and 24 respectively in Research center of genetic and breeding of coldwater fishes, Yasouj, Iran and the genetic structure of them was investigated using 6 microsatellite markers. Then 19 morphometric and 5 meristic of broodstock were measured and compared in two populations. Along with broodstock maturation, fertilization 1:1(female:male) were randomly assigned and occurred in 25 of 12 Iranian and French treatment respectively. Reproductive parameters were recorded for the whole family. Average number of observed alleles in Iranian and French stocks was 6.68 and 6.83, respectively. Average number of effective alleles in Iranian and French stocks was 3.13 and 3.45 respectively. Fixation index Fst was calculated based on allelic frequency between two stocks was 0.058 with significant difference between 2 stocks. Morphometric analysis showed significant difference between two stocks in 8 characteristics. Meristic characters was without significant difference in broodstock groups. Eyed percentage for french broodstock calculated zero and deleted. Fertilization rate (100-0), the eyed percentage (98- 0), The hatch rate (98-0), the average fecundity 4114.708, the average eggs size 4.88 mm, Survival in the first three months 19-73% calculated for Iranian broodstocks. Considering the quality of eggs and larvae at different stages and selection between the different family and the within family remained 10 treatments and are kept as future broodstocks. The relationship between fecundity - egg size, fecundity - weight , fecundity - length, egg size- weight was performed using regression. The results showed that Fecundity was influenced more by weight and productive length. The research is beginning to ID the broodstock in our country.

Biological characteristics and determination of population dynamics parameters of Nemipterus japonicus in the coastal waters of Province Bushehr, Persian Gulf

This study was conducted to determine biological characteristics and population dynamics parameters of threadfin bream (Nemipterus japonicus) in Persian Gulf (Bushehr Province), during November 2006 and October 2007. The minimum and maximum specimens were 75-273 mm FL and their weight was 7.6 - 351.9 g. Based on the exponential relationship between fork length and weight, slope (b) for individuals, males and females was 2.987321, 2.992546 and 3.007314, respectively. The emptiness value (V) was 45.6% and it shows that N. japonicus is a moderate feeder. The results of Fp indicates that crustacean with 78.2% are main foods, mollusca (27.7%), fishes (20.7%), polychaeta (19.2%) and Foraminifera (11.7%) were identified as minor foods and phytoplanktons (9.9%), nematoda(8.0%), echinodermata (2.3%) and sea weeds (0.3%) were random foods. The reproduction studies showed the spawning season extended within 2 peaks, from April- May and September and main spawning occurs in spring season.The mean absolute and relative fecundities were 472388±42633 and 3817±293 (X±SE), respectively. The maximum, minimum and mean of oocyte diameter were 0.448, 0.022 and 0.221mm (SE=0.071), respectively.The fork length at 50% maturity estimated to be 20.25 cm for females. The growth coefficient (K) , length infinity (L∞ ) and ɸ' was estimated 0.42/yr , 34.17 cm and 2.69, respectively. The coefficient of total mortality, fishing mortality, natural mortality and E was 1.37, 0.43, 0.94 and 0.31, respectively.

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.

The study of biological characteristics and heavy metal determination of Cd, Pb, Fe, V, Ni in the muscle, liver and gonad tissues of kingfish Scomberomorus commerson in the coastal waters of the Hormozagan Province (Persian Gulf)

Biological studies and heavy metal (Ni, v, Fe, Pb,Cd) determination in liver, gonad and muscle of Scomberomorus commerson were carried out from Oct 2006 to Sept 2007 in Hormozgan coastal waters. 599 Samples were gutted for reproduction and nutrition studies, fork length and weight were measured to nearest cm and g respectively. Meanwhile 40 samples were also investigated for heavy metal studies. All specimens were collected from two major landing sites (Bandar Lengeh & Bandar Abbas). Minimums & maximum fork length & weight were 29, 128 cm & 235 and 15350 g respectively.Isometric growth was shown according to our study and b was estimated 2.9 (overall), 2.91(male) & 2.89 (female). The average relative gut length was 0.52± 0.007 and it was determined that S. commerson is a carnivorous. More than 99 percent of gut content was different teleost fishes. Gastro somatic index had two peaks in Nov & Jan (before spawning) and with a decreased trend in July, the spawning period. Occurrence of empty stomach was estimated % 65.77. Maximum amount of condition factor was in Dec. Spawning season was started from June. The average of Absolute & relative fecundity (to weight unit) was 1217149±179315 and 178.2±15.58 respectively. Lm50% was estimated 75 cm for females. Sex ratio was 0.97: 1 (male: female). Chi- Square test showed no significant difference (p>0.05). Maximum amount of hepatosomatic index was estimated in March.Metal concentrations were determined using either Flame Atomic Absorption Spectroscopy (for Fe) or Graphite Furnace Atomic Absorption Spectroscopy (for Pb,Cd,Ni and V). The mean concentration (μg/g dry weight)of Pb,Cd,Ni,V and Fe in the liver were 0.0309, 0.0268, 0.0672, 0.0077, 2.5159 in the gonad 0.0440 ,0.0295, 0.1096, 0.0000, 1.4449 and in the muscle 0.0244, 0.0324, 0.0656, 0.0128, 1.6138 respectively. The maximum metal concentrations were below the maximum permissible limits for human consumption recommended by the USEPA, WHO and the UK. The results of Kendall's Tau-b correlation coefficient were as follows: The Liver tissue: There were significant positive linear relationships between accumulation of V, Fe, and Pb with Fork length, Pb and Fe with weight, GSI with Pb, Cd, V and 109 Fe, and a negative linear relationships between HSI with accumulation of V and Fe, Fork length, weight and GSI. The Gonad tissue: There were significant positive linear relationships between GSI with accumulation of Pb, Cd, Fe, Fork length and weight, a negative linear relationship between HSI with Fork length, weight and GSI. The Muscle tissue: There were significant positive linear relationships between accumulations of V, Fe with Fork length and weight factors and as well as GSI with Cd, V, Fe, Pb, Fork length and weight,a negative linear relationship between HSI with Fork length, weight, Cd, Fe and GSI. The results of Mann-Whitney U tests (P≤0.05) show that there were significant differences between summer and autumn from heavy metal contents in the studied tissues point of view. The only exceptions were for Ni in the liver, gonads and muscle and as well as there were significant differences between male and female from heavy metal contents in the studied tissues. The only exceptions were for Pb in muscle, Ni in liver, gonad and muscle, V in muscle, and Cd and Fe in gonads.

Identification and determination of muscle's fatty acids and amino acids in Cobia and Indian squid

The present study aims to find the effect of freezing Time on the quality of Cobia (Rastrelliger kanagurta) and Indian Squid in commercial scale during freezing and subsequent frozen storage (−18◦C). Total time for freezing was significantly different (P<0.05) between the Cobia and Indian squid samples. The difference in the freezing time could be attributed to the varied quality of the 2 samples. Upon freezing, the moisture content decreased in Indian Squide samples compared to Cobia freezer where protein content decreased in both the samples. Upon freezing and during frozen storage, lipid oxidation products (peroxide value, and free fatty acid value) and volatile bases (total volatile base nitrogen) showed an increasing trend in both the samples with values slightly higher in Indian squid samples compared to cobia frozen samples. The total plate counts showed a significantly (P<0.05) decreasing trend in both the samples. K value did not show any significant (P<0.05) difference between the samples whereas the histamine formation was significantly (P<0.05) increased in Indian squid frozen samples compared to cobia samples. The taste and overall acceptability was significantly different (P<0.05) in cobia samples compared to Indian squid frozen samples on 5th month. Both samples were in acceptable condition up to 5 month but the Cobia frozen samples quality was slightly better than the air blast frozen samples.

The study of biological characteristics and determination of population dynamic parameters of sawtooth barracuda (Sphyraena putnamae) in the Persian Gulf (Hormozgun Province waters)

The biological characteristics and population dynamisms of Sphyraena putnamae, were studied in the northern Persian Gulf and Oman Sea restricted to Hormuzgan province waters within 13 months period, from November 2006 up to November 2007. Biometrical and anatomical measurements were carried out, and biological surveys were conducted on 486 specimens. On the other hand, the growth and mortality parameters were estimated by using 3096 samples. These samples were collected from 3 landings, namely Bandar Abbas, Bandar Lengeh and Bandar Jask. The measurements of the minimum and maximum Fork lengths and weights were 11.7 to 8.03 cm and 135.0 to 4140.0 g, respectively. The results indicated that this species, having the Relative Length of Gut, RLG=0.34±0.002, is strongly carnivorous (often fish-eater), proven by the fact that more than 98% of its stomach contents were fish pieces. Examining the changes in the index of stomach emptiness by the percentage of CV = 0.47% indicates that this fish is Moderate feeder. The level of feeding increased in March, before spawning and decreased in June and September, simultaneously with the spawning season. There are 2 peaks of reproduction or spawning seasons during the months of April-May and September, of which the prior is assumed as the main spawning. The sex ratio (M:F) was calculated 0.5:1.0(X2 =2.11), which did not show a significant difference with expected level of 1:1 (P>0.05). The average absolute and relative reproduction rates of Sphyraena putnamae is respectively as follows: 1866827.1±255448.9 and 1097.7±94.3. The highest and the lowest diameter of matured egg are from 200 to 750 μ, and its average diameter is 402.10 ± 0.190 μ. A parameter for Saw-tooth barracuda length measurement, Lm50, based on the Fork-length, was calculated as 54.01 cm. In other words, as far as the fisheries management is concerned, the fish whose lengths are less than 54.01 cm should not be caught. The calculated level of (R2) (correlations of total length & weight), indicated strong correlations between length and weight of this fish, and the obtained formula included W =0.007100 FL 2.9295 and reinforced this assumption. The “K” Index for this fish in 3 above mentioned landings (Jask, Bandar-Abbas and Bandar-Length) were 1.24, 0.37 and 0.46 per year, respectively and the FL index for the same landings were estimated as 129, 110 and 134 cm, respectively. The growth coefficient (MONRO) for the above mentioned regions were calculated as 3.601, 3.647 and 3.917, respectively; and in the surveyed regions there were no significant differences in populations. The Total mortality coefficient (Z) was calculated 0.76, 1.12 and 1.07 per year, the Natural mortality coefficient was 0.46, 0.63 and 0.70, and the Fishing mortality coefficient (rate) (F) was found to be 0.30, 0.49 and 0.37 per year. The value of the exploitation rate (E) is equal to 0.39 per year, indicating that this species is an under-exploited resource, and there is no excessive fishing pressure on the fish supply of this species in the afore-said regions. The highest level of exploitation was found for ‘Bandar Abbas’ fishing region and the lowest level of exploitation is in ‘Bandar Lengeh’ waters.

Physiological and biochemical study of over-ripened oocyte in the cultivated Caspian brown trout (Salmo trutta caspius) and determination of egg quality markers

To determine the best time for egg stripping after ovulation and over-ripened oocyte in the Caspian brown trout (Salmo trutta caspius), the eggs were retained in the parental abdominal cavity for 40 days post-ovulation (DPO) at 7±0.6°C. Eggs were stripped every 10-day interval in 4 treatment and were fertilized with a pool of semen obtained from 8 males. Also, the physiology and biochemistry of the eggs and ovarian fluids were studied. Results showed that the level of eyed eggs and hatched alevins declined with over-ripening time: that is, the expected amounts (90.65 ± 6.28% for eyeing and 86.33 ± 6.82% for hatching) in newly ovulated eggs (0–10 DPO) decreased to 0.67 ± 1.34% and 0.49 ± 0.98%, respectively, in over-ripened eggs (30–40 DPO). However, larval abnormalities remained constant for 30-days after ovulation. During the course of oocyte over-ripening, the pH of the ovarian fluid significantly decreased and the concentration of glucose, protein, calcium, iron, and aspartate aminotransferase activity significantly increased. Moreover, the concentration of protein, triglycerides, and aspartate aminotransferase activity in the eggs also changed. In the newly ovulated egg, the yolk consisted of homogenous tissue and its perivitelline space diameter had no considerable differences. With over-ripening, the yolk became heterogeneous, while chorion diameter and micropyle did not change. The perivitelline space diameter varied among different areas. The present study demonstrated that the best time to take Caspian brown trout eggs after ovulation at 7± 0.6°C was up to 10 DPO. Among the studied parameters of the egg and ovarian fluid, egg quality was related to both ovarian fluid parameters (e.g., pH, protein, aspartate aminotransferase, glucose, cholesterol, triglycerides, calcium, iron) and egg parameters (e.g., cholesterol, triglycerides, iron, aspartate aminotransferase). Thus, these parameters can be used as a egg quality markers in this species.

Determination of biological characteristics and blood groups of hilsa shad (Tenualosa ilisha) in the north of Persian Gulf (Bushehr coastal waters)

Hilsa shad, Tenuolosa ilisha, belongs to Clupeidae family and Alsinae subfamily is an euryhaline pelagic and anadromous species living in marine and freshwater waters. Regarding to study the biological characteristics of this species, this study was carried out in the Northern Persian Gulf within Bushehr province waters during years 2006-7. A total of 344 specimens were collected and transported to the laboratory for further different biological measurements consist of: reproduction (GSI, fecundity, maturity stages), feeding (stomach contents, food preference ...), aging; 58 morphometric and ٧ meristic measurements. The results indicate that minimum, maximum and mean body weight are 203, 953 and 481.35±147.64 g, respectively and this values for total length are 26.5, 45.1 and 35.99±3.98 cm. For food regimes, the most abundant phytoplankton foods belong to Bacillariophyta (83.74٪) and zooplanktons of Arthropoda (51٪). The mean RLG was measured about 1.53±0.22 and the mean feeding intensity was about 51.79±38.13. Also, the aging of studied fishes showed that they have 2 to 4 years and most of the samples had 2+ years. The reproduction studies showed that Indian shad is a batch spawner species. The average gonadosomatic index (GSI) is 1.64±1.382 and the main spawning season was determined for April-May.