Study of gill and kidney tissue changes in Tenualosa ilisha during migration from sea to the Karun and Bahmanshir Rivers

In this research, histological changes of gill and kidney, as a basic and important organ in osmoregulation, during migration from Musa creek to Karun and Bahmanshir Rivers was studied. Ten fish from each station of Persian Gulf, Karun and Bahmanshir with similarity in biometric of size and weight were collected. After fixation in Bouin's solutions, routine procedure of tissue preparation was done and 6 μm sections stained with Hematoxylin and Eosin, studied by optical microscope equipped with a daynolit lenses. The greatest number and area of chloride cells were seen in sea samples but they decreased during migration to Karun and Bahmanshir Rivers (p0.05). In kidney, during migration from the sea to the river, the diameter of proximal and distal lumen increased. But in Karun samples, diameter of lumen of distal tubules were higher than others station. Also, there was no significant difference in other tubule between in Karun and Bahmanshir samples. Therefore according to this study, gill and kidney have active participation in osmoregulation. This fish can be a good model for study of osmoregulation in different salinities because of adaptation and response to different environmental salinities in gill that associated with histologic changes

Investigation on genetic structure of cobia (Rachycentron canadum) using microsatellite markers

The genetic diversity of Cobia, Rachycentron canadum populations in the Persian Gulf and Oman Sea were assessed using microsatellite technique. We removed about 3-5g of pectoral and dorsal fin tissue from 184 samples in winter 2006 and spring 2007, and stored it in pure ethylic alcohol (96%). Polymerase chain reactions (PCR) were conducted on the target DNA using 10 paired microsatellite primers. The dendrogram was constructed and drawn using MEGA software package version 4. Based on the analysis of molecular variance, the highest Fst (0.063) was observed when comparing specimens from Dayer Port and Pozm zones. Significant differences (P<0.01) were not observed between R5t recorded for the specimens studied in the same region but were observed between RSA recorded for different regions. The dendrogram of genetic distance showed two major clusters: the Bushehr and Dayer populations were in one cluster, and the remaining four populations in the other. The second cluster was further separated into two sub-clusters: the Lengeh and Bandar Abbas populations composed one cluster and the Pozm and the Beris populations were in the other cluster. The present study showed that at least three different populations of R. canadum are living in the Persian Gulf and Oman Sea. The populations include Bushehr, Bandar Abbas and Chabahar populations

Antimicrobial activities of semi polar-nonpolar and polar secondary metabolites of sponge Dysidea pallescens from Hengam Island, Persian Gulf

Sponges are the simplest multicellular animals that lack defense mechanisms and rely on chemical defense that have been used by mankind to develop antimicrobial drugs against diseases. The present study was designed to demonstrate the antibacterial and antifungal activities of marine sponge Dysidea pallescens semipolar and nonpolar extracts. In this study, D. pallescens were collected from Hengam Island in the Persian Gulf. The extracts were produced by Bligh and Dyer method. Broth Dilution Methods were used to check the antimicrobial activity of D. pallescens extracts against Escherichia coli (ATCC 15224), Pseudomonas aeruginosa (ATCC 25619), Staphylococcus aureus aureus (ATCC 1764), Bacillus subtilis pizizenii (ATCC 6633), Candida albicans (ATCC 10231) and Aspergillus fumigates (PTCC 5009). The results showed diethyl ether extract has bactericidal activity against S. aureus aureus (MBC=10mg/mL) and B. subtilis spizizenii (MBC=20mg/mL). D. pallescens diethyl ether extract showed a very weak antifungal activity but methanol extract showed fungicidal activity against A. fumigates (MFC=5mg/mL) and C. albicans (MFC=1.5 mg/mL). Therefore nonpolar-semipolar secondary metabolites of D. pallescens solutions in diethyl ether have shown significant antibacterial activity and polar-secondary metabolites solutions in methanol have shown significant antifungal activity

Genetic characterization of Sillago sp. from Hormozgan coastal waters using microsatellite markers

Using 5 microsatellite loci Sillago sp. From Hormozgan North Persian Gulf were investigated through 68 specimens in two stations: Minab (31 specimens) and Bandar Lengeh (38 specimens). DNA was extracted using modified CTAB (Hexadecyl trimethyl-ammonium bromide) protocol. 5-17 alleles were observed in 5 loci with 8.4 mean allelic frequency. Observed and expected heterozygosity values was calculated (0.115<Ho<0.792), (0.598<He<0.902). Genetic distance and identity according to Nei was estimated 0.520 and 0.595 respectively which shows genetic distance of genus level. Significant deviation from Hardy-Weinberg equilibrium (p<0.001) was observed at all loci at two stations. Analysis of Molecular Variance (AMOVA) of Genetic differentiation index (Fst) revealed mean significant (p=0.001) differentiation between two studied regions and 3.90 gen flow. Despite low distance of two studied regions (180km) there are probably two distinct populations of Sillago sp. which should be considered in management efforts

Assay of genetic diversity of cuttlefish (Sepia pharaonis; Ehrenberg, 1831) populations using microsatellite markers in Bandar Abbas and Bushehr regions

In this present study genetic diversity of cuttlefish (Sepia pharaonis) populations were investigated using microsatellite markers. Total 51 samples were collected from Bandarabass and Bushehr regions. Tissue sample of arm tips (tentacle) were preserved in 96% ethanol alcohol until using in biotechnology laboratory of Khorramshahr University of Marine Science and Technology. Genomic DNA was extracted with CTAB method. The quality and quantity of extracted DNA was assessed by 1% agarose gel electrophoresis and spectrophotometry, respectively. Polymerase chain reaction conducted with 6 pairs of microsatellite primers. PCR products were electrophoresed on 8% polyacrylamide gel and stained with silver nitrate. These primers were shown 4 pairs of polymorph and 2 pairs of monomorp. Allele Sizes were measured in populations then genetic parameter were calculated using Arlequin and Gen Alex Programs and phylogenetic relationship was determinated and drawn using TFPGA Program. Result obtained showed genetic distance and resemblance distance is 0.282 and 0.754, respectively and Weak but significant genetic differentiation was present 0.031 between of populations

Study of morphological characteristics and frequency of Dendrobranchiata larval shrimp in artificial reefs of Khuzestan coasts, Bahrekan area

This study was done to identify larval stages and biodiversity of Dendrobranchiata planktonic communites in artificial reefs of Khuzestan coasts(Bahrekan area in Northwest Persian gulf) for one year from May 2011 to April 2012 monthly basis. Sampling was conducted using a plankton net with a spring 300 micrometers. Totally, from two superfamily: Penaeoidea and Sergestoidea, 3 families: Sergestidae, Luciferidae, Penaeidae were identified.From Penaeidae family, Metapenaeus affinis ,Penaeus indicus and Parapenaeopsis stylifera species, from Luciferidae family, Lucifer hanseni specie and from Sergestidae family, Acetes sp specie were identified and introduced. Also larval density of identified species in different months and stations are calculated and determined Metapenaeus affinis specie (With the average of number per cubic meter in 1029±830) from Penaeidae family account for larval density is maximum in September. Finally, two abundance peak was observed that the first peak was in September and the second peak was in November. The maximum of relative frequency percentage in Penaeoidea superfamily related to mysisI stage and 62 percentage. The results obtained indicate that the artificial structures are well managed nursery area for Penaeidae shrimp. The maximum average value of Shanon-wiener index was in November (1.36) due to the presence of all species in the same month. Simpson index was lowest in November, which confirms the above conclusion

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and it’s expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Study of genetic diversity of mudskipper (Periophthalmus waltoni) using RAPD markers in the Persian Gulf

We collected 69 samples of Mudskipper from dominant spices periophthalmus waltoni for genetic study from Hendijan, Khore Zangi and Delvar. A specimen of fin was fixed in Ethanol alcohol (96%) and moved to Biotechnology lab in center study of Persian Golf in Bushehr. Total genomic DNA was extracted by proteinase-K digestion, phenol: chloroform: isoamyl alcohol extraction and ethanol precipitation. The quality and quantity of DNA was assessed using 1% agarose gel electrophoresis and spectrophotomeric method. Polymerase chain reactions (PCR) were conducted on the target DNA using 6 primers RAPD. PCR products were separated by electrophoresis on polyacrylamid gels (8%) that were stained using silver nitrate. Allele frequency, the effective and real allele, expected and observed heterozygosis, genetic diversity, Shannon Information index, Gst and gene based on analysis of molecular variance (AMOVA) were calculated using the GENALEX and Pop Gene software. The average of Shannon index for Hendijan, Khore zangi and Delvar was respectively 0.3887, 0.2860 and 0.3509. Highest of Gst observed between Hendijan and Khore zangi (0.260) and lowest between Khore zangi and Delvar (0.195). Highest of gene flow (Nm) was observed between Delvar and Khore zangi (1.033) and lowest between Hendijan and Khore zangi (0.687). Highest of Genetic identity observed between Delvar and Khore zangi (0.924) but lowest was between Hendijan and Khore zangi community (0.833). Eventually this study show that three community of our research are separate and belong to two clusters

Inheritance of microsatellite markers in the hybrid of female Chalcalburnus chalcoides and male Vimba vimba persa Chalcalburnus chalcoides (Gueldenstaedt, 1772) ×Vimba vimba persa (Pallas, 1814)

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