Transgene expressions in seabass (Lates calcarifer) following muscular injection of plasmid DNA: a strategy for vaccine development?

Muscular injection has become one of the direct methods for transferring foreign DNA into organisms. The technique has been recently introduced in the development of vaccines and gene therapy. Vaccine development, in particular, would be desirable in managing viral diseases in farmed fish. In this study, the technique was performed on seabass (Lates calcarifer) and was found that the foreign gene could be transferred successfully through injection into the muscles.

Quantification of drag and lift imposed by pop-up satellite archival tags and estimation of the metabolic cost to cownose rays (Rhinoptera bonasus)

The recent development of the pop-up satellite archival tag (PSAT) has allowed the collection of information on a tagged animal, such as geolocation, pressure (depth), and ambient water temperature. The success of early studies, where PSATs were used on pelagic fishes, has spurred increasing interest in the use of these tags on a large variety of species and age groups. However, some species and age groups may not be suitable candidates for carrying a PSAT because of the relatively large size of the tag and the consequent energy cost to the study animal. We examined potential energetic costs to carrying a tag for the cownose ray (Rhinoptera bonasus). Two forces act on an animal tagged with a PSAT: lift from the PSATs buoyancy and drag as the tag is moved through the water column. In a freshwater flume, a spring scale measured the total force exerted by a PSAT at flume velocities from 0.00 to 0.60 m/s. By measuring the angle of deflection of the PSAT at each velocity, we separated total force into its constituent forces — lift and drag. The power required to carry a PSAT horizontally through the water was then calculated from the drag force and velocity. Using published metabolic rates, we calculated the power for a ray of a given size to swim at a specified velocity (i.e., its swimming power). For each velocity, the power required to carry a PSAT was compared to the swimming power expressed as a percentage, %TAX (Tag Altered eXertion). A %TAX greater than 5% was felt to be energetically significant. Our analysis indicated that a ray larger than 14.8 kg can carry a PSAT without exceeding this criterion. This method of estimating swimming power can be applied to other species and would allow a researcher to decide the suitability of a given study animal for tagging with a PSAT.

Application of pop-up satellite archival tag technology to estimate postrelease survival of white marlin (Tetrapturus albidus) caught on circle and straight-shank (“J”) hooks in the western North Atlantic recreational fis

Short-duration (5- or 10-day) deployments of pop-up satellite archival tags were used to estimate survival of white marlin (Tetrapturus albidus) released from the western North Atlantic recreational fishery. Forty-one tags, each recording temperature, pressure, and light level readings approximately every two minutes for 5-day tags (n= 5) or four minutes for 10-day tags (n= 36), were attached to white marlin caught with dead baits rigged on straight-shank (“J”) hooks (n =21) or circle hooks (n=20) in offshore waters of the U.S. Mid-Atlantic region, the Dominican Republic, Mexico, and Venezuela. Forty tags (97.8%) transmitted data to the satellites of the Argos system, and 33 tags (82.5%) transmitted data consistent with survival of tagged animals over the deployment duration. Approximately 61% (range: 19−95%) of all archived data were successfully recovered from each tag. Survival was significantly (P<0.01) higher for white marlin caught on circle hooks (100%) than for those caught on straight-shank (“J”) hooks (65%). Time-to-death ranged from 10 minutes to 64 hours following release for the seven documented mortalities, and five animals died within the first six hours after release. These results indicate that a simple change in hook type can significantly increase the survival of white marlin released from recreational fis

Hardware paralelo reconfigurável para identificação de alinhamentos de sequências de DNA.

Amostras de DNA são encontradas em fragmentos, obtidos em vestígios de uma cena de crime, ou coletados de amostras de cabelo ou sangue, para testes genéticos ou de paternidade. Para identificar se esse fragmento pertence ou não a uma sequência de DNA, é necessário compará-los com uma sequência determinada, que pode estar armazenada em um banco de dados para, por exemplo, apontar um suspeito. Para tal, é preciso uma ferramenta eficiente para realizar o alinhamento da sequência de DNA encontrada com a armazenada no banco de dados. O alinhamento de sequências de DNA, em inglês DNA matching, é o campo da bioinformática que tenta entender a relação entre as sequências genéticas e suas relações funcionais e parentais. Essa tarefa é frequentemente realizada através de softwares que varrem clusters de base de dados, demandando alto poder computacional, o que encarece o custo de um projeto de alinhamento de sequências de DNA. Esta dissertação apresenta uma arquitetura de hardware paralela, para o algoritmo BLAST, que permite o alinhamento de um par de sequências de DNA. O algoritmo BLAST é um método heurístico e atualmente é o mais rápido. A estratégia do BLAST é dividir as sequências originais em subsequências menores de tamanho w. Após realizar as comparações nessas pequenas subsequências, as etapas do BLAST analisam apenas as subsequências que forem idênticas. Com isso, o algoritmo diminui o número de testes e combinações necessárias para realizar o alinhamento. Para cada sequência idêntica há três etapas, a serem realizadas pelo algoritmo: semeadura, extensão e avaliação. A solução proposta se inspira nas características do algoritmo para implementar um hardware totalmente paralelo e com pipeline entre as etapas básicas do BLAST. A arquitetura de hardware proposta foi implementada em FPGA e os resultados obtidos mostram a comparação entre área ocupada, número de ciclos e máxima frequência de operação permitida, em função dos parâmetros de alinhamento. O resultado é uma arquitetura de hardware em lógica reconfigurável, escalável, eficiente e de baixo custo, capaz de alinhar pares de sequências utilizando o algoritmo BLAST.

Development of Beluga, Delphinapterus leucas, Capture and Satellite Tagging Protocol in Cook Inlet, Alaska

Attempts to capture and place satellite tags on belugas, Delphinapterus leucas, in Cook Inlet, Alaska were conducted during late spring and summer of 1995, 1997, and 1999. In 1995, capture attempts using a hoop net proved impractical in Cook Inlet. In 1997, capture efforts focused on driving belugas into nets. Although this method had been successful in the Canadian High Arctic, it failed in Cook Inlet due to the ability of the whales to detect and avoid nets in shallow and very turbid water. In 1999, belugas were successfully captured using a gillnet encirclement technique. A satellite tag was attached to a juvenile male, which subsequently provided the first documentation of this species’ movements within Cook Inlet during the summer months (31 May–17 September).

DNA extraction from formalin-fixed Franciscana tissues

The present paper reports the extraction of DNA from formalin-fixed Pontoporia blainvillei tissues. Following the Vachot and Monerot (1996) protocol, fragmented DNA (300-700bp) was extracted from more than 95% of liver and muscle samples. DNA yield in liver samples was significantly higher than in muscle samples (4.574 ± 1.169mg DNA/mg versus 0.808 ± 0.297mg DNA/mg). Similar results were obtained from nine other species of cetaceans and five species of pinnipeds. It is of special interest to have a method that allows the utilisation of museum specimens not originally preserved for genetic studies, which may include rarely available, declining or extinct species. SPANISH: El presente trabajo reporta la extracción de ADN a partir de tejidos formolizados de Pontoporia blainvillei. Siguiendo el protocolo de Vachot y Monerot (1996) se pudo extraer ADN degradado (300-700pb) en más del 95% de las muestras de hígado y músculo analizadas. El rendimiento en ADN fue significativamente mayor en muestras de hígado que en muestras de músculo (4.574 ± 1.169mg DNA/mg tejido húmedo versus 0.808 ± 0.297mg DNA/mg tejido húmedo). Resultados similares se obtuvieron en otras nueve especies de Cetáceos y cinco de Pinnípedos. Resulta de gran interés contar con un método que permita la utilización de especímenes depositados en museos y que no hayan sido originalmente colectados para estudios genéticos, incluyendo especies de difícil obtención, en franca declinación o extintas.