Venomics and biological analysis of Scatophagus argus argus (Spotted scat) venom isolated in Persian Gulf, extraction and purification of hemolytic protein

Green scat namely as Scatophagus argus argus is a venomous aquarium fish belonging to Scatophagidae family. It can induce painful wounds in injured hand with partial paralysis to whom that touch the spines. Dorsal and ventral rough spines contain cells that produce venom with toxic activities. According to unpublished data collected from local hospitals in southern coastal region of Iran, S. argus is reported as a venomous fish. Envenomation induces clinical symptoms such as local pain, partial paralysis, erythema and itching. In the present study green scat (spotted scat) was collected from Persian Gulf coastal waters. SDS-PAGE indicated 12 distinct bands in the venom ranged between 10-250 KDa. The crude venom had hemolytic activity on human erythrocytes (1%) with an LC100 (Lytic Concentration) of about 1.7 μg. The crude venom can release 813 μg proteins from 0.5% casein. Phospholipase C activity was recorded at 3.125 μg of total venom. Our findings showed that the edematic activity remained over 48 h after injection. The purification of the venom was done by HPLC and 30 peaks were obtained within 80 min but only one peak in 68 min retention time showed hemolytic activity at 90% acetonitril was isolated. The area percentage of the hemolytic protein showed that this hemolytic protein consist of 32 percent of total proteins and its molecular weight was 72 KDa in SDS_PAGE. The results demonstrated that crude venom extracted from Iranian coastal border has different toxic and enzymatic activities.

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.

The study of biological characteristic and population dynamic of Caspian vimba (Vimba vimba persa), in coastal waters of Caspian Sea (Mazandaran Province)

This study was conducted to determine reproduction characteristics, diet regime, age structure and population dynamics parameters of the vimba vimba persa (Pallas, 1811) in Mazandaran waters of the Caspian Sea, from October 2008 to September 2009. A total of 994 specimens were monthly collected by beach seine and cast net from six fish landings of Ramsar, Tonekabon, Chaloos, Mahmood Abad, Sari and Behshahr. Biometric characters were measured for each specimen at the laboratory. Scales were used for age determination. Sex determination and fecundity were determined. Population dynamic parameters as well as stock assessment including cohort analysis were estimated using FISAT software. The finding showed that the mean of fork length and body weight of the Caspian Vimba were 168.4±2.6 mm and 71.94±32.24 g respectively. Strong correlation was found between these two variables (a= 0.012; b = 3.047; r2 = 0.955). 92 specimens were studied from the fecundity point of view. This species was found to have more abundance in spring (esp. Apr-May). The samples composed of 397(42.6%) male, 537(57.4%) female; Overall sex ratio (M: F =1: 1.35) was significantly different from the expected 1:1 ratio (p ≤0.05). The advanced stages of maturity (4th & 5th) were found in April and May. The highest Gonadosomatic Index in female was in May and the lowest one was in July. This fish is therefore a spring spawner. The maximum absolute and relative fecundities were 34640 and 260.9, respectively; the minimum absolute and relative fecundities were 5400 and 94.5 respectively. The averages of absolute and relative fecundities were 17198±7710 and 171.85±48.8, respectively. Coefficient vacuity index was 59.2% which indicates that this fish is mesophagous. Among of living creature consumes by Caspian Vimba mollusks, 76 arthropods, worms, plants, detritus and fishes were found 32.9% , 26.7% , 13.4% , 17% , 4.4% and 1.6% respectively. The infinite fork lengths were 261 mm for females, 25mm for males and 261 mm for both sexes respectively. For population growth and mortality parameters; K ( 0.28 per year for both sexes, 0.3 per year for males, 0.33 per year for females); t0 ( -0.65 year for both sexes, -0.23 year in females, -0.51 year in males ); Φ' ( 2.28 ); Z ( 0.98 per year ); M ( 0.59 per year); F ( 0.39 per year) and Exploitation coefficient was 0.4. The analysis showed that total biomass and MSY were 1336 and 528.8 tonnes respectively.

Evaluation of some biological indices and population dynamic of "Saurida tumbil" in the northern Persian Gulf and Oman Sea

The Great lizardfish is one of the commercial demersal fishes in the Persian Gulf and Oman Sea. In this study the food preference index (FPI), Fullness index (FI) and stomach contents of Saurida tumbil were evaluated to assess the quantity and kind of food which this species consumes in the study area. The samples were collected monthly from commercial bottom trawl fisheries in main fishing grounds of eastern Strait of Hormuz. A total of 346 specimens were collected during years 2009-2010 of which 269 specimens were female and 77 specimens were male. The minimum and maximum total length were 161 and 590 mm, respectively with the highest frequency in length group of 350-370mm. Results showed that 20.9% of stomachs were full, 18.9% were semi-full and 60.2% were empty. The main stomach contents (FPI=80.2%) were fishes and minor stomach contents (FPI=25.7%) were crustacean (Specially crab and shrimp) and cephalopods were found as random food. The minimum and maximum FIs were 2.5 and 42.5 in December and September respectively; and the overall CV was 60.2. The results indicate that this fish is a relatively frugal species which consumes fishes as main food and there is no difference in feeding of male and female.

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.

Biomonitoring of polycyclic aromatic hydrocarbons (PAH) and heavy metals Ni, V in sediments and anodonts of the Anzali Lagoon and applicability of netural red retention assay (NRR) as biomarkers of these pollutants

Biochemical ecotoxicology and biomarkers using are a new sciences that are used for biomonitoring in aquatic environment. Biomonitoring plays a vital role in strategies to identify, assess, and control contaminants. On the other hands in recent year's attention to polycyclic Aromatic Hydrocarbons (PAHs) and heavy metals increased in aquatic environments because of their carcinogenic and mutagenic properties combined with their nearly ubiquitous distribution in depositional environments by oil pollution or industrial waste waters. The present research aimed to assess PAHs and Ni, V levels in surface sediments and bivalves (Anodonta cygnea)and the effects of PAHs and heavy metals (Ni,V) on the hemocyte of the Anodonta cygnea were investigated in 2 stations (Mahrozeh, Selke in Anzali Lagoon, North of Iran). Samples were collected during at 2 different periods of the year, Dry and rain seasons, (June & September) and to confirm our first observations, Cage station is added. The bivalves hemocytes were monitored for membrane injury by NRR methods (neutral red retention assay). Heavy metal (Ni, V) concentrations were determined by Atomic Absorption in Anodonta cygnea and the sediments in Anzali Lagoon. The vanadium concentration in bivalves and sediments was ND(not detect )-0.4231 μg/g and 1.4381-306.9603 μg/g dry weight respectively. Nickel concentration in bivalves and sediments was 0.0231-1.3351, 0.4024-19.3561 μg/g dry weight respectively. PAHs concentrations were determined by GC-Mass in Anodonta cygnea and the sediments. Average concentration of PAHs is 115-373.788 ng/g dry weight in bivalves and average concentration of PAHs is 34.85-1339.839 ng/g dry weight in sediments. Bioaccumulation sediments factor(BASF) is high about PAHs (>1) and BASF is low for Ni, V (<1) . Internal Damage mechanisms of bivalves hemocytes (cell mortality, dye leakage, decreased membrane stability, are observed (Lowe Methods). Statistical analysis was used to explore the relationship between altered cellular and above contaminants. There are power and negative correlations between PAHs and NRR method for hemocytes in Anodonta cygnea (P<0.0005), but good correlation is not observed between Ni, V and NRR method for hemocytes in every time. This research indicates that the NRR assay is a useful screening technique able to discriminate polluted sites and at first we announce that Anodonta cygnea hemocytes are efficient biomarker for PAHs pollutants in fresh water.

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.

CORRELATION OF ZONA-BINDING WITH OOCYTE MATURATION AND SPERM MOTILITY IN RHESUS-MONKEYS BY HEMIZONA ASSAY

The hemizona assay (HZA) in Rhesus monkeys was employed to study the correlation of zona-binding ability with sperm motility or with naturally developing oocytes at various maturational stages. Oocytes from unstimulated ovaries were retrieved within 2 hr from monkeys sacrificed for vaccine production (in reproductive season, but with their menstrual cycles not determined). Oocytes were divided into four groups based on their morphological maturation: 1) Oocytes surrounded by more than one cumulus layer (MC); 2) Oocytes retaining intact germinal vesicle nuclei (GV); 3) Oocytes with germinal vesicle breakdown showing distinct perivitelline space (PVS); and 4) Oocytes extruding the first polar body (PB1). The mean numbers of sperm bound to hemizona for PBI, PVS, GV, and MC groups were 132.9 +/- 12.0, 71.5 +/- 10.1, 36.1 +/- 4.0, and 20.1 +/- 2.9 (Mean +/- SE), respectively. The four groups showed significant differences from each other in sperm/egg binding ability (P < 0.01). The number of bound sperm significantly increased with oocyte maturation. The present study also showed that zona-binding ability was also affected by sperm motility. For sperm with 67.7% motility and sperm with 31.2% motility, the average numbers of bound sperm were 43.5 +/- 2.2 and 25.3 +/- 2.9 (Mean +/- SE), respectively. There was significantly higher binding ability for sperm with higher motility (P < 0.01). The results suggest that: 1)The rhesus monkey model can serve as a very sensitive model for studying sperm/egg interaction by HZA; 2) Sperm motility positively correlated with sperm/egg binding; and 3) Sperm/egg binding ability increases with oocyte maturation. The binding ability is highest when oocytes matured to the PB1 stage, which is also the best opportunity for fertilization. This is strong evidence for the ''zona maturation'' hypothesis. (C) 1994 Wiley-Liss, Inc.

Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

The effect of spirulina (fresh and dry) on some biological factors in Penaeus semisulcatus larvae

Spirulina is a filamentous cyanobacteria with many applications in food and drug industries, as a food in human, aquaculture, vet and poultry industries… . Semi and mass culture of Spirulina carries out in different countries. This study was carried out in five phases in order to produce this microalga in Iran. The first phase, Spirulina pure stock was imported from Indonesia. After identification of species, it was cultured in laboratory until we took 20 liters of biomass. The semi-mass culture was carried out in green house. Cell concentration and size of Spirulina were recorded during culture daily and their growth rates were calculated. After two weeks, when the size of Spirulina was suitable, biomass of Spirulina was harvested then accumulated Spirulina weighted and dried in 24 hours in laboratory. In order to microbiological study, the samples of Spirulina (dry and fresh) were cultured on blood agar medium and coliforms were counted. The chemical composition of produced Spirulina was measured by standard methods. Fatty acid and amino acid profiles were acquired by GC and HPLC instruments, respectively. The amount of chlorophyll in Spirulina was determined by spectroscopy method. Also astaxanthin pigment as an important carotenoid was measured by HPLC in Spirulina and Penaeus semisulcatus larvae fed on Spirulina. At final phase of this project, larva fed on produced Spirulina (biomass and dry powder) was compared to Z plus, microencapsulated Spirulina (M.C.F) and Chaetoceros algae as control. This experiment was carried from zoa to early post larvae stage then survival and growth rate of larvae were recorded. The growth rate of larvae was evaluated with ANOVA test and survival rate of treatments was assessed by Log Rank (Mantel –Cox) test. Also during larvae stage, two parameters of water such as nitrate and nitrite were measured in zoa, mysis and post larvae stages. The results of this study were shown that colifom counts were 1.85×106 and 92.3×105 coliform per ml in fresh and dry spirulina, respectively. Protein percent of dry spirulina was 50.93 % (dry weight) and the amount of astaxanthin in spirulina and larvae fed on spirulina were 0.21 and 0.01 mg/kg, respectively. The most survival rate of larvae were observed in zoa III (88.8%) with Z plus supplement treatment, in mysis III (76.5%) combination of Z plus and dry spirulina in comparative between treatments. Larvae growth (4.5mm) of control in early post larvae was the best.

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.

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.