Adsorption of minerals on the isolated mantle of Patinopecten yessoensis

The isolated mantle of scallop ( Patinopecten yessoensis ) was shown to adsorb minerals (Zn, Ca, Mg, Fe, Cu, Ni and Co) to more than 1% upon treatment with mineral solution. The amounts of adsorbed Zn, Ca and Mg were 44.3, 47.4 and 15.5 mg/g dry weight, when the mantle was treated with 5% ZnSO₄, CaCl₂ and MgSO₄ solutions at room temperature, respectively. The amounts of adsorbed Zn, Ca and Mg were dependent on the concentration of mineral solution used. This is the first report showing that the isolated mantle adsorbs nutritionally important minerals such as Ca, Mg and Fe. This finding may lead to the development of new mineral-enriched food supplements.     

Purification and characterization of transglutaminase from squid gill

ABSTRACT: The present study used squid gill as a source of transglutaminase (TGase) because it has extremely high TGase activity compared with other tissues. The enzyme was purified using successive chromatographies of Sephacryl S-300 and hydroxyapatite columns. The yield and purification-fold of the enzymatic activity was 12.6% and 14.1-fold, respectively. The molecular mass of the purified enzyme was estimated to be 94 kDa by using sodium dodecylsulfate–polyacrylamide gel electrophoresis analysis. Enzyme activity was enhanced 15-fold with an increase in NaCl concentration. Although the activity was dependent on Ca²⁺ concentration, it was not sufficiently activated even by 50 mM CaCl₂ in the absence of NaCl, but could be fully activated with 10 mM CaCl₂ in 0.7 M NaCl. However, in the absence of substrates, the enzyme was rapidly inactivated. The pH and temperature optima of the enzyme were approximately pH 8.0 and 20°C, respectively. It was stable in the absence of Ca²⁺ at pH 7.5–9.0 and had a rate constant (K _D ) of 1.6 × 10^–5 s^–1 for thermal inactivation at 50°C. These results in which squid gill TGase could be activated at higher concentrations of Ca²⁺ and NaCl than at a physiological concentration, suggest that contact with seawater or body fluid seems to activate the enzyme if the tissue is disrupted.     

Effect of different diets on larval production, quality and fatty acid profile of the marine ornamental shrimp Lysmata amboinensis (de Man, 1888), using wild larvae as a standard

The present work evaluates the influence of broodstock diets [Marine Cuisine^®– MC, MC supplemented with highly unsaturated fatty acid- (HUFA) enriched Artemia biomass – MC + AB, and MC supplemented with squid – MC + S] on larval production, newly hatched and early zoeal stage survival and fatty acid profile of newly hatched larvae of Lysmata amboinensis . These parameters are compared with those from larvae hatched from embryos spawned in the wild. The number (±SE) of larvae produced with MC and MC + S (1077 ± 219 and 1103 ± 184, respectively) was similar to that in broodstock carrying embryos spawned in the wild (1224 ± 111), while those fed MC + AB displayed significantly lower values (1044 ± 161). Larvae produced with MC + AB displayed lower survival for all starvation periods, while larvae spawned in the wild displayed the highest survival. No larvae resisted 144 h of starvation and none moulted to zoea II. The fatty acid comparison revealed that larvae from embryos spawned in the wild displayed the highest levels of DHA, as well as higher DHA/EPA and n -3/ n -6 ratios. These results suggest that broodstock diets commonly used to promote ornamental shrimp's maturation (based on mixed frozen components) are far from being optimal.     

Role of sodium bicarbonate on the initiation of sperm motility in the Japanese eel

ABSTRACT:   In order to find out the role of sodium bicarbonate (NaHCO₃) on the initiation of sperm motility in the Japanese eel Anguilla japonica , interactions were investigated between NaHCO₃ and various reagents (K⁺ channel blocker 4-aminopyridine [4-AP], ammonium chloride [NH₄Cl], sodium acetate and calcium chloride [CaCl₂]) that could regulate internal factors (intracellular K⁺, intracellular pH [[pH]_i] and intracellular Ca²⁺) in sperm motility. Contradictory effects of NaHCO₃ were observed (i.e. an inhibitory effect when 4-AP was absent and a promoting effect when 4-AP was present). Sodium bicarbonate inhibited the initiation of sperm motility in the Japanese eel. However, NaHCO₃ restored the motility of immotile sperm that 4-AP inhibited. The inhibitory effect of NaHCO₃ disappeared with the addition of NH₄Cl, which raised [pH]_i, but the promoting effect was not affected by [pH]_i. Although NaHCO₃ recovered motility in the presence of 4-AP, this recovery was also observed with the addition of CaCl₂ instead of NaHCO₃. In the initiation of sperm motility in the Japanese eel, two roles for NaHCO₃ are suggested: an inhibitory role relating to the regulation of [pH]_i and a promoting role relating to the uptake of another initiation factor, which could be Ca²⁺.     

The effects of cadmium and copper on embryonic and larval development of ide Leuciscus idus L.

The effects of Cd and Cu on embryos and larvae of the ide Leuciscus idus were evaluated. The ide is an European cyprinid fish, natural populations of which tend to decrease. The ide is also used as a bioindicator organism to evaluate water quality. However, sensitivity of ide early developmental stages to heavy metal intoxication is not known. Fish were exposed to Cd or Cu (100 μg/L) during embryonic, larval or both developmental periods. Survival of the embryos, time of hatching, size and quality of newly hatched larvae were evaluated at the end of embryonic period. Correctly developed larvae from the control and Cd or Cu-exposed groups were transferred to clean water, Cd or Cu solutions (100 μg/L) immediately after hatching. Larval development was observed, and the larvae were photographed. Time of yolk sac resorption, onset of active feeding and swim bladder inflation were evaluated, and the measurements were done on body and swim bladder size. The results showed that exposure of embryos to Cd and Cu significantly reduced embryonic survival and increased frequency of body malformations and death in newly hatched larvae and delayed hatching. Exposure to Cd and Cu during larval period reduced larval survival, growth and delayed development (yolk utilization, beginning of active feeding and swim bladder inflation). Cadmium was more toxic to the ide embryos and larvae than copper. Exposures to metals during embryonic period alone caused adverse impact on larval performance even when larval development took place in clean water. However, exposure of embryos to Cu reduced toxic impact of metal on larvae in continuous Cu exposure compared to the non-preexposed fish, but no such an effect occurred in case of Cd exposure. The results show that even a short-term exposure to Cd or Cu during early development of ide may adversely affect recruitment of this species. Among the measured endpoints, quality of newly hatched larvae (frequency of body malformations and larvae dead immediately after hatching) and swim bladder size were the most sensitive to intoxication with both metals. Embryos were more sensitive to Cu intoxication than larvae, while in case of Cd, sensitivity of both stages was similar.     

五种重金属对早繁鮸鱼胚胎和仔鱼的毒性效应

在水温27.5℃、盐度29条件下,研究了Cu2+、Cd2+、Zn2+、Cr6+和Pb2+5种重金属对早繁鮸鱼胚胎发育和初孵仔鱼的影响。结果表明:重金属在胚胎发育过程中引起各种畸形,如胚胎尾巴弯曲、初孵仔鱼不能出膜、胚胎死亡等;胚胎和初孵仔鱼上出现黑色颗粒,初孵仔鱼脊柱弯曲成L、V、S形或尾部没有展开呈逗号状,褶皱边缘被破坏等畸形。Cu2+、Cd2+、Zn2+、Cr6+和Pb2+5种重金属均延迟胚胎发育速率。综合孵化率、胚胎畸形和仔鱼畸形率等指标得出5种重金属对胚胎的毒性大小依次为Cu2+>Cd2+>Zn2+>Pb2+>Cr6+。计算了5种重金属对早繁鮸鱼初孵仔鱼的LC50和安全浓度,得出5种重金属对早繁鮸鱼初孵仔鱼毒性大小顺序为:Cu2+>Cd2+>Pb2+>Zn2+>Cr6+。     

Survival and Growth of Red Drum Sciaenops ocellatus in Saline Groundwaters of West Texas, USA

Abstract.— West Texas saline groundwaters were assessed as potential media for culturing red drum, Sciaenops ocellatus . In 30-d bioassays with juvenile red drum (0.4–3.0 g), highest survival was 85% in a 5-ppt (parts per thousand = g/L) salinity, high-sulfate (1,723 mg/L SO₄^-2), high-calcium (427 mg/L Ca^±2) groundwater from a windmill catchment pond in Pecos County. The lowest survival was 0% in a 35-ppt saline groundwater from a gravel pit, also in Pecos County. In complementary bioassays of low-salinity (<5 ppt) groundwaters from the same region, calcium chloride (CaCl₂) addition to a 3-ppt. Reeves County ground-water low in Ca^±2 resulted in the greatest increase in survival (0–93). It was unclear whether the beneficial effect of CaCl₂ was a result of increasing Ca^±2 concentration (36–336 mg/L), increasing Cl^- concentration (639–1,296 mg/L). or both. The concentration of total dissolved solids ("salinity") appears to be the single most important consideration in estimating the red drum aquaculture potential of a groundwater, with the optimum salinity being 5–15 ppt. Concentrations of arsenic (As), cadmium (Cd), chromium (Cr), mercury (Hg), lead (Pb), and selenium (Se) tended to be higher in west Texas groundwaters than in typical U. S. surface waters. However, based on current maximum tolerable daily intake recommendations, red drum cultured for 30 d in west Texas groundwaters contained whole-body As, Cd, Cr, Hg, Pb, and Se concentrations that would not pose a substantial health risk to adult human consumers.     

Characterization of exotoxin produced by a shellfish-pathogenic Vibrio sp.

Abstract. Broth filtrate of a Vibrio species, isolated from spontaneously occurring epizootics among cultured oyster larvae, Crassostrea virginica (Gmelin), was shown to contain an exotoxin. Bioassays demonstrated that LC₅₀ value was less than 46.6 μg of toxin/l culture of oyster embryos. This quantity was produced by 2.9 × 10⁹ colony-forming units. Purified toxin showed neither proteolytic nor amylase activity; it did, however, demonstrate bacteriostatic capability. Studies showed that the toxin was heat-labile and that exposure to heat-produced toxoid had a beneficial effect on oyster embryonic development. Data revealed that although toxin was inactivated by heating, the bacteriostatic capability of the metabolite was not lost. Molecular weight of the toxin was estimated to be 68000.     

A study on Flexibacter columnaris infection in loach, Misgurnus anguillicaudatus (Bleeker, Günther)

Abstract. Infectivity of Flexibator columnaris to loach, Misgurnus anguillicaudatus (Blecker, Günther), was studied with variation conditions and the presence of other bacteria in the water medium. Invasion of fish by the poathogen varied with different water conditions. The highest prevalence of infection was observed in an experimentally formulated water containing 0·03% NaCl 0·01% KCl, 0·002% CaCl₂2H₂O and 0·004% MgCI₂ 6H₂O. The percentage of infection in tap water was similar to that of the formulated water. The infeetion rate was low in the individual salt solutions, and no infection occurred in distilled water. Even in the formulated water, the pathogen failed to infect the fish when Citrobacter freundii, Aeromonas hydrophila or Pseudomonas fluorescens were present at densities 100 times that of F. columnaris. However, in such cases, the numbers of F. columnaris in the mucus of the fish body surface did not increase much throughout the experimental period. In contrast, F. columnaris suceessfully invaded the fish in the presence of either Streptococcus sp., Flavobacterium flavescens or Flavobacterium fuscum , even when the numbers of these bacteria were 1000 times that of F. columnaris. In this ease, F. columnaris gradually increased its numbers on the fish body surface.     

Effects of Environmental Sodium Chloride on Percent Hatch, Yolk Utilization, and Survival of Channel Catfish Fry

Abstract.— Only limited research has addressed the effect of salinity on hatching of channel catfish Ictalurus punctatus eggs, and no studies have evaluated the effect of salinity on fry development and survival. This study was undertaken to determine the effect of environmental sodium chloride (0, 1, 2, and 4 g/L. NaCl) on percent hatch, yolk utilization, and survival of channel catfish fry. Experiments were conducted in recirculating systems using seven egg masses (1–2 d old). Each egg mass was divided into smaller portions which remained undissociated or were dissociated with sodium sulfite (NaSO₃). Eggs were incubated until hatching. Wet and dry weights were obtained for sacfry at 1 and 5 d post-hatch to determine wet weight gain and dry weight loss, and fry were sampled 7 d after initiation of exogenous feeding to determine survival. Percent hatch, yolk utilization, and survival of fry hatched from undissociated eggs were greatest at 1 g/L NaCl. In addition, treatment of eggs with NaSO₃ significantly reduced percent hatch at all NaCl levels. Although our results indicate that addition of NaCI to hatchery water supplies can increase production of channel catfish fry, additional research is needed before this practice can be recommended on a commercial basis.     

The effects of Roundup on gametes and early development of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L)

To determine the effects of Roundup, a commercial formulation of glyphosate, gametes, and embryos of common carp (Cyprinus carpio L) was exposed to wide range of herbicide concentrations (0.0, 0.1, 0.5, 2.0, 5.0, 10.0, 20.0, and 50.0 mg/l). The obtained results showed different effects of Roundup on common carp gametes. Herbicide reduced swelling of eggs (but the effect was not concentration-related), while sperm showed low sensitivity to Roundup (time of spermatozoa motility was reduced in a significant way only at 20 mg/l, and at remaining concentrations, only a slight tendency was observed). During the embryonic development, Roundup caused a decrease of common carp embryonic survival (and the effect was concentration-related); however, it had no effect on development rate. During the embryogenesis, three types of embryo body malformation were observed: yolk sac edema, spine curvature, and shortening of body, but their frequencies were not associated with the presence or concentration of herbicide. However, Roundup affected quality of newly hatched larvae of common carp by increasing their mortality. No effect of herbicide on percentage of deformed larvae was observed but larvae hatched in water with Roundup tended to show more complex anomalies compared to those from the control. Obtained data showed that even low concentrations of this herbicide in waters can significantly reduce egg swelling, survival of embryos, and quality of fish larvae.     

Piscivory in larval perch (Perca fluviatilis): mechanisms structuring larval roach (Rutilus rutilus) cohorts

Abstract – Perch ( Perca fluviatilis ) can act as a piscivore from larval stage VI (body size 10.3 mm) on newly hatched larval roach ( Rutilus rutilus ), bream ( Abramis brama ) and smaller siblings of its own cohort. Consumption rates at this stage were approx. 0.5 prey/perch*h at 21°C. Larval perch predation was strongly gap-limited, and the maximum size of roach consumed by perch (perch length interval 10.3–62.0 mm) under experimental conditions followed the linear regression, P_prey-max.=0.478*L_Pred.+1.829 ( r ²=0.99, P <0.001, n =12). Under experimental conditions, predatory 0⁺ perch substantially affected the size distributions of 0⁺ roach prey cohorts, since smaller prey individuals were predated more frequently than larger ones. In both unimodal and bimodal size distributions of prey roach, the distributions changed according to the maximum prey size consumed by the added predatory perch. Unimodal prey distributions were positively skewed when piscivorous perch were added compared with controls without predators. According to the size distributions of lake-living 0⁺ roach and 0⁺ perch and the relative size difference between prey and predator, the vulnerability of 0⁺ roach cohort to 0⁺ perch predation changed from June to September. Prey vulnerability was extremely sensitive to the relative size difference between predator and prey. Therefore differences in hatching time and growth rates between the two species will strongly influence the potential for predator-prey interactions. ^Note     

Ingestion of Artemia nauplii by Chinese mitten crab Eriocheir sinensis zoea larvae

A series of ingestion trials were conducted to determine the ingestion rate of Artemia nauplii by Eriocheir sinensis zoea larvae with increasing densities of Artemia and with or without rotifers as a co-feed. At each zoeal stage, 10 groups of 10 larvae were reared individually in glass beakers and fed with increasing densities of newly hatched Artemia nauplii (0.5, 2.5, 5, 10 and 20 individual (ind.) mL⁻¹) with or without rotifers (15–25 ind. mL⁻¹) as a co-feed. The average number of ingested Artemia was measured over 24 h. In addition, the average larval development rate (Larval Stage Index, LSI) over a longer period (time needed for the best treatment to reach 100% moult or metamorphosis to the next larval stage) was compared. The results showed that Artemia ingestion rate of E. sinensis larvae increased with increasing prey densities and larval development, and had a significantly negative correlation with rotifer consumption for all zoeal stages. Rotifers as an alternative prey significantly affected the intake of Artemia at early larval stages (Z1 and Z2) and promoted LSI at a lower Artemia density. Further experiments are needed to clarify the effect of prey density on survival and larval development when larvae are reared communally.     

Hormones and Egg/Larval Quality in Fish

Recent evidence suggests that hormones are passed on to eggs by broodfish. This store of maternal hormones may fill the regulatory needs of fish larvae for growth, development, osmoregulation, stress response and other physiological functions prior to the functional development of their own endocrine glands. Thus the hormonal levels in eggs/larvae may be an important determinant of egg/larval quality.
Thyroid hormones have been best studied in this regard. Both thyroxine (T₄) and triiodothyronine (T₃) are present in fish eggs and in most cases, the levels decrease as development proceeds until the onset of endogenous thyroid hormone production, which usually occurs before or at around yolk-sac resorption. Enhancement of T₄/T₃ levels in newly-hatched larvae through immersion or maternal injection has been shown to promote larval growth, development and/or survival in several fish species, although overdose causes thyrotoxicosis.
Other hormones such as corticosteroids, sex steroids, growth hormone and prolactin are also being studied, but to a much lesser extent.
The paper reviews the evidence and discusses the practical implications of the various findings for larval rearing in aquaculture.     

Thermal denaturation and autolysis profiles of myofibrillar proteins of mantle muscle of jumbo squid Docidicus gigas

ABSTRACT:    Jumbo squid was very similar to Japanese common squid in terms of myofibrillar Ca²⁺-, Mg²⁺- and K⁺(EDTA)-ATPase activities. Myofibrils of jumbo squid were significantly stabilized upon addition of Ca²⁺ and destabilized by increasing KCl concentration for heating. Incubation of muscle homogenate of jumbo squid induced a selective cleavage of myosin into two major fragments and the cleavage was inhibited by EDTA. Autolysis was prominent at and above 0.3 M NaCl where myosin filaments dissolve. The enzyme involved in the autolysis was proved to be unstable showing maximal autolysis rate at 25°C. Washing the homogenate partially reduced the autolysis activity.     

盐度对暗纹东方鲀胚胎发育的影响

研究了不同盐度对暗纹东方鲀胚胎发育的影响，盐度为4和8的试验组中受精卵的发育历期、孵化率及初孵仔鱼24小时的存活能力和淡水相比均无显著差异。在盐度12及以上，绝大部分受精卵死亡，仅有极少数个体孵化出膜，且全为畸形并都在24小时内死亡；发育速度随着盐度的升高而变慢。因此，暗纹东方鲀胚胎正常发育所能耐受的盐度范围是0～8。     

The glycerinated body wall of the sea cucumber as a suitable preparation for electron microscopic and physiological studies of ‘catch mechanism’

ABSTRACT:   The body wall of the sea cucumber changes its stiffness by ionic environments. The stiff state can be held for a long time, and the mechanism concerned is known as 'catch mechanism'. In the present study, the direct effects of ions on the mechanism using the glycerinated body wall treated with 50% glycerin to clarify how the ions effect changes of stiffness were examined. The glycerinated body walls contained collagen fibers and some broken cells in the connective tissue ultrastructurally. Cell membranes were not clearly present in the broken cells, and cell organelles were dispersed around the cells. The glycerinated body walls went into a limp state during addition of 10 mM ethylenediaminetetraacetic acid (EDTA), and showed height elongation rate in this study's experimental system. In contrast, the elongation rate decreased by the addition of 10 mM CaCl₂, that is, the body wall came to a stiff state. This stiff state could be considered as equivalent to 'catch state' of glycerinated body wall. Collagen fibers in those samples showed more compact arrangements at 10 mM CaCl₂ treatment than the one of 10 mM EDTA ultrastructurally. These features and physiological results suggested that EDTA and/or CaCl₂ from outside affect directly to the main part of the 'catch' mechanism in the glycerinated body wall.     

Effect of Oxygen Saturation in Water on Reproductive Performances of Pacu Piaractus brachypomus

Broodstock pacu Piaractus brachypomus as well as their eggs during their embryonic development were exposed to either normoxia (5.5–7.5 mg O₂/L) or hypoxia (2.0–4.5 mg O²/L) conditions. The plasma concentrations of 11-ketotestosterone in males and estradiol-17β in females, as well as that of their precursor testosterone (T) were significantly ( P < 0.01) higher in fish maintained under normoxic conditions than in fish exposed to hypoxia. After ovulation and spermiation induced by hormonal treatments, the plasma concentrations of T and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) significantly ( P 0.05) increased in both sexes under both normoxic and hypoxic conditions. The plasma levels of T and 17,20βP achieved under normoxic conditions were higher than the ones recorded under hypoxia, except for those of 17,20βP in males. Males responded positively to the hormonal treatments, and the concentration of spermatozoa was 10.5 ± 0.8 10⁹/mL under both oxygen conditions. Hypoxia resulted in significantly lower survival of embryos (17.3 ± 28%) in comparison to normoxic conditions (68.5 ± 25%). Moreover, larval deformities were found when exposed to hypoxia (91.6 ± 6%). During embryonic development of this species 4 mg/L of oxygen is tolerated at 26–27 C without negative impact. We conclude that despite the highly adaptable nature of adult pacu to environmental hypoxia, oxygen concentrations below 4 mg/L severely impacted survival of embryos.     

Effects of Heavy Metals on the Hatching Rates of Brine Shrimp Artemia salina Cysts

The effects of heavy metals (Hg²⁺, Cd²⁺, Co²⁺, Cr³+, Pb²⁺, Cu²⁺, Fe²⁺. Ni²⁺, Zn²⁺, Mn⁷⁺, Mn⁴⁺ and Mn²⁺) on the relative hatching percentage of brine shrimp, Artemia salina were assessed. Artemia salina cysts (San Francisco Bay Brand) hatched in seawater containing various heavy metal concentrations. After 48 hours, the number of hatched nauplii were counted. A negative linear relationship between hatching rate of Ariemia salina cysts and heavy metal concentration occurred, except copper which conformed to a cubic model. The relative toxicities of heavy metals on Artemia salina cysts were as follows (Cu²⁺, most toxic; Mn²⁺, least toxic): Cu²⁺ > Zn²⁺ > Ni²⁺ > Co²⁺ > Mn⁷⁺ > Cr³⁺ > Cd²⁺ > Fe²⁺ > Hg²⁺ > Mn⁴⁺ > Pb²⁺ > Mn²⁺.     

Conditions for the induction of some selective enzymes from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> and their hydrolysis ability against mackerel and gracilar (asparagus, <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis>)

ABSTRACT:   This study was conducted to optimize the cultivation conditions for Bacillus subtilis to produce proteases, amylases and cellulase, and to further investigate the hydrolysis ability against mackerel and asparagus. The extracellular enzymes from B. subtilis after 2 and 4 days incubation in a modified medium, containing 1% skim milk, 1% soya meal, 0.25% starch, 0.25% K₂HPO₄, 0.5% NaCl and 0.05% MgCl₂ were collected for the hydrolysis of asparagus and minced mackerel, respectively. Except for the α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging ability of the hydrolyzed asparagus, the trolox equivalent antioxidation capacity and DPPH scavenging ability of both samples increased significantly ( P  < 0.05) after 1 h hydrolysis and further increased during elongated hydrolysis at 50°C. Sodium dodecylsulfate–polyacrylamide gel electrophoresis indicated severe degradation of muscle proteins during hydrolysis. Changes in reducing sugar, soluble proteins and peptides before/after hydrolysis suggested the extracellular enzymes from B. subtilis could effectively hydrolyze the mackerel or asparagus, and subsequently improve their antioxidation ability.