Haemoglobin and oxygen transport of the West Australian dhufish, Glaucosoma hebraicum Richardson, and other species

To better understand the possible role of deoxygenation of haemoglobin in the pathogenesis of exophthalmos in the West Australian dhufish, Glaucosoma hebraicum Richardson, some oxygen transport properties of haemoglobin from this species were determined and compared with haemoglobin from black bream, Acanthopagrus butcheri (Munro), snapper, Pagrus auratus (Bloch & Schneider) and King George whiting, Sillaginodes punctata (Cuvier). Dhufish had a single haemoglobin with a pronounced Root effect. Snapper and black bream blood had six and five types of haemoglobin, respectively. The magnitude of the Root effect of haemoglobins in the haemolysate was demonstrated by comparing oxygenation and total deoxygenation at pH 8 with relative deoxygenation at pH 6.9. Dhufish haemoglobin demonstrated a large Root effect, with the Root effect of King George whiting, snapper and black bream being of lesser magnitude. Deoxygenation of all haemolysates was more pronounced in the presence of adenosine triphosphate (ATP). Seasonal changes in haematological parameters and ATP content of dhufish blood were not evident. The Root effect is discussed in the context of its possible role in oxygen supply to the retina and adaptation to the fish's habitat and the environment.     

Pathogenesis and epidemiology of spontaneous exophthalmos in the West Australian dhufish, Glaucosoma hebraicum Richardson

The West Australian dhufish, Glaucosoma hebraicum Richardson (family Glaucosomatidae) is a potentially valuable aquaculture species, but spontaneous exophthalmos is common in freshly caught and cultured dhufish. To investigate the epidemiology and pathogenesis of exophthalmos in tanks of wild-caught and captive-bred dhufish, records of culture conditions, the prevalence of lesions and pathological examinations were used to study the disease and associated environmental and management factors. Naturally occurring cases of exophthalmos that were examined were mostly unilateral and occurred more frequently in summer months during periods of increasing water temperature. In 15 affected eyes that were examined histologically gas bubbles and haemorrhages were consistently present in the choroid. In some, gas bubbles and haemorrhage were also present in retrobulbar tissues associated with perforation of the sclera. Oxygen concentrations were measured in 12 exophthalmic eyes and concentrations of 50–73% were recorded in gas bubbles in the anterior chamber of three of these that were acute cases. Very low oxygen tensions were recorded at the retinal–vitreal junction of four eyes with retrobulbar haemorrhages indicating that there may have been disruption of arterial blood supply to the choroid in chronic lesions or with perforation of the sclera. The results of experiments to determine the significance of exercise, high water temperature, sudden increases in light intensity, fish handling or tank cleaning indicate that exercise and/or high water temperature (25.5 °C) may be important risk factors for the development of exophthalmos in dhufish. Possible mechanisms for the formation of gas bubbles and haemorrhage in the choroid are discussed.     

Production of bactericidal substances by a marine vibrio isolated from cultures of the scallop Argopecten purpuratus

A marine vibrio (strain C33) having inhibitory effects on the growth of the pathogen Vibrio anguillarum-VAR was isolated from seawater used in mass culture of the north-Chilean scallop Argopecten purpuratus. This bacterial isolate demonstrated broad inhibitory activity on several bacterial strains, including some pathogenic vibrios. Ethyl acetate extracts of extracellular products of strain C33 were separated by thin layer chromatography (TLC), and three fractions thus obtained were found to have antimicrobial activity when tested using microplate bioassays. One of the fractions (A2) having marked antimicrobial activity, was further purified using TLC and analyzed by IR spectrophotometry and NMR'H and was characterized on a preliminary basis as an aliphatic hydroxyl ether. This compound demonstrated bacteriostatic activity against the important marine pathogens Vibrio parahaemolyticus and V. splendidus, and as discussed in the paper, may be useful in developing natural strategies for the control of pathogens in mass cultures of Argopecten purpuratus and possibly other molluscs affected by these bacteria.     

The effect of dietary protein source on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 12 weeks on juvenile Australian freshwater crayfish (Cherax destructor) (mean weight (SE) 0.82 (0.02)g) maintained on five isoenergetic diets with a protein content of 30%. Diets differed in the primary source of protein used, with meat, snail, soybean, yabby, and zooplankton meals comprising the major protein ingredient, varying from 56–60% of total protein. Mean percentage weight gain per day ranged from 7.57% (yabby meal diet), to 9.42% (snail meal diet). No significant difference occurred in mean weight, percentage weight gain, specific growth rate (%), or survival among diets. A maximum size of 16.44g was attained on the yabby meal diet. Largest mean weight was 8.27g on the snail-based diet. Food conversion ratios were all good, with a minimum value of 0.95 recorded for the snail-based diet. Initial food consumption per day was approximately 5% of mean animal weight, decreasing to around 2.4%, and is collectively described by a power curve. Protein retention ranged from 29.57% in the zooplankton meal diet to 41.15% in the snail-based diet. Carcass composition was influenced by feed type, with the most marked difference occurring in carapace colour. Animals fed the zooplankton-based pellets developed the strongest pigmentation. Recommendations are made for including certain protein-based ingredients in manufactured yabby diets.     

南海大眼金枪鱼和黄鳍金枪鱼的群体遗传结构

测定了南海西沙和南沙群岛附近海区(11～12°N,15°N;110～112°E)黄鳍金枪鱼61尾(17尾成鱼、44尾幼鱼)和大眼金枪鱼26尾(22尾成鱼、4尾幼鱼)的线粒体基因组控制区部分序列(D-loop),结合GenBank数据库中印度洋、太平洋和大西洋群体的同源数据,分析结果:(1)黄鳍金枪鱼与大眼金枪鱼均具极高的单倍型多样性(Hd>99%),聚类树及群体间分化指数(FST和Snn)表明大眼金枪鱼群体分化程度明显高于黄鳍金枪鱼群体;(2)大眼金枪鱼和黄鳍金枪鱼的南海群体与印度洋群体之间基因流最强(Nm=51.638和261.280 10),其次为太平洋群体(Nm=10.868 8和-50.801 81);(3)黄鳍金枪鱼和大眼金枪鱼都基本服从群口扩张模型,而mismatch分布分别呈单、双峰,其中大眼金枪鱼的南海群体扩张较晚(Tau=7.902)且最为明显(θ1/θ0=99 999/14.752)。     

Comparative digestive enzyme ontogeny in two marine larval fishes: Pacific threadfin (Polydactylus sexfilis) and bluefin trevally (Caranx melampygus)

The specific activity of digestive enzymes; aspartic and serine protease, collagenase, lipase, acid and alkaline amylase, acid and alkaline phosphatase, and chitinase was assayed throughout early development in two species of marine fishes: the Pacific threadfin (Polydactylus sexfilis) and bluefin trevally (Caranx melampygus). Specific enzyme activities were determined on whole larval extracts sampled at selected stages of development, from day 0 to day 30 post-hatching. Similar developmental patterns of enzyme specific activity were observed in the two species, although differences in timing, amplitude and effects of first feeding were noted. Amylase activity increased prior to first feeding, peaking at the middle of the larval period, and becoming nearly undetectable by the time of larval-to-juvenile metamorphosis. Serine protease, collagenase, lipase and alkaline (and acid for threadfin) phosphatase activities increased gradually, followed by sharp increases to a plateau during the second half of larval development. Aspartic protease and chitinase activities, in addition to acid phosphatase (for trevally), were low to undetectable in the first half of development, increasing through metamorphosis. In the trevally only, this group of enzymes exhibited high activity levels at the time of hatching, followed by a precipitous drop. Species-dependent differences in enzyme specific activity at first feeding may have been a result of differences in yolk utilization. These results, taken in the context of earlier reports, lead us to conclude that carbohydrate utilization may play a significant role in the earlier phases of development among some marine larvae, followed by a shift to protein and lipid utilization.     

Genetic variation in mitochondrial genes and intergenic spacer region in harmful algae <Emphasis Type="Italic">Chattonella</Emphasis> species

In this study, nuclear ribosomal RNA gene internal transcribed spacer regions and the cox2-cox1 fragment of the mitochondrial (mt) genome were sequenced in 24 strains of Chattonella spp. Variability in both regions showed that the mt genome sequences of Chattonella spp. have a higher evolutionary rate than the nuclear rRNA gene sequences. A maximum likelihood tree based on the mt sequence grouped the Japanese Chattonella strains into two groups (Groups A and B), although no correlation was observed amongst the phylogenetic groups, their morphologies, or the isolated areas. Groups A and B were clearly identified by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay using Fokl, without the need for a sequencing experiment. The PCR-RFLP assay revealed that Chattonella cells obtained from sea water in Oita, Japan, in 2004 and 2005 belonged to Group B. This is the first report showing the genetic variation in Chattonella spp. using a PCR-RFLP identification protocol.     

Recent advances in European sea bass and gilthead sea bream nutrition

European sea bass (Dicentrarchus labrax) and gilthead sea bream(Sparus aurata) are amongst the most important finfish speciescultured in the Mediterranean region. Production of these species isnowadays a well-controlled process, but knowledge of their nutritionalrequirements is still very limited. Nevertheless, a considerable amountof data has been accumulated in recent years, and the purpose of thispaper is to review the recent advances on the nutritional requirementsof sea bass and sea bream. The optimum protein to energy ratio of thediets of sea bass and sea bream seem to be higher than for salmonids,and there is some evidence that high dietary lipid levels have nobeneficial effects on fish performances. Although the essential aminoacid requirements were estimated by the ideal protein method, data basedon the dose-response method is only available for a few amino acids.Essential fatty acid requirements were estimated for sea bream juvenilesbut data is lacking for sea bass. Vitamin and mineral requirements ofthese species are practically unknown. Although the importance ofbroodstock nutrition on gonadal development, spawning and egg quality isrecognized, few studies were done to elucidate these aspects. The recentdevelopment of microparticulate diets for larvae will contribute to theaccurate evaluation of their nutritional requirements.     

Profitability evaluation of resource allocation for growing redtail and Chinese shrimp

A farming survey was conducted to evaluate the profitability of resource allocation for producing redtail shrimp, Penaeus penicillatus (Alock) and Chinese shrimp, P. chinensis (Osbeck). Data on production costs and total revenue according to different harvest sizes of shrimp species were collected based on 108 individual ponds operated by 25 surveyed shrimp farmers. The results indicated that growing Chinese shrimp to a marketable size of 12 g was the most profitable management strategy. The profits could be maximized by integrating small-scale farms to a 12.41 ha optimal farming area when the available resources remained the same as those for a 10 ha farm.     

Effects of replacement of animal protein by soybean meal on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 59 days with juvenile Cherax destructor, mean weight (se) 0.61 (0.01) g, reared communally and maintained on 16 isoenergetic diets containing crude protein levels of 15, 20, 25, and 30%. For each protein level the fish meal component was replaced by soybean meal to produce diets in which 0, 20, 40, or 60% of the protein originated from soybean meal. Mean percentage weight gain per day ranged from 2.98% (15% protein, 60% soybean meal diet), to 11.75% (30% protein, 40% soybean meal diet). When soybean meal was included at a level of 40–60%, growth rate was reduced relative to that achieved with control diets at 15% and 20% protein levels. In no case did a 20% substitution significantly affect growth over that achieved with controls. A two-way interaction occurred between dietary protein and the level of dietary soybean meal. Feeds of higher protein content appeared to permit higher soybean meal inclusion levels without significantly affecting growth. Increases of 5% protein produced a significant improvement in growth when soybean meal contributed from 40–60% of the total protein. This effect was less pronounced in the control diets and the 20% soybean meal series. The percentages of protein increased and lipid decreased in the carcass as the level of dietary protein increased. A similar effect occurred by increasing the soybean meal substitution level to 60%. An obvious trend in carcass moisture, energy, and ash did not occur. A protein requirement of 30% is apparent when fish meal and soybean meal are included in diets at levels of 20% and 24% respectively. A maximum weight of 14.13 g was recorded for an individual fed the 30% protein, 20% soybean meal diet.     

Growth and age at first maturity in turbot and halibut reared under different photoperiods

The effect of extended photoperiods on growth and age at first maturity was investigated in 166 (79 females and 87 males) individually tagged Atlantic halibut Hippoglossus hippoglossus and in 114 (50 females and 64 males) individually tagged turbot Scophthalmus maximus. The halibut were reared at 11 °C on four different light regimes from 10 February to 6 July 1996: simulated natural photoperiod, (LDN), continuous light (LD24:0), constant 8 h light and 16 h darkness (LD8:16) and LD8:16 switched to continuous light 4 May 1996 (LD8:16–24:0). From 6 July 1996 to 9 February 1998 the LD24:0 and LD8:16–24:0 were reared together under continuous light and the LDN and LD8:16 together under natural photoperiod. The turbot were reared at 16 °C on three different light regimes: constant light (LD24:0), 16 h light:8 h darkness (LD16:8), or simulated natural photoperiod (LDN). After 6 months on the different photoperiods, the turbot was reared together on LDN for approximately 12 months until first maturation. Juveniles subjected to continuous light (halibut) or extended photoperiods (halibut and turbot) exhibited faster growth than those experiencing a natural photoperiod or a constant short day. Moreover, when the photoperiod increased naturally with day-length or when fish were abruptly switched from being reared on short-day conditions to continuous light, a subsequent increase in growth rate was observed. This growth enhancing effect of extended photoperiods was more apparent on a short time scale in Atlantic halibut than in turbot, but both species show significant long-term effects of extended photoperiods in the form of enhanced growth. In both species lower maturation of males was seen in groups exposed to extended or continuous light compared to LDN and this could be used to reduce precocious maturation in males leading to overall increase in somatic growth. This revised version was published online in August 2006 with corrections to the Cover Date.     

厚壳贻贝MyD88-4基因的生物学特性及其对沙氏弧菌的免疫应答

为理解髓样分化因子88(myeloid differentiation factor 88, MyD88)基因的生物学特性以及对细菌胁迫的响应,本研究克隆了厚壳贻贝MyD88基因(命名为McMyD88-4)cDNA全长序列,其全长3 930 bp,开放阅读框2 607 bp,编码868个氨基酸。其中,13～109位的氨基酸序列为死亡结构域(death domain,DD),347～481位的氨基酸序列为TIR(toll/interleukin-1 receptor)结构域,TIR结构域包含3个高度保守的区域Box 1、Box 2和Box3;McMyD88-4蛋白的空间结构包含6个α螺旋(α-helix)和4个β折叠(β-sheet)。同源性分析显示,McMyD88-4蛋白序列与长牡蛎MyD88最相似,其一致性和相似性分别为60%和77%;其次,与虾夷扇贝、海湾扇贝和菲律宾蛤仔相似度较高,其一致性和相似性分别为40%～51%和58%～67%。系统进化树结果显示,McMyD88-4先与长牡蛎和扇贝聚为一支,然后与黑腹果蝇聚为一支,脊椎动物单独聚为一支。实时荧光定量PCR(qRT-PCR)检测发现,McMyD88-4基因在厚壳贻贝各组织和器官中均有表达,其中在外套膜和鳃中的表达量最高,而血细胞中表达量最低。厚壳贻贝经沙氏弧菌感染后,McMyD88-4基因表达量在免疫相关组织中急剧上升,分别在感染后3和6 h达到峰值,且在消化腺中的上调水平显著高于鳃和外套膜。研究表明,McMyD88-4在厚壳贻贝抵御外界病原体侵染过程中,尤其是弧菌感染方面发挥重要作用。     

Early morphological predictors of maturity in one-sea-winter Atlantic salmon

Early separation of maturing from immature fish would be very valuable in Atlantic salmon (Salmo salar) culture. Possible morphometric predictors of maturation in one sea-winter Atlantic salmon were investigated from measurements of body weight, length and depth and adipose fin length undertaken at 14 day intervals from May until August in a population of 80 individually tagged fish, 61 (male and female) of which were subsequently found to be maturing (i.e. grilse). Stepwise discriminant analysis revealed that a combination of forklength and condition factor proved to be the strongest and most consistent predictor of maturation (over 88% accurate) until August. In August when maturing fish could be easily distinguished by shape and colour, forklength and condition factor no longer had any discriminatory power, but body depth at the anal and dorsal fins were good predictors. A second trial was performed on a separate stock of fish for which the same predictors proved robust. The discriminant functions derived from one stock of fish did not, however, produce the same discriminatory power when applied to the other stock. Hence there does not appear to be a simple mathematical means for the general discrimination of maturing and immature salmon. However the morphological predictors described above could be used for visual discrimination when ‘grilse grading’.     

Increased winter temperature did not affect completion of smolting in Atlantic salmon

Four groups of 1+ year-old Atlantic salmon, Salmo salar, pre-smolts were reared under various temperature regimes: constant 10 °C from November onwards; ambient temperature until either 15 December or 1 February, then 10 °C; or ambient temperature throughout (control; 2–3 °C November–March). From 20 May onwards, temperature in all groups was ambient, increasing from 10 °C to 17 °C in late July. Rearing temperature had no significant effect on either the timing of completion of smolting, or the duration of the smolt-window. Mean gill Na⁺-K⁺ ATPase activity was not significantly affected by temperature regime; it increased in all groups from < 2.0 mol Pi mg protein^–1 h^–1 (units) in January to 5–7 units in mid-April, then back to < 2.0 units in July. Survival in 96 h, 37) salinity (S) tolerance tests was similar in all groups, increasing from < 10% in early March, to > 90% from mid-April to mid-June, then decreasing to < 20% by early July. Increased winter temperature significantly increased growth and condition factor compared with the control, but during April–May all four groups exhibited similar temporary decreases in condition factor in association with the completion of smolting.     

Tilapia wild spawning control through predator fishes: Israelitrial with red-drum and hybrid bass

Tilapia wild spawning is a nuisance in warm freshwater aquaculture growout ponds. To cope with this problem two experiments were carried out with predatory fish that do not reproduce in fresh water. One experiment tested the capacity of hybrid bass (Morone saxatilis × M. chrysops) and red-drum (Sciaenops ocellatus) as predators of wild spawning of hybrid tilapia (Oreochromis niloticus × O.aureus), and the other compared predation effectiveness of red-drum of different sizes and stocking densities.Both hybrid bass and red-drum effectively reduced tilapia wild spawning and improved by 15–20% tilapia performance and food conversion ratio. These effects were obtained stocking small red-drum (20 g) or large red-drum (60–80 g) or bass (135 g) at stocking densities of 500–1000 predators/ha, together with 15000 tilapia/ha of 65–75 g. Hybrid bass stocked at 750/ha and large red-drum stocked at 500/ha presented over 90% survival. Red-drum at higher stocking density and/or lower stocking weight presented reduced survival (40–60%). Red-drum of all examined stocking weights presented better growth rates when stocked at 500/ha than at higher densities.     

Particle clearance and selection in three species of juvenile scallops

Juvenile scallops (<2 mm shell height) of three species (Placopecten magellanicus, Patinopecten yessoensis, Argopecten irradians) were fed mixed, unialgal cultures. Scallops were fed a total of six algal clones simultaneously and clearance rates were monitored using flow cytometric techniques. In another experiment, scallops were presented with natural assemblages of particulate matter as a food source. Data are presented on differences in clearance rates for the individual algal species as well as size-related differences of algal clones, and uptake of chlorophyll vs. non-chlorophyll cells, both within and between scallop species. Significant differences in clearance rates of individual algal species have been found within and between scallop species. Particle selection does not appear to be based upon size alone and is apparently based on other characteristics of the algae as well. The results demonstrate pre-ingestive sorting.     

Size heterogeneity can reduce aggression and promote growth in Atlantic salmon parr

Aggression in groups of 0+ Atlantic salmon (Salmo salar) was monitoredat weekly intervals in two tanks containing 100 fish each. Three 1 + salmonparr were added to one of these. After 5 weeks, fish weights were measuredin both tanks and the conditions reversed. At ten weeks, weights of fish inboth tanks were measured again. In both populations, levels of aggressionamong the smaller fish were significantly lower and growth ratessignificantly higher when the large fish were present. Although the largefish attacked the small ones, the rate at which they did so was an order ofmagnitude lower that the rate at which small fish attacked each other in theabsence of larger conspecifics. This raises the possibility that levels ofaggression among farmed salmon might be reduced by the addition of a fewlarge conspecifics.     

Vitellogenin in tilapia (Oreochromis mossambicus): Induction of two forms by estradiol,quantification in plasma and characterization in oocyte extract

Two forms of vitellogenin were isolated by DEAE agarose ion-exchange chromatography from plasma of the tilapia, Oreochromis mossambicus. The monomers have apparent molecular masses of 200 and 130 kDa, as indicated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and a total amount of phosphorus of 1.7 and 0.1%, respectively. Antibodies specific to the two forms, designated tVTG-200 and tVTG-130, were generated in rabbits and used to develop enzyme-linked immunosorbent assays (ELISAs) and in Western blot analyses of plasma and oocyte extract. SDS-PAGE of the oocyte extract showed a major protein band at 106.6, minor bands at 26.6, 24.2, and 23.7 kDa, and very faint bands at 83.4 and 17.5 kDa. Western blots of the oocyte extract revealed that the antiserum to tVTG-200 recognized strongly the protein bands at 24.2 and 23.7 kDa, and less strongly the bands at 25.1 and 22.6 kDa, whereas the antiserum to tVTG-130 recognized mainly the protein band at 106.6 kDa. The presence of both VTGs in untreated male tilapia was detected with the ELISAs using relatively high plasma volumes. Their presence in males was confirmed by VTG-like immunoreactive materials eluting from the ion-exchange column at the same positions as tVTG-200 and tVTG-130. The concentrations of the VTGs in males were several orders of magnitude lower than in vitellogenic females. Treatment of male tilapia with estradiol-17β (E₂) induced both VTGs within 24h. After 7 days, tVTG-130 reached a maximum concentration in plasma, whereas tVTG-200 continued to increase. Our findings demonstrate that the two vitellogenins are biochemically distinct, possibly differentially regulated, and made by both sexes.