Effect of Temperature Fluctuation and Nutritional Status on Starry Flounder, Platichthys stellatus, Survival and Adaptive Physiological Response

Starry flounder (Platichthys stellatus) is a commercially important cold-water fish. Our aim was to investigate the effects of fluctuating water temperature on flounders after periods of starvation and feeding. Fish were divided into starvation and feeding groups. The water temperature was increased stepwise in experiment 1; more focused variations, based on the results of experiment 1, were studied in experiment 2. At temperatures ≤27 °C, there was no significant difference observed in survival. At 28 °C, mortality increased, survival was lower (21%) in the starvation group than in the feeding group (46%), and weight loss was the highest (15%) in the starvation group. In experiment 2, survival was ≥86%, and there was no significant difference between the starvation/feeding groups. However, when the water temperature was increased to 27 °C after being decreased to 12 °C, weight loss was the highest (11%). Glucose, cortisol, superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels increased with increasing water temperature, and then gradually decreased. Glutamic pyruvic transaminase (GPT)/glutamic oxaloacetic transaminase (GOT) levels showed large variations among individuals. Triglyceride, cholesterol, and protein levels gradually decreased with long-term starvation. Survival was not affected by water temperature drop ≤27 °C after starvation/feeding. These results indicate that 27 °C is the upper limit of tolerable water temperature for the survival of starry flounders. Therefore, aquaculture farms should ensure maintaining water temperatures at ≤27 °C during high-temperature periods.

Origin of the type I antifreeze gene in flounders in response to Cenozoic climate change

Antifreeze proteins (AFPs) inhibit ice growth within fish and protect them from freezing in icy seawater. Alanine-rich, alpha-helical AFPs (type I) have independently (convergently) evolved in four branches of fishes, one of which is a subsection of the righteye flounders. The origin of this gene family has been elucidated by sequencing two loci from a starry flounder, Platichthys stellatus, collected off Vancouver Island, British Columbia. The first locus had two alleles that demonstrated the plasticity of the AFP gene family, one encoding 33 AFPs and the other allele only four. In the closely related Pacific halibut, this locus encodes multiple Gig2 (antiviral) proteins, but in the starry flounder, the Gig2 genes were found at a second locus due to a lineage-specific duplication event. An ancestral Gig2 gave rise to a 3-kDa "skin" AFP isoform, encoding three Ala-rich 11-a.a. repeats, that is expressed in skin and other peripheral tissues. Subsequent gene duplications, followed by internal duplications of the 11 a.a. repeat and the gain of a signal sequence, gave rise to circulating AFP isoforms. One of these, the "hyperactive" 32-kDa Maxi likely underwent a contraction to a shorter 3.3-kDa "liver" isoform. Present day starry flounders found in Pacific Rim coastal waters from California to Alaska show a positive correlation between latitude and AFP gene dosage, with the shorter allele being more prevalent at lower latitudes. This study conclusively demonstrates that the flounder AFP arose from the Gig2 gene, so it is evolutionarily unrelated to the three other classes of type I AFPs from non-flounders. Additionally, this gene arose and underwent amplification coincident with the onset of ocean cooling during the Cenozoic ice ages.

Dead-End (dnd) Gene Cloning and Gonad-Specific Expression Pattern in Starry Flounder (Platichthys stellatus)

dnd is a germline-specific maternal RNA expressed in various vertebrate classes, which encodes an RNA-binding protein that is essential for PGC migration. The purpose of this study is fundamental research about starry flounder dnd gene for germ cell marker development. In this study, we cloned and analyzed the expression levels of Platichthys stellatus dead end (psdnd) in various tissues and embryonic stages. The psdnd gene was isolated from starry flounder ovaries, cloned into a pGEM-t vector, and sequenced. Full-length of psdnd cDNA was 1495 bp long, encoding 395 amino acids. psdnd expression levels were investigated by real-time polymerase chain reaction (qRT-PCR) in various tissues and embryo developmental stages. psdnd transcripts were detected in the testes and ovaries, but not in somatic tissues. Embryonic psdnd expression levels were higher during early embryo development stages than during late embryogenesis; psdnd expression was highest at the 1 cell stage, then gradually decreased throughout the subsequent developmental stages. The spatial expression pattern was analyzed by whole-mount in situ hybridization (WISH). The psdnd transcripts migration pattern was similar with zebrafish (Danio rerio). Our results suggest that psdnd may function as a germ cell-specific marker.

Assessment of the efficacy of using taurine supplements to improve growth and feed utilization of juvenile starry flounder (Platichthys stellatus) given diets based on soy-protein

A feeding trial was conducted to assess the feasibility of supplementing taurine in soy-based diets for juvenile starry flounder Platichthys stellatus. The basal diet (Crude protein 66.5%, crude lipid 8.5%) was supplemented with 0 (control), 0.5%, 1.0%, 1.5%, 2.0% and 2.5% taurine to formulate six test diets. Each diet was fed to 40 juvenile fish (22.25 g) in triplicate tanks (120 L) attached to a sea water circulation-system. Fish were fed twice daily by hand to apparent satiation during the 56-d trial. At the end of the trial, fish were counted and weighed for the analyses of growth performance, diet utilization and survival after a 24-h fast. Blood, intestines and muscles were collected for the analyses of serum oxidation resistance, digestive enzymes and body compostion. Livers were collected from the remaining fish at 4 h post-feeding for metabolic enzymes analyses. The results showed that fish fed diets supplemented with 1.0–2.5% taurine grew from 22.25–22.26 g to 47.88–50.40 g with higher average weight gain (25.62–28.12 vs 23.07 g ), specific growth rate (1.37–1.46 vs 1.27%/d ), feed intake (1.04–1.06 vs 1.00%/d), protein efficiency (2.50–2.61 vs 2.44) and lower feed conversion rate (0.84–0.83 vs 0.89) than the control treatment. Diets supplemented with 1.5–2.5% taurine significantly elevated the activities of pepsin (2.47–2.55 vs 2.22, U mg−1 prot), trypsin of distal intestine(14.55–15.24 vs 11.94, U mg−1 prot), hepatic glucokinase (126.62–129.42 vs 105.56, U mg−1 prot) and fatty acid synthetase (125.56-136.89 vs 108.45, U mg−1 prot). All diets supplemented with taurine increased the activities of lipase (32.23–36.67 vs 29.53, U g−1 prot) and trypsin (35.85–37.89 vs 33.54, U mg−1 prot) of proximal intestine, hepatic aspartate transaminase (736.990–832.38 vs 699.24, U mg−1 prot), alanine aminotransferase (477.40–551.86 vs 373.97, U mg−1 prot) and glycogen synthase (2.16–2.59 vs 1.97, U mg−1 prot), as well as serum superoxide dismutase (4.33–4.59 vs 4.07, U mg−1 prot ) and glutathione peroxidase (42.23–50.25 vs 39.17, mol mg−1 prot). Therefore, taurine supplementation benefits juvenile starry flounder growth, digestion, nutrients metabolism and oxidation resistance. The optimal taurine requirement for starry flounder is 1.75%, and the recommended supplementation level is at least 1.6% for maximizing growth of fish fed a low-fishmeal diet (13.6%).