Isolation and sequence of cDNA encoding a 3-methylcholanthrene-inducible cytochrome P450 from wild red sea bream,Pagrus major

We have isolated a cDNA clone of mRNA for the cytochrome P450 from a 3-methylcholanthrene (MC)-treated red sea bream,Pagrus major, using a cDNA fragment for rat P4501A2 as a probe. The cloned cDNA is ca. 1.8 kb long and contains an open reading frame of 1545 nucleotides for polypeptides of 515 amino acids. The deduced N-terminal amino acid sequence of the cDNA is very similar to that for purified cytochrome P450 protein from the marine fish scup, which was reported previously (Klotz et al. 1983). A conserved amino acid sequence containing a putative heme-binding cysteine is present in the equivalent position proximate to the C-terminus of the molecules. The deduced amino acid sequence shows more than 50% positional identity with known members of the mammalian aromatic hydrocarbon-inducible P450 family. RNA blot analysis indicates that P450 mRNA (s) is expressed in the liver, kidney, gill and gut of the MC-treatedP. major.     

Sexual maturity and hermaphroditism of the red porgy Pagrus pagrus (Teleostei: Sparidae)

Gonadal changes related to sexual maturity were examined by histology in reared populations of the red porgy Pagrus pagrus L., aged 0.5 to 6.5 years. More than 50% of red porgies were mature at the age of 4 years although some individuals were already mature at the age of 3, as indicated by changes in the gonadosomatic index, plasma levels of vitellogenin, 17β-estradiol and testosterone in females and males. Sex frequency distribution analysis in relation to the age and histological analysis of the gonads indicated the presence of males originated from the sex change of adult functional females. P. pagrus is a protogynous species. Some females had not changed sex up to the age of 6.5 years. Received: 16 July 1998 / Accepted: 29 April 1999     

Feeding of laboratory-reared larvae of the sea bream Archosargus rhomboidalis (Sparidae)

Feeding by larvae of the sea bream Archosargus rhomboidalis (Linnaeus) was investigated from late September, 1972 to early May, 1973 using laboratory-reared larvae. Fertilized eggs were collected from plankton tows in Biscayne Bay, and the larvae were reared on zooplankton also collected in plankton nets. Techniques were developed to estimate feeding rate, food selection, gross growth efficiency, and daily ration. Daily estimates of these were obtained through 16 days after hatching at rearing temperatures of 23°, 26°, and 29°C. Feeding rate increased exponentially as the larvae grew, and increased as temperature was raised. At 23°C larvae began feeding on Day 3, at 26° and 29°C larvae began feeding on Day 2. Feeding rates at initiation of feeding and on Day 16 were, respectively: 23°C, 7.16 food organisms per larva per hour (flh) and 53.78 flh; 26°C, 7.90 flh and 168.80 flh; 29°C, 17.62 flh and 142.07 flh. Sea bream larvae selected food organisms by size. At initiation of feeding they selected organisms less than 100 m in width. As larvae grew they selected larger organisms and rejected smaller ones. The major food (more than85% of the organisms ingested) was copepod nauplii, copepodites, and copepod adults. Minor food items were barnacle nauplii, tintinnids, invertebrate eggs, and polychaete larvae. Mean values for gross growth efficiency of sea bream larvae ranged from 30.6% at 23°C to 23.9% at 29°C. Mean values for daily ration, expressed as a percentage of larval weight, ranged from 84% at 23°C to 151% at 29°C and tended to decline as the larvae grew.This paper is a contribution from the Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, Florida, USA     

Structure of monogenean populations,ectoparasites of the gilthead sea bream Sparus aurata

In the Western Mediterranean, the gilthead sea bream Sparus aurata Linnaeus, 1758 is parasitized by two gill monogeneans, Furnestinia echeneis (Wagener, 1857) Euzet and Audouin, 1959 (Monopisthocotylea: Diplectanidae) and Sparicotyle chrysophrii (van Beneden and Hesse, 1863) Mamaev, 1984 (Polyopisthocotylea: Microcotylidae). The spatial structure of these parasitic populations were studied on the French coast in the Gulf of Lions in 1984 and 1985. The relations between the two species forming this guild have been demographically analyses. Their distribution profiles are very similar and display some stability with time, although some increasing trends are perceptible in the spring. These two monogeneans are globally frequent, but always present in only moderate intensities in infrapopulations. Recruitment of F. echeneis is independent of that of S. chrysophrii. This mode of contamination limits massive and simultaneous infestations of a single host, and interspecific competition risks are consequently considerably reduced. Our results suggest these parasites can affect the size of a natural gilthead population to only a limited degree.     

Energy metabolism during development of eggs and larvae of gilthead sea bream (Sparus aurata)

Developing eggs and larvae of laboratory-reared gilthead sea bream (Sparus aurata) maintained in filtered seawater (40 ppt) at 18°C, were measured for oxygen uptake, ammonia excretion, contents of free amino acids (FAA), protein, fatty acids (FA) accumulated ammonia, and volumes of yolk-sac and oil globule. Absorption of the yolk coincided with the consumption of FAA and was complete ca. 100 h post-fertilisation. Amino acids from protein were mobilised for energy in the last part of the yolk-sac stage. Absorption of the oil globule occurred primarily after hatching following yolk absorption, and correlated with catabolism of the FA neutral lipids. Overall, FAA appear to be a significant energy substrate during the egg stage (60 to 70%) while FA from neutral lipids derived from the oil globule are the main metabolic fuel after hatching (80 to 90%).     

Utilization of yolk reserves during post-embryonic development in three teleostean species: the sea bream Sparus aurata,the sea bass Dicentrarchus labrax,and the pike-perch Stizostedion lucioperca

A comparative study of the use of yolk reserves during post-embryonic development was performed on three teleostean fishes of aquacultural interest: the sea bream Sparus aurata, the sea bass Dicentrarchus labrax, and the pike-perch Stizostedion lucioperca. The first part of the work was devoted to the structural development of the periblast, the vitellus and the oil globule during the endotrophic and endo-exotrophic phases of larval life. Observations using transmission electron microscopy revealed a perivitelline circulation system closely related to the liver, and suggested an important function of the periblast in the utilization of reserves. In the second part of the study, cytochemical reactions revealed the way in which the reserves are resorbed. Glycogen contained in the periblast at hatching was utilized during the endotrophic period. The successive appearance of lipoprotein particles in the periblast endoplasmic reticulum, Golgi apparatus, and perivetelline circulation zone is consistent with the mobilization of yolk and oil globule components. Comparison of fed and fasting individuals revealed that fasting tended to slow down rather than increase the utilization of reserves.     

Seasonal cycle of gonadal development and serum levels of vitellogenin of the red porgy, Pagrus pagrus (Teleostei: Sparidae)

Histological examination of gonads of female and male red porgy, Pagrus pagrus, reared in captivity, was carried out in order to describe the main gonadal changes related to gametogenesis and the seasonal changes of environmental factors. The gonadosomatic index (GSI) and the concentration of serum vitellogenin (VTG) were also determined. The frequency distribution of gonad development stages and the GSI and vitellogenin concentration during the annual cycle indicated the separation of the female and male reproductive cycles into three main periods. The autumn period when gametogenesis begins (October-November), the period of exogenous vitellogenesis (December-March) or spermiation (December-March), and the spawning season (March-May). The spawning period coincided with an increase in temperature (15-19°C) and daylength. Serum levels of vitellogenin rose significantly in January, reached a peak at the beginning of the spawning period (March, 405.5 µg ml^-1) and remained high until the end of the spawning period (May).     

Effect of low dissolved oxygen concentrations on behavior and predation rates on red sea bream Pagrus major larvae by the jellyfish Aurelia aurita and by juvenile Spanish mackerel Scomberomorus niphonius

A shift in outcomes of predator-prey interactions in plankton community may occur at sublethal dissolved oxygen concentrations that commonly occur in coastal waters. Laboratory experiments were conducted to investigate how a decline in dissolved oxygen concentration alters the predation rate on fish larvae by two estuarine predators. Behavior and consumption of larval fish by moon jellyfish Aurelia aurita (103.1±12.4 mm in bell diameter) and by a juvenile piscivore, Spanish mackerel Scomberomorus niphonius (30.1±2.1 mm in standard length: SL), were observed under four oxygen concentration treatments (1, 2 and 4 mg l^–1 and air-saturated: 5.8 mg l^–1). Larvae of a coastal marine fish species, red sea bream Pagrus major (7.21±0.52 mm SL), were used as prey for the experiment. Bell contraction rate of the jellyfish did not vary among the oxygen concentrations tested, indicating a tolerance to low oxygen concentration. Gill ventilation rate of the Spanish mackerel increased and swimming speed decreased as the oxygen concentration decreased, indicating that oxygen concentrations 4 mg l^–1 are physiologically stressful for this species. The number of larvae consumed in 15 min. by jellyfish increased whereas those consumed by Spanish mackerel decreased with the decrease in oxygen concentration. Low oxygen concentrations that are commonly observed in coastal waters of Japan during summer have the potential to increase the relative importance of jellyfish as predator of fish larvae and to change the importance of alternative trophic pathways in estuarine ecosystems.Communicated by T. Ikeda, Hakodate     

Element levels in cultured and wild sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) from the Adriatic Sea and potential risk assessment

In this study, the role of aquaculture activity as a source of selected metals was analyzed. Significant differences in element content between cultured (Dicentrarchus labrax, Sparus aurata) and wild fishes as well as between fish muscle and their feed were detected. Higher concentrations of trace elements (i.e., As, Cu, Hg, Se) in wild fish tissues in comparison with cultured ones indicate additional sources of metals beside fish feed as natural and/or anthropogenic sources. Generally, mean Cd, Cu, Pb, Se, and Zn concentrations in cultured (0.016, 1.79, 0.14, 0.87, and 34.32 μg/g, respectively) and wild (0.011, 1.97, 0.10, 1.78, and 23,54 μg/g, respectively) fish samples were below the permissible levels, while mean As (2.57 μg/g in cultured, 4.77 μg/g in wild) and Cr (5.25 μg/g in cultured, 2.92 μg/g in wild) values exceeded those limits. Hg values were lower in cultured (0.17 μg/g) and higher in wild (1.04 μg/g) fish specimens. The highest elemental concentrations were observed in almost all fish samples from Kor?ula sampling site. The smallest cultured sea basses showed As (4.01 μg/g), Cr (49.10 μg/g), Pb (0.65 μg/g), and Zn (136 μg/g) concentrations above the recommended limits; however, values decreased as fish size increased. Therefore, the majority of metal concentrations in commercial fishes showed no problems for human consumption. Also calculated Se:Hg molar ratios (all >1) and selenium health benefit values (Se-HBVs) (all positive) showed that consumption of all observed fishes in human nutrition is not risk.     

Genetic heterogeneity among adult and recruit red sea urchins, Strongylocentrotus franciscanus

Allozyme electrophoresis was used to characterize genetic variation within and among natural populations of the red sea urchin Strongylocentrotus franciscanus. In 1995 to 1996, adult urchins were sampled from twelve geographically separated populations, seven from northern California and five from southern California (including Santa Rosa Island). Significant population heterogeneity in allelic frequencies was observed at five of six polymorphic loci. No geographic pattern of differentiation was evident; neighboring populations were often more genetically differentiated than distant populations. Northern and southern populations were not consistently distinguishable at any of the six loci. In order to assess within-population genetic variation and patterns of recruitment, large samples were collected from several northern California populations in 1996 and 1997, and were divided into three size classes, roughly representing large adults (>60 mm), medium-sized individuals (31 to 60 mm, “subadults”) and individuals <2 yr of age (≤30 mm test diam, referred to as “recruits”). Comparisons of allelic counts revealed significant spatial and temporal differentiation among size-stratified population samples. Recruit samples differed significantly from adult samples collected at the same locale, and showed extensive between-year variation. Genetic differentiation among recruit samples was much higher in 1997 than in 1996. Between-year differences within populations were always greater for recruits than for adults. Potential explanations for the differentiation of recruit samples include pre- and post-settlement natural selection and high interfamily variance in reproductive success or “sweepstakes” recruitment. Unless recruit differentiation can be attributed to an improbable combination of strong and spatially diverse selection, such differentiation across northern California populations indicates that the larval pool is not well mixed geographically (even on spatial scales <20 km), despite long planktonic larval duration. Received: 6 July 1999 / Accepted: 25 January 2000     

Diurnal vertical migration of Cochlodinium polykrikoides during the red tide in Korean coastal sea waters

The diurnal vertical migration of Cochlodinium polykrikoides (C. polykrikoides), which caused a red tide in the Korean coastal waters of the East Sea/Sea of Japan in September 2003, was examined by determining the time-dependent changes in the density of living cells in relation to the depth of the water column. The ascent of this species into the surface layer (depth of water 2 m) occurred during 1400-1500. The descent started at 1600 and a high distribution rate (86%) at 15-20 m was observed at 0300. During the ascent, the cells were widely distributed at each depth level from 0600 hr and at 0800-1100, the cells were primarily distributed in the middle layer (0-6 m). The concentration of dissolved inorganic nitrogen was generally < or = 2.86 micromol l(-1), but at 1400-1500, the concentration in the surface layer reduced to < or = 0.14 micromol l(-1). Moreover, the concentration gradually increased as the depth increased to > or = 5 m. These results showed that the nutrient-consumption rate associated with the proliferation of C. polykrikoides during a red tide is more influenced by the inorganic-nitrogen resources ratherthan the inorganic-phosphorus compounds.     

Movement and feeding activity of red sea urchins (Strongylocentrotus franciscanus) adjacent to a kelp forest

Movement and feeding were studied in a population of red sea urchins, Strongylocentrotus franciscanus (Agassiz, 1863), found within and immediately seaward of a kelp forest offshore from Santa Cruz, California, USA. Mean sea urchin movements varied from 7.5 cm/day inside the kelp forest to over 50 cm/day at 15 and 100 m outside the kelp forest. The percentage of sea urchins feeding decreased from 66% inside the kelp forest to 16 and 15% at 15 and 100 m outside the kelp forest. These data indicate that movement by these sea urchins is a response to a low food supply.     

Population structure of red porgy, <Emphasis Type="Italic">Pagrus pagrus</Emphasis>, in the Atlantic Ocean

The red porgy, Pagrus pagrus (L.), is a protogynous sparid associated with reefs and hard bottom habitat throughout the warm-temperate Atlantic Ocean. In this study, the degree of geographic population differentiation in Atlantic populations was examined with microsatellite and mitochondrial DNA markers (mtDNA). Six microsatellite loci were amplified and scored in 690 individuals from the eastern North Atlantic (Crete, Madeira, and Azores), western North Atlantic (North Carolina to Florida, and the eastern Gulf of Mexico), and Brazil. At two loci, fixed allelic differences were found among the three major geographic areas, while frequency differences were observed at three other loci. The DNA of 371 individuals was amplified at the mtDNA control region, and 526 bp were sequenced. Tamura–Nei’s D was used as a measure of nucleotide diversity and divergence: diversity averaged 0.011 within samples, while the corrected divergence averaged 0 between samples within the same area and 0.061 between samples from different areas. Transversion haplotype minimum spanning networks, nucleotide divergence, and F _ST values all show that the western Atlantic samples were more closely related to each other than any was to samples from the eastern North Atlantic. Within the western North Atlantic, no significant population differentiation was observed, and within the eastern North Atlantic, only the Azores sample showed detectable differences from Crete and Madeira. These data indicate general homogeneity within large areas, and deep divisions between these areas. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Interaction of short-term testosterone treatment with osmotic acclimation in the gilthead sea bream <Emphasis Type="Italic">Sparus auratus</Emphasis>

To assess the interaction between testosterone (T) treatment and acclimation to different salinities, seawater-acclimated gilthead sea bream (Sparus auratus) were implanted with slow-release coconut oil implants alone (control) or containing T (5 μg/g body mass). After 5 days, eight fish of control and T-treated groups were sampled. The same day, eight fish of each group were transferred to low salinity water (LSW, 6 ppt, hypoosmotic test), seawater (SW, 38 ppt, control test) and high salinity water (HSW, 55 ppt, hyperosmotic test) and sampled 9 days later. Gill Na⁺, K⁺-ATPase activity increased in HSW-acclimated fish with respect to SW- and LSW-acclimated fish in both control and T-treated groups. Kidney Na⁺, K⁺-ATPase activity was also enhanced in HSW-acclimated fish, but only in T-treated group. From a metabolic point of view, most of the changes observed can be attributed to the action of salinity and T treatment alone, since few interactions between T treatment and osmotic acclimation to different salinities were observed. Those interactions included in treated fish: in the liver, decreased capacity in using glucose in fish acclimated to extreme salinities; in the gills, decreased capacity in using amino acids in HSW; in the kidneys increased capacity in using amino acids in extreme salinities; and in the brain, decreased glycogen and acetoacetate levels of fish in LSW.     

Energy investment in the annual reproduction cycle of female red porgy, <Emphasis Type="Italic">Pagrus pagrus</Emphasis> (L.)

Seasonal cycle in energy content of gonad, perivisceral fat, liver, and muscle was used to estimate the energy cost of spawning of female red porgy (Pagrus pagrus) from Buenos Aires coasts, Argentina. Spawning was observed primarily between November and January, although a low-intensity spawning was also observed at the beginning of this period. The energy required for gonad development of female red porgy (the difference between the maximum and minimum energy content of ovaries along the year) was 2,652 J g⁻¹ dry weight or 558 kJ. Female red porgy expend 22.8% of their total body energy (3,481 kJ) during the spawning period. Of this energy, relative contributions from different sources were as follows: ovaries 62.5% (496 kJ), perivisceral fat 17.8% (142 kJ), liver 10.3% (82 kJ), and muscles 9.4% (75 kJ). Female spawners lost about 7.2% (119 g) of their total body weight (1,654 g) in the spawning period. From the 7.2% of the total weight lost, female gonad represented a relative reduction of 73.8% (87.6 g), the perivisceral fat declined by 5.7% (6.8 g), the liver declined by 10.6% (12.5 g), and muscle tissue exhibited a 10% loss in total weight (11.9 g). Reproductive effort represented 7.6% of food energy, while resting metabolism accounted for almost 38% of food energy. Despite the fact that red porgy females feed throughout the entire spawning period, it is shown that between pre-spawning (September, water temperature 15°C) and mid-spawning (December, water temperature 19°C) they lost 45 and 38% of their muscle and liver lipids, respectively, and 89% of the perivisceral fat associated with the gut and body cavity. Red porgy used both their energy reserves and food energy during ovarian maturation. This asynchronous species have a strategy of a lower energy allocation and a higher dependence of food during the spawning time (late-spring and summer) for successful reproduction.     

淫羊藿果实成熟与生境的关系

以四川省南充市金城山3个不同海拔高度(562 m, 640 m和726 m)的淫羊藿为研究对象,对其果实成熟的时间及数量进行了研究;同时也对其果实成熟量与大气湿度、温度、光照强度之间的关系进行了研究.相关的数据分析表明,淫羊藿果实成熟过程可分为初始期、高峰期和末期3个阶段.果实成熟主要受湿度,光照和温度的影响.气温的突变可能是促进淫羊藿果实成熟的机制之一;果实数与相对光照、温度之间都存在极显著的相关性.随着海拔高度的变化,淫羊藿果实成熟时间也有相应变化.由于淫羊藿果实成熟后易自裂,所以研究不同生境淫羊霍果实成熟时间的差异对于淫羊霍种子的采收也相当重要.     

Nutritional requirements of the gilthead bream Chrysophrys aurata. Quantitative protein requirements

Six semi-synthetic diets with a casein and corn-starch base, containing 10 to 60% protein were fed for 4 months to young gilthead breams (Chrysophrys aurata, of average initial weight 2.6 g). The object of the study was to determine the optimum amount of protein necessary for growth. The daily intake of the gilthead breams was 5.2 to 2.3% of their live weight. The less protein the diet contained, the more feed the fish consumed. The conversion factor decreased regularly as protein percentage increased. The most favourable conversion rate was 1.4, noted for the group receiving 60% protein. The less protein in the diets, the better the protein efficiency for weight gain and body protein retention. Growth of gilthead breams was proportional to protein content of the diet up to an incorporation rate of 40%. Beyond this, no significant difference in final average individual weight was observed. These results are discussed in relation to those reported for other species of fishes. They show that the gilthead bream requires about 40% protein in its diet, as do most fresh-water species.     

Determinants of group size in the red colobus monkey (Procolobus badius): an evaluation of the generality of the ecological-constraints model

The ecological-constraints model proposes that increased group size increases within-group feeding competition, necessitating increased travel and, consequently, constraining group size. Previous studies have supported the model for frugivores, but its applicability to folivores remains untested. This study evaluated the generality of the model by re-examining the relationship between day range and group size for a folivorous species for which published accounts have not found a relationship between these factors. This study differs from earlier studies by accounting for variation in food availability, which may drive changes in day range. We quantified the relationships among food availability, day range, and group size for two red colobus groups at Kibale National Park, Uganda. Mean day range and home range were significantly greater for the group of 48 individuals compared to the group of 24 individuals. The large group traveled more and rested less than the small group. The large group also traveled more rapidly than the small group. Food availability significantly predicted mean day range for the large group, but not for the small group. Analyses of covariance demonstrated that the large and small group responded differently to changes in food availability. These observations suggest that the large red colobus group experiences greater levels of within-group feeding competition than the small group. This study provides added support for the generality of the ecological-constraints model and contributes toward an understanding of the mechanisms controlling feeding competition and social organization in primates.     

Bioassay of biotin and its distribution in the sea

A simple, rapid and sensitive method of microbiological assay for the determination of biotin in seawater has been developed by using a marine bacterium, Achromobacter sp. (Strain yH-51) as an assay organism. When the organism was incubated at 25°C for 48 h in a synthetic medium with constant shaking, a standard reference curve of bioassay was obtained. The organism is sensitive in the range 0.1 to 8.0 ng/l biotin. The distribution of biotin has been determined in the East China Sea, the North Pacific Ocean, and the bays and inlets along the Pacific coast of Japan. Biotin concentrations in surface waters were in the range from less than 0.1 ng/l up to 57.9 ng/l; generally high in coastal waters and low in deep open waters. The concentrations were generally high in surface waters, and rapidly decreased with depth down to 50 m, below which values were generally low. The vertical distribution of biotin followed closely the concentration of chlorophyll a, with high concentrations of biotin restricted to the euphotic layer of high chlorophyll a contents at most stations observed. Biotin concentrations in surface waters were generally high during summer, but in autumn and winter the concentrations decreased markedly.