Use of the Nematode Panagrellus redivivus as an Artemia Replacement in a Larval Penaeid Diet

Penueus vannumei larvae (P₂ through M₃) and early postlarvae (<24 h old postlarva) were fed diets consisting of algae-only, nematodes ( Panugrellus redivivus ) plus algae or Artemia plus algae. Growth (dry biomass gain) of second and third stage protozoea larvae fed the nematodealgae diet was significantly better than that of larvae fed the Artemia-algae diet. From the first mysis through the postlarval substage (<24 h old), growth of shrimp fed the nematodealgae diet equalled that of larvae fed the Artemia-algae diet. All larval substages fed nematodes plus algae accumulated significantly greater biomass than those fed a diet of only algae. Survival and percent metamorphosis of larvae fed nematodes plus algae did not differ significantly from that of larvae fed either Artemia plus algae or algae alone. A nematodealgae feeding regime, which potentially yields growth, survival and metamorphosis equal to that obtained on the standard Artemia plus algae regime, is proposed for P. vannamei .     

THE EFFECTS OF EDTA (ETHYLENEDINITROTETRAACETIC ACID) ON THE SURVIVAL AND DEVELOPMENT OF SHRIMP NAUPLII (Penaeus stylirostris STIMPSON) AND THE INTERACTIONS OF EDTA WITH THE TOXICITIES OF CADMIUM,CALCIUM, AND PHENOL

To examine the hypothesis that EDTA increases hatching rates and survival of penaeid shrimp larvae by decreasing the toxicities of heavy metals through chelation, the toxicity of cadmium, a highly toxic metal, was compared to the toxicities of calcium and phenol in both the presence and absence of EDTA. In addition, the toxicity of EDTA at higher concentrations was examined. The toxicities of EDTA, cadmium, calcium, and phenol were evaluated in terms of the percentage of nauplii surviving after 24 hours of exposure and the percentage of nauplii which metamorphosed to protozoea. The toxicities of cadmium, calcium, and phenol were also determined in the presence of EDTA. EDTA concentrations of 1.34 mM were lethal to nauplii. At 0.67 mM, EDTA reduced the percentage of nauplii which metamorphosed to protozoea but below 0.3 mM neither survival nor metamorphosis were affected. Cadmium, phenol, and calcium were lethal to nauplii at concentrations of 20 μM, 7 mM, and 400 mM, respectively. Metamorphosis was blocked by concentrations of 1 μM, 0.9 mM, and 200 mM, respectively. However, in the presence of 0.3 mM EDTA, the toxicities of cadmium and calcium were reduced. Cadmium concentrations of 20 μM did not affect either survival or metamorphosis in the presence of EDTA. In calcium concentrations of 50 and 100 mM, the percentage of nauplii that metamorphosed to protozoea was increased by the addition of EDTA. The interaction of EDTA and phenol toxicity was not significant.     

中国对虾糠虾幼体病原哈氏弧菌的鉴定及毒力基因检测

2011年春季对江苏连云港某对虾育苗场中国对虾Fenneropenaeus chinensis病死糠虾幼体分离到优势生长菌,对分离菌进行致病性、形态与生理生化特征及16S rRNA和gyrB基因同源性与系统发育分析。结果显示,引起糠虾幼体大量死亡的病原为哈氏弧菌Vibrio harveyi,菌株kx1对中国对虾仔虾和日本对虾仔虾的半数致死量LD50分别为2.0×106CFU/ml和7.0×105CFU/ml。为进一步明确分离菌株毒力基因的携带情况,进行了分离鉴定的4株病原菌对群体效应调节基因(luxR)、毒力调控基因(toxR)、溶血素基因(vhhA和vhhB)、金属蛋白酶基因(vhpA和vhpB)、毒力相关基因(toxS)、鞭毛结构基因(flaA)及锌金属蛋白酶基因(pap6)共9种毒力基因的检测,结果表明,4株病原菌均可检测到luxR、toxR、vhhA、vhhB和pap6毒力基因,扩增片段大小分别为679、390、1324、216和355 bp,其他4种毒力基因未检测到。分离鉴定的4株病原哈氏弧菌携带相同的毒力基因,这些毒力基因可作为检测致病性哈氏弧菌的生物学标记。     

Trypsin enzyme activity during larval development of Litopenaeus vannamei (Boone) fed on live feeds

Larval stages of the Pacific white shrimp, Litopenaeus vannamei (Boone) were fed standard live diets of mixed microalgae from the first to the third protozoea (PZ1 to PZ3), followed by Artemia nauplii until post‐larvae 1 (PL1). Trypsin enzyme activity for each larval stage was determined using N‐α‐p‐toluenesulphonyl‐l ‐arginine methyl ester (TAME) as a substrate. Results were expressed as enzyme content to assess ontogenetic changes during larval development. Tissue trypsin content (IU µg^?1 DW for each larval stage) was significantly highest at the PZ1 stage and declined through subsequent stages to PL1. This contrasts with previously observed patterns of trypsin development in Litopenaeus setiferus (Linnaeus) and other penaeid genera, which exhibit a peak in trypsin activity at the third protozoea/first mysis (PZ3/M1) larval stage. Litopenaeus vannamei larvae transferred to a diet of Artemia at the beginning of the second protozoea (PZ2) stage were significantly heavier on reaching the first mysis stage (M1) than those fed algae, while survival was not significantly different between treatments. At both PZ2 and PZ3 stages, trypsin content in larvae feeding on Artemia was significantly lower than in those feeding on algae. The rapid decline in trypsin content from PZ1 and the flexible enzyme response from PZ2 suggest that L. vannamei is physiologically adapted to transfer to a more carnivorous diet during the mid‐protozoeal stages.     

Differences in larval growth among families of Penaeus stylirostris Stimpson and P. vannamei Boone

Abstract. Full-sib families of Penaeus stylirostris Stimpson and Penaeus vannamei Boone were grown under controlled conditions in five experiments. Total length of shrimp larvae was measured at the following substages: protozoea one, mysis one and postlarva one. Analysis of variance showed that families differed significantly in size in all experiments at all substages tested. The average size possessed by a group of larvae early in development was significantly correlated with the size obtained in the later stages in P. siylirosiris , but not in P. vannamei . The component of variance in size due to family differences was determined and used to estimate heritabilities of size at the three larval substages in both species. Estimated heritabilities were higher for P. stylirostris than P. vannamei . In P. vannamei . the hcritability of size is greater for the protozoea and postlarval stages than the mysis stage.     

Predation and energetics of Penaeus indicus (Decapoda: Penaeidae) larvae feeding on Brachionus plicatilis and Artemia nauplii

Brachionus plicatilis and Artemia nauplii were fed to a number of larval stages of the penaeid prawn Penaeus indicus to determine ingestion rates, larval energy requirements and to establish at which stage larval predation commenced. The raptorial feeding rates were then contrasted on an energy basis with filter feeding rates for P. indicus larvae to compare the relative efficiency of these two feeding mechanisms. Brachionus was first eaten as early as protozoea 1 to protozoea 2, while the maximum ingestion rate of 300 rotifers larva^?1 d^?1 (1.06 J larva^?1 d^?1) was obtained during protozoea 3 to mysis 1. Artemia were effectively ingested by P. indicus protozoea 3 (4.1 J larva^?1 d^?1) to post-larva (8.2 J larva^?1 d^?1). Daily energy intake rate from filter feeding increased from 1.1 J larva^?1 d^?1 during protozoea 1 to reach a peak of 5.32 J larva^?1 d^?1 during mysis 3 after which it declined to 2.66 J larva^?1 d^?1 during the post-larval stage. This decline in energy intake from filter feeding with a concomitant increase in energy intake from Artemia predation demonstrates a predominant feeding mode changeover point during mysis 3. Energy intake was consistently low with Brachionus, indicating that it may be unnecessary for commercial culture purposes.     

Characteristics of divalent minerals extracted from liver of Japanese common squid Todarodes pacificus under various experimental conditions

In relation to the problem of how to utilize the waste from squid processing, we determined the proximate and mineral compositions of liver of the Japanese common squid Todarodes pacificus, and characterized the behavior of divalent minerals, focusing in particular on cadmium removal. The squid liver contained fat (44.0 g/100 g dry matter), protein (13.5 g/100 g dry matter), and ash (2.11 g/100 g dry matter). It also contained the macrominerals sodium, potassium, magnesium, and calcium, as well as the trace minerals iron, zinc, cadmium, and copper. Low- and high-pH treatments (pH 2 and 12) removed the cadmium entirely, but the trace minerals zinc, iron, and copper, as well as soluble protein, which are all important for proper functioning of the human organism, were also removed in large quantities. High percentages of soluble minerals and soluble protein were found in the low molecular weight (MW < 10,000) fraction. The solubility of zinc and copper were strongly related to the solubility of cadmium, but there was no relationship between the solubilities of magnesium and cadmium. On the other hand, while treatment with either 0.9 % NaCl or 100 mM phosphate buffer (pH 7.4) also removed the cadmium completely, it left some useful compounds (trace minerals and soluble protein) in the solid part, so these treatments may represent appropriate methods for removing cadmium from squid processing waste.     

重金属离子除去装置在中国对虾育苗生产中的应用

自行研制的高分子吸附剂装置，在两处中等规模的中国对虾育苗场进行了应用实验。结果表明，在其他条件一致的情况下，应用装置与使用自然海水育苗对比，卵子的孵化率平均提高１１．８％，无节幼体到蚤状幼体变态率平均提高１０．２％；应用装置与使用ＥＤＴＡ（２．０ｍｇ／Ｌ）比较，卵子的孵化率平均提高７．５％，无节幼体到蚤状幼体的变态率平均提高３．６％。     

The Effect of Live and Artificial Diets on Growth, Survival, and Trypsin Activity in Larvae of Penaeus indicus

In this study, partial and total replacement of live diets (microalgae and Artemia nauplii) with microencapsulated diets (MED) are demonstrated for larval culture of P. indicus . Slower growth and lower survival rate of larvae fed experimental MED were significantly improved by a supplement of 15 cells/μL frozen mixed algae (1:2, Tetrtaselmis and Skeletonema ) during protod stages (PZ1–PZ3). This low level of algal supplement to MED resulted in survival (85–92%) equal to that obtained from control live diets (91%) during protod stages. These significant improvements in larval growth and survival are likely to be due to higher larval digestive enzyme activities and hence more efficient digestion of the artificial diet by the larvae. Like other penaeids, P. indicus larvae show high total and tissue trypsin activities during PZ stages, with a peak at mysis stage 1 (M1), and a decrease during subsequent stages when fed on conventional live diets of algae followed by Artemia during mysis stages. Larvae fed 15 ceUs/μL mixed frozen algae in addition to MED demonstrated a significantly higher trypsin activity throughout herbivorous larval stages in comparison to larvae fed solely on MED. A freeze dried alga Rhinomonas reticulata incorporated into a MED at 23Vo (v/v) induced larval trypsin activity equal to that produced by live algae. Hence, the algal substances, which trigger digestive enzyme production, may be retained within the microcapsules. At mysis stages, however, addition of live prey (one Artemia/ mL) to cultures fed with MED significantly improved growth and survival although it depressed trypsin activity. For mysis stages it appears that the use of predigested ingredients is necessary to improve the digestibility of formulated diets.     

Induction of metallothionein synthesis in rainbow trout,Salmo gairdneri,during long-term exposure to waterborne cadmium

Rainbow trout were exposed to 200 g cadmium/l in the water during four months at 6–10°C. The liver, kidney and gills were analyzed for cadmium, copper, zinc, metallothionein and metallothionein mRNA. Cadmium accumulated in all three organs and reached the highest concentration in the kidney. The tissue zine and copper concentrations showed no major alterations during the experiment. The cytosolic distribution of cadmium, copper and zinc was followed during four months of exposure by Sephadex G-75 chromatography. It was found that cadmium was predominantly associated with proteins of an apparent molecular vieght of 10,000 daltons. These proteins were further identified as metallothioneins after fast protein liquid chromatography on a Mono-Q column. The metallothionein concentration was significantly higher in liver of exposed fish than in control fish after only one month. The kidneys reached significantly elevated levels of metallothionein in the exposed group after three months. In the gills, elevated metallothionein concentrations were observed after four months of exposure. After four months of exposure, the metallothionein mRNA content of liver and kidney was analyzed using a rainbow trout anti-sense RNA probe. Elevated MT mRNA levels were observed in both kidney and liver. These results demonstrate thatde novo synthesis of metallothionein is induced by cadmium in rainbow trout after exposure to the metalvia water.     

Effect of Lead on Survival,Osmoregulation, and Histological Changes of the Gills of the White Shrimp,Litopenaeus vannamei,Boone, 1931

The purpose of this study was to evaluate the acute toxicity of lead for different life stages of shrimp, Litopenaeus vannamei, and to determine the effect of its sublethal concentrations on osmoregulatory capacity (OC) as well as the possible histological alteration in the gills of juvenile shrimp. The 24‐, 48‐, and 96‐h LC₅₀ values for lead to L. vannamei increased progressively with increasing life stage, from nauplii < zoeae < mysis < postlarvae < juvenile. After 15‐d exposure to 3.25, 6.50, and 9.75 mg Pb/L, the OC values of exposed shrimp were reduced by 39, 73, and 157%, respectively compared to control animals. Compared to controls, the lead concentrations in gill tissues increased significantly by 127,500, 137,500, and 141,000% in shrimps exposed to 3.25, 6.50, and 9.75 mg Pb/L, respectively. After lead exposure, hemocytic congestion in efferent vessels and multiple hyperplasia were observed in gill filaments, resulting in narrowed hemolymphatic lacuna. The histopathological effects increased with increasing Pb concentration.     

Specific accumulation of heavy metals in squid collected from offshore Korean waters: preliminary results for offshore biomonitoring and food safety assessment

ABSTRACT:   Japanese common squid Todarodes pacificus collected from the East Sea/Sea of Japan were analyzed for cadmium, zinc and copper to elucidate the specific accumulation of heavy metals in various organs of squid of varying size and sex. There was no relationship between the size and sex of squid and the concentrations of heavy metals in various organs. Amongst organs, the liver contained the highest concentrations of all metals and the mantle had the lowest concentrations. In the liver and gills, concentrations of copper were higher than the other metals because of the existence of copper in hemocyanin in the blood. Cadmium showed the highest ratios (L/M) of the concentrations in the liver and in the mantle, even though cadmium in liver had lower concentrations than copper and zinc. This indicates that cephalopods constitute an important source of cadmium for cephalopod predators. The results show that the Japanese common squid is a useful bioindicator of metal pollution in offshore Korean waters.     

The Larval Development,Metamorphosis and Juvenile Growth of the Turban Snail Lithopoma (Astraea) undosa (Wood, 1828) (Gastropoda:Turbinidae)

This work presents the embryonic, larval, and postlarval development of Lithopoma undosa (L. undosa) (20 ± 2 C and the effect of natural inducers on the larval metamorphosis. Embryonic development until hatching of the trochophore larva took 15 h. Early veliger larvae with well‐developed shell were after 24 h. At 48 h, the operculum was completely formed. Cephalic tentacles were visible after 72 h. Larvae reached competence 6 to 7 d after fertilization, and was characterized by the presence of branched cephalic tentacles. The effect of natural inducers was evaluated. The percentage of metamorphosis with diatoms and adult mucus was 94% and 58% respectively, differing from the 25% without inducer. When was repeated, the metamorphosis was 100%, and 16% with benthic diatoms. The initial mean size of the postlarvae was 0.316 ± 0.02 mm. After 264 d the juveniles had attained a mean size of 5.98 ± 0.16 mm. These results will not only be of use in the evaluation of the culture of L. undosa, but also in studies on the characterization of inducers.     

Susceptibility of steelhead trout Salmo gairdneri Richardson to redmouth infection Yersinia ruckeri following exposure to copper

Abstract. Steelhead trout were exposed to sublethal concentrations of copper and inoculated with Yersinia ruckeri. Copper exposure at 7 and 10 μg/1 for 96 h caused more fish to die of infection than control fish (no copper). Infection susceptibility increased with time of exposure to a single dosage of copper (10 μg/1), reaching a maximum at 48 h. Lowering the copper concentration to 5 μg/1 caused the infection susceptibility to occur at 24 h. The infectious dose of Y. ruckeri was lower in fish exposed to 10 μg/1 copper for 48 h than control fish.     

Evaluation of the Dietary Zinc Requirement of Penaeus vannamei and Effects of Phytic Acid on Zinc and Phosphorus Bioavailability

Experiments were conducted to determine the dietary zinc requirement of Penaeus vannomri and evaluate the effects of phytate on zinc bioavailability. Prior to initiation of the growth trial, 20-day-old P. vonnamei postlarvae (mean weight 0.0032 g) were fed a casein-gelatin based semi-purified diet lacking zinc supplementation but containing 18 mg Zn/kg diet for one week. Subsequently, juveniles (mean weight 0.058 g) were fed one of seven diets containing either supplemental zinc (0, 15, 30, 60 mg/kg diet) without phytate or supplemental zinc (0, 60, 200 mg/kg diet) with 1.5% phytate for 33 days. Weight gain was greatest in shrimp fed 15 mg supplemental Zn/kg diet. In the absence of dietary phytate, zinc concentrations in the hepatopancreas of shrimp were maximized when zinc was supplemented at levels greater than or equal to 15 mg Zn/kg diet (33 mg total Zn/ kg). Supplementation of 1.5% phytate to the diet did not have a significant effect on growth or zinc concentrations in the carapace; however, it did depress zinc levels in the hepatopancreas. Supplementation of 200 mg Zn/kg diet was required to overcome the depressed bioavailability of zinc caused by the presence of dietary phytate and return zinc levels of the hepatopancreas to that observed when phytate was not present. Based on apparent digestibility values phytate phosphorus was unavailable to the shrimp and the presence of phytate depressed the bioavailability of phosphorus and zinc.     

DECREASED TOXICITY OF COPPER AND MANGANESE IONS TO SHRIMP NAUPLII (Penaeus stylirostris STIMPSON) IN THE PRESENCE OF EDTA

Although ethylenediaminetetraacetic acid (EDTA) is used routinely in the intensive culture of penaeid shrimp larvae to increase both the percentage of eggs that hatch and the survival of larvae, the mode of action is not known. To explain EDTA's beneficial effect this study compares the toxicities of copper and manganese divalent ions in the presence and absence of 10 mg EDTA/liter seawater. Toxicity was evaluated in terms of the percentage of Penaeus stylirostris nauplii surviving after 12 and 24 hours of exposure to either copper or manganese and in terms of the percentage of nauplii that metamorphose to the protozoeal stage. In the absence of EDTA, copper and manganese ions were toxic to nauplii with copper causing 100% mortality at a much lower concentration (20 μM) than manganese (20,000 μM). In the presence of EDTA, survival of nauplii exposed to copper was increased but survival of nauplii exposed to manganese was not affected. At concentrations lower than the levels that reduce survival, copper ions (0.2 μM) and manganese ions (2 μM) reduce the percentage of nauplii that metamorphose to protozoea. In the presence of EDTA, the percentage of nauplii that metamorphose to protozoea is increased with both nauplii exposed to copper ions and nauplii exposed to manganese ions. The beneficial effects of EDTA are probably due to the chelation of copper and manganese ions by EDTA. Chelation lowers the free concentration of these ions and thus reduces their toxicities.     

Ultrasound Enhanced Immersion Protocols for Masculinization of Nile Tilapia, Oreochromis niloticus

Androgen immersion protocols have been unsuccessful in consistently producing all-male tilapia at a high enough ratio for them to be commercially viable. This study explored the use of ultrasound to improve on the results of previous immersion studies. Variables tested include two hormones (trenbolone acetate-TBA and 17α-methyldihydrotestoterone-MDHT) at two concentrations (100 and 250 μg/L) and with or without ultrasound (cavitation level). All hormone treatments with ultrasound and non-ultrasound resulted in significantly higher masculinization than the appropriate controls ( P < 0.05). Among pairs of treatments of the same hormone at the same dose, all ultrasound treatments resulted in significantly higher number of males compared with non-ultrasound treatments with the exception of MDHT 250 μg/L ( P 0.05). Comparing across all ultrasound treatments, TBA 250 μg/L with ultrasound had higher masculinization than all the other ultrasound treatments ( P 0.05). Comparing across all non-ultrasound treatments, TBA 250 μg/L had higher ( P 0.05) masculinization than MDHT 100 μg/L and TBA 100 μg/L with non-ultrasound. Two of the three replicates of TBA 250 μg/L ultrasound treatment resulted in 100% males and the highest mean percentage (98%) of males. This study thus demonstrated the potential of a short-term immersion protocol using ultrasound to more predictably produce all-male, commercially viable tilapia seed.     

栉孔扇贝对铜、铅、镉的累积效应

试验结果表明:当常规理化指标一致时,随着Cu、Pb、Cd质量浓度的升高,栉孔扇贝各组织的蓄积量明显升高;Cu、Cd在扇贝组织中蓄积量是内脏团>鳃>肌肉,Pb在扇贝组织中的蓄积量是鳃>内脏团>肌肉;当Cu、Pb、Cd质量浓度(0.1 mg/L)相同时,其鳃、内脏团、肌肉中重金属蓄积量的上升速率是Cu>Cd>Pb。     

The Toxicity of Erythromycin, Minocycline, Malachite Green, and Formalin to Nauplii of the Shrimp Penaeus stylirostris

The toxicities of erythromycin, minocycline, malachite green, and formalin to nauplii of the shrimp Penaeus stylirostris were determined in a static bioassay. Toxicity was assessed on the basis of survival of nauplii after 12 and 24 hours of exposure to the compounds and metamorphosis of the nauplii to protozwa. The results suggested that metamorphosis to protozoea is more susceptible to toxic effects than is naupliar survival. Metamorphosis was either reduced or completely inhibited by lower concentrations of erythromycin, minocycline, and malachite green than was naupliar survival at 12 or 24 hours. Metamorphosis was either reduced or completely inhibited by erythromycin, minocycline, malachite green, and formalin concentrations of 80, 100, 0.08, and 27 mg/ L, respectively. Toxic effects were not observed at erythromycin, minocycline, malachite green, and formalin concentrations of 16, 62.5, 0.016, and 2.7 mg/L, respectively. The results suggest that formalin may be toxic at therapeutic levels frequently recommended for post larvae and older penaeids, but that erythromycin, minocycline and malachite green are not.     

The Effect of Depuration on Metals,Bacteria, and Nutrition in Venus verrucosa from the Aegean Sea: Benefit and Risk for Consumer Health of Warty Venus

ABSTRACT

In this study, the effect of the depuration period on proximate composition, metals, fatty acids, and bacteria in the edible tissue of warty venus (Venus verrucosa) collected from ?zmir Bay in the Aegean Sea was investigated. In addition, benefit and risk for human health associated with the consumption of warty venus were determined by atherogenic index (AI), thrombogenic index (TI), polyene index (PI), fish lipid quality (FLQ), target hazard quotient (THQ), and hazard index (HI). The depuration process did not show a significant (P > .05) effect on proximate composition, except for ash. The depuration process was successful in reducing bacteria growth and the concentrations of all metals, except for copper. Total aerobic mesophilic bacteria count of 1.94 log CFU/g in the fresh samples was uncountable at 24 h of depuration. The concentrations of cadmium, lead, copper, iron, and zinc were below the limit values allowed for safe consumption. The analyzed metals were safe for human consumption based on provisional tolerable weekly intake (PTWI), THQ, and HI. Significant variations were also observed in the fatty acid values depending on the depuration periods (P < .05). The n3/n6 and polyunsaturated/saturated fatty acid ratios were greater than those recommended by the FAO/WHO. Moreover, the AI and TI of samples were under established limits.