Induction of moulting in hatchery‐reared mangrove crab Scylla serrata juveniles through temperature manipulation or autotomy

The effects of temperature and autotomy of chelipeds on survival, growth and moulting of mangrove crab (Scylla serrata) juveniles were investigated under laboratory conditions for 60 days. Hatchery‐produced crabs with 2.0–2.3 cm internal carapace width (1.7–2.2 g body weight) at intermoult stage were exposed to one of four temperature treatments (constant 29, 32 or 35°C, or ambient [24–31°C]) or subjected to cheliped autotomy. All crabs held at 35°C had 100% mortality due to incomplete moulting during first moult. The mean survival of crabs at termination was 58%, 64% and 50% for ambient temperature, 29 and 32°C respectively. Specific growth rate (SGR) of crabs in the ambient and 29°C were comparable but significantly lower than those at 32°C. The moult interval of the crabs was significantly shorter in treatments with constant water temperature of 29 and 32°C compared with ambient temperature. The survival of crabs with intact chelipeds was comparable with those with one or two autotomized chelipeds. Crabs with intact or one autotomized chelipeds had significantly higher SGR than crabs with both chelipeds autotomized in the first moult. On the second moult, however, high SGR was observed in crabs with two chelipeds autotomized. The moult interval was significantly shorter in the autotomized crabs compared with crabs with intact chelipeds. The results suggest that the optimum water temperature for rearing S. serrata juveniles ranges from 29 to 32°C. Likewise, autotomy of chelipeds can promote moulting without adversely affecting survival of crabs.     

Autolysis and Characterization of Sarcoplasmic and Myofibril Associated Proteinases of Oxeye Scad (Selar boops) Muscle

ABSTRACT

The autolytic profile of oxeye scad mince was characterized. Mince showed higher proteolytic activity than washed mince. The highest autolysis was observed at 65 and 60°C for mince and washed mince, respectively. Both mince and washed mince showed the optimum pH for autolysis at pH 9.0, and their activities decreased with increasing NaCl concentration (0–3.5%). Autolysis of washed mince was markedly inhibited by soybean trypsin inhibitor (SBTI), suggesting that myofibril-associated proteinase was serine proteinase. Sarcoplasmic proteinase was characterized to be heat-activated alkaline proteinase having the optimal pH and temperature of 9.0 and 60°C, respectively. The activities were stable at pH range of 8.0–11.0 at 20–40°C. The crude proteinase was inhibited by N-p-tosyl-L-lysine chloromethyl ketone, SBTI, and phenylmethanesulfonyl fluoride, suggesting the predominance of serine proteinases, especially trypsin. NaCl suppressed the activity while β-mercaptoethanol, dithiothreitol, and CaCl₂ activated the activity. Therefore, trypsin-like proteinase is a major endogenous proteinase responsible for autolysis in oxeye scad muscle. The present results can be used as scientific guidelines to predict the gel strength of surimi made from oxeye scad muscle.     

Characterization and growth conditions of Vibrio parahaemolyticus strains with different virulence degrees that cause acute hepatopancreatic necrosis disease in Litopenaeus vannamei

The phenotypic characteristics and growth kinetics at several temperatures, salinities, and pH values of three Vibrio parahaemolyticus (Vp) strains with different virulence and one nonpathogenic strain were evaluated. Independent of the virulence of the strain, a high metabolic diversity was found, which yielded different colored phenotypes on the CHROMagar? Vibrio. All strains were resistant to ampicillin and carbenicillin, and Vp AHPND+ organisms were the most sensitive to enrofloxacin. The exponential growth of Vp strains started at 1–2 hr of incubation, although no relationship was observed between the bacterial density and degree of virulence. Moreover, the growth of the most virulent strain was independent of the nutrients in the incubation media during the initial hour postinoculation. No strain grew at 4°C in 0% NaCl and pH 4, but only Vp AHPND+ grew at 44°C. For all strains, the lag phase was proportional to the NaCl concentration, and the growth was better at pH 8–9. However, the Vp AHPND? strain displayed a greater variability, was more sensitive to extreme conditions, and showed a lag phase of 9 hr independent of the pH.     

Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch)

Two new cell culture systems namely epitheloid cells of Lates (LCE) and fibroblastic cells of Lates (LCF) have been developed from fry and fingerling of the economically important brackishwater fish Lates calcarifer. Primary cultures were initiated by explant technique using caudal fin of fingerling and whole body tissue of the fry. The nutritional requirements and the growth pattern in response to different culture environment were similar for the two cell cultures. The culture medium used was Leibovitz‐15 supplemented with 20% fetal bovine serum (FBS) and 1% fish serum. The LCE comprised of epithelioid cells and LCF cells were fibroblastic. With a split ratio of 1:2, the confluency of cells was attained in 8–10 days at an incubation temperature of 28°C. The cells were found to grow well in a wide range of temperature (24–32°C) and stable at 20 and 36°C. The growth rate of LCF and LCE cells increased proportionately with the concentration of FBS from 5% to 20%. A decrease of serum level to 10% after eight subcultures produced no apparent change in cell morphology and growth rate. The viability of cells was found to be 70% when revived after a month of storage in liquid nitrogen (?196°C).     

Effect of temperature on the development, growth, survival and settlement of green mussel Perna viridis (Linnaeus, 1758)

The effect of temperature on the development, growth, survival and settlement of Perna viridis was studied under controlled conditions to provide information needed for the development of commercial hatchery technology for green mussel P. viridis. Total mortality of the larvae occurred after 24 h at temperatures of 33°C and 35°C. At 24°C, larvae took longer to settle than at temperatures of 27°C, 29°C and 31°C. For optimum larval development (8–13 h), growth (17.2±0.84 μm day^–1) and survival (55.2±0.84%), a hatchery rearing temperature of 31°C is required. For settlement no significant difference was seen between the percentage settlement at 29°C (49.3±3.34%) and 31°C (45.8±1.76%). However, the process of settlement began and ended earlier at 29°C (from 15 to 18 days) than at 31°C (from 18 to 20 days). Thus for larval settlement a temperature of 29°C is recommended.     

Temperature preference and oxygen consumption of the largemouth bass Micropterus salmoides (Lacépède) acclimated to different temperatures

The behavioural and metabolic responses of the largemouth bass Micropterus salmoides (Lacépède) to temperature were determined to define optimal thermal conditions. The final preferendum of largemouth bass juveniles determined with acute and gravitation methods was independent of the method (29.0–28.1 °C). The displacement velocity in the horizontal thermal gradient of bass juveniles was 22.4 cm h⁻¹ in the light phase and 22.6 cm h⁻¹ in the dark phase. Oxygen consumption rates in the largemouth bass increased significantly (P<0.05) from 48.8 to 69.4 mg O₂ Kg⁻¹ h⁻¹ with an increase in the acclimation temperature from 20 to 33 °C. The temperature quotient (Q₁₀) in the juveniles was 1.37–2.00 in the range of acclimation temperatures of 26–29 and 29–32 °C. The optimum temperature range for growth calculated using Jobling's equation was 28.1–28.6 °C and for Q₁₀ values 26–29 °C. The results are discussed in relation to the use of this information in aquaculture.     

Effects of chronic upper temperature regimes on growth of two geographical strains of channel and hybrid catfish

Climate change is a growing concern for pond culture of catfish, due to possible exacerbation of temperature fluctuations and increased maximum daily temperatures. Because channel catfish (Ictalurus punctatus) have a broad natural distribution from Canada to Mexico, it was hypothesized that natural differences in thermal tolerance and seasonal growth may be attributed to different geographical strains. Furthermore, it was hypothesized that these differences would be observed in hybrid catfish (I. punctatus x [blue catfish] I. furcatus). Chronic thermal tolerance and growth rate were quantified in two geographically distinct strains of channel catfish and a corresponding hybrid catfish from one of these strains with a cross to an industry standard blue catfish strain. In a 6‐week growth experiment, catfish were subjected to daily cycling temperatures of either 27–31°C or 32–36°C, mimicking pond fluctuations. Hybrid catfish had the highest survival at both temperatures, and both strains of channel catfish had greater growth in weight or length at 27–31°C than at 32–36°C. Therefore, these results indicate that physiological performance, in terms of growth, decreases in channel catfish at temperatures greater than 27–31°C regardless of geographical origin of strain, whereas hybrid catfish did not show a decrease in weight under the same temperature conditions.     

Aspects of the growth of cultured cuttlefish Sepia officinalis (Linnaeus 1758)

Feeding rates, growth rates and feed efficiency ratios were studied in experimentally reared juvenile cuttlefish Sepia officinalis which had been hatched from eggs collected from three different locations, Plymouth, North Wales and Southampton. Groups of newly hatched cuttlefish were either maintained at 19°C and well fed (experiment 1) or were maintained at ambient seawater temperature (7–16°C) with little food for 6 months so that their development was delayed and then transferred to optimum conditions (experiment 2). In the first investigation (expt 1), no significant differences in growth rates (3.72±0.08%, 3.75±0.04% and 3.55±0.04% body weight (BW) day^?1 respectively), feeding rates (9.53±0.36%, 9.28±0.36% and 8.95±0.37% BW day^?1 respectively) and feed efficiency ratios (38.11±1.67%, 40.52±1.78% and 39.96±1.78% respectively) were observed between cuttlefish from the 3 locations. During the second investigation (expt 2), cuttlefish, whose development was initially delayed after hatching and then were stimulated to grow under optimum conditions (19°C and fed), showed growth rates (3.46±0.08% BW day^?1) similar to those held under optimum conditions of seawater temperature (19°C) and food supply shortly after hatching. Feeding rates and feed efficiency ratios were however significantly higher in cuttlefish maintained at 19°C compared to 11°C (8.27±0.14% BW day^?1, 41.25±0.52% and 2.75±0.09% BW day^?1, 24.87±1.87% respectively).     

The combined effects of temperature and salinity on embryos and larvae of the black-lip pearl oyster, Pinctada margaritifera (L.)

This paper reports on a 4 × 4 factorial design experiment conducted to examine the combined effects of temperature and salinity on embryonic development and growth and survival of black-lip pearl oyster, Pinctada margaritifera (L.) larvae. The temperatures used were 20 °C, 25 °C, 30 °C and 35 °C, and the salinities were 25°/_oo, 30°/_oo, 35°/_oo and 40°/_oo. Response surface contour diagrams were generated from the survival and growth data to estimate optimal conditions. Normal development of embryos occurred only from 25 °C to 30 °C. The optimal conditions for maximum survival and growth were 26–29 °C and 28–32°/_oo. Temperatures of 35 °C or greater were lethal for larvae and, at all temperatures tested, larval growth and survival were lowest at a salinity of 40°/_oo.     

Establishment,characterization of a new cell line from heart of half smooth tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>)

A new cell line was established from the heart of a cultured marine fish, half smooth tongue sole (Cynoglossus semilaevis), designated as CSH (Cynoglossus semilaevis heart cell line). The CSH cells grow over 400 days in minimum essential medium (MEM) supplemented with 10% fetal bovine serum (FBS) and 2 ng/ml basic fibroblast growth factor (bFGF). The suitable temperature for the cell growth was 24–30°C with the optimum growth at 24°C and a reduced growth at 12 and 30°C. FBS and bFGF concentration were the two important components for CSH cells proliferation. Twenty percent FBS in the medium was found to be the optimum concentration and bFGF promoted the growth of CSH cells. The double time of the cells at 24°C was determined to 73.39 h. Chromosome analysis revealed that 44% of the cells maintained a normal diploid chromosome number (2n = 42) in the CSH cells at Passage 58. The fluorescent signals were observed in CSH after the cells were transfected with green fluorescent protein (GFP) reporter plasmids. CSH cells showed the cytopathic effect (CPE) after infection with lymphosystis disease virus (LCDV). Moreover, the LCDV particles can be observed in the cytoplasm of virus-infected cells by electron microscopy, and a segment of MCP gene for major capsid protein of LCDV was found by PCR amplification DNA of virus-infected cells.     

Effect of dissolved oxygen and temperature on growth, survival and body composition of juvenile Farfantepenaeus californiensis (Holmes)

Survival, growth and proximal body composition of juvenile Farfantepenaeus californiensis (Holmes) were examined at two dissolved oxygen concentrations (mean 5.8 and 2.6 mg L^?1) and three temperatures (19 °C, 23 °C and 27 °C) for 50 days. Three replicate experiments were performed with a 12 h light/dark photoperiod. Survival was 68–85% at the lower dissolved oxygen level and 77–82% at the higher level and was not significantly affected by the treatments (P > 0.01). Growth at the lower oxygen concentration was significantly less (P < 0.01) than at the higher concentration, with growth rates of 2, 12 and 24 mg day^?1 from the lowest to the highest temperature and 4, 17 and 26 mg day^?1 for the three temperatures at the higher dissolved oxygen level. Body lipids were 1.2% wt/wt at the higher temperature and 2.1% wt/wt at the lower temperature, while proteins were 15.1% wt/wt at the higher temperature and 12.5% wt/wt at the lower temperature. These results indicate that low oxygen levels and low temperature significantly depress growth at this particular stage of life.     

Effect of high water temperature on the survival, moulting and food consumption of Penaeus (Marsupenaeus) japonicus (Bate, 1888)

The kuruma shrimp, Penaeus (Marsupenaeus) japonicus (Bate, 1888), is a valuable aquaculture species in Queensland, Australia. The shrimp is supplied live to the Japanese market and must survive emersed transport for up to 36 h. In-transit mortality after harvest from high water temperatures (> 30 °C) has been reported by the industry, and a knowledge of the effects of high water temperature may provide important information for producers on grow-out management, timing of production and farm location. Experiments were conducted to determine the effect of high water temperature on survival, moulting and food consumption in P. japonicus. Replicated groups of 15.6 ± 0.2 g shrimp were acclimated and exposed to five temperatures, between 28 and 36 °C, for up to 28 days. Mortality was highest at 36 °C and equally lowest between 28 °C and 32 °C. Intermoult period was not significantly different for temperatures between 28 and 32 °C (19.8–15.5 days) but was significantly greater above 32 °C (27.4 days at 34 °C and > 104 days at 36 °C). There was evidence of moulting synchrony at 28 °C. Mean daily food consumption was highest at 32 °C at 2.34% of body weight, but decreased to 1.56% at 28 °C and 1.33% at 36 °C. Over the range of water temperatures examined, survival, moulting rate and food consumption were highest at 32 °C.     

Sperm cryopreservation of tiete tetra Brycon insignis (Characiformes): effects of cryoprotectants,extenders, thawing temperatures and activating agents on motility features

The aim of this study was to test the effects of cryoprotectants [dimethyl sulphoxide (DMSO) and methylglycol], extenders (0.9% NaCl, 5% glucose, Beltsville Thawing Solution^? and Merck III^?), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) on the cryopreservation process of tiete tetra Brycon insignis sperm. Sperm was loaded in 0.5 mL straws, frozen in nitrogen vapour at ?170 °C and stored in liquid nitrogen. Post‐thaw sperm quality was evaluated in terms of subjective motility rate, quality motility score (0=no movement; 5=rapidly swimming spermatozoa), duration of motility and vitality (eosin–nigrosin staining). Post‐thaw sperm motility rate was greater in methylglycol (76–88%), compared with DMSO (23–59%). In general, the highest quality motility scores were observed when sperm was thawed at 30 °C and triggered in 1% NaHCO₃ (3.5–4.3). Duration of motility was longer when triggered in 1% NaHCO₃ (95–120 s) compared with 0.29% NaCl (69–107 s). Sperm vitality was not affected by any of the parameters tested and varied from 51% to 69% intact sperm. Brycon insignis sperm frozen in methylglycol combined with any of the extenders tested and using the methods described above yields motility above 57% and that should last long enough to fertilize oocytes.     

Effects of culture conditions on larval growth and survival of stalked barnacles (Pollicipes pollicipes)

Pollicipes pollicipes (Crustacea: Scalpelliformes) is a highly prized food in Portugal and Spain and consequently a species of considerable interest to aquaculture. Surprisingly, however, larval culture conditions for this barnacle have not been optimized. This study investigated the effects of temperature, diet, photoperiod and salinity on the growth and survival of P. pollicipes larvae. Temperature had a significant effect on specific growth rate (2.6–5.9% total width per day, from 11 to 24°C), reducing mean development time to the cyprid from 25 days at 11 °C to 10 days at 24°C, although this was accompanied by a significant increase in mortality to over 90% above 22°C. Mid‐range temperatures (15–20°C) maximized total survival (19–31% respectively). Algal diets of Tetraselmis suecica, T. suecica/Skeletonema marinoi and S. marinoi/Isochrysis galbana did not affect specific growth rate significantly, but survival (on average 39% in 15 days) and the proportion of high‐quality healthy cyprids was significantly higher on the latter two diets (11–15% of initial number of larvae). Photoperiod did not significantly affect the survival, although specific growth rate was significantly higher at 24:0 and 16:8 L:D. Salinity (20–40 g L^?1 range) did not affect growth and survival significantly. The best growth and survival were accomplished using rearing temperatures of 15–20°C, daily feeding with T. suecica/S. marinoi or I. galbana/S. marinoi and a photoperiod of 24:0 L:D.     

Characteristics of a self-assembled fibrillar gel prepared from red stingray collagen

A translucent collagen gel was formed from a transparent acidic solution of red stingray collagen by adjusting to physiological ionic strength and pH in phosphate buffer and then incubating at 25–37°C. During fibril formation from red stingray collagen, the turbidity increased when the NaCl concentration was increased at constant pH and the rate of fibril formation was accelerated by higher pH or lower NaCl concentration. The T _m of red stingray collagen fibrillar gel was estimated as 44.3 ± 3.5°C, which was higher than that of the collagen solution, 33.2°C. In addition, red stingray collagen gel maintained its shape without melting and was suitable for culture of mouse stromal cells at 37°C.     

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C_maxe; J day^?1) and the specific growth rate in terms of energy (SGRe; % day^?1) in animals of three body sizes (mean±SE) – large (134.0±3.5 g), medium (73.6±2.2 g) and small (36.5±1.2 g) – were determined at water temperatures of 10, 15, 20, 25 and 30°C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20°C. This trend, however, was not apparent at 25 and 30°C, which could be influenced by aestivation. High water temperatures (above 20°C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15°C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3–139.3 g) was 24.5?25.5°C, while that for small animals (28.9–40.7 g) was between 25.5 and 30.5°C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.     

Tetraploid induction in tropical oysters,Crassostrea belcheri (Sowerby) and Crassostrea iredalei (Faustino)

Tetraploid induction has been conducted on temperate oysters but not on tropical oysters. In this study, different heat shocks (32, 35 and 38°C) and cold shocks (1, 4 and 7°C) were used to induce tetraploidy in two tropical oyster species, Crassostrea belcheri and Crassostrea iredalei, through meiosis I inhibition. Temperature shocks were applied on the newly fertilized eggs at 8–10 min post fertilization and terminated when second polar bodies began to form in the control eggs. The ploidy of the larvae and spat was determined via direct chromosome count. The percentage of larval survival until Day 20 was low (between 0.4% and 42.9%) for both temperature shocks and oyster species. No surviving larva was recorded for induction at 1, 4 and 38°C. Tetraploid spat was only recorded in C. iredalei but the percentage is low through heat shock induction of 32 and 35°C. This study shows that the tetraploid induction success rate was slightly higher in C. iredalei compared to C. belcheri. No surviving tetraploid spat were recorded for both oyster species through the cold shock method. This study shows that heat shock can be used to inhibit meiosis for the production of tetraploids but more experiments need to be conducted to determine the optimum temperature when dealing with tropical oysters.     

EXPERIMENTAL CULTURE OF THE OCEAN QUAHOG,Arctica islandica

Larvae and early postlarvae of the ocean quahog, Arctica islandica, were reared under experimental hatchery conditions. Mature eggs were stripped from ripe adults and exposed to a dilute solution of ammonium hydroxide for various lengths of time prior to addition of stripped sperm. The larval clams were reared through settlement and metamorphosis using the Wells-Glancy (centrifuged, incubated seawater) method of algal culture and/or modifications of standard hatchery techniques developed by Loosanoff and Davis. Experimental cultures were maintained at various temperatures ranging from 8.5° to 14.5°C. At temperatures of approximately 13°C, the minimum time to settlement was 32 days, while settlement was not observed in a culture maintained between 8.5° and 10.0°C until approximately 55 days after fertilization. Larval growth rates were significantly lower in the culture maintained at 8.5–10.0°C than in cultures maintained at 11.0–14.5°C. An optical micrograph sequence of larval stages from the straight-hinge stage through metamorphosis is presented to facilitate identification of Arctica islandica specimens isolated from plankton samples. While various workers have reported exceedingly low growth rates of juvenile and adult Arctica, growth rates of larval Arctica appear to be fairly “typical” of rates encountered within the class Bivalvia.     

Thermal challenge of the Indian shrimp Penaeus indicus

This study investigated short‐term effects of increasing water temperature from 27 to 41°C on survival and feed consumption of Penaeus indicus at three different ages: PL₂₅ (postlarvae 25 days old), PL₅₀ and PL₉₀. For each age group, water temperature was maintained at 27°C in the control, but increased to 32, 35, 38 and 41°C at a rate of 1°C every eight hours. The temperature was then kept stable until the end of the 7‐day experiment. Results showed that increasing water temperature affected both survival and feed consumption of the experimental shrimps (p < .01). Survival was highest at 32 and 35°C ranging from 93.8% to 100%, but significantly reduced to 40.0%–81.6% at 38°C. No shrimp survived the 41°C treatment. PL₂₅ were more tolerant to 38–41°C than PL₅₀ and PL₉₀ in terms of survival. Increasing water temperature had no effects on feed consumption of PL₂₅ (p > .05). For PL₅₀ and PL_90, feed consumption significantly increased at 38 and 41°C (p < .01) and was similar within the range of 27–35°C. This study suggests that P. indicus in tropical areas can tolerate water temperatures of at least 35°C and should be considered for farming during the summer time.