Surface mucous as a source of genomic DNA from Atlantic billfishes (Istiophoridae) and swordfish (Xiphiidae)

Procedures for sampling genomic DNA from live billfishes involve manual restraint and tissue excision that can be difficult to carry out and may produce stresses that affect fish survival. We examined the collection of surface mucous as a less invasive alternative method for sourcing genomic DNA by comparing it to autologous muscle tissue samples from Atlantic blue marlin (Makaira nigricans), white marlin (Tetrapturus albidus), sailfish (Istiophorus platypterus), and swordfish (Xiphias gladius). Purified DNA from mucous was comparable to muscle and was suitable for conventional polymerase chain reaction, random amplified polymorphic DNA analysis, and mitochondrial and nuclear locus sequencing. The nondestructive and less invasive characteristics of surface mucous collection may promote increased survival of released specimens and may be advantageous for other marine fish genetic studies, particularly those involving large live specimens destined for release.

Applications in adaptive cluster sampling of Gulf of Alaska rockfish

Adaptive cluster sampling (ACS) has been the subject of many publications about sampling aggregated populations. Choosing the criterion value that invokes ACS remains problematic. We address this problem using data from a June 1999 ACS survey for rockfish, specifically for Pacific ocean perch (Sebastes alutus), and for shortraker (S. borealis) and rougheye (S. aleutianus) rockfish combined. Our hypotheses were that ACS would outperform simple random sampling (SRS) for S. alutus and would be more applicable for S. alutus than for S. borealis and S. aleutianus combined because populations of S. alutus are thought to be more aggregated. Three alternatives for choosing a criterion value were investigated. We chose the strategy that yielded the lowest criterion value and simulated the higher criterion values with the data after the survey. Systematic random sampling was conducted across the whole area to determine the lowest criterion value, and then a new systematic random sample was taken with adaptive sampling around each tow that exceeded the fixed criterion value. ACS yielded gains in precision (SE) over SRS. Bootstrapping showed that the distribution of an ACS estimator is approximately normal, whereas the SRS sampling distribution is skewed and bimodal. Simulation showed that a higher criterion value results in substantially less adaptive sampling with little tradeoff in precision. When time-efficiency was examined, ACS quickly added more samples, but sampling edge units caused this efficiency to be lessened, and the gain in efficiency did not measurably affect our conclusions. ACS for S. alutus should be incorporated with a fixed criterion value equal to the top quartile of previously collected survey data. The second hypothesis was confirmed because ACS did not prove to be more effective for S. borealis-S. aleutianus. Overall, our ACS results were not as optimistic as those previously published in the literature, and indicate the need for further study of this sampling method.

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

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