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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. Iranian Fisheries Science Research Institute Published

In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected which indicates protein expression of luxB, luxA.

In this study genetic characterization of Barbus brachycephalus caspius, Lucioperca lucioperca , Rutilus rutilus caspius , Rutilus frisi kutum and Salmo trutta caspius were examined by 611 samples from regions in East (Guilan province), Middle (Mazandaran province) and west (Golestan province) of southen part of the Caspian Sea. DNA was extracted from fin tissue by phenol - chlorophorm method and then PCR was performed using special primers. Statistical analysis of data was performed by Gene Alex, MEGA and Arlequin softwares. - Rutilus frisi kutum: The results showed that nine of ten primers were polymorphic loci. The mean of effective and observed alleles were 7.26±0.49 and 4.37± 0.35 respectively. Also, the mean of expected and observed heterozygosity were 0.55±0.03 and 0.69±0.02 respectively. Of the analysed loci, all of the samples (except Tajan and samples in LOC4 and Gilan samples in MFW2) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium(P

This study was conducted for the first time and the effects of probiotic bacteria isolated from the gut of Rainbow trout (Oncorhynchus mykiss). In the first step, after the purification of the isolated bacteria, probiotic properties in vitro was assessed with injection to healthy fish and challenge with three species of pathogenic bacteria in culture media. The second phase of the bacteria isolated in 5 treatments (log 7, 8 and 9 of the lactic bacteria (LAB), Vibrio sp and Pseudomonas sp) and a control treatment on growth and survival, hematological parameters (RBC and WBC cells), immunology and physiology parameters tested during the 60 days of the above parameters after 30 and 60 days and finally at the end, the final assessment was Streptococcosis resistance in fish. The results showed that the isolated bacteria were able to enhance the growth parameters (weight, feed conversion, feed fat, protein efficiency ratio and specific growth rate) and survival. The results showed that log 8 LAB was significant difference with other treatments and control. When using log 8 LAB and Vibrio, the MCV, MCH and MCHC were decreased with no significant difference. The log 8 has the greatest effect on the amount of liver enzyme (AST), IgM and complement component C3, and had significant difference with other treatments. The challenge examination to Streptococcus iniae showed the highest survival in treatments with log 8 (96.66%), and Vibrio (93.33%) and then subjected to other treatments and control are also the least survival (25.38%). The conclusion of that study is the first probiotic properties of bacteria isolated from trout to changes in quality indicators are in particular the LAB bacteria and the second log 8 of LAB had significant positive of changes development and safety, and the fish are resistanted against Streptococcosis.

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.

This study was conducted for the first time and the effects of probiotic bacteria isolated from the gut of Rainbow trout (Oncorhynchus mykiss). In the first step, after the purification of the isolated bacteria, probiotic properties in vitro was assessed with injection to healthy fish and challenge with three species of pathogenic bacteria in culture media. The second phase of the bacteria isolated in 5 treatments (log 7, 8 and 9 of the lactic bacteria (LAB), Vibrio sp and Pseudomonas sp) and a control treatment on growth and survival, hematological parameters (RBC and WBC cells), immunology and physiology parameters tested during the 60 days of the above parameters after 30 and 60 days and finally at the end, the final assessment was Streptococcosis resistance in fish. The results showed that the isolated bacteria were able to enhance the growth parameters (weight, feed conversion, feed fat, protein efficiency ratio and specific growth rate) and survival. The results showed that log 8 LAB was significant difference with other treatments and control. When using log 8 LAB and Vibrio, the MCV, MCH and MCHC were decreased with no significant difference. The log 8 has the greatest effect on the amount of liver enzyme (AST), IgM and complement component C3, and had significant difference with other treatments. The challenge examination to Streptococcus iniae showed the highest survival in treatments with log 8 (96.66%), and Vibrio (93.33%) and then subjected to other treatments and control are also the least survival (25.38%). The conclusion of that study is the first probiotic properties of bacteria isolated from trout to changes in quality indicators are in particular the LAB bacteria and the second log 8 of LAB had significant positive of changes development and safety, and the fish are resistanted against Streptococcosis. Iranian Fisheries Science Research Institute Published

Genetic knowledge helps to protect biodiversity and optimal harvest resources by several ways. This knowledge can help to reduce the risk of extinction to those populations that have high genetic diversity or diminished by detected them. Also, studies of population genetics can present guidelines for improving the structure of the population and understanding the biology of species. One of the most important applications of DNA database is detecting species, fishing offense, diagnose of anemia and genetic classification of animals. DNA bank has helped to identify the species that are hunted and were discovered only parts of their meat and texture. Optimal Storage of aquatic genetic resources and the conservation of biological diversity, as the national capital by using biotechnology methods are the most important goals of gene bank and database reserves of the country creation. Collecting biological specimens of aquatic species, preparation and long-term maintenance of them, genetic registration of endangered and threatened species and the use of biotechnology techniques for the protection, conservation and management of aquatic genetic resources is one of the other functions of the gene bank. Gene bank of inland waters of Iran (Gorgan) is planned and implemented to identify, collect, maintain and protect species at the level of genes, cells, tissues and live fish research and commercial exploitation. Despite the predicted projected funding for this project, unfortunately funding sources have been considered, does not allocated. Therefore some parts of the aims of this project has been organized by Golestan province credits to build molecular laboratory and aquaculture pond in Alang farm and exploitation of potential Gharasou station and Sijoval center restocking and the conditions for the bulk of the project goals is provided. Some parts like tissue culture laboratory and bioinformatics as well as parts and equipment related to the freezing of sperm and embryo development are the items that should be considered in the development of the this bank.

Goals: Determine of barcode of DNA in green tiger prawn, Penaeus semisulcatus, in the Gen bank of the species. Material and methods: In these study 30 specimens of Penaeus semisulcatus from each region in the Persian Gulf and Sea of Oman were sampled and preserved in ethanol 96%. 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. Result and discussion: Molecular analysis revealed some degree of interpopulation differences within two areas. Also for population study molecular data of species Penaeus semisulcatus were analysed base on COI RFLP and 16SrRNA sequences respectively. The results indicated that COI gen is a good marker for shrimp species differentiation that would be helpful to protect shrimp species.

Goals: Determine of barcode of DNA in green tiger prawn, Penaeus semisulcatus, in the Gen bank of the species. Material and methods: In these study 30 specimens of Penaeus semisulcatus from each region in the Persian Gulf and Sea of Oman were sampled and preserved in ethanol 96%. 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. Result and discussion: Molecular analysis revealed some degree of interpopulation differences within two areas. Also for population study molecular data of species Penaeus semisulcatus were analysed base on COI RFLP and 16SrRNA sequences respectively. The results indicated that COI gen is a good marker for shrimp species differentiation that would be helpful to protect shrimp species. Iranian Fisheries Science Research Institute Published

In this study genetic characterization of Barbus brachycephalus caspius, Lucioperca lucioperca, Rutilus rutilus caspius, Rutilus frisi kutum and Salmo trutta caspius were examined by 611 samples from regions in East (Guilan province), Middle (Mazandaran province) and west (Golestan province) of southen part of the Caspian Sea. DNA was extracted from fin tissue by phenol -chlorophorm method and then PCR was performed using special primers. Statistical analysis of data was performed by Gene Alex, MEGA and Arlequin softwares. -Rutilus frisi kutum: The results showed that nine of ten primers were polymorphic loci. The mean of effective and observed alleles were 7.26±0.49 and 4.37±0.35 respectively. Also, the mean of expected and observed heterozygosity were 0.55±0.03 and 0.69±0.02 respectively Of the analysed loci, all of the samples (except Tajan and samples in LOC4 and Gilan samples in MFW2) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium (P

In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected. , which indicates protein expression of luxB, luxA. Iranian Fisheries Science Research Institute Published

S. iniae is an important pathogen in both marin animals and humans, causing systemic infections in these hosts. The symptoms of infections are very similar to symptoms of infections caused by human¬specific streptococcal pathogens. The enzyme phosphoglucomutase (PGM) has recently been discovered to play an important role in polysaccharide capsule production and virulence in s. iniae. We aim to isolate S. iniae and cloning phosphoglucomutase gene. S. iniae grew on sheep blood agar as mucoid and beta- hemolytic colonies after 24 h of incubation at 37C. Then confirmed it by sensitive and rapid method of PCR. The pgm gene was amplified successfully and cloned in pTZ57R cloning vector.The recombinant plasmid was sub cloned into pETDuet-l expression vector by rectriction enzymes and confirmed by PCR . Although S. iniae is a pathogen of economic importance, relatively little is known regarding mechanisms of its pathogenesis and few specific virulence determinants have been established. These include the enzyme phosphoglucomutase which contributes to cell wall integrity and resistance to antimicrobial peptides. Thus, this enzyme is the best choice to product the vaccine and more investigations. Iranian Fisheries Science Research Institute Published

Nowadays, medical plants extract is used in aquaculture industry to enhance growth, immune system and fish resistance to different pathogens. In this study, some non-specific immune responses, hematology and biochemical parameters in rainbow trout juveniles (16 gr mean weight) and their resistance to Streptococcus iniae were evaluated following oral administration of Echinacea purpurea and Astragalus sp. extract. Three concentrations (0.5, 1 and 1.5 gr/kg of feed) of Echinacea purpurea and three concentrations (2, 3 and 5 gr/kg of feed) of Astragalus sp extract and control group were used for 60 days. Parameters evaluated included: levels of C4, C3 (complement components), free oxygen radicals (respiratory burst), Lysozyme, numbers of lymphocytes, monocytes and neutrophils. In the end, the relative survival rate (RSR) of fish was evaluated against S. iniae. The results showed that the levels of C3, lysozyme, oxygen free radicals, the number of neutrophils and lymphocytes in the experiment groups (the highest dose) compared to control group were increased significantly (p 0.05). The relative survival rates of fish following challenge with Streptococcus iniae, were 91/11, 93/33 and 44/44 percent in experiments (Echinacea purpurea and Astragalus sp.) and control groups, respectively. In conclusion, it may be state that Echinacea purpurea and Astragalus sp. extracts enhance the non-specific immune system and fish resistance against Streptococcusis, and it seems it can be used as immunostimulant in feed. Iranian Fisheries Science Research Institute Published

Nowadays, medical plants extract is used in aquaculture industry to enhance growth, immune system and fish resistance to different pathogens. In this study, some non-specific immune responses, hematology and biochemical parameters in rainbow trout juveniles (16 gr mean weight) and their resistance to Streptococcus iniae were evaluated following oral administration of Echinacea purpurea and Astragalus sp. extract. Three concentrations (0.5, 1 and 1.5 gr/kg of feed) of Echinacea purpurea and three concentrations (2, 3 and 5 gr/kg of feed) of Astragalus sp extract and control group were used for 60 days. Parameters evaluated included: levels of C4, C3 (complement components), free oxygen radicals (respiratory burst), Lysozyme, numbers of lymphocytes, monocytes and neutrophils. In the end, the relative survival rate (RSR) of fish was evaluated against S. iniae. The results showed that the levels of C3, lysozyme, oxygen free radicals, the number of neutrophils and lymphocytes in the experiment groups (the highest dose) compared to control group were increased significantly (p 0.05). The relative survival rates of fish following challenge with Streptococcus iniae, were 91/11, 93/33 and 44/44 percent in experiments (Echinacea purpurea and Astragalus sp.) and control groups, respectively. In conclusion, it may be state that Echinacea purpurea and Astragalus sp. extracts enhance the non-specific immune system and fish resistance against Streptococcusis, and it seems it can be used as immunostimulant in feed.

In order to have a sustainable management on Persian sturgeon (Acipenser persicus) as a highly commercial species in the South Caspian Sea, we need to identify its population structure and the level as well as its conservation status in their natural habitat. To develop a conservation program for this all Caspian Sea' sturgeon species it requires knowledge of its genetic diversity using reliable molecular marker to study population genetic structure. For these purposes, an enriched library was prepared based on a modified biotin-capture method. Approximately 1800 positive clones were screened for microsatellites in an Acipenser persicus genomic library. Of these 350 positively hybridizing clones were sequenced, and 81 clones were identified as having microsatellites with adequate flanking regions. We developed and tested 68 microsatellite primer pairs for Persian sturgeon. Out of 68 primer pairs developed, 11 pairs resulted in poor or no amplification, 13 were ambiguous, 6 were monomorphic, 20 were tetrasomic and 18 were octosomic in Persian sturgeon. While none of the markers showed disomic inheritance in Persian sturgeon and Russian sturgeon (A. gueldenstaedtii). Several of the markers appeared useful for studies stellate sturgeon (A. stellatus), ship sturgeon (A.nudiventris) and beluga (Huso huso). Nearly all the polymorphic pattern for ship, stellate and beluga displayed the simple banding patterns characteristic of disomic loci, while those for Russian sturgeon displayed banding patterns characteristic of tetraploid or higher polyploid levels. These markers may prove useful in a variety of future sturgeon population genetic studies in the Caspian Sea. Iranian Fisheries Science Research Institute Published

In this study, some non-specific immune responses and hematological parameters in rainbow trout juveniles (16g mean weight) and their resistance to Streptococcusis were investigated following dietary administration of 3 concentrations (0.5, 1, 1.5g/kg of feed) of Echinacea purpurea extract. The non-specific immune response and hematological parameter compared with control group for 60 days. Evaluated parameters included were of C3, C4, (complement components), superoxide ions (respiratory burst), lysozyme activity, number of WBC, percentage of blood lymphocytes, monocytes and neutrophils. At the end of trial, the relative survival rate (RSR) of fish was evaluated against S. iniae. The results showed that the levels of C3, lysozyme activity, superoxide ions, number of WBC and percentage of neutrophils in the experiment groups (the highest concentration, 1.5g/kg of feed) were increased significantly compared to the control group. Whereas, no significant difference was found in the value of C4 and the percentage of monocytes and lymphocytes in comparison to the control group. The relative survival rates of fish following challenge with Streptococcus iniae, were 91.11 and 44.44 percent in experiment (Echinacea purpurea, concentration of 1.5g/kg of feed) and control group, respectively. In conclusion, the results of the present study demonstrated that Echinacea purpurea extract enhanced the non-specific immune system and fish resistance against streptococcusis, suggesting that this extract might be used as immunostimulant in fish feed.

In order to do this study, 150 Persian sturgeon having 75g mean weight transferred to tanks with 500L volume. After dividing the fishes into two groups (experimental and control group), the fingerlings fed by concentrate food that contained different levels of Zataria multiflora essential oil (15g/kg, 25g/kg, 50g/kg, 100g/kg) for 8 weeks. Finally, sampling carried out on blood and serum of fishes. After preparing the samples based on standard methods, haematology and serology studies, performed. The results showed significant difference in mean and standard deviation of monocytes between experimental and control groups (P 0.05). This research also shows that Zataria multiflora essence has an anesthetic effect but because of its unwanted severe irreparable breathing (respiration) side effects and also the deformation of the fish which is against the fisheries’ ethics (death accompanied with extreme pain) it would be better not to be used as anesthetics. One of the advantages of Zataria multiflora essence as anesthetics is its low concentration (25 ppm) for anesthetizing the fish and also its economical cost. In addition, this essence has antimicrobial and disinfecting effects and, therefore, it can remove the infection from the external surface of the body of the fish. One of the most noticeable defects of Zataria multiflora essence is that at the time of anesthetizing it causes a state of excitement and irritability in the fish which manifests itself in the speed of swimming and the possibility of jumping out of water. As a result, although Zataria multiflora essence has anesthetizing effects, it is recommended not to be used for anaesthetizing the fish, because it posses unwanted side effects that may even cause the death of the fish; however, it may be used as a sedative during manipulating the fish.

Streptococcosis is an acute infectious disease that causes mortality in marine and freshwater aquacultures. One of the most important hosts is susceptible to the disease of rainbow trout and during the past decade its industrial production has been growing in Iran. According to FAO, Iran is among the 10 countries producing fish in the world. The most important bacteria causing Strptococcosis includes Strptococcus, Vagococcus and Enterococcus genera. But in all cases, the clinical sings are the same. More than a decade has passed since the first report of this disease in Iran and unfortunately, this disease has become the most important problems of trout production in Iran. Therefor, 72 farms were selected in 8 provinces including Mazandaran, Gilan, Lorestan, CharmahalBakhtiyari, Fars, Kermanshah, Tehran and Kokiloye-Boyer Ahmad and a total 520 moribund and apparently healthy rainbow trout (weight 50 – 200gr) were collected during summer 2008 to 2009. Fish kidney, spleen and liver samples were culture aseptically and finally 206 isolates were identified as gram positive cocci. Using conventional biochemical test, S.uberis, S.agalactiae, S.dysgalactiae, S.faecium and S.inia. Additionally, 172 from 206 isolates were confirmed as S.uberis، S.agalactiae، S.dysgalactiae، S.faecium and S.inia using a PCR assay ased on the results, the most prevalent is belong to S.uberis (isolated from 5 of 7 province), S.dysgalactiae (isolated from 4 of 7 province), S.agalactiae (isolated from 2 of 7 province) and S.faecium and S.inia (from 1 of 7 province) respectively. It is necessary to mention, S.inia was identified just to Fars and any gram positive cocci was isolated from Tehran.

Streptococcosis is an acute infectious disease that causes mortality in marine and freshwater aquacultures. One of the most important hosts is susceptible to the disease of rainbow trout and during the past decade its industrial production has been growing in Iran. According to FAO, Iran is among the 10 countries producing fish in the world. The most important bacteria causing Strptococcosis includes Strptococcus, Vagococcus and Enterococcus genera. But in all cases, the clinical sings are the same. More than a decade has passed since the first report of this disease in Iran and unfortunately, this disease has become the most important problems of trout production in Iran. Therefor, 72 farms were selected in 8 provinces including Mazandaran, Gilan, Lorestan, CharmahalBakhtiyari, Fars, Kermanshah, Tehran and Kokiloye-Boyer Ahmad and a total 520 moribund and apparently healthy rainbow trout (weight 50 – 200gr) were collected during summer 2008 to 2009. Fish kidney, spleen and liver samples were culture aseptically and finally 206 isolates were identified as gram positive cocci. Using conventional biochemical test, S.uberis, S.agalactiae, S.dysgalactiae, S.faecium and S.inia. Additionally, 172 from 206 isolates were confirmed as S.uberis ، S.agalactiae ، S.dysgalactiae ، S.faecium and S.inia using a PCR assay. Based on the results, the most prevalent is belong to S.uberis (isolated from 5 of 7 province), S.dysgalactiae (isolated from 4 of 7 province), S.agalactiae (isolated from 2 of 7 province) and S.faecium and S.inia (from 1 of 7 province) respectively. It is necessary to mention, S.inia was identified just to Fars and any gram positive cocci was isolated from Tehran. Iranian Fisheries Science Research Institute Published

In order to have a sustainable management on Persian sturgeon (Acipenser persicus) as a highly commercial species in the South Caspian Sea, we need to identify its population structure and the level as well as its conservation status in their natural habitat. To develop a conservation program for this all Caspian Sea' sturgeon species it requires knowledge of its genetic diversity using reliable molecular marker to study population genetic structure. For these purposes, an enriched library was prepared based on a modified biotin-capture method. Approximately 1800 positive clones were screened for microsatellites in an Acipenser persicus genomic library. Of these 350 positively hybridizing clones were sequenced, and 81 clones were identified as having microsatellites with adequate flanking regions. We developed and tested 68 microsatellite primer pairs for Persian sturgeon. Out of 68 primer pairs developed, 11 pairs resulted in poor or no amplification, 13 were ambiguous, 6 were monomorphic, 20 were tetrasomic and 18 were octosomic in Persian sturgeon. While none of the markers showed disomic inheritance in Persian sturgeon and Russian sturgeon (A. gueldenstaedtii). Several of the markers appeared useful for studies stellate sturgeon (A. stellatus), ship sturgeon (A.nudiventris) and beluga (Huso huso). Nearly all the polymorphic pattern for ship, stellate and beluga displayed the simple banding patterns characteristic of disomic loci, while those for Russian sturgeon displayed banding patterns characteristic of tetraploid or higher polyploid levels. These markers may prove useful in a variety of future sturgeon population genetic studies in the Caspian Sea.

The Mendelian Inheritance of produced hybrids from Chalcalburnus chalcoides and Vimba vimba persa was investigated using Microsatellite Markers. Genomic DNA from the produced larva and also small piece of dorsal and pectoral fin of their parents were extracted by Phenol-Chlorophorm method and PCR reaction was accomplished. 10 loci were amplified, in which 7 of them were amplified with reasonable polymorphism and 3 were monomorphism. In order to study the segregation of the produced F1 hybrids from female Chalcalburnus chalcoides and male Vimba vimba persa, seven loci (LCO3, LCO1, Lid11, Rru2, Z1-2, Z7-8, and Z9-10) were used. According to the results it showed that parents and F1 larva‘s allelic distribution were almost segregated based on Mendelian inheritance in 7 loci. The present study showed that Microsatellite markers are useful marker in study of inheritance.

In order to study the Inheritance of Microsatellite Markers in the hybrid of Chalcalburnus chalcoides and Vimba vimba persa in the Southern part of the Caspian Sea, hybridization between the species was carried out during spring (2009) in Shahid Ansari reproduction and culture workstation. 30 samples from the produced larva and also small piece of dorsal and pectoral fin of their parents were collected and stored in pure ethanol (96%) and transferred to the biotechnology of the Caspian Ecology Academy. DNA was extracted from fin samples of the parents, whilst the whole F1 progeny was used to isolate the DNA by Phenol-chloroform method. Quantity and quality of extracted DNA was determined using 1% Agarose electrophorus. DNA was used for PCR ampilification of 10 microsatellite loci. The amplifications were run on 6% Polyacrylamid electrophorus. The present study showed that 7 out of 10 markers were polymorphic and variated between the parents. Goodness of fit to the Mendelian inheritance ratio was determined by X2 analysis. 4 primers were segregrated according to Mendelian expectations at a significance level of P 0.05. Based on UPGMA dendrogram (Nei, 1978) two different clusters were obtained in which Chalcalburnus chalcoides and hybrid were in the same cluster but Vimba vimba persa was in the other. It seems that Vimba vimba persa is a joint ancestor for these two. High heterozygosity presence in parents and their F1 shows the high potential of genetic variety in Chalcalburnus chalcoides and Vimba vimba persa which from the aquaculture management and conservation view is important and considering the high genetic variety in their hybrid, the hybrid has the sufficient potential for race improvement. Iranian Fisheries Science Research Institute Published

Study of Mahisefid population using mtDNA with PCR Mahisefid diversity was studied in four rivers including Lamir, Sefidrood, Shirrood and Tajan. Total sample was collected from four rivers when fish migrat to river for spawning, 100 samples from Sefidrood, 98 samples from Lamir, 48 samples from Shirrood and 48 samples from Tajan. DNA was extracted with phenolcholorophorm. Samples were used for RFLP, the PCR product were digested by 20 restriction enzymes as follow: TasI, HaeIII, HinfI, HincII, SalI, DraI, AccI, AvaII, XhaI, BshNI, AvaI, BclI, BshII, MspI, PstI, RsaI, SdnI, TaqI, TruI, VspI. The four restriction enzymes including: TasI, HaeIII, HinfI, HincII showed diversity, 6 enzyme didn t have any restriction and 14 enzyme showed monomorphic. Total 20 haplotype studied that haplotype AAAA and BAAA had most frequency. The average haplotype frequency of AAAA was 29.93% and the average haplotype frequency of BAAA was 27.55%. 2- Study of population of Mahisefid with microsatellite markers 120 specimens of R.frissi kutum were used from four rivers to test thirty primers (30 samples from each river) of which 8 primers showed polymorphism. A large variation in heterozygosity average over all samples was observed among loci, that ranged from 0.13 to 0.91. For a given locus, observed heterozygosity varied greatly among the samples. For example, in Lamir 0.07 at SPY5 and 1 in Sefid Rud at CA1. Tajan had an observed heterozygosity of only 0.17, whilst the Lamir had an observed heterozygosity of 0.07 at locus SPY5. At locus SPY4, Tajan and Sefid Rud are the same and (0.53) and Shir Rud is 0.83 and Lamir is 0.47. However, despite these differences, there was clear difference in average heterozygosity observed between the samples. To investigated of Hardy-Weinberg Equilibrium in all locus and all rivers deviate showed from Hardy-Weinberg Equilibrium was significantly different ( P 0.05) between all samples except Lamir River and Sefid Rud River. The highest Fis was in Lamir River (0.84) in locus SPY5 and in Sefid Rud River in locus SPY5 (0.71), in locus CA12 (0.67) in Shir Rud River and SPY5 (0.63) in Tajan River. The lowest Fis is in locus CA1 (-0.55) in Sefid Rud River, in locus CA1 (-0.47) in Tajan River, in locus CA1 (-0.28) in Lamir River and in locus SPY6 (0.16) in Shir Rud River.

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 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. Iranian Fisheries Science Research Institute Published

Study of Mahisefid population using mtDNA with PCR Mahisefid diversity was studied in four rivers including Lamir, Sefidrood, Shirrood and Tajan. Total sample was collected from four rivers when fish migrat to river for spawning, 100 samples from Sefidrood, 98 samples from Lamir, 48 samples from Shirrood and 48 samples from Tajan. DNA was extracted with phenolcholorophorm. Samples were used for RFLP, the PCR product were digested by 20 restriction enzymes as follow: TasI, HaeIII, HinfI, HincII, SalI, DraI, AccI, AvaII, XhaI, BshNI, AvaI, BclI, BshII, MspI, PstI, RsaI, SdnI, TaqI, TruI, VspI. The four restriction enzymes including: TasI, HaeIII, HinfI, HincII showed diversity, 6 enzyme didn t have any restriction and 14 enzyme showed monomorphic. Total 20 haplotype studied that haplotype AAAA and BAAA had most frequency. The average haplotype frequency of AAAA was 29.93% and the average haplotype frequency of BAAA was 27.55%. 2- Study of population of Mahisefid with microsatellite markers 120 specimens of R.frissi kutum were used from four rivers to test thirty primers (30 samples from each river) of which 8 primers showed polymorphism. A large variation in heterozygosity average over all samples was observed among loci, that ranged from 0.13 to 0.91. For a given locus, observed heterozygosity varied greatly among the samples. For example, in Lamir 0.07 at SPY5 and 1 in Sefid Rud at CA1. Tajan had an observed heterozygosity of only 0.17, whilst the Lamir had an observed heterozygosity of 0.07 at locus SPY5. At locus SPY4, Tajan and Sefid Rud are the same and (0.53) and Shir Rud is 0.83 and Lamir is 0.47. However, despite these differences, there was clear difference in average heterozygosity observed between the samples. To investigated of Hardy-Weinberg Equilibrium in all locus and all rivers deviate showed from Hardy-Weinberg Equilibrium was significantly different ( P 0.05) between all samples except Lamir River and Sefid Rud River. The highest Fis was in Lamir River (0.84) in locus SPY5 and in Sefid Rud River in locus SPY5 (0.71), in locus CA12 (0.67) in Shir Rud River and SPY5 (0.63) in Tajan River. The lowest Fis is in locus CA1 (-0.55) in Sefid Rud River, in locus CA1 (-0.47) in Tajan River, in locus CA1 (-0.28) in Lamir River and in locus SPY6 (0.16) in Shir Rud River. Iranian Fisheries Science Research Institute Published

Neogobius caspius is a small benthic fish, native to the Caspian Sea. The fish is highly important as it comprises the main food item of the Caspian Sturgeons. The genetic diversity of N. caspius populations in the Caspian Sea was studied using microsatellite technique. In the study, 115 specimens of N. caspius from two regions (Turkmen Bandar and Anzali Bandar) in south Caspian Sea were collected. DNA was extracted using 12 pairs of microstellite primers for which 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, of which 9 were polymorphic, 2 monomorphic and one showed smear. The average observed and expected hetrozygosity was 0.749 and 0.638, respectively. Significant genetic differences between the two regions were observed (P 0.01). Deviations from Hardy-Weinberg equilibrium were in all specimens. These results indicate that at least two populations of N. caspius exist in the south Caspian Sea.

The project was carried out between March of 2007 and November of 2009.Five laboratories of research center (Clinical pathology, Plankton, Molecular genetics, Pollutants, Analysis Instrument were selected for accreditation. The main stages for establishment of the system consisted of: 1-Conducting a gap analysis to compare the present state of the laboratories with ISO/IEC 17025 Training 2 General requirements for the competence of testing and calibration laboratories Validation of methods Estimation of uncertainty Internal audits 3- Performing of technical and management requirements 4-Submit of quality manual to Iran Accreditation System (IAS) in order to accredit In January of 2010 the laboratories were accredited by Iran Accreditation System (IAS). The main results were including: 1-Increase the accuracy of measurement 2-Improvement of the Repeatability and Reproducibility of the test methods 3-Traceability and standardization of test methods 4- Calibration of measurement instruments 6- Updating of test methods 7-Standardization of physical condition of the laboratories 8- Getting the certification from Iran Accreditation System (IAS).

Genetic diversity of Neogobios gorlap population was examined on 133 samples from regions in southern parts of the Caspian Sea (Golestan 28 samples, Mazandaran 53 samples and Guilan 52 samples). Genomic DNA was extracted from fin tissue by phenol-chlorophorm method. PCR was performed using 6 microsatellite primers. The PCR products of samples were accompanied with standard marker (50 by ladder). To measure fragment size, samples were run on a 6% polyacrylamid gel. Allele count and frequency, the effective and real allele, expected heterozygosity and observed heterozygosity, genetic diversity; Shannon Information Index, genetic identity and genetic distance, Fst, Rst, gene flow and Hardy-Weinberg Equilibrium based on Analysis of Molecular Variance (AMOVA) were calculated using the Gene Alex software. All studied loci were polymorph. The mean observed and effective allele number was 11.44 and 7.28 respectively. Also, the maximum observed and expected heterozygosity was 0.964 and 0.932 respectively. Specimens from all regions (with the exception of locus NG71 and NG111 in Golestan region) were not in Hardy-Weinberg Equibrium in all of the loci (P

The molecular structure in populations of Mnemiopsis leidyi were examined for 200 samples from the south (Guilan, Mazandaran and Golestan provinces) and north of the Caspian Sea. DNA was extracted from M leiydi tissue by phenol-chiorophorm method and its concentration was found at 50 to 10Ong. We used the PCR method using 19 random primers (10bp). The PCR products of samples were accompanied with standard marker (50bp DNA). To measure fragment size, samples were run on a 1% agarose gel. Ten of the ninteen primers showed polymorphism. Statistical analysis of data was performed using POPGENE software. The avarage genetic variation was 0.189 in total samples and the maximum variation was found in samples from north of the Caspian Sea. Also, the maximume genetic distance was between north of the Sea and Golestan coasts in the south (0.089). The minimum genetic distance was between Mazandaran and Guilan coasts (0.001). The UOGMA dendogram showed two clusters. The samples of Mazandaran, Guilan and Golestan coasts were placed in one cluster and samples of the north area in another. The genetic diversity was significantly different between samples of the north area and Golestan coasts (P

Report of the project entitled "determination of the global population structure of the great sturgeon (Huso huso) with emphasis on the Golestan Province" is in front of your attention. Outcome is the results of study on the samples collected from the total catch in the landing sites namely Bandar-e-Anzali, Babolsar, and Ashooradeh, in the three Northern provinces. 224 specimens were collected according to the sampling manual during the fishing seasons in 2005 and 2006. Three methods were applied in this study to find out the population structure of the great sturgeon in the southern Caspian Sea. These were as follows: Morpho-metric meristic Population dynamics Population genetics (microsatellite loci) Results indicate that the range of different age groups of the great sturgeon in the Golestan Province is larger than the other two provinces. Results show that the older age groups are found in the samples from the Golestan Province rather than the other two provinces. Results demonstrate that the sample size for the two methods (morpho-metric meristic and population dynamics) is not enough. In order to obtain precise results with less uncertainty one needs to collect more specimens. Therefore, according to the few number of the great sturgeon in the total sturgeon landing, it is recommended that researchers apply methods other than the two methods which are mentioned above in order to make sure that the results are most reliable. Population genetics methods including microsatellite loci are among the methods which could produce good results with minimum bias in this regard. Results indicate that there are two distinguishable populations of the great sturgeon (Huso huso) in the southern part of the Caspian Sea off the shores of the three Northern provinces. Results also show that these two populations are scattered in the two east end (Golestan Province) and west end (Guilan Province) of the southern Caspian Sea. The two populations have overlapped each other in the Mazandaran Province. Iranian Fisheries Science Research Institute Published

Report of the project entitled "determination of the global population structure of the great sturgeon (Huso huso) with emphasis on the Golestan Province" is in front of your attention. Outcome is the results of study on the samples collected from the total catch in the landing sites namely Bandar-e-Anzali, Babolsar, and Ashooradeh, in the three Northern provinces. 224 specimens were collected according to the sampling manual during the fishing seasons in 2005 and 2006. Three methods were applied in this study to find out the population structure of the great sturgeon in the southern Caspian Sea. These were as follows: Morpho-metric meristic Population dynamics Population genetics (microsatellite loci) Results indicate that the range of different age groups of the great sturgeon in the Golestan Province is larger than the other two provinces. Results show that the older age groups are found in the samples from the Golestan Province rather than the other two provinces. Results demonstrate that the sample size for the two methods (morpho-metric meristic and population dynamics) is not enough. In order to obtain precise results with less uncertainty one needs to collect more specimens. Therefore, according to the few number of the great sturgeon in the total sturgeon landing, it is recommended that researchers apply methods other than the two methods which are mentioned above in order to make sure that the results are most reliable. Population genetics methods including microsatellite loci are among the methods which could produce good results with minimum bias in this regard. Results indicate that there are two distinguishable populations of the great sturgeon (Huso huso) in the southern part of the Caspian Sea off the shores of the three Northern provinces. Results also show that these two populations are scattered in the two east end (Golestan Province) and west end (Guilan Province) of the southern Caspian Sea. The two populations have overlapped each other in the Mazandaran Province.

The project was carried out between March of 2007 and November of 2009.Five laboratories of research center (Clinical pathology, Plankton , Molecular genetic, Pollutants , Analysis Instrument were selected for accreditation. The main stages for establishment of the system consisted of: 1-Conducting a gap analysis to compare the present state of the laboratories with ISO/IEC 17025 Training 2 General requirements for the competence of testing and calibration laboratories Validation of methods Estimation of uncertainty Internal audits 3- Performing of technical and management requirements 4-Submit of quality manual to Iran Accreditation System (IAS) in order to accredit In January of 2010 the laboratories were accredited by Iran Accreditation System (IAS). The main results were including: 1-Increase the accuracy of measurement 2-Improvement of the Repeatability and Reproducibility of the test methods 3-Traceability and standardization of test methods 4- Calibration of measurement instruments 6- Updating of test methods 7-Standardization of physical condition of the laboratories 8- Getting the certification from Iran Accreditation System (IAS). Iranian Fisheries Science Research Institute Published

The population genetic structure of common carp (Cyprinus carpio) was examined on 260 specimens from Tajan and Gorgan Rivers, Gorgan Gulf, Anzali Lagoon and other regions in east, middle and west of south Caspian Sea. DNA was extracted from fin tissue by phenol-chlorophorm method with a concentration of 50-100 nanograms. PCR was performed using ND-3/4 and ND-5/6 genes. The PCR products of samples were digested by 15 restriction endonuclease enzymes. The digested products accompanied with standard marker (50 pb). To measure fragment size, samples were run on a 6% vertical poyacrylamide gel. The fragments were visualized by silver staining of the polyacrylamide gel. Statistical analysis of data was performed by Reap software. We detected 14 and 12 different haplotypes in ND-3/4 and ND-5/6 genes of common carp. The mean values of haplotype diversity among populations were 0.59 and 0.48 and the average nucleotide diversity was 0.06 and 0.03 for ND3/4 and ND5/6 genes. Also, the mean values of nucleotide divergence among populations were 0.05% and 0.02%, respectively. The haplotype distribution was not significantly different between Mazandaran and Guilan coasts, Mazandaran and Golestan coasts, Golestan coast and Gorgan Gulf and Gorgan River (P

Over the past few years, the syndrome of streptococcosis has been associated with outbreaks in rainbow trout (Oncorhynchus mykiss, Walbaum) and caused significant economic losses in the aquaculture industry in Iran. The main purpose of this work was molecular identification of some causative agents of streptococcosis in rainbow trout. A total of 520 samples were collected from the head kidney of diseased fish (weight, 50_200g) in 72 farms of 8 provinces in Iran, during 2008 to 2009. Bacterial isolates representing morphology and biochemical profiles of Streptococcus spp. were further confirmed by polymerase chain reaction (PCR). DNA extraction was carried out from a single colony by using the extraction promega kit following the conditions described by the supplier. The PCR assay was developed based on the 16S rRNA and glucose kinase genes of Streptococcus spp. for the rapid and specific detection and identification of this pathogen from different sources. Approximately 40% of specimens were infected to Streptococcus spp. Consequently, five pathogenic species have been identified, including S. iniae in Fars province, S. faecium in Mazandaran province, S. agalactiae in Gilan and Mazandaran provinces, S. dysgalatiae in Lorestan, Kohgiluyeh and Boyerahmad, Gilan and Kermanshah provinces and S. uberis, which was common in all provinces (except Mazandaran and Lorestan). The dominant species (based on important species index) were S. uberis, S. dysgalactiae and S. agalactiae, respectively.