Isolation and characterization of polymorphic microsatellite loci from a dinucleotide-enriched genomic library of starry flounder (Platichthys stellatus) and cross-species amplification

Starry flounder (Platichthys stellatus) is a rare fish species in China. Here, we reported 12 polymorphic microsatellite loci isolated from a dinucleotide-enriched genomic library of starry flounder (P. stellatus). The number of alleles, observed and expected heterozygosity per locus in 30 individuals ranged from two to six, from 0.2500 to 1.0000 and from 0.4512 to 0.7667, respectively. One locus significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium between pairs of loci was found. Cross-species amplification of these microsatellite loci in additional three fish species was performed. These polymorphic microsatellite loci would be useful for investigating genetic population structure and construction of genetic linkage map in P. stellatus. Guidong Miao and Changwei Shao have contributed equally.     

石鲽细菌性败血感染症及其病原细菌研究

2001—2002年,分别对两起养殖的石鲽(Stone flounder,Kareius bicoloratusL.)病害进行了检验,均表现为败血症感染特征。经以20尾病(死)石鲽(每起病例各10尾)做病变组织中细菌检查、细菌分离与鉴定、人工感染试验等的病原学检验,表明两起被检病例均为同种的杀鲑气单胞菌(Aeromonas salmonicida)感染;另在其中1起病例所检10尾石鲽的2尾中,同时检出了呈继发感染的不动杆菌(Acinetobacter)。通过对所分离后做纯培养的60株杀鲑气单胞菌6、株不动杆菌分别进行较系统的表观分类学指征(Phenotypic information)及代表菌株16S rRNA基因序列测定与系统发育学分析,表明其中的杀鲑气单胞菌为一个新亚种,不动杆菌为琼氏不动杆菌(Acinetobacter junii)的一个新形态型;又择代表菌株送中国典型培养物保藏中心(CCTCC)进行了表观性状、DNA中C Cmol%的测定等的复核鉴定,并将其中的杀鲑气单胞菌新亚种定名为杀鲑气单胞菌杀鲽亚种(Aeromonas salmonicidasubsp.flounderacidasubsp.no...     

Five new microsatellite loci for Oliver flounder (Paralichthys olivaceus) from an expressed sequence tag (EST) library and cross‐species amplification

Five microsatellite loci have been isolated and characterized from a cDNA library of Oliver flounder, Paralichthys olivaceus. All loci were found to be polymorphic and had between four and 10 alleles. Observed and expected heterozygosities varied from 0.70 to 0.90 and from 0.52 to 0.80, respectively. Five additional fish species assessed for cross‐species amplification revealed between zero and three positive amplifications and between zero and two polymorphic loci per species.     

Freshwater adaptation during larval, juvenile and immature periods of starry flounder Platichthys stellatus, stone flounder Kareius bicoloratus and their reciprocal hybrids

The freshwater tolerance of starry flounder Platichthys stellatus , stone flounder Kareius bicoloratus and their reciprocal hybrids, produced by artificial insemination, were examined in the larval, juvenile and immature phases. Survival rate on being transferred to fresh water in the pre-settlement phase was 0% in stone flounder and hybrids and 16·7% in starry flounder. This rate in the post-settlement phase was elevated to >50% in starry flounder and hybrids but was still 0% in stone flounder and similarly in the immature period starry flounder and hybrids survived in fresh water, although stone flounder did not. The lamella chloride cells of the gill epithelium increased in starry flounder and hybrids in fresh water in all periods. Densities of lamella chloride cells increased from 1·6 ± 0·4 (mean ± s . e . number of cells per 1 mm filament) before the transferral (day 0) to 60·3 ± 6·2 on 14 days after the transfer to fresh water (day 14) in starry flounder in the immature period. These densities in hybrids were 0·6 ± 0·3 and 1·0 ± 0·3 on day 0, and, 35·3 ± 2·8 and 23·2 ± 4·6 on day 14, respectively. Stone flounder did not show a substantial change in chloride cell densities throughout the experimental period. These results suggest that low salinity tolerance was well developed in the settlement period in starry flounder and hybrids, and hybrids were also adapted to fresh water sufficiently regardless of the cross type.     

A pore-forming protein,perforin, from a non-mammalian organism,Japanese flounder,<Emphasis Type="Italic"> Paralichthys olivaceus</Emphasis>

A perforin cDNA of Japanese flounder, Paralichthys olivaceus, was cloned from a cDNA library of kidney stimulated with ConA/PMA. The full-length cDNA is 2,157 bp, which encodes 587 amino acids. The Japanese flounder perforin gene consists of five exons and four introns, with a length of approximately 3 kb. The amino acid sequence of the Japanese flounder perforin is 36% identical to that of rat perforin and 37% identical to amino acid sequences of mouse and human perforin. The Japanese flounder perforin also showed low homology to human and mouse complement components (C6, C7, C8 and C9), ranging from 19% to 24%. However, the membrane attack complex/perforin domain is conserved. A phylogenetic analysis placed the Japanese flounder perforin in the same cluster with other known mammalian perforins. RT-PCR analysis revealed that the perforin gene was expressed in the peripheral blood leukocytes, head kidney, trunk kidney, spleen, heart, gill and intestine of healthy fish. Recombinant perforin produced in insect cells using the baculovirus expression system showed calcium-dependent hemolytic activity.     

Protein synthesis, nitrogen excretion and long-term growth of juvenile Pleuronectes flesus

This study aimed to measure protein synthesis using a stable isotope method, investigate protein-nitrogen flux in a flatfish Pleuronectes flesus , and use the data to test the hypothesis that individual differences in growth efficiency were related to individual differences in protein-nitrogen flux mediated through differences in protein synthesis and degradation. Three measurements of protein-nitrogen flux via consumption, protein synthesis and nitrogenous excretion were made for individual flounder during a 212-day period and fractional rates of protein-nitrogen flux were scaled for a 50–g flounder to provide mean values for protein consumption (2·11 ± 0·21% day⁻¹), protein synthesis (2·08±0·23% day⁻¹), protein growth (0·71±0·06% day⁻¹) and protein degradation (1·37±0·24% day⁻¹). Mean rates of nitrogenous excretion were 0·142 mg N g⁻¹ day⁻¹ and 0·047 mg N g⁻¹ day⁻¹ for ammonia and urea, respectively. Individual flounder had different protein growth efficiencies and this was correlated negatively and significantly with mean rates of protein synthesis ( r - 0·70; P <0·05) and degradation ( r - 0·67; P < 0·05) and correlated positively and significantly with the efficiency of retaining synthesized protein ( r +0·63, P <0·05). This supported the proposed hypothesis that flounder which grow more efficiently achieve this through adopting a low protein turnover strategy.     

Distribution and ontogeny of digestive enzymes in larval yellowtail and winter flounder

The distribution of digestive enzymes was studied using histoenzymological methods in yellowtail and winter flounder larvae reared on three different diets: live food, weaned at day 15, or starved. Alkaline phosphatase, dipeptidyl peptidase IV, aminopeptidase M and esterase were present at 3 days post-hatch and became differentially distributed coinciding with morphological development. For larvae fed a live diet, activity of these enzymes was present in the intestine of both species and rectum of yellowtail flounder. Alkaline phosphatase was also present in the post-oesophageal swelling (stomach anlage) of winter flounder. Activity of all enzymes was absent in starved winter flounder larvae and a decrease in aminopeptidase M and esterase activity occurred in starved yellowtail flounder larvae. Acid phosphatase was not identified in either species. The eVect of weaning on enzymatic activity was not evaluated fully as the larvae did not survive long enough after the introduction of the artificial diet to complete experimentation.     

The effect of feeding regime on growth, locomotor activity pattern and the development of food anticipatory activity in greenback flounder

The mean final weight of juvenile greenback flounder Rhombosolea tapirina fed a single daily meal during the scotophase was lower than that of groups fed continuously or fed twelve, six, three or one meal during the photophase. Final weight did not differ among the latter five groups. Size variation in all groups of fish decreased during the course of the trial over 120 days. Groups assigned to a higher frequency (and hence a longer total daily meal duration) of daily meals were less active during mealtime. A distinct food anticipatory effect was observed in the groups of fish receiving a single daily meal during either photophase or scotophase, and also developed before at least one meal in the group of fish fed three meals per day.     

On the age and growth of flounder Paralichthys orbignyanus (Jenyns, 1842) in Bahía Blanca Estuary, Argentina

Estimates of age and growth of flounder Paralichthys orbignyanus (Jenyns,1842) were made by analysing fish from commercial catches in Bahía Blanca estuary (39° LS). A total of 823 fish caught by fishing fleets operating in the estuary were collected between February 1997 and January 1998. Age was determined via scale reading and growth estimation parameters using von Bertalanffys equation. Annulus formation occurred in August. The age for the total population, which ranged from 7.7 to 87.5 cm TL was 0–7. Age ranged from 0 to 6 years (21.5–71.9 cm TL) in males and from 0 to 7 years (24.9–87.5 cm TL) in females. Length–weight relationships were W(g)=0.0093 L(cm) exp. 3.03 for the total population, W(g)=0.0147 L(cm) exp. 2.91 for females, and W(g)=0.0168 L(cm) exp. 2.87 for males, respectively. As from age three, females were longer and heavier than males. The growth parameters estimated for the total population, for females and for males were L=83.29, 79.66, and 46.11 cm, respectively; k values were 0.18, 0.23, and 0.92, respectively, and t₀ values were –1.87; –1.54, and –0.62, respectively. Similarly to other flounders,P. orbignyanus is a typical inhabitant of estuaries and coastal regions. Adults stay in the study area mainly during spring and summer, they go outside or offshore during the spawning period and then they return to the estuary for feeding and recovering.