Liver cytochrome oxidase P4501A1 purification of Acipenser persicus and designing ELISA for measuring

Moon oxygenases related to cytochrome P450s are the molecular Biomarkers which have important role in Biotransformation of endogenous and exogenous compounds and catalazyin of many biological reactions. One of the important isoenzyme is cytochrome P4501A. This isoenzyme involved in metabolism of environment pollutnts such as PAHs. Because of its inducibility, it has a key tool for impact assesment of contaminants in aquatic environment. In this study, at first, that fractions containing Acipenser persicus and Huso huso isoenzyme were purified, and after that Antibodies against them were prepared. For isolation of isoenzyme fraction, Microsomes were prepared from fish liver using differential centrifugation at high speeds. microsomes were solubiized by cholat sodium and Emulgen. Extraction of this isoenzyme was done with the combinatuion of ionexchange chromatography and gelfiltration or chromatofocusing chromatography. Ion exchange chromatography and gel filtration were applied in DEAE sepharose fast flow and sephacryl S200 respectively and chromatofocusing was done at poly buffer 74 and 94 exchanger. The results of SDS-PAGE Showed that the molecular weight of isoenzyme was about 58±1 KDa. Furthermore the inmunoblotting results confirmed this subject. Isoenzyme activigy based on EROD (Ethoxyresorofin o-deethylase) reaction showed about 20-26 fold increase in enzyme activity of treated fish than control fish. The results of Elisa, Using monoclonal anti cod P4501A demonstrated the inducibility and highly elevated of its activity in treated sample more than the control fish. Mean while, the fish sample were showed the strong reaction to polyclonal antibody against beluga P4501A1 prepared in our Lab compared to monoclonal anti body.

A molecular genetic investigation of the population structure of Atlantic menhaden (Brevoortia tyrannus)

Atlantic menhaden (Brevoortia tyrannus), through landings, support one of the largest commercial fisheries in the United States. Recent consolidation of the once coast-wide reduction fishery to waters within and around Chesapeake Bay has raised concerns over the possibility of the loss of unique genetic variation resulting from concentrated fishing pressure. To address this question, we surveyed variation at the mitochondrial cytochrome c oxidase subunit I (COI) gene region and seven nuclear microsatellite loci to evaluate stock structure of Atlantic menhaden. Samples were collected from up to three cohorts of Atlantic menhaden at four geographic locations along the U.S. Atlantic coast in 2006 and 2007, and from the closely related Gulf menhaden (B. patronus) in the Gulf of Mexico. Genetic divergence between Atlantic menhaden and Gulf menhaden, based on the COI gene region sequences and microsatellite loci, was more characteristic of conspecific populations than separate species. Hierarchical analyses of molecular variance indicated a homogeneous distribution of genetic variation within Atlantic menhaden. No significant variation was found between young-of-the-year menhaden (YOY) collected early and late in the season within Chesapeake Bay, between young-of-the-year and yearling menhaden collected in the Chesapeake Bay during the same year, between YOY and yearling menhaden taken in Chesapeake Bay in successive years, or among combined YOY and yearling Atlantic menhaden collected in both years from the four geographic locations. The genetic connectivity between the regional collections indicates that the concentration of fishing pressure in and around Chesapeake Bay will not result in a significant loss of unique genetic variation.

A new species of western Atlantic lizardfish (Teleostei: Synodontidae: Synodus) and resurrection of Synodus bondi Fowler, 1939, as a valid species from the Caribbean with redescriptions of S. bondi, S. foetens (Linnaeus, 1766), and S. intermedius (Agassiz, 1829)

Western Atlantic synodontid species were studied as part of an ongoing effort to reanalyze Caribbean shorefish diversity. A neighbor-joining tree constructed from cytochrome c oxidase I (COI) data revealed 2 highly divergent genetic lineages within both Synodus intermedius (Agassiz, 1829) (Sand Diver) and S. foetens (Linnaeus, 1766) (Inshore Lizardfish). A new species, Synodus macrostigmus, is described for one of the S. intermedius lineages. Synodus macrostigmus and S. intermedius differ in number of lateral-line scales, caudal pigmentation, size of the scapular blotch, and shape of the anterior-nostril flap. Synodus macrostigmus and S. intermedius have overlapping geographic and depth distributions, but S. macrostigmus generally inhabits deeper water (>28 m) than does S. intermedius and is known only from coastal waters of the southeastern United States and the Gulf of Mexico, in contrast to those areas and the Caribbean for S. intermedius. Synodus bondi Fowler, 1939, is resurrected from the synonymy of S. foetens for one of the S. foetens genetic lineages. The 2 species differ in length and shape of the snout, number of anal-fin rays, and shape of the anterior-nostril flap. Synodus bondi and S. foetens co-occur in the central Caribbean, but S. bondi otherwise has a more southerly distribution than does S. foetens. Redescriptions are provided for S. intermedius, S. foetens, and S. bondi. Neotypes are designated for S. intermedius and S. foetens. A revised key to Synodus species in the western Atlantic is presented.

Bathyraja panthera, a new species of skate (Rajidae: Arhynchobatinae) from the western Aleutian Islands, and resurrection of the subgenus Arctoraja Ishiyama

We provide morphological and molecular evidence to recognize a new species of skate from the North Pacific, Bathyraja panthera. We also resurrect the skate subgenus Arctoraja Ishiyama, confirming its monophyly and the validity of the subgenus. Arctoraja was previously recognized as a distinct subgenus of Breviraja and later synonymized with Bathyraja (family Rajidae). Although the nominal species of Arctoraja have all been considered synonyms of Bathyraja parmifera by various authors, on the basis of morphometric, meristic, chondrological, and molecular data we recognize four species, including the new species. Species of Arctoraja are distributed across the North Pacific Ocean and adjacent seas from southern Japan to British Columbia. Bathyraja parmifera is abundant in the eastern Bering Sea, Aleutian Islands, and northern Gulf of Alaska; B. smirnovi is a western Pacific species found in the Sea of Okhotsk and Sea of Japan; B. simoterus is restricted to waters around the northern and eastern coasts of Hokkaido, Japan; and the new species B. panthera is restricted to the western Aleutian Islands. Bathyraja panthera is diagnosed by its color pattern of light yellow blotches with black spotting on a greenish brown background, high thorn and vertebral counts, chondrological characters of the neurocranium and clasper, and a unique nucleotide sequence within the mitochondrial cytochrome oxidase gene. Furthermore, the species presently recognized as Bathyraja parmifera exhibits two haplotypes among specimens from Alaska, suggesting the possibility of a second, cryptic species.

Intraspecific variation of isozymes in Arenicola marina (Polychaeta: Annelida)

Six enzyme systems, namely acid phosphatase, leucine aminopeptidase, phosphoglucose isomerase, tetrazolium oxidase, esterases and malate dehydrogenase were studied electrophoretically in Arenicola marina from various localities in United Kingdom. Out of 13 presumed loci, ten were found monomorphic. The three loci which appeared to be polymorphic are LAP-1, EST-2 and TO-1. Due to small sample size allele frequencies and genetic identity were not calculated. However, results indicate genetic difference among the population of A. marina.

Marine bacteria as indicators of upwelling in the Sea

The “oxidase reaction” (using p-amino-dimethyl-aniline oxalate as the reagent) has been used to distinguish oxidase-negative from oxidase-positive bacteria from the sea, when grown on membrane filters. By this means, it has been shown (a) that under conditions of stable stratification of the sea as in the tropics, a relationship exists between the percentage incidence of oxidase negative bacteria in the flora and the depth of the water; (b) that the maximum value for this percentage incidence (100) is reached at or immediately below the upper limit of the oxygen minimum layer; (c) that this percentage value (expressed as Oxⁿvalues) may be used to demonstrate the movements of water masses during upwelling. Such upwelling as indicated by theoretical findings and by temperature determinations along two transects off the west coast of Ceylon during the north east monsoon, has been confirmed by the distribution of Oxⁿvalues at these transects.

A Comparison of the bacterial flora of surface and sub-surface water of the sea off Ceylon

Under stable conditions of stratification of the sea, evidence of generic differences of the associated bacterial flora of the water masses has been obtained, between surface and sub-surface water. Gram negative rods, especially pseudomonads and achromobacters were more frequent at the surface. The fermentative and oxidase negative flora was more frequent in sub-surface water. The surface water in general had a greater variety of bacterial types while the sub-surface water had a flora with a greater range of biochemical activity. These results are discussed in relation to the hydrological condition of the water masses and the bacterial flora of freshly caught fish.

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.

A survey on relation of morphological, molecular and phylogenetic structure of zoanthids of the islands located in the Hormoz Strait (Hormoz, Qeshm, Larak, Hengam)

The order Zoantharia (Zoanthids) is one of the most neglected orders of cnidarians in the Persian Gulf. The present study aims to investigate the biodiversity of this order with morphological and molecular examination in the Persian Gulf. For this purpose, 123 colonies of zoanthids with variety of shape and colors have been collected from intertidal and shallow water zone of four islands, i. e. Hengam, Qeshm, Larak and Hormoz. After sampling, morphological characteristics of each specimen were recorded based on in situ photographs. Then DNA was extracted using the cetyl trimethyl ammonium bromide (CTAB) method. Both mitochondrial 16S ribosomal DNA (mt 16S rDNA) and cytochrome oxidase subunit I (COI) gene fragments were amplified and sequenced. The results of preliminary morphological identification integrated with two mitochondrial markers sequencing demonstrated the presence of five different species in this region; Zoanthus sansibaricus, Palythoa mutuki, Palythoa cf. mutuki, Palythoa tuberculosa and Neozoanthus persicus?. Although at first sight, morphological properties were not successful to delineate zoanthid species, they become reliable criteria to identify and delineate species in field studies after molecular identification.

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.