Identification of Sex and Female’s Reproductive Stage in Commercial Fish Species through the Quantification of Ribosomal Transcripts in Gonads

The estimation of maturity and sex of fish stocks in European waters is a requirement of the EU Data Collection Framework as part of the policy to improve fisheries management. On the other hand, research on fish biology is increasingly focused in molecular approaches, researchers needing correct identification of fish sex and reproductive stage without necessarily having in house the histological know-how necessary for the task. Taking advantage of the differential gene transcription occurring during fish sex differentiation and gametogenesis, the utility of 5S ribosomal RNA (5S rRNA) and General transcription factor IIIA (gtf3a) in the molecular identification of sex and gametogenic stage was tested in different economically-relevant fish species from the Bay of Biscay. Gonads of 9 fish species (, Atlantic, Atlantic-chub and horse mackerel, blue whiting, bogue, European anchovy, hake and pilchard and megrim), collected from local commercial fishing vessels were histologically sexed and 5S and 18S rRNA concentrations were quantified by capillary electrophoresis to calculate a 5S/18S rRNA index. Degenerate primers permitted cloning and sequencing of gtf3a fragments in 7 of the studied species. 5S rRNA and gtf3a transcript levels, together with 5S/18S rRNA index, distinguished clearly ovaries from testis in all of the studied species. The values were always higher in females than in males. 5S/18S rRNA index values in females were always highest when fish were captured in early phases of ovary development whilst, in later vitellogenic stages, the values decreased significantly. In megrim and European anchovy, where gonads in different oogenesis stages were obtained, the 5S/18S rRNA index identified clearly gametogenic stage. This approach, to the sexing and the quantitative non-subjective identification of the maturity stage of female fish, could have multiple applications in the study of fish stock dynamics, fish reproduction and fecundity and fish biology in general.

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.

Quantification and reduction of unobserved mortality rates for snow, southern Tanner, and red king crabs (Chionoecetes opilio, C. bairdi, and Paralithodes camtschaticus) after encounters with trawls on the seafloor

Unobserved mortalities of nontarget species are among the most troubling and difficult issues associated with fishing, especially when those species are targeted by other fisheries. Of such concern are mortalities of crab species of the Bering Sea, which are exposed to bottom trawling from groundfish fisheries. Uncertainty in the management of these fisheries has been exacerbated by unknown mortality rates for crabs struck by trawls. In this study, the mortality rates for 3 species of commercially important crabs—red king crab, (Paralithodes camtschaticus), snow crab (Chionoecetes opilio) and southern Tanner crab (C. bairdi)—that encounter different components of bottom trawls were estimated through capture of crabs behind the bottom trawl and by evaluation of immediate and delayed mortalities. We used a reflex action mortality predictor to predict delayed mortalities. Estimated mortality rates varied by species and by the part of the trawl gear encountered. Red king crab were more vulnerable than snow or southern Tanner crabs. Crabs were more likely to die after encountering the footrope than the sweeps of the trawl, and higher death rates were noted for the side sections of the footrope than for the center footrope section. Mortality rates were ≤16%, except for red king crab that passed under the trawl wings (32%). Herding devices (sweeps) can expand greatly the area of seafloor from which flatfishes are captured, and they subject crabs in that additional area to lower (4–9%) mortality rates. Raising sweep cables off of the seafloor reduced red king crab mortality rates from 10% to 4%.

Redundancy quantification in the safety assessment of existing concrete beam-and-slab bridges

In current practice the strength evaluation of a bridge system is typically based on firstly using elastic analysis to determine the distribution of load effects in the elements and then checking the ultimate section capacity of those elements. Ductility of the components in most bridge structures permits local yield and subsequent redistribution of the applied loads from the most heavily loaded elements. As a result a bridge can continue to carry additional loading even after one member has yielded, which has conventionally been adopted as the "failure criterion" in bridge strength evaluation. This means that a bridge with inherent redundancy has additional reserves of strength such that the failure of one element does not result in the failure of the complete system. For these bridges warning signs will show up and measures can be undertaken before the ultimate collapse is happening. This paper proposes a rational methodology for calculating the ultimate system strength and including in bridge evaluation the warning level due to redundancy. © 2004 Taylor & Francis Group, London.

The identification and quantification of highly stable 'common hairpin' in the dynamic process of co-transcriptional mRNA folding

In our studies, 88 human mRNA samples were collected from the Integrated Sequence-Structure database and then the dynamic process in co-transcriptional mRNA folding was simulated using the RNAstructure version 4.1 program. Through statistical analyses of the frequencies of occurrence of hairpins, a group of special folding structures-the 'common hairpins'-were identified. These 'common hairpins' have lower energies and occur in all the subsequent folding units that formed in the dynamic folding process. By applying the formulas (1)-(4) of the 'common hairpins' statistical model, 163 'common hairpins' were found, to make up about 7% of the total of 2286 hairpins. Classified studies further show that the 'common hairpins' that were studied may oscillate in the dynamic folding process. However, the hairpin loops of the 'common hairpins' and stems proximal to those 'common hairpins' loops maintain topologically stable structures, while other loops and stems distal to the 'common hairpins' loops are shown to be alterable structures. Strikingly, further studies indicate that the stable structures of these 'common hairpins' may have unbeknown effects on controlling the formation of protein structures in the translation process (unpublished results). (c) 2005 Elsevier B.V. All rights reserved.

Quantification of the effects of antibodies on the extra- and intracellular dynamics of Salmonella enterica.

Antibodies are known to be essential in controlling Salmonella infection, but their exact role remains elusive. We recently developed an in vitro model to investigate the relative efficiency of four different human immunoglobulin G (IgG) subclasses in modulating the interaction of the bacteria with human phagocytes. Our results indicated that different IgG subclasses affect the efficacy of Salmonella uptake by human phagocytes. In this study, we aim to quantify the effects of IgG on intracellular dynamics of infection by combining distributions of bacterial numbers per phagocyte observed by fluorescence microscopy with a mathematical model that simulates the in vitro dynamics. We then use maximum likelihood to estimate the model parameters and compare them across IgG subclasses. The analysis reveals heterogeneity in the division rates of the bacteria, strongly suggesting that a subpopulation of intracellular Salmonella, while visible under the microscope, is not dividing. Clear differences in the observed distributions among the four IgG subclasses are best explained by variations in phagocytosis and intracellular dynamics. We propose and compare potential factors affecting the replication and death of bacteria within phagocytes, and we discuss these results in the light of recent findings on dormancy of Salmonella.