Determination of the age of young American eels, Anguilla rostrata, in fresh water, based on otolith surface area and microstructure

The time elvers of the American eel, Anguilla rostrata , spend in an estuary prior to their migration into fresh water was assessed. A distinct mark was formed on elvers' otoliths during their first 2 to 3 weeks in the river estuary. This mark was used to distinguish between growth in fresh water and in salt water. Migrating eels collected at a falls 4 km from the estuary exhibited bimodal length and weight distributions. Frequency distributions showed that eels collected in the estuary were smaller and had smaller otoliths than eels collected at the falls, indicating that elvers do not reach the falls in the same year as they enter the estuary. The three modal groups most probably represent three age–classes. However, the otoliths of elvers collected in the estuary had only the mark of transition whereas eels in the first and second mode at the falls usually had two rings (1–4) and four rings (3–6) per otolith, respectively, in addition to the mark of transition, as viewed under SEM. The possibility that ring formation is not annual means that the use of otoliths for the age determination of eels in this study has not been validated.     

Protein kinase C is regulated in vivo by three functionally distinct phosphorylations

Background: Protein kinase Cs are a family of enzymes that transduce the plethora of signals promoting lipid hydrolysis. Here, we show that protein kinase C must first be processed by three distinct phosphorylations before it is competent to respond to second messengers.Results We have identified the positions and functions of the in vivo phosphorylation sites of protein kinase C by mass spectrometry and peptide sequencing of native and phosphatase-treated kinase from the detergent-soluble fraction of cells. Specifically, the threonine at position 500 (T500) on the activation loop, and T641 and S660 on the carboxyl terminus of protein kinase C βII are phosphorylated in vivo. T500 and S660 are selectively dephosphorylated in vitro by protein phosphatase 2A to yield an enzyme that is still capable of lipid-dependent activation, whereas all three residues are dephosphorylated by protein phosphatase 1 to yield an inactive enzyme. Biochemical analysis reveals that protein kinase C autophosphorylates on S660, that autophosphorylation on S660 follows T641 autophosphorylation, that autophosphorylation on S660 is accompanied by the release of protein kinase C into the cytosol, and that T500 is not an autophosphorylation site.Conclusion Structural and biochemical analyses of native and phosphatase-treated protein kinase C indicate that protein kinase C is processed by three phosphorylations. Firstly, trans-phosphorylation on the activation loop (T500) renders it catalytically competent to autophosphorylate. Secondly, a subsequent autophosphorylation on the carboxyl terminus (T641) maintains catalytic competence. Thirdly, a second autophosphorylation on the carboxyl terminus (S660) regulates the enzyme's subcellular localization. The conservation of each of these residues (or an acidic residue) in conventional, novel and atypical protein kinase Cs underscores the essential role for each in regulating the protein kinase C family.     

Reconstructing Native American Migrations from Whole-Genome and Whole-Exome Data

There is great scientific and popular interest in understanding the genetic history of populations in the Americas. We wish to understand when different regions of the continent were inhabited, where settlers came from, and how current inhabitants relate genetically to earlier populations. Recent studies unraveled parts of the genetic history of the continent using genotyping arrays and uniparental markers. The 1000 Genomes Project provides a unique opportunity for improving our understanding of population genetic history by providing over a hundred sequenced low coverage genomes and exomes from Colombian (CLM), Mexican-American (MXL), and Puerto Rican (PUR) populations. Here, we explore the genomic contributions of African, European, and especially Native American ancestry to these populations. Estimated Native American ancestry is in MXL, in CLM, and in PUR. Native American ancestry in PUR is most closely related to populations surrounding the Orinoco River basin, confirming the Southern America ancestry of the Taíno people of the Caribbean. We present new methods to estimate the allele frequencies in the Native American fraction of the populations, and model their distribution using a demographic model for three ancestral Native American populations. These ancestral populations likely split in close succession: the most likely scenario, based on a peopling of the Americas thousand years ago (kya), supports that the MXL Ancestors split kya, with a subsequent split of the ancestors to CLM and PUR kya. The model also features effective populations of in Mexico, in Colombia, and in Puerto Rico. Modeling Identity-by-descent (IBD) and ancestry tract length, we show that post-contact populations also differ markedly in their effective sizes and migration patterns, with Puerto Rico showing the smallest effective size and the earlier migration from Europe. Finally, we compare IBD and ancestry assignments to find evidence for relatedness among European founders to the three populations.     

A spatial approach to matching marine fish diversity and abundance with habitat features

Current issues in marine resource management have in common a geospatial component and a need to integrate both biotic and abiotic data from various sources. We propose a practical approach to address these issues looking at the American plaice (Hippoglossoides platessoides) and the demersal fish fauna in the Gulf of St. Lawrence (Canada). Central to our approach was the use of a common spatial grid and three different methods to match biotic and abiotic features at a broad regional scale, (1) matching plaice distribution with habitat categories determined a priori on the basis of abiotic features (cluster analysis), (2) habitat categories determined taking into consideration both plaice density and abiotic features (simple regression tree), and (3) habitat categories determined taking into consideration demersal fish species density (70 fish species) and abiotic features (multivariate regression tree, MRT). Hot spots and cold spots of plaice abundance in summer were described and matched with specific habitats. The spatial distribution of habitats was similar whether biotic variables were used in the classification or not. The MRT, however, identified 56 different fish species in the plaice habitat (median species richness by 100 km² cell = 12), pointing to potential interactions with other fish species.     

The effect of energetic condition on growth dynamics and health of Atlantic cod (Gadus morhua)

Prolonged starvation resulting in sublethal condition factor values was hypothesized to have a detrimental effect on short‐term growth capacity upon refeeding. Cod (Gadus morhua) were food‐deprived and their length and mass measured before refeeding and after 3, 6, 9 and 12 weeks of ad libitum feeding. Total mass increase during the first 3 weeks of feeding was greatest in fish with a higher initial condition factor. The reverse situation was observed during the last 3 weeks of feeding. Specific growth rate peaked in the period from week 4 to week 6, except in cod with the highest condition factor for which a steady decline in specific growth rate was observed, and was not influenced by the condition factor at the start of the feeding period. Total mass increase over 12 weeks was also not influenced by initial condition factor. Thus by the end of the experiment, condition factors were lowest in fish with initially low condition factors. The hepatosomatic index and gonadosomatic index did not differ at the end of the experiment, but the proportion of mature cod increased with increasing initial condition factor. A disease outbreak caused significant mortalities among fish shortly after the start of the feeding period. Forty‐one percent of the fish had died after 84 days. No mortality was observed among fish that had started the experiment with the highest condition factor. Mortality was inversely related to initial condition factor. Growth was examined for survivors exclusively. Poor condition in wild fish may increase vulnerability to diseases and compensatory growth may not allow cod with low condition factors to fully recover unless food availability remains high over a long period of time.     

Phenotypic flexibility of digestive system in Atlantic cod (Gadus morhua)

This study examined the restoration of the digestive capacity of Atlantic cod (Gadus morhua Linnaeus) following a long period of food deprivation. Fifty cod (48 cm, 1 kg) were food-deprived for 68 days and then fed in excess with capelin (Mallotus villosus Müller) on alternate days. Ten fish were sampled after 0, 2, 6, 14 and 28 days and the mass of the pyloric caeca, intestine and carcass determined. Two metabolic enzymes (cytochrome c oxidase and citrate synthase) were assayed in white muscle, pyloric caeca and intestine, and trypsin activity was measured in the pyloric caeca. A delay of 14 days was required before body mass started to increase markedly, whereas most of the increase in mass of both the pyloric caeca and intestine relative to fish length occurred earlier in the experiment. By day 14, the activities of trypsin and citrate synthase in the pyloric caeca as well as citrate synthase in the intestine had reached maxima. The growth of the digestive tissues and restoration of their metabolic capacities thus occur early upon refeeding and are likely required for recovery growth to take place. The phenotypic flexibility of the cod digestive system is therefore remarkable: increases in trypsin activity and size of pyloric caeca resulted in a combined 29-fold increase in digestive capacity of the fish during the refeeding period. Our study suggests that Atlantic cod are able to cope with marked fluctuations in food availability in their environment by making a rapid adjustment of their digestive capacity as soon as food availability increases.     

Two-dimensional gel electrophoresis based technologies for potential biomarkers identification in amniotic fluid: a simple model

To assess the efficiency of two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and of two-dimensional electrophoresis and ammoniacal silver staining (2D-E), different amounts of soybean trypsin inhibitor and horse myoglobin were added to amniotic fluid. In this model, a minimum of 5 to 10 ng of "artificial" biomarkers was detected.     

Detection of Legionella spp. by fluorescent in situ hybridization in dental unit waterlines

AIMS: To confirm the presence of viable Legionella spp. in dental unit waterlines (DUWL) using fluorescent in situ hybridization (FISH) and compare this method with culture approach and also to validate the utility of an enrichment to increase FISH sensitivity. METHODS AND RESULTS: Water samples from 40 dental units were analysed. Three different techniques for detecting Legionella spp. were compared: (i) culture approach, (ii) direct FISH and (iii) FISH with a previous R2A medium enrichment (R2A/FISH). The FISH detection was confirmed by PCR. The use of the direct FISH does not improve significantly the detection of legionellae when compared with the culture. On the contrary, when R2A/FISH was performed, sensitivity was, respectively, two- and threefold higher than that with the direct FISH and culture approach. Using R2A/FISH, 63% of water samples analysed showed a contamination by legionellae. CONCLUSIONS: Legionellae detection by direct FISH and R2A/FISH in dental unit water is possible but is more rapid and more sensitive (R2A/FISH) than the culture approach. SIGNIFICANCE AND IMPACT OF THE STUDY: R2A/FISH showed that several pathogens present in DUWL are viable but may not be culturable. Unlike PCR, R2A/FISH is designed to detect only metabolically active cells and therefore provides more pertinent information on infectious risk.     

Systematics of the grey mullets (Teleostei: Mugiliformes: Mugilidae): molecular phylogenetic evidence challenges two centuries of morphology-based taxonomy

The family Mugilidae comprises mainly coastal marine species that are widely distributed in all tropical, subtropical and temperate seas. Mugilid species are generally considered to be ecologically important and they are a major food resource for human populations in certain parts of the world. The taxonomy and systematics of the Mugilidae are still much debated and based primarily on morphological characters. In this study, we provide the first comprehensive molecular systematic account of the Mugilidae using phylogenetic analyses of nucleotide sequence variation at three mitochondrial loci (16S rRNA, cytochrome oxidase I, and cytochrome b) for 257 individuals from 55 currently recognized species. The study covers all 20 mugilid genera currently recognized as being valid. The family comprises seven major lineages that radiated early on from the ancestor to all current forms. All genera that were represented by two species or more, except Cestraeus, turned out to be paraphyletic or polyphyletic. Thus, the present phylogenetic results generally disagree with the current taxonomy at the genus level and imply that the anatomical characters used for the systematics of the Mugilidae may be poorly informative phylogenetically. The present results should provide a sound basis for a taxonomic revision of the mugilid genera. A proportion of the species with large distribution ranges (including Moolgarda seheli, Mugil cephalus and M. curema) appear to consist of cryptic species, thus warranting further taxonomic and genetic work at the infra-generic level.