Aquaculture Asia, Vol.13, No.3, pp.1-60, July-September 2008

Comments on possible improvements to carp culture in Andhra Pradesh. Aquaculture and environmental issues in the region of Nai Lagoon, Ninh Hai district,Ninh Thuan province, Viet Nam. Climate change impacts on fi sheries and aquaculture. New initiatives in fisheries extension. Selection potential for feed efficiency in farmed salmonids. Freshwater prawn hatcheries in Bangladesh: Concern of broodstock. Production of Cirrhinus molitorella and Labeo chrysophekadion for culture based fisheries development in Lao PDR 2: Nursery culture and grow-out. Mussel farming: alternate water monitoring practice. Benefit-cost analysis for fi ngerling production of kutum Rutilus frisii kutum (Kamensky, 1901)in 2005 in Iran. The effects of feeding frequency on FCR and SGR factors of the fry of rainbow trout,Oncorhynchus mykiss. Asia-Pacific Marine Finfish Aquaculture Network Magazine: The use of poultry by-product meals in pelleted feed for humpback grouper. Production update – marine finfish aquaculture in the Asia-Pacific region. Crustacean parasites and their management in brackishwater finfish culture. NACA Newsletter

Management of wetland resources in the lower Mekong Basin: issues and future directions

The Lower Mekong Basin has extensive wetlands and these are being threatened by numerous problems. Most of these problems are interdependent and interact with one another. The lack of an appropriate definition of wetlands applicable to the region, pervasive inefficiencies and chronic lack of funds among riparian governments, and the poor appreciation of the true economic importance of wetlands and its resources are among the most prominent. The current definition, based on the Convention on Wetlands (Ramsar, Iran, 1971), is too broad when compared to the understanding of wetlands as being swamps, marshes and the like, and was developed specifically for wetlands with international importance as waterfowl habitats. Furthermore, wetlands are composed of different types of resources, which require different modes of management. Often, institutional competition, overlapping mandates and sometimes jealousies occur between government departments when they try to assert their authority on a particular wetland resource and use, and put forward their development plans without considering how these may conflict with other wetlands uses. Finally, effective wetland management requires reliable statistics or information on rate of harvest of natural resources such as fish and others, fishing/harvesting methods over time in order to determine the level of exploitation, and the status of the natural resources. This information is needed to identify opportunities for expansion, to establish historical trends, and to determine when management interventions are necessary to protect the resources from being overused by other developments. In order to address these issues, ICLARM - The World Fish Center has launched a project, the aim of objectives of which are described in this paper.

A new method for venom extraction from venomous fish, green scat

Scatophagus argus argus (Green Scat) is a pretty aquarium fish. Its hard spines are venomous and can cause painful injury. In this study 60 specimens of Green Scat were collected periodically from coastal waters of Boushehr (south of Iran) from May 2011 to April 2012. Anatomical features of venomous spines were investigated. Scat venom was extracted from the spines in a new manner for keeping the specimens alive. The nature of venom was tested by SDS-PAGE. Ethical issues and animal welfare principles such as rapid and instantaneous anesthetizing, post operation disinfection and fast recovery of the specimens was practiced in order to minimize the complications. This method enhanced the purity and quantity of venom as demonstrated by 12 separated proteins in electrophoresis. New ethical issues were developed to surviving the specimens and prolong viability as well.

Study morphologic, phylogenic relationships DNA barcoding of of identified decapod group (Crustacean) of Persian Gulf and Oman Sea.

Phylogenetic relationships among all described species (total of 12 taxa) of the decapoda, were examined with nucleotide sequence data from portions of mitochondrial gene and cytochrome oxidase subunit I (COI). The previous works on phylogeny proved that the mitochondrial COI gene in crustacean is a good discriminative marker at both inter- and intra-specific levels. We provide COI barcode sequences of commertial decapoda of Oman Sea, Persian Gulf, Iran. Industrial activities, ecologic considerations, and goals of the decapoda Barcode of Life campaign make it crucial that species of the south costal be identified. The reconstruction of evolut phylogeny of these species are crucial for revealing stock identity that can be used for the management of fisheries industries in Iran. Mitochondrial DNA sequences were used to reconstruct the phylogeny of the Penaeus species of marine shrimp. For this purpose, DNA was extracted using phenol- chloroform well as CTAB method. The evolutionary relationships among 12 species of the decapoda were examined using 610 bp of mitochondrial (mt) DNA from the cytochrome oxidase subunit I gene. Finally the cladograms were compared and the resulted phylogenetic trees confirmed that the Iran's species origin is Indo-west pacific species. Iran's species, which were not grouped with the other decapoda taxa seem to always form a sister clade with Indo-west pacific species with strong bootstrap support 100%. The result completely agrees with the previously defined species using morphological characters.However, we still lack any comprehensive and clear understanding of phylogenetic relationships in this group.

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.

Study of seasonal plankton communities changes in the Karoun dam reservoir and their relationship with water quality parameters

This study was under taken at Karoun Lake Dam No.4. (Southwest of Iran). Water samples were collected from March 2012 to February 2013 in three selected silts. Environmental parameters and chlorophyll a concentration were measured, as well as identification and abundance of Phytoplankton communities were studied. According to this study, 30 species of Phytoplankton were identified at four seasons. Most abundance was related to the phyla Bcillariphyta (17 species), Chlorophyta (6 species), Crysophyra (4 species), Dinophyta (2 species) and Cyanophyta (1 species) respectively. The results showed, the maximum rate of chlorophyll a concentration was measured in the warm with minimum level measured in the cold months. The rate of chlorophyll a concentration showed an oligotrophic condition in the lake of karoon 4 dam. positive significant correlation were seen between the parameters of COD,NO3,temperature, pH, turbidity, chlorophyll a and phytoplankton abundance (P<0.01). The chlorophyll a concentration and phytoplankton community had a significant negative correlation with transparency (-P < 0.01). According to this research, 4 phyla of zooplankton was identified, include Rotifera, Protozoa, Cladocera and Copepoda. Overal 43 species were identified at four seasons. Most abundance was related to the phyla Rotifera (27 species), Copepoda (7 species), Cladocera (5 species) and Protozoa (4 species) respectively. The chlorophyll a concentration, amount of phosphate and zooplankton indicator spesies, showed an oligotrophic condition in the lake of karoon 4 dam. A positive significant correlation was seen between all groups of zooplanktons abundance and temperature, as well as chlorophyll a concentration. (P<0.01) , whereas, there was negative correlation whith no significant between DO and zooplankton communities (P>0.05).

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.

Species identification and authentication of some of the most important seafood products in Iranian markets using biotechnological methods based on DNA

This study describes the molecular identification of sixteen fish species present in processed products imported into Iran for human consumption. DNA barcoding using direct sequencing of about 650 bp of the mitochondrial Cytochrome Oxidase subunit I gene revealed incorrect labeling (31.25%). Substitution of fish species constitutes serious economic fraud, and our results increase concern regarding the trading of processed fish products in Iran from both health and conservation points of view.

Effects of different levels of energy and protein sources on the growth performance, feeding, survival rate and the chemical body composition of juvenile Pacific white shrimp (Litopenaeus vannamei)

The present study was carried out in order to establish an economical effective diet for the pacific white shrimp in the southern part conditions of Iran. With the consideration of three dietary energy levels (E1=262, E2=312, E3=362 kcal 100 g-1 diet) and six ratios of fish meal (FM) to soybean meal (SBM) [(P1=100%FM+0%SBM, P2=80%FM+20%SBM, P3=60%FM+40%SBM, P4=40%FM+60%SBM, P5=20%FM+80%SBM, P6=0%FM+100%SBM)], 18 experimental diets (with 36% crude protein) were prepared. Completely randomized design was used to assign 54 polyethylene 300 litre round tanks provided by aeration and flow through water system and was stocked by 19 juvenile as 3 replicates to each treatment. Shrimps average weight was about 0.77 grams at the start. After 56 days culture period, maximum growth and nutritional performances were observed in the P6E1 treatment (containing 100% soybean meal and 262 kcal 100 g-1 diet) and P5E1 treatment (containing 80% soybean meal and 262 kcal 100 g-1 diet). Also the highest survival rate of the shrimps was observed in the P1E1, P1E2, P3E3 and P5E3 treatments. Additionally interactive effect of different protein ratios and energy levels had significant difference on body protein, fat, fiber and ash contents (P<0.05). Results of the present study suggest the possibility replacement of at least 80% of dietary fish meal by soybean meal in the diet of pacific white shrimp in the conditions of southern part of Iran.

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.

Population genetic structure of Rutilus frisii kutum in Caspian Sea using biometry and microsatellite molecular technique

In order to carry out Biometric studies, 75 samples were caught from 3 locations ( Tajan river, Sefidrud and Shirud) using Salic and the length (±1 mm) and weights (± 5 gr) of samples were determined. Using One-way ANOVA by SPPSS software, there wasn’t significant difference between locations in length and fecondity (P ≥0.01(, but there was significant difference between Shirud and tajan samples with sefidrud in weight ) P≤0.01(. In order to carry out genetic variation studies, 210 fish were caught from 3 different regions of the Iranian coastline (Khoshkrud, Tonekabon, Gorganrud) and 1 region in Azerbaijan (Waters of the Caspian Sea close to Kura River mouth) during 2008-2009 . Genomic DNA was extracted of fin using the phenol-chloroform. The quantity and quality of DNA from samples were assessed by spectrophptometer and 1% agarose gel electro-phoresis. PCR was carried out using 15 paired microsatellite primers. PCR products were separated on 8% polyacrylamide gels that were stained using silver nitrate. Molecular weight calculate using UVTech software. The recorded microsatellite genotypes were used as input data for the GENALEX software version 6 package in order to calculate allele and genotype frequencies, observed (Ho) and (He) expected heterozygosities and to test for deviations from Hardy-Weinberg equilibrium. Genetic distance between two populations was estimated from Nei standard genetic distance and genetic similarity index (Nei, 1972). Genetic differentiation between populations was also evaluated by the calculation of pairwise estimates of Fst and Rst values. From 15 SSR markers were used in this investigation, 9 of them were polymorph. Average of expected and observed heterozygosity was 0.54 and 0.49 respectively. Significant deviations from Hardy-Weinberg expectations were observed in all of location except Anzali lagoon- autumn in AF277576 and EF144125, Khoshkrud in EF144125 and Gorganrud and Kura in AF277576. Using Fst and Rst there was significant difference between locations ) P≤0.01(. According to Fst , the highest population differentiation (Fst= 0.217) was between Gorganrud and Khoshkrud that have the lowest Nm and the lowest (Fst= 0.086) was between Gorganrud and Tonekabon that have the highest Nm. Using Rst the highest population differentiation (Rst= 0.271) was between Tonekabon and spring Anzali lagoon and the lowest (Rst= 0.026) was between Tonekabon and Autumn Anzali 159 lagoon. Also the difference between Spring Anzali lagoon and Autumn Anzali lagoon was noticeable (Fst=0.15). AMOVA analysis with consideration of 2 sampling regions (Iran and Azerbaijan) and 7 sampling locations (Iran: Khoshkrud, Tonekabon, Gorganrud, Spring Anzali lagoon and Autumn Anzali lagoon ; Azerbaijan: the Kura mouth) revealed that almost all of the variance in data namely 83% )P≤0.01( was within locations, Genetic variances among locations was 14% )P≤0.01( and among regions was 3% )P≤0.01(. The genetic distance was the highest (0.646) between Gorganrud and Autumn Anzali lagoon populations, whereas the lowest distance (0.237) was between Gorganrud and Tonekabon River. Result obtained from the present study show that at least 2 different population of Rutilus frissi kutum are found in the Caspian sea,which are including the kura river population and the southern Caspian sea samples and it appears that there is more than one population in southern Caspian sea that should be attantioned in artifical reproduction Center and stoke rebilding.

The survey on growth and survival rate parameters with slurry and comparison by common method

This project was on “Study of Slurry, as an enrichment compound, on growth and survival parameters of Rutilus firisii kutum Kamensky 1901 larva and compare with routine condition according to hypothesis that use of Slurry, fertile organic compound, to increase the efficiency of survivorship and growth of Rutilus firisii and in other hand adaptation of natural food was performance.” The object of this project is to compare growth of Rutilus firisii and in usual condition and the condition of use Slurry The experiences performed in culture and propagation center Dr. Yousefpour decent (Associan of Dr. Beheshti culture and propagation) in Siahkal village, 32 Km far from southeast, in North of Iran. In this plan, three different treatments and related these nine pools , in 1.7 hec area (in same condition) were determined so that the density of storage was 1.7 million/hec larva. In this research at first treatment m we use slurry as enrichment compound during larval within a period of 13 days, and it used all the days of during larval in second treatment. Then the result of this study compared to control treatment. The results have shown the level of this study compared to control treatment. The results have shown the level of survivorship in pools were nutrition by slurry was 1.7 million times or twice than pools were nutrition with usual concentrate. In addition, growth coefficient such as daily weight growing (DWG) index, daily length growth (DLG), specific growth ratio (SGR) were measured in this case. All of these parameters in slurry treatment were shown noticeable enhancement in first treatment Thereof in the first week there was significant difference (P<0.001) between the average of length and weight in first week , and in the second week there was a significant difference (P<0.05) between them.

Study of inter species diversity and population structure by molecular genetic method in Iranian Artemia

Artemia is a small crustacean that adapted to live in brine water and has been seen in different brine water sources in Iran. Considering the importance of genetic studies manifest inter population differences in species, to estimate genetic structure, detect difference at molecular level and separate different Artemia populations of Iran, also study of phylogenic relationships among them, samples of Artemia were collected from nine region: Urmia lake in West Azerbaijan, Shoor and Inche-Borun lakes in Golestan, Hoze-Soltan and Namak lakes in Qom, Maharloo and Bakhteghan lakes in Fars, Nough pool in Kerman and Mighan pool in Markazi and DNA extracted by phenol-chloroform method. Primers designed on a ribosomal fragment (16s rRNA) of mt DNA sequence and PCR was done. Digestion of the 1566 bp segment PCR product by 10 restriction endonuclease (Alu I, EcoR I, Eco47 I, Hae III, Hind III, Hinf I, Mbo I, Msp I, Rsa I, TaqI) showed 25 different haplotypes: 9 in Urmia, 4 in Shoor and Inche- Borun, 1 in Namak and Hoze-Soltan, 3 in Mighan, 1 in Bakhtegan Maharlo, 3 in Maharloo and 4 in Nough. Measurement of haplotype and nucleotide diversity intra population and nucleotide diversity and divergence inter populations and evolutionary distance between haplotypes showed a high diversity in mitochondrial genome of Artemia in studied regions whose results are similar to those explained for highly geographic expansion organism. In addition, results showed considerable heterogeneity between different populations and there are enough evidences in haplotypic level for separation of studied samples and division of Iranian Artemia to seven populations including Urmia, Shoor and Inche-Borun, Hoze-Soltan and Namak, Maharloo, Bakhteghan, Nough and Mighan. Phylogenetic analysis of the 16S rRNA data set resulted strict consensus and neighbor joining distance trees, demonstrated that all samples were monophyletic and parthenogenetic form derivation from bisexual populations and genetically high resemblance to those of A. urmiana. Study of 270 specimens from different region showed the genus Artemia in Iran clustered into three clades including: 1- Shoor, Inche-Burun, Hoze-Soltan, Namak, Bakhtegan and Maharloo 2- Nough and Mighan 3- Urmia. Totally, obtained results indicated to ability of used techniques for study of inter species diversity, population structure, reveal of phylogenic relationship and dividing of different populations of Artemia in Iran.

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.