三聚氰胺对花鲈的急性毒性实验研究

本文以花鲈(Lateolabrax japonicus)为实验对象,采用接触、腹腔注射及口服三种致毒方式,进行了非蛋白氮物质-三聚氰胺对花鲈的急性毒性实验研究。口服急性毒性实验中,通过在饲料中添加不同浓度的三聚氰胺（0、500、2000、5000及10000 mg/kg）,进行了21d的花鲈养殖实验。实验结果表明：三聚氰胺溶解度较低,其水溶液没有表现出急性毒性,LC50 > 3500 mg/L;而在腹腔注射致毒方式下,三聚氰胺对花鲈的半致死剂量LD50 = 890.07 mg/kg•w;LD5095%可信限为：778.63－1017.45 mg/kg•w;在口服急性毒性实验中,10000 mg/kg组三聚氰胺降低了花鲈的摄食与生长(p<0.05),饲料系数显著升高 (p<0.05);三聚氰胺对花鲈的存活率、肥满度、肝体比及脏体比均无显著影响 (p>0.05)。饲料中添加三聚氰胺没有显著影响花鲈血清的丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总蛋白、葡萄糖、胆固醇、甘油三酯及尿素氮 (p>0.05),但显著影响了花鲈血清的碱性磷酸酶活性,10000 mg/kg组的碱性磷酸酶活性显著高于其他各处理组 (p<0.05),其他各组间无显著差异(p>0.05)。三聚氰胺对花鲈21d的最大未观察到有害作用剂量（NOAEL, no-observed-adverse-effect-level）为131.99 mg/kg•w•d。     

Differential population structuring and demographic history of two closely related fish species, Japanese sea bass (Lateolabrax japonicus) and spotted sea bass (Lateolabrax maculatus) in Northwestern Pacific

The Quaternary cold periods in the Northwestern Pacific are thought to have heavily influenced the amount and distribution of intraspecific genetic variation in marine fishes. To estimate the demographic history and genetic structure of Lateolabrax maculatus and L. japonicus in the Northwestern Pacific, 256 individuals were sampled from 19 localities throughout the distribution range of the two species. Mitochondrial DNA variation was analyzed using DNA sequence data from the cytochrome b gene and control region. Nucleotide diversity was much higher in L. japonicus (0.030) than in L. maculatus (0.012). The demographic history of the two species was examined using neutrality tests and mismatch distribution analyses and results indicated Pleistocene population expansion in both species. Estimates of population expansion time suggested earlier population expansion in L. japonicus than in L. maculatus. Molecular variance analyses showed differential genetic structuring for these two closely related species. The results indicated that L. japonicus is panmictic throughout its range. In contrast, populations of L. maculatus showed statistically significant levels of genetic structuring. Pattern of isolation by distance was observed in L. maculatus, suggesting that L. maculatus is in genetic equilibrium. In contrast, L. japonicus did not exhibit isolation by distance.     

Biochemical characterization of trypsins from the hepatopancreas of Japanese sea bass (Lateolabrax japonicus)

A cationic trypsin (trypsin A) and an anionic trypsin (trypsin B) were highly purified from the hepatopancreas of the Japanese sea bass (Lateolabrax japonicus) by ammonium sulfate precipitation, column chromatographies of DEAE-Sepharose and Sephacryl S-200 HR. Purified trypsins revealed single band on SDS-PAGE and their molecular masses were 21 kDa and 21.5 kDa, respectively. Trypsins A and B exhibited maximal activity at 40°C, and shared the same optimal pH at 9.0 using Boc-Phe-Ser-Arg-MCA as the substrate. The two trypsins were stable up to 45°C and in the pH range from 7.0 to 11.0. Trypsin inhibitors such as Pefabloc SC, PMSF and benzamidine are effective to these two enzymes and their susceptibilities were similar. Apparent K(m)s of trypsins A and B were 1.12 and 0.7 μM and k(cat)s of them were 72.08 and 67.79 S(-1) for Boc-Phe-Ser-Arg-MCA, respectively. The N-terminal amino acid sequences of the two trypsins were determined to the 24th residues, which were highly identical to trypsins from other species of fish while trypsins A and B only shared 45.8% identity. The digestive effect of the two trypsins on native shrimp muscular proteins indicated their effectiveness in the degradation of food proteins.     

饥饿对于鲈肌肉、肝脏和血清主要生化组成的影响

于 2 2 93± 2 15℃条件下 ,在室外水泥池 (3m× 2m× 1m)中对正常鲈 (2 85 2 6± 6 5 4 g)和患脂肪肝病鲈 (4 6 4 71± 5 4 2 2g)进行为期 9周的饥饿处理。分别在实验开始后第 0周、 3周、 5周、 7周和 9周取样 ,以观察饥饿对于鲈内脏相对重量、肌肉肝脏和血清主要生化指标的影响。研究表明 ,鲈对饥饿耐受能力较强 ,在饥饿时首先快速动用肠系膜脂肪和肌肉脂肪作为能量供应 ,而在整个饥饿阶段则主要以肌肉蛋白质作为能量来源 ,肝脏中能源物质在饥饿中并无明显减少 ,故不是鲈饥饿时的主要供能物质。饥饿时 ,肌肉和肝脏中的水分和脂肪含量呈现负相关 ,尤其在肝脏中表现明显。鲈血清中脂肪酶、甘油三酯、胆固醇、低密度脂蛋白和高密度脂蛋白在饥饿中表现出周期性和阶段性的变化 ,其中正常鲈表现出有规律的波浪状图形 ,而脂肪肝病鲈则表现出山峰状图形 ,说明脂肪肝病鲈代谢机制不如正常鲈灵敏 ,9周的饥饿并不能减轻或消除鲈的脂肪肝病.     

Lunar synchronization of spawning in sea bass, Lates calcarifer (Bloch): effect of luteinizing hormone-releasing hormone analogue (LHRHa) treatment

Based on egg collection records, spontaneous spawning activity of sea bass, Lates calcarifer (Bloch), reared in floating net cages followed a semilunar cycle. The peak of multiple spawnings coincided with declining spring tides of quarter moon periods. Maximum diameter of intra-ovarian, ripe oocytes (0.51–0.55 mm) occurred in synchrony with the quarter moon periods. Smaller oocytes (0.44–0.47 mm) were sampled during the new and full moon periods. Two structural analogues of luteinizing hormone-releasing hormone (LHRHa) (des-Gly¹⁰, d -Ala⁶-LHRH ethylamide and d -Ala⁶, Pro⁹-Nethylamide-LHRH), in pelleted or dissolved form, induced mature female sea bass with a mean egg diameter of at least 0.40 mm to spawn at any day during the lunar cycle. The onset of spontaneous and LHRHa-induced spawnings occurred during low tides in the evening until dawn (from 19.00 to 05.00 hours). These results demonstrate that LHRHa can effectively induce mature sea bass to spawn independent of the highly predictable semilunar spawning rhythm. In addition, the occurrence of both spontaneous and hormone-induced spawnings at a precise period of the day suggest a tidal and diurnal cue entraining spawning of mature female sea bass.     

Microsatellite DNA polymorphism of Japanese sea bass (Laterolabrax japonicus) inhabiting Chinese and Japanese coasts

Twenty‐two microsatellite DNA markers were developed for Japanese sea bass (Laterolabrax japonicus), of which 19 were independent from each other and at Hardy–Weinberg equilibrium across the three populations of Japanese sea bass inhabiting Chinese coasts (defined as China group) and the five populations inhabiting Japanese coasts (defined as Japan group). These 19 markers were used to determine the number of alleles and the expected heterozygosity across the eight populations. The majority of individuals (93.8–98.8%) of the three populations of the pre‐defined China group were assigned to an inferred cluster, and 90.9–94.6% of the individuals of the five populations of the pre‐defined Japan group were assigned to the other. The average number of alleles across the 19 loci was significantly lower in the China group than in the Japan group (10.3 vs 15.4), however, the average expected heterozygosity across the 19 loci of the China group was similar to that of the Japan group (0.743 vs 0.750). An effective population size reduction (i.e. bottleneck effect) was detected in the China group (P = 0.00357), which may have resulted from either over‐catching or glaciations or both. The pairwise F_ST among populations of the China group (0.019–0.029) and among populations of the Japan group (0.003–0.021) were lower than those between the populations of the China group and the populations of the Japan group (0.076–0.101). The average pair‐wise F_ST between the populations of the China group and those of the Japan group reached 0.075, and the variation between the China group and the Japan group accounted for 7.16% of the total. Nei’s original measures of genetic distances among the populations of the China group and the Japan group ranged from 0.123 to 0.145 and from 0.055 to 0.123, respectively, while that between the populations of the China group and the populations of the Japan group ranged from 0.326 to 0.450. Japanese sea bass is able to disperse over a long distance; however, our observations demonstrated that it cannot migrate across a possible barrier existing between Chinese and Japanese coasts. Most individuals of the Zhoushan population of the China group were assigned to two inferred clusters, and most individuals of Ariake Sea, Tokyo Bay and Ishikawa populations of the Japan group were assigned to three inferred clusters, indicating that these locations were the gathering grounds of Japanese sea bass.     

Morphological and genetic characteristics of sea bass,Lateolabrax japonicus, from the Ariake Sea, Japan

Sea bass,Lateolabrax japonicus, from the Ariake Sea, characterized by black dots on the lateral body region as in the Chinese sea bass,L. sp., were examined and compared morphologically and genetically withL. japonicus andL. sp. Some meristic characters of the Ariake form tended to fall midway between values for the two former species. Genetic features, evaluated by isozyme analyses, indicated that the Ariake form as represented a simple Mendelian population, there being no significant differences from a Hardy-Weinberg equilibrium according to chi-square tests. Although some extreme differences in allelic frequencies were found at some loci betweenL. japonicus andL. sp., the Ariake form possessed many heterozygotes at thePROT-1 ^* locus, in addition to allelic frequencies at some loci conforming to those ofL. sp. Average allele numbers per locus, rate of polymorphic loci and average heterozygosity of the Ariake form were higher than for eitherL. japonicus orL. sp., indicating high genetic variation in the former. The results suggested that the Ariake population is genetically independent of other populations ofL. japonicus, but might be genetically influenced byL. sp.     

Cloning of Wap65 in sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata) and expression in sea bass tissues

The complementary DNA encoding WAP65 protein was cloned from the liver of two fish species sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata). Full-length cDNA sequences were obtained from reverse transcribed total RNA, followed by 5′ and 3′ rapid amplification of cDNA end (RACE) experiments. The full-length cDNA sequence of D. labrax is 1709 bp and the coding sequence is flanked by a 67 bp 5′-UTR and a 358 bp 3′-UTR. The full-length cDNA sequence of S. aurata is 1599 bp, and the coding sequence is flanked by a 48 bp 5′-UTR and a 273 bp 3′-UTR. The deduced amino acid putative primary sequences are composed of 427 and 425 amino acid residues for D. labrax and S. aurata, respectively. They display high homologies with previously described fish WAP65 and other hemopexin-like proteins from rabbit (Oryctolagus cuniculus). Expression of Wap65 has proved to be a natural physiological adaptive answer of teleost fish to warm temperature acclimation. In all fish species studied to date, Wap65 was found expressed mainly by the liver, although other tissues seem able to express Wap65 in response to a warm temperature acclimation, in a specie specific manner. Here, we investigate the tissue specific expression of Wap65 in D. labrax and S. aurata in response to a warm temperature acclimation, by RT-PCR analysis.     

Litterfall in two Canadian deciduous woods: quality, quantity and timing

Litterfall in two deciduous woods in Ontario, Canada, was caught in bag traps and by new screen traps for two years. The beech-maple wood produced 818 g m⁻² (8.2 t ha⁻¹) and the poplar wood produced only 450 g m⁻² (4.5 t ha⁻¹). Of these totals, canopy leaves alone made up 386 g m⁻² for the beech-maple wood and 270 g m⁻² for the poplar wood leaving 47% and 34%, espectively, for other litter components. Data are presented for seven types of litter in addition to canopy leaves.
Redistribution of litter on the ground causes spatial heterogeneity of substrate and of habitats for decomposers and hence of decomposition.
Temporal distributions of various types of litterfall differ between the two woods in relation to their vegetation structure. The diversity of litter types combines with extensive temporal distribution of litterfall in the beech-maple woodland to give that wood the potential for nearly continuous decomposition activity. In contrast, the poplar woodland has low diversity of litter and short duration of litterfall.     

Feeding ecology of larval and juvenile black seabream (Acanthopagrus schlegeli) and Japanese seaperch (Lateolabrax japonicus) in Tolo Harbour, Hong Kong

Black seabream, Acanthopagrus schlegeli, and Japanese seaperch, Lateolabrax japonicus, are important commercial species in the coastal waters of western Pacific Ocean, including Japan, Korea and China. In Hong Kong, larvae and juveniles of these two species occur in bays and estuaries during late winter and spring. This study reports on the ontogenetic changes in food habits in larvae and juveniles of these species in an artificial rocky shore area. Copepods and cladocerans were the most numerous food items for black seabream. There was a shift to larger and benthic prey as the fishes grew. Japanese seaperch <2.1cm fed predominantly on copepods and cladocerans, while larger prey were added as fish size increased. Japanese seaperch >6.0cm were piscivorous. Maximum prey width increased with fish standard length and mouth gape width in both species. Overall, black seabream showed greater diet breadth than did Japanese seaperch. In black seabream, diet breadth increased with fish size. In Japanese seaperch, diet breadth increased with size for fishes <4.0cm, then decreased as the fishes became piscivorous. Prey selectivity in black seabream was determined using information on prey availability in plankton samples. In general, preference was stronger for cypris larvae, Penilia avirostris and decapod larvae than for copepods and podonids. In recent years, overfishing and environmental degradation have led to the decline of fish populations in Tolo Harbour. Absence of fishes with empty gut indicates that inner Tolo Harbour is still an important nursery area for these two commercial species.     

Genetic differences between wild and hatchery populations of Korean spotted sea bass (Lateolabrax maculatus) inferred from microsatellite markers

The spotted sea bass, Lateolabrax maculatus, is popular in recreational fishing and aquaculture in Korea. Its natural population has declined during the past two decades; thus, beginning in the early 2000s stock-enhancement programs were introduced throughout western and southern coastal areas. In this study, genetic similarities and differences between wild and hatchery populations were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 96 alleles were identified. Although many unique alleles were lost in the hatchery samples, no significant reductions were found in heterozygosity or allelic diversity in the hatchery compared to the wild population. High genetic diversity (He = 0.724–0.761 and Ho = 0.723–0.743), low inbreeding coefficient (F _IS = 0.003–0.024) and Hardy–Weinberg equilibrium were observed in both wild and hatchery populations. However, the genetic heterogeneity between the populations was significant. Therefore, genetic drift likely promoted inter-population differentiation, and rapid loss of genetic diversity remains possible. Regarding conservation, genetic variation should be monitored and inbreeding controlled in a commercial breeding program.     

Multiple habitat use of Japanese sea bass Lateolabrax japonicus in the estuary region of the Tone River system,implied by stable isotope analysis

Ichthyological Research - Japanese sea bass Lateolabrax japonicus is a euryhaline fish species. The present study aimed to elucidate their habitat use in the estuary region of the Tone River system...     

The immunohistochemical study of nodavirus changes in larval, juvenile and adult sea bass tissue

Owing to its quickness and sensitivity, and the minimal financial effort required, immunohistochemistry is a leading technique for nodavirus diagnosis. The aim of the study was to give detailed results of immunohistochemical changes in the various tissues of larvae, juvenile and adult sea bass (Dicentrarchus labrax L.). Histological sections were treated with polyclonal anti‐noda rabbit IgG, while diaminobenzidine‐peroxidase was used for the enzyme–substrate complex. The development from first changes to vacuolating necrosis was recorded in the brain, spinal cord and retina. For the first time, immunolabelling was noted in the liver endothelium. The other sampled tissue was immunonegative.     

The efficacy of eugenol and tricaine methanesulphonate as anaesthetics for juvenile Chinese sea bass (Lateolabrax maculatus) during simulated transport

The purpose of this study was to evaluate the anaesthesia effects of eugenol and MS‐222 sedatives applied on juvenile Lateolabrax maculatus during simulated transport. In experiment 1, the juveniles were divided into two groups, with seven concentrations tested on each group (eugenol [4, 6, 8, 10, 12, 14 and 16 mg/L] and MS‐222 [20, 30, 40, 50, 60, 70 and 80 mg/L]). Induction and recovery times were recorded. The time for anaesthesia was shortened, and the time for complete recovery was prolonged as the anaesthetic concentration increased. The optimal transport concentration for each anaesthetic tested was 6 mg/L of eugenol and 30 mg/L for MS‐222. In experiment 2, the 5‐hr simulated transport test showed that the survival rate of L. maculatus juveniles with anaesthesia was 100%, and without anaesthesia, survival was 60%. After 24 hr of recovery following transport, the fish showed 100% survival for the group with added anaesthetic and 40% for the group without added anaesthetic. Compared to the non‐anaesthetized groups, the anaesthetized transport groups showed significant increases in the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) (p < 0.05). The levels of AST, ALT and alkaline phosphatase (AKP) were significantly higher in the MS‐222 transport group than in the eugenol transport group (p < 0.05). The levels of AKP were significantly higher in the non‐anaesthetized transport group than in the anaesthetized group (p < 0.05). According to the present experiment results, eugenol was an efficient anaesthetic in L. maculatus, and we recommend eugenol instead of MS‐222 as an anaesthetic for the short‐time transport of L. maculatus.     

Age-linked changes in salinity tolerance of larval spotted seatrout (Cynoscion nebulosus, Cuvier)

Acute salinity tolerance limits for the estuarine spawning spotted seatrout, Cynoscion nebulosus (Cuvier). were evaluated by examining 18 h survival of larvae in an extensive range of salinity treatments (0 to 56 ppt). Larvae from eggs spawned in two different salinities (24 and 32 ppt) as well as larvae acclimated in hypersaline and brackish waters were compared. Both upper and lower salinity tolerance limits showed an age-linked pattern, decreasing to a minimum tolerance range (6.4 to 42.5 ppt) at age 3 days after hatching (at 28 o C) and increasing to the widest range tolerated (1.9 to 49.8 ppt) on the last day tested (age 9 days). Acclimation to hyposaline conditions was demonstrated by larvae spawned at 32 ppt although significant hypersaline acclimation could not be demonstrated. Altered upper limits to the range tolerated by larvae from different spawning salinities indicated parental and/or early acclimation effects are important. Consistently greater vulnerability to both hyper- and hyposaline conditions at age 3 days after hatching was observed in all tests conducted. Exposures related to the onset of feeding at this time are likely explanations for this reduced tolerance.     

Population genetic structure of the sea bass (Lateolabrax japonicus) in Korea based on multiplex PCR assays with 12 polymorphic microsatellite markers

Population genetics has been recognized as a key component of policy development for fisheries and conservation management. In this study, natural sea bass (Lateolabrax japonicus) populations in three ocean basins in Korea were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 203 alleles and similarly high levels of genetic diversity [mean number of alleles (N_A) = 14.43, mean expected heterozygosity (He) = 0.84] were detected. All populations showed significant heterozygote deficiency at four loci, which could be explained by the presence of null alleles. The genetic population subdivision was low and was significantly different according to F-statistics (overall F _ST = 0.003, R _ST = 0.005). However, this substructure was not supported by an analysis of molecular variance test, analyses of isolation by distance or Bayesian analysis. The passive dispersal of eggs/larvae via the main currents appears to facilitate gene flow. The possibility of a recent genetic bottleneck was observed in all three populations of L. japonicus, indicating that overfishing and degradation of the environment in recent years has led to a decline in the sea bass populations in Korea. Our study demonstrates that sea bass in Korea do not appear to be genetically partitioned and should be managed as a single unit; however, the potential for a rapid loss of genetic diversity remains. Information regarding the genetic characteristics of Korean sea bass populations has important implications for fishery management and conservation efforts and will aid in the sustainable exploitation of fishing resources and the preservation of biodiversity.     

Temperature effects on cranial deformities in European sea bass, Dicentrarchus labrax (L.)

The present study investigates the effect of water temperature on the development of deformities during embryonic and larval stages of European sea bass, Dicentrarchus labrax (L.). Two temperature conditions were examined in duplicate by studying the presence of skeletal deformities in a sample of 45 to 51 fish taken from each population at the end of the rearing trials [20–23 mm total length (TL)]. The results indicated that water temperature during the embryonic and larval phase has significant effects on deformation of the branchiostegal rays (P < 0.001), but not of the mouth and the fins (P > 0.05). At 15°C, 27.2–33.4% of the examined fish had branchiostegal rays of abnormal shape and/or orientation, whereas at 20°C this deformity had a frequency of only 4.0–4.1%. The frequency distribution graph of branchiostegal counts demonstrated a significant deviation in the deformed fish from the normal (seven rays on each side of the body) phenotype at both temperatures tested. This deviation was mainly expressed as a lack of one to four rays (56.7% of deformed fish), or the formation of one extra ray (26.7% of deformed fish) (P < 0.001, G‐test). The results are discussed in respect to the possible mechanisms of temperature effects on the development of skeletal deformities.