Geir Dahle

The population genetics of Calanus finmarchicus in the North Atlantic

Abstract DNA sequence variation for a 350 base-pair portion of mitochondrial 16S rRNA gene was determined for the copepod, Calanus finmarchicus, collected from eight sites in the western N. Atlantic Ocean and seven sites in the Norwegian Sea. Among 104 individuals for which sequence data were obtained there were 30 mitochondrial haplotypes. One haplotype occurred in 59 individuals and was found in all but two of the samples, while 21 haplotypes were unique. Both nucleotide diversity (π = 0.0061) and haplotype diversity (h= 0.67) were moderate. There were significant differences in the frequencies of the haplotypes among the samples (P < 0.001 by chi-square test by Monte Carlo simulation). One sample collected from the Norwegian Sea and one sample from the northern Gulf of St. Lawrence were distinct from all other samples, perhaps as a result of advection from different oceartographic regions. Chi-square tests resolved four groups: one consisting of the three Georges Bank, two Gulf of Maine, and two (of th...

The vgll3 Locus Controls Age at Maturity in Wild and Domesticated Atlantic Salmon(Salmo salar L.) Males

Wild and domesticated Atlantic salmon males display large variation for sea age at sexual maturation, which varies between 1–5 years. Previous studies have uncovered a genetic predisposition for variation of age at maturity with moderate heritability, thus suggesting a polygenic or complex nature of this trait. The aim of this study was to identify associated genetic loci, genes and ultimately specific sequence variants conferring sea age at maturity in salmon. We performed a genome wide association study (GWAS) using a pool sequencing approach (20 individuals per river and phenotype) of male salmon returning to rivers as sexually mature either after one sea winter (2009) or three sea winters (2011) in six rivers in Norway. The study revealed one major selective sweep, which covered 76 significant SNPs in which 74 were found in a 370 kb region of chromosome 25. Genotyping other smolt year classes of wild and domesticated salmon confirmed this finding. Genotyping domesticated fish narrowed the haplotype region to four SNPs covering 2386 bp, containing the vgll3 gene, including two missense mutations explaining 33–36% phenotypic variation. A single locus was found to have a highly significant role in governing sea age at maturation in this species. The SNPs identified may be both used as markers to guide breeding for late maturity in salmon aquaculture and in monitoring programs of wild salmon. Interestingly, a SNP in proximity of the VGLL3 gene in humans (Homo sapiens), has previously been linked to age at puberty suggesting a conserved mechanism for timing of puberty in vertebrates.

Mixed stock analysis and the power of different classes of molecular markers in discriminating coastal and oceanic Atlantic cod (Gadus morhua L.) on the Lofoten spawning grounds, Northern Norway

Atlantic cod (Gadus morhua) encompasses many different populations or stocks, which in part are managed separately. In the northeast Atlantic cod is divided into two main management units; northeast Arctic cod and coastal cod. These two groups have traditionally been separated by otolith classification. In this study, the power of different classes of genetic markers in separating the two cod groups was investigated. The variation in thirteen genetic markers, including allozymes, haemoglobin, the scDNA locus Pantophysin (Pan I) and a number of microsatellites was investigated, and mixed stock analysis and individual assignment tests were performed on samples comprising a mixture of individuals of putative coastal and oceanic type cod. The genetic analyses showed a large genetic differentiation between outer stations and stations located closer to the mainland shore. Mixed stock analysis and individual assignment tests used for estimation of stock proportions gave results similar to those obtained by otolith classification.

Genetic diversity within and among Atlantic cod (Gadus morhua) farmed in marine cages: a proof-of-concept study for the identification of escapees.

This study presents a molecular genetic characterization of Atlantic cod reared in commercial marine farms. Samples consisted of approximately 47 fish collected from nine cages located on four farms throughout Norway. In addition, 28 farmed escapees were recaptured in the sea (443 fish in total). Nine microsatellite loci and the Pan I gene were analysed, revealing a total of 181 alleles. Each sample contained 43–63% of total allelic variation. Comparing variation with published data for wild cod indicates that lower genetic variation exists within single cages than in wild populations. Significant linkage disequilibrium was observed amongst pairs of loci in all samples, suggesting a low number of contributing parental fish. Global FST was 0.049, and the highest pairwise FST value (pooled loci) was 0.085. For single loci, the Pan I gene was the most diagnostic, displaying a global FST of 0.203. Simulations amongst the samples collected on farms revealed an overall correct self-assignment percentage of 75%, demonstrating a high probability of identifying individuals to their farm of origin. Identification of the 28 escapees revealed a single cage as the most likely source of origin for half of the escapees, whilst the remaining fish were assigned to a mixture of samples, suggesting more than one source of escapees.

An anaesthetic protocol in the young domestic pig allowing neuromuscular blockade for studies of cardiac function following cardioplegic arrest and cardiopulmonary bypass

Background:  Neuromuscular blockade should, for ethical reasons, not be allowed in animal experiments unless the use is strongly motivated. Beforehand, the anaesthetic protocol must be documented without muscle relaxation in the species studied. Documentation is difficult to obtain from the scientific literature. When focusing on cardiac function over time, in particular, the ideal anaesthetic protocol should cause no or minor alterations in cardiac variables.

Multidose Cold Oxygenated Blood Is Superior to a Single Dose of Bretschneider HTK-Cardioplegia in the Pig

BACKGROUND A single-dose strategy for cardioplegia is desired in minimal invasive approaches to valve surgery and aortic arch repairs. We hypothesized that a single infusion of Bretschneider HTK solution offers myocardial protection comparable to repeated cold oxygenated blood. METHODS Sixteen pigs on bypass with 60 minutes of aortic cross-clamping were randomized to a single dose of Custodiol (HTK group) or repeated oxygenated blood cardioplegia (CBC group). Left ventricular function and perfusion were evaluated by conductance catheter, echocardiography, and microspheres. Myocardial injury was assessed with serum troponin-T. RESULTS Baseline values showed no group differences. One hour after declamping cardiac index was reduced in the HTK group, 3.5 +/- 0.2 L x min(-1) x m(-2) (mean +/- standard error of the mean) compared with 4.7 +/- 0.4 L x min(-1) x m(-2) in the CBC group (p < 0.0005), decreasing to 4.0 +/- 0.2 and 3.9 +/- 0.2 L x min(-1) x m(-2) after 2 and 3 hours, respectively (p < 0.005 versus 1 hour). In the HTK group cardiac index remained low and unchanged. In the CBC group preload recruitable stroke work was 72.6 +/- 1.2 mm Hg 1 hour after declamping, decreasing to 65.2 +/- 2.5 and 60.3 +/- 3.9 mm Hg after 2 and 3 hours, respectively (p < 0.05 versus 1 hour). In the HTK group corresponding values after 1, 2, and 3 hours were low at 47.2 +/- 4.4, 48.4 +/- 4.2, and 50.7 +/- 4.3 mm Hg, respectively (p < 0.025 versus CBC for all). Subendocardial radial peak systolic strain averaged 80.5% +/- 4.8% after declamping in the CBC group versus 53.4% +/- 5.5% in the HTK group (p = 0.002). Serum troponin-T release was lower in the CBC group. CONCLUSIONS Repeated oxygenated blood cardioplegia provides better myocardial protection and preservation of left ventricular function than a single dose of HTK during the early hours after declamping.

The frequency of spontaneous triploidy in farmed Atlantic salmon produced in Norway during the period 2007–2014

BackgroundSpontaneous triploidy has been reported in a number of fish species, and is often linked with in vivo or in vitro ageing of eggs post ovulation. Here, we provide the first investigation into the frequency of spontaneous triploidy in farmed Atlantic salmon by analysing more than 4000 fish from 55 farms, and approximately 1000 recaptured escapees, all sampled in the period 2007–2014. In addition, we compare microsatellite genotyping against flow cytometry and red blood cell diameter in a set of 45 putatively diploid and 45 putatively triploid Atlantic salmon.ResultsThe three methods implemented for ploidy determination gave consistent results, thus validating the methods used here. Overall, 2.0% spontaneous triploids were observed in salmon sampled on farms. The frequency of spontaneous triploids varied greatly among sea cages (0-28%), but they were observed in similar frequencies among the three primary breeding companies (1.8-2.4%). Spontaneous triploids were observed in all farming regions in Norway, and in all years sampled. Spontaneous triploids were also observed among the escapees recaptured in both the marine environment and in rivers.ConclusionsSpontaneous triploidy in commercially produced Atlantic salmon is likely to be a result of the practices employed by the industry. For logistical reasons, there is sometimes a pause of hours, and in some cases overnight, between killing the female broodfish, removal of her eggs, and fertilization. This gives the eggs time to age post ovulation, and increases the probability of duplication of the maternal chromosome set by inhibition of the second polar body release after normal meiosis II in the oocyte.

The influence of acute unloading on left ventricular strain and strain rate by speckle tracking echocardiography in a porcine model

Noninvasive measurements of myocardial strain and strain rate by speckle tracking echocardiography correlate to cardiac contractile state but also to load, which may weaken their value as indices of inotropy. In a porcine model, we investigated the influence of acute dynamic preload reductions on left ventricular strain and strain rate and their relation to the pressure-conductance catheter-derived preload recruitable stroke work (PRSW) and peak positive first derivative of left ventricular pressure (LV-dP/dtmax). Speckle tracking strain and strain rate in the longitudinal, circumferential, and radial directions were measured during acute dynamic reductions of end-diastolic volume during three different myocardial inotropic states. Both strain and strain rate were sensitive to unloading of the left ventricle (P < 0.001), but the load dependency for strain rate was modest compared with strain. Changes in longitudinal and circumferential strain correlated more strongly to changes in end-diastolic volume (r = -0.86 and r = -0.72) than did radial strain (r = 0.35). Longitudinal, circumferential, and radial strain significantly correlated with LV-dP/dtmax (r = -0.53, r = -0.46, and r = 0.86), whereas only radial strain correlated with PRSW (r = 0.55). Strain rate in the longitudinal, circumferential and radial direction significantly correlated with both PRSW (r = -0.64, r = -0.58, and r = 0.74) and LV-dP/dtmax (r = -0.95, r = -0.70, and r = 0.85). In conclusion, the speckle tracking echocardiography-derived strain rate is more robust to dynamic ventricular unloading than strain. Longitudinal and circumferential strain could not predict load-independent contractility. Strain rates, and especially in the radial direction, are good predictors of preload-independent inotropic markers derived from conductance catheter.

Multi-layer radial systolic strain vs. one-layer strain for confirming reperfusion from a significant non-occlusive coronary stenosis

AIMS The aim of this study was to investigate whether multi-layer radial strain and strain rate analysis is superior to one-layer strain analysis for confirming reperfusion following a non-occlusive coronary stenosis. METHODS AND RESULTS In 10 anaesthetized pigs, an extracorporeal shunt was inserted from the brachiocephalic to the left anterior descending coronary artery. Microspheres were injected and left ventricular (LV) short- and long-axis echocardiographic views were recorded with the open shunt, during the 120 min of severe stenosis and 20 min (early) and 100 min (late) after reperfusion. The anterior wall was analysed for radial one-layer and three-layer tissue Doppler imaging (TDI) strain and strain rate, in addition to radial, circumferential, and longitudinal speckle-tracking echocardiography (STE) strain. During stenosis, perfusion was reduced in the two inner wall layers (P< 0.01). All peak systolic strain and strain rate parameters were reduced, whereas post-systolic longitudinal strain and post-systolic strain in the two inner layers increased (P< 0.001). At early reperfusion, hyperaemia was evident in all layers (P< 0.01). Peak systolic TDI strain and strain rate increased in the mid- and subendocardial layer, whereas post-systolic strain decreased (P< 0.05). Peak systolic STE strain increased in the circumferential and longitudinal direction, whereas post-systolic longitudinal strain decreased (P< 0.05). At late reperfusion, strain and strain rate were unchanged while perfusion returned to baseline values in the mid- and subendocardium. CONCLUSION Both multi-layer radial TDI strain and strain rate and one-layer STE strain measurements in the circumferential and longitudinal direction can confirm reperfusion early after a non-occlusive coronary stenosis. An advantage of multi-layer analysis was not evident.

Discrimination among species of the genus Sebastes in the North Atlantic by random amplified polymorphic DNA

Genetic variation among and within species of the genus Sebastes [ S. viviparus , S. fasciatus , S. mentella (including deep-sea and oceanic types) and S. marinus (both ordinary and giant types)] from the North Atlantic was investigated by polymerase chain reaction (PCR)-based random amplification of polymorphic DNA (RAPD). Initial analyses were based on 80 different 10 base pair primers applied to a small number of individuals of each of the four species. Four of the most variable primers were selected for analysis of the total samples from Norway, Iceland and Canada. Only one primer, OPA20, could be used to diagnostically distinguish among species. Statistical analysis clustered all samples in relation to known species structure. No genetic distance was found between ordinary and giant-type S. marinus and minor differences were observed between S. viviparus from Iceland and Norway. Sebastes mentella from the different locations clustered together, but some structuring was shown within this species. Twelve untyped Sebastes and S. fasciatus from Gulf of St. Lawrence clustered closer to S. viviparus than to S. marinus and S. mentella .