National plan for studying on how can develop aquaculture in Sistan and Baluchistan by carried out research projects such as: Twice a year cultured of shrimp, propagation and breeding of Hamoon fish (Shizothorax zarudnyi) and propagation and breeding of three Indian carp species

Hamoun fish, Schizothorax zarudnyi, is an indigenous species of the eastern waters of Iran. 331 Spawners weighing 800-2450 g were collected from the Chahnimeh reservoirs in early autumn, 2006. from 5/3/2007 (the project is supported and communicated on 2010) , Ovulation was stimulated with three stimulators; pituitary extract (3-6 mg kg-1 body weight), GnRH-A (20-30 mg kg-1 body weight) and anti dopamine (10-15 mg kg-1 body weight) that was given in 2-3 doses to breeders. Of 169 injected breeders , some were injected On mid March of 2007 (12-13 ºC water temperature) responded to the injection 25% ,while the rest were injected On April of 2008 (14-16 ºC water temperature) responded to the injection 65%. In the present project of 167 breeders 82 were female and 87 male. Totally 30 female breeders released their eggs in different stages. 20 female breeders released their eggs completely, 3 breeders released half of their eggs and 7 released 1/3 of their eggs. Larvae were fed with a mixture of powdered milk and egg yolk in this stage followed by decapsulated Artemia cysts and nauplii of Artemia and then on formulated starter diets used for carps. Because the ponds were not ready, larvae were maintained in troughs for about ten days before they were transferred to two 1200 m^2 earthen ponds where they reached a body weight of about 1 g. Larvae were fed with the starter feed SFCO in the earthen ponds. About 350 thousand larvae were stocked in two earthen ponds. Based on the results of present study and other studies we may conclude that artificial breeding in Schizothorax can be successfully achieved at 14-16 ºC in flow through systems using hormone therapy (combination of GnRha and anti dopamine) and larvae could be easily cultured in earthen ponds. However this species exhibits lower growth rates as compared to carps its high expenses could have an important role in economical feasible. Farming of Indian white Shrimp P.indicus in 4 private farm was done in 2008. Surveillance and monitoring of these farms, the possible obstacles to the harmful effects of management strengths and weaknesses and develop in the future was done in corporation of Offshore fisheries research center of Chabahar and fisheries of Sistan and Balouchestan. First crop was successful, but the shrimp of Nursery pond and second crop due to the occurrence of white spot disease (WSSV) disease and casualties were died and did not actually do the work. The average minimum and maximum feed conversion in Culture period 1.46 and 1.96 respectively, in C2 _31 and C2 _ 14 farms were observed. Maximum production was 41,376 kg in farm C2 _31. The rainfall on 14 August severe flooding and water supplying cussed suddenly fall down of Inland channel water salinity at day 16-20th August to 4-5PPT and the PH go up to 8.8-9. After 2 days the white spot disease in South of site was confirmed and was transferred immediately to the north of site. (The pilot’s farm). The results showed that shrimp farming can be done in two periods in year with a predetermined .In this study the only factor that could have adverse effects on the project was Feed supply problems during the growing period and the consequences that it caused low average body weight and final harvested Shrimp resulted to delay in daily growth. 30000 juvenile Indian majour carps (LabeoRohita, Cirhinusmerigala, Catlacatla) imported (2007 and 2008) and transferred to the earthen ponds in Astaneh Fisheries Research Station (Gilan ) and SheibanResaerch center (Khozestan) , In order to assess the viability of rearing fry up to 1 gram fingerlings under the climatic condition of north and south part of Iran . Growth parameters from the larval stage to 1 g were studied also .The fry with 300 mg weight, were released in to 3ponds. Results have indicated adaptations and considerable growth in this species, with an increase from the initial weight of 300 mg to an average of 13.5±1.6 mg in 50 days period until the end of November for fingerlings. Catla was gain 1 g. during 15 day while Roho and Merigal were gain 1 g. during 14 and 10 days respectively. Results revealed that the Indian major carps compatibility with the condition in Gilan province was successful and the combination of species and optimization of their compatibility specially in other provinces like Sistan and Baluchestan were remind for further studies

Population genetic study on two commercial shrimp species and determining of DNA-Barcoding of 6 – 8 species by partial sequencing of mtDNA using molecular method (PCR-Sequencing)

Barcodes are short segments of DNA that can be used to uniquely identify an unknown specimen to species, particularly when diagnostic morphological features are absent. These sequences could offer a new forensic tool in plant and animal conservation especially for endangered species. Ideally, barcodes could be used to positively identify illegally obtained material even in cases where diagnostic features have been purposefully removed or to release confiscated organisms into the proper breeding population. In the present study molecular data were used besides the morphological ones to introduce a specific barcode for six selected species of Shrimp. Based on the previous phylogenetics study, it has been proved that the mitochondrial COI gene in crustacean is a good discriminative marker at both inter- and intra-specific levels. In this regard, six species of shrimp (Fenoro penaeus indicus, Fenoro penaeus merguensis, penaeus semisulcatus, Metapenaeus affinis, Marsupenaeus japonicas, Fenoro penaeus penicillatus were collected from the Persian Gulf and Oman sea. After identification, the total DNA was extracted; COI gene was first amplified and then sequenced for each species. Finally the collected data were analyzed with the specific phylogenetic software. Molecular analysis revealed some degree of interpopulation differences but six good species were recognizable based on COI sequences. Also for population study molecular data of two species Penaeus semisulcatus and Metapenaeus affinis were analysed base on COI RFLP and 16SrRNA sequences respectively. The results would be helpful to protect shrimp species

Population genetic structure of Neogobius caspius (Eichwald, 1831) in the east and central of the south Caspian Sea using microsatellite markers

Neogobius caspius is a small benthic fish that is native to the Caspian Sea. The importance of this fish is because of it is role as a main food resource of the Sturgeon fish. The genetic diversity of N.caspius populations in the Caspian Sea was studied using microsatellite technique. In the study 95 Samples of Neogobius caspius from two regions (Torkman port and Chalus) in south Caspian Sea were collected. DNA was extracted using 12 pairs of microstellite primers polymerase chain Reaction (PCR) was conducted. DNA Bands were analyzed using uvdoct and GenAlEx software package. Out of 12 microsatellite primers, 11 loci were produced, in which 5 of them were polymorphic, 6 monomorph and one of them showed smear. Average of observed and expected hetrozygosity was 0.798 and 0/843 respectively. Significant genetic differences between 2 regions was observed (P<0.01). Deviations from hardy- Weinberg equilibrium were in all cases. These results indicate that at least, 2 population of Neogobius caspius exist in the south Caspian Sea

Genetic national plan study in order to shrimp breeding and developing of shrimp culture technology

Sampling was done using 90 post larvae which were produced by reproduction of some broodstock of Penaeus indicus in one day and reared in the same situation for 4 month. Samples were classified in 3 group high growth, medium and low (according to their weight and length). Genomic DNA was extracted from a 1cm^2 piece of muscle using the phenol-chloroform method. The polymerase chain reaction (PCR) was done using 21 RAPD loci. And PCR products were separated on 3% Agarose gels. From 21 studied loci, 12 produced polymorphic bands. The most polymorphic band produced using OPAQ 9 and the least by OPAQ 7. According to Nei 1972, the highest distance (0.457) was between low growth group and medium and the lowest (0.091) between high growth group and medium, therefore the highest identity (0.912) was between high growth group and medium and the lowest (0.633) between low growth group and medium. Consensus neighbour-joining tree using Nei (1972 and 1978) resulted in two clades, the first including high and medium growth groups and the second low growth group, it appears that low growth group are depended on separated population of the two others. With considering of mean weight of F1(16.25±1.5), mean weight of 15 ±1.2 and mean weight of parent 31.6, response to selection (R) and heritability for growth in this species were estimated 1.2±0.2 and 0.07±0.01 respectively. In another part of this study Sequencing of specific bands and primer design were done and examining of them on the same age specimens is necessary in following

Genetic characterization of Vimba vimba persa in southern part of the Caspian Sea using microsatellite marker

Population genetic structure of Vimba vimba persa was investigated using microsatellite markers from 4 regions along the Iranian coastline of Southern the Caspian Sea (Anzali lagoon & havigh River in Gilan province, BabolRoud River in Maz&eran province & GorganRoud River in Golestan province). Genomic DNA from 121 specimens was extracted from fin tissue by phenol-Chlorophorm method & PCR reaction was accomplished with 17 microsatellite primers, out of 17 microsatellite primers 14 loci were amplified, in which 10 of them were amplified with reasonable polymorphism & 4 were monomorphism. Totally 302 alleles were identified on average 7.5 Observed & expected heterozygosity averages were 0.80 & 0.77 respectively. Most cases significantly deviated from Hardy-Weinberg equilibrium (p 0.01). The estimation of Fst (p 0.01) revealed significant population structuring & estimate four population of Vimba vimba persa is identified in the Caspian Sea. These studies were to apply & develop population genetic approaches to assist conservation, sustainable harvest & restocking of these populations

Population genetic study on croaker species Atrobucca nibe in Persian Gulf and Oman Sea using molecular methods

Black Mouth Croaker (Atrobucca nibe) is of significant value from the economics point of view. In the following study, the genetic diversity of Atrobucca nibe has been investigated using the PCR-sequencing method from three different regions of the Persian gulf and Oman Sea, namely Chabahar in the Sistan and Baluchestan and Jask in Hormozgan Province of Iran and Fujairah in the Fujairah Emirate of the United Arab of Emirates. The purpose of the following exploration has essentially beencoined to further the study of Atrobucca nibe genetic diversity and its population structure in the Persian Gulf and Oman Sea. For this purpose, a number of 54 fish have been collected from the aforementioned regions. The DNA of the fish had been extracted through the Ammonium Acetate method and the quality of the DNAs were analyzed via the spectrophotometry and furthermore the quantity of the extracted DNA were assessed viaelectrophoresis. Polymerase Chain Reaction (PCR) was conducted on the targeted DNAs and thenceforth DNA sequencing was carried out. Subsequently, through sequencing it was discovered that the D-loop region in the mitochondrial DNA (mtDNA) of Black Mouth Croaker,contained 270 base pairs (bp). For the purpose of analyzing the genetic diversity of Atrobucca nibe and its population structure in the Persian Gulf and Oman Sea, the following softwares have been conducted, namely theBioEdit, Arlequin, Dnasp and Mega5 software. Furthermore, the Kimura 2-parameter was used for the purpose of genetic distance analysis. Through observations of average levels of FST between the regions, it can be summed up that genetic differences do exist among the present populations. On the grounds of the obtained results from the Persian Gulf and Oman Sea, it can be concluded that there currently are three different populations of Atrobucca nibe residing in the waters of Sistan & Baluchestan, Hormozgan and Fujairah

Study on propagation and breeding of Hamoun fish (shizothoraxzarudnyi) to one gram weight in earthen pond

Hamoun fish, Schizothorax zarudnyi, is an indigenous species of the eastern waters of Iran, which is exclusively found in this region. However recently drought occurrence in the Chahnimeh reservoirs (a semi natural water body) making them vulnerable to extinction. As an appropriate action to address this problem and according to the 3 side contract between Italian Government – UNDP - Islamic republic of Iran Government and the Italian government financial support to reduce poverty in the Province of Sistan-Baluchestan through the reinforcement and dissemination of aquaculture activities the project was developed at the Hamoun Research Unit by the Chahbahar Fisheries Research Center to record the breeding normative of 1 g weight larvae for restocking and other researching purposes. 331 broodstocks of the indigenous species Shizothorax zarudnyi weighing 800-2450 g were collected from the Chahnimeh reservoirs in early autumn, 2006. From 5/3/2007 (the project is supported and communicated on 2010) , Ovulation was stimulated with three stimulators; pituitary extract (3-6 mg kg-1 body weight), GnRH-A (20-30 mg kg-1 body weight) and anti dopamine (10-15 mg kg-1 body weight) that was given in 2-3 doses to breeders. Of 169 injected breeders , some were injected On mid March of 2007 (12-13 ºC water temperature) responded to the injection 25% ,while the rest were injected On April of 2008 (14-16 ºC water temperature) responded to the injection 65%. In the present project of 167 breeders 82 were female and 87 male. Totally 30 female breeders released their eggs in different stages. 20 female breeders released their eggs completely, 3 breeders released half of their eggs and 7 released 1/3 of their eggs. The male breeders just injected in the final dose of hormontrapy and all were ready for releasing sperms however the ovulation in female breeders occurs between 353-428 h ºC and after the final dose of injection. Ripe eggs were stripped from the females and fertilization was done by the dry method. Fertilized eggs were transferred to veis incubators and troughs. Incubation period for eggs differs and larvae hatch out after about 910 days at an average water temperature of 12.5 °C. Maintained at 13-14 °C, complete absorption of yolk sac in Shizothorax zarudnyi larvae occurred after 5-8 days. Larvae were fed with a mixture of powdered milk and egg yolk in this stage followed by decapsulated Artemia cysts and nauplii of Artemia and then on formulated starter diets used for carps. Because the ponds were not ready, larvae were maintained in troughs for about ten days before they were transferred to two 1200 m2 earthen ponds where they reached a body weight of about 1 g. They were then handed over to the Iranian Fisheries department in the region. Larvae were fed with the starter feed SFCO in the earthen ponds. About 350 thousand larvae were stocked in two earthen ponds. Based on the results of present study and other studies we may conclude that artificial breeding in Schizothorax can be successfully achieved at 14-16 ºC in flow through systems using hormone therapy (combination of GnRha and anti dopamine) and larvae could be easily cultured in earthen ponds. However this species exhibits lower growth rates as compared to carps its high expenses could have an important role in economical feasible

Molecular genetics study on population of Salmo trutta caspius spring and autumn forms in Southern Caspian Sea using microsatellite marker method basin and establish an alive gene bank

Caspian brown trout (Salmo trutta caspius) is an important from the point of economy. In this study genetic diversity of (Salmo trutta caspius) was investigated using microsatellite markers from two regions of the Iranian coastline of Southern Caspian Sea (Cheshmeh Kileh River in Mazandaran Province and Karganrud River in Gilan Province). The purpose of this research was the study of Salmo trutta caspius possible populations related to genetic diversity and population structure in the Caspian Sea and introducing the useful genetic markers. For this purpose the number 225 fishes from two Rivers were collected. DNA was extracted and amplified with 16 pairs of microsatellite primers using Polymerase Chain Reaction (PCR). The amplified products were loaded on polyacrylamide gels and bands were analyzed using Gene Alex and Pop Gene soft wares. Observed and expected heterozygosity averages were 0.46 ± 0.09 and 0.60 ± 0.10, respectively. According to the results from this study, Genetic polymorphism has been seen in this population

Genetic variation study on the grey mullet (Mugilcephalus) stocks from Mediterranean Sea, the Pacific Ocean and Oman Sea

Two of the three objectives of project was carried out success that one of them is following: The genetic diversity of Liza salien(Risso,1810) in the south part of Caspian sea using the Mitochondrial DNA sequencing (mtDNA) was carried out as first objective of project that based on the mitochondrial DNA sequencing (mtDNA) of 16S rRNA was used in order to clarify genetic structure and genetic diversity of lizasaliens in three western (Anzali), middle (sari), and eastern (Gomishan lagoon) of south part of Caspian sea. As a result we obtained 552base pairs of 16SrRNA sequence. A total of 6 different haplotypes and 29 variable sites were identified .The average nucleotide diversity(π) and haplotype diversity(h) in samples of all regions were 0.29 , and 0.004 respectively. The results obtained from genetic distance showed low rate in that of 3 regions. Estimates of gene flow indicated there is no reproductive isolation between three regions and also there was not significant genetic differentiation between differentregions (p>0.05). the findings from the present study suggest that there is equal population of Liza saliens in the studied regions: Genetic differences and phylogenic relationships among six Mugilidae species (Mugilcephalus, M. capito, Liza subviridis, L. saliens, L. aurata, Valamugilbuchanani) were determined using PCR-sequencing as second objective of project. M. cephalus, L. subviridis and V. buchanani from the Persian Gulf and Oman Sea, and L. aurata and L. saliens from the Caspian Sea were collected. Samples of an imported, Egyptian species M. capito (this species was mixed with the main imported species as M. cephalusfingerling) were obtained from the Gomishan Research Center in Gorgan. Total DNA from the samples were extracted according to phenol-chloroform method Mithochondrial DNA ,16s RNA was amplified using thermo cyclermachine with universal primers and thensequenced by sending to Takapoozist Company and thereafter to France. Analysis of the sequences showed great differences between Mugil species and the other studied species. The phylogenetic tree obtained through Neighbor-Joining method revealed that L. saliensand L. aurata were in the same branch while L. subviridis was in a separate branch. In contrast, Maximum Parsimony tree located L. subviridis and L. aurata in a single branch and assigned L. saliens to a distinct branch. This result brings in the question of monophyletic origin of the genus Liza.Also

Determining of DNA-barcoding of 6 – 8 species by partial sequencing of mtDNA (CO1) using molecular method (PCR-Sequencing)

Barcodes are short segments of DNA that can be used to uniquely identify an unknown specimen to species, particularly when diagnostic morphological features are absent. These sequences could offer a new forensic tool in plant and animal Conservation-especially for endangered species. It was proved that a small fragment of mitochondrial DNA from the 5′-end of cytochrome c oxidase subunit 1 (COI) gene as a reliable, quick and costeffective identification system for most Crustacea like shrimp. Take a look in DNA barcoding website show that there is a little data about Iranian shrimps which live in Persian Gulf and Oman Sea. In this Study six species of shrimp: Fenoro penaeus indicus, Fenoro penaeus merguensis, penaeus semisulcatus, Metapenaeus affinis, Marsupenaeus japonicas, Fenoro penaeus penicillatus were collected from different stations in Persian Gulf and Oman Sea. All materials were preserved in 70% ethanol and were shipped to the laboratory for taxonomic studies. After identification, the total DNA was extracted; COI gene was first amplified and then sequenced for each species. Finally the collected data were analyzed with the specific phylogenetic software. The results were amazing and the interesting part was that analytical methods for showing species relationship suggested that Fenoro penaeus penicillatus is closer to Marsupenaeus gnus than penaeus gnus. This finding needs more investigation to be proved. We suggest a workflow for DNA barcoding, including database generation and management, which will ultimately be necessary if we are to succeed to join universal DNA barcode for Crustacea