Effects of stocking density and water conditioners on yolk‐sac larvae of two marine finfish during simulated air transport

As marine finfish aquaculture expands, there is an increasing interest in the ability to ship early life stages from breeding centres to hatcheries so that each hatchery does not have to maintain its own broodstock. Here, we conducted 24 h air‐shipping simulations with yolk‐sac larvae of California yellowtail (CYT; Seriola lalandi) and white seabass (WSB; Atractoscion nobilis) to help fill in the informational gaps for shipping marine fish larvae. We examined the effects of a pH buffer on water quality, post‐shipping larval survival and subsequent survival to first feeding at larval densities of 1000, 3000, 6000 and 9000 larvae L^?1. The pH buffer, 8.3 Trizma^®, was tested at varying concentrations of zero (NT = 0.00 g L^?1), low (LT = 0.75 g L^?1), medium (MT = 1.5 g L^?1) and high (HT = 3.0 g L^?1). Trials were conducted using replicate 2 L aquarium bags filled with 500 mL of seawater and held in a water bath at 19–20°C. Results showed an interspecific difference in survival at the highest shipping densities under these experimental conditions. Shipping densities up to 6750 CYT larvae L^?1 or 3000 WSB larvae L^?1 consistently yielded >90% survival immediately after simulated shipment and >85% survival 48 h after the simulations. Furthermore, at these densities, pH was maintained at ~8.0 when buffered at 1.5 g L^?1. The highest tested densities of 9580 CYT larvae L^?1 and 9940 WSB larvae L^?1, yielded lower survival 69–79% and 0.0–1.3% respectively after 24 h. Final pH in the high density CYT trials were unsatisfactory (below 7.0), regardless of the buffer concentration; however pH in the WSB high density trials improved with increasing buffer concentration. On the basis of the results from these air‐shipping simulations, we recommend CYT and WSB larvae be shipped in seawater with 1.5 g L^?1 Trizma^® at densities not greater than 6750 larvae L^?1 for CYT and 3000 larvae L^?1 for WSB. We believe this represents an important step in improving long distance transport protocols for these species and provides useful guidance in air transport of other economically and ecologically important marine species. Additional research is warranted to compare these simulation results with those from actual air shipments, as we did not account for factors that may vary in flight like temperature and pressure variations, and physical agitation.     

Efficacy of formalin,iodine and sodium chloride in improvement of egg hatching rate and fry survival prior to the onset of exogenous feeding in yellow perch

Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L^?1) and 25 mg L^?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L^?1), iodine at three concentrations (50, 100 and 250 mg L^?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L^?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L^?1 formalin for 30 min, 50 mg L^?1 iodine for 15 min and 500 mg L^?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L^?1), iodine (250 mg L^?1) and sodium chloride (1000 and 3000 mg L^?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L^?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.     

Chemical surface disinfection of eggs of Baltic cod,Gadus morhua L.

The effect of two disinfectants on eggs and larvae of Baltic cod, Gadus morhua, was investigated. The eggs were disinfected for 10 min using various concentrations of either glutaraldehyde (100, 200, 400, 600 and 800 mg L^?1) or iodophor (10, 50, 100 and 150 mg L^?1), 1–4‐days post‐fertilization. Bactericidal effect of disinfection, survival to hatching, hatching success and larval abnormalities were assessed. Larval survival was recorded at 5‐, 10‐ and 15‐days post‐hatch (dph). Although Baltic cod eggs have an unusually thin chorion, they could tolerate surface disinfection. A reduction in bacterial growth was observed with increased concentrations of disinfectant (3.0 × 10⁷–1.6 × 10¹ CFU mL^?1). Abnormalities in newly hatched larvae were not related to disinfection. Survival of the yolk sac larvae was significantly better for eggs treated with 400 mg L^?1 glutaraldehyde for 10 min at 10 and 15 dph. Effective disinfection was also recorded using 100 mg L^?1 Actomar K30. Egg batch effect rather than initial bacterial concentration, disinfectant type or incubation method determined the survival of the eggs to hatching and survival of larvae. Because of the carcinogenic effect of glutaraldehyde, iodophor is recommended for routine disinfection of cod eggs.     

Light intensity effects on early life stages of black sea bass, Centropristis striata (Linnaeus 1758)

The effects of four light intensities on growth and survival of first‐feeding stage black sea bass larvae Centropristis striata were investigated in a controlled‐environment laboratory. Fertilized eggs, obtained from LHRHa‐induced spawning of captive broodstock, were stocked (72 eggs L^?1) into twenty 15 L black tanks under light intensities of 100, 500, 1000 and 1500 lx, with five replicate tanks per treatment. The photoperiod was 12L:12D, the temperature was 20°C and the salinity was 35 g L^?1. Larvae were fed rotifers Brachionus rotundiformis from day 2 post‐hatching (d 2ph) at 5–10 rotifers mL^?1. Microalgae Nannochloropis oculata and Isochrysis sp. were added (1:1) daily to maintain a density of 300 000 cells mL^?1. Hatching success and larval growth and survival from d 2ph through d 15ph were monitored. Hatching success was 28–38% under all light intensities, and notochord length at hatching ranged from 2.8 to 3.0 mm, with no significant differences among treatments. By d 15ph, growth (mg wet weight) was significantly higher in the 1000 lx (0.914) and 1500 lx treatments (0.892) than in 100 lx (0.483), and a highly significant trend (P<0.01) towards increased survival with increasing light intensities was observed, from 1.3% at 100 lx to 13.9% at 1500 lx. Higher light intensities within the range of 100–1500 lx improved growth and survival of early larval black sea bass, suggesting that even higher light intensities may improve culture performance. This is consistent with conditions in shallow, near‐shore locations where eggs and larvae are distributed in nature.     

Disinfection of almaco jack (Seriola rivoliana Valenciennes) eggs: Evaluation of three chemicals

Almaco jack (Seriola rivoliana Valenciennes) is an excellent candidate for aquaculture due to its fast growth rate and high market value. While S. rivoliana have adapted well to captivity, survival at early life stages can be improved to increase profitability during production. A wide range of variables cause larval mortalities but high bacterial loads in rearing tanks are often correlated with these losses. The aim of this study was to investigate the effect of egg disinfection on bacterial load and hatch rate of S. rivoliana. Disinfectants tested included formalin (F100 and F200; 100 and 200 mg/L, respectively, for 60 min), hydrogen peroxide (HPO; 300 mg/L for 10 min) and peracetic acid/hydrogen peroxide (PAA/HPO; 15.7 mg/L/39.6 mg/L for 1 min). Concentrations and contact times were determined based on current use in marine aquaculture and preliminary trials. Eggs treated with HPO and F100 had significantly higher hatch rates than the untreated control group. All treatments significantly decreased total Vibrio counts compared to untreated eggs; however, total bacterial counts were only decreased following treatments with PAA/HPO and F200. To prevent egg mortality due to bacterial overgrowth, consideration should be given to the use of surface disinfection using HPO or F100. Future studies should investigate the use of peracetic‐based products at lower doses.     

Efficacy of clove oil and ethanol against Saprolegnia sp. and usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss (Walbaum) eggs

Inhibitory concentrations of clove oil and ethanol against growth of Saprolegnia sp. hyphae were screened by a modification of the hemp (Cannabis sativa L.) seed MicroPlate (HeMP) method and their usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss eggs was tested. In vitro experiment showed that in continuous static exposure, clove oil at 100 mg L^?1 significantly inhibited the growth of Saprolegnia, whereas in bath exposures, clove oil at 500 mg L^?1 had no significant effect at any exposure time tested (15, 60 and 240 min), but clove oil at 10 000 mg L^?1 significantly inhibited growth at all exposure times. Clove oil and ethanol treatments had no visible effects on the onset or spread of the fungus during incubation of rainbow trout eggs. Clove oil at 1000 mg L^?1 resulted in 95–100% mortality before the eyed stage was reached. Sublethal concentrations of clove oil and ethanol had no effects on the development rate of the embryo or growth and yolk utilization efficiency after hatching. This study suggests that clove oil and ethanol may not be options in controlling aquatic fungi infestations during incubation of rainbow trout eggs.     

In vitro and in vivo efficacy of drugs against the protozoan parasite Azumiobodo hoyamushi that causes soft tunic syndrome in the edible ascidian Halocynthia roretzi (Drasche)

It was discovered recently that infection by a protozoan parasite, Azumiobodo hoyamushi, is the most probable cause for soft tunic syndrome in an edible ascidian, Halocynthia roretzi (Drasche). In an attempt to develop measures to eradicate the causative parasite, various drugs were tested for efficacy in vitro and in vivo. Of the 20 antiprotozoal drugs having different action mechanisms, five were found potent (24‐h EC₅₀ < 10 mg L^?1) in their parasite‐killing effects: formalin, H₂O₂, bithionol, ClO₂ and bronopol. Moderately potent drugs (10 < 24‐h EC₅₀ < 100 mg L^?1) were quinine, fumagillin, amphotericin B, ketoconazole, povidone‐iodine, chloramine‐T and benzalkonium chloride. Seven compounds, metronidazole, albendazole, paromomycin, nalidixic acid, sulfamonomethoxine, KMnO₄, potassium monopersulphate and citric acid, exhibited EC₅₀ > 100 mg L^?1. When ascidians were artificially infected with A. hoyamushi, treated using 40 mg L^?1 formalin, bronopol, ClO₂, or H₂O₂ for 1 h and then monitored for 24 h, very low mortality was observed. However, the number of surviving parasite cells in the ascidian tunic tissues was significantly reduced by treating with 40 mg L^?1 formalin or ClO₂ for 1 h. The data suggest that we might be able to develop a disinfection measure using a treatment regimen involving commonly available drugs.     

Inhibition of biofilm bacteria and adherent fungi from marine plankton cultures using an antimicrobial combination

The presence of organic matter in plankton cultures will lead to 10- to 1000-fold increases in bacterial density in less than 24 h. The presence of bacteria and fungi can damage cultivated phytoplankton and zooplankton. These microorganisms can also inhibit experiments investigating the role of these microorganisms in the community and in biological and ecological laboratory studies. The aim of this study was thus to evaluate the effect of penicillin?+?streptomycin?+?neomycin (antibiotics) in combination with nystatin (antifungal) to select an antimicrobial combination for the inhibition of biofilm bacteria and adherent fungi that is effective and also non-toxic to marine phytoplankton and zooplankton. Acartia tonsa was exposed to different antimicrobial treatments and application routes (culture medium, culture food, both) to evaluate the survival and egg and fecal pellet production endpoints. The same treatments were also applied to measure Amphibalanus improvisus survival and settlement and Conticribra weissflogii growth endpoints. We selected the most sensitive experimental organism and exposed it to some novel antimicrobial combinations. To evaluate the inhibition potential, biofilm bacteria and adherent fungi were exposed to the treatments that were safe for the bioindicator species. A tonsa was considered the most sensitive of all tested organisms. The treatment composed of 0.025 g L^?1 penicillin G potassium?+?0.08 g L^?1 streptomycin sulfate?+?0.04 g L^?1 neomycin sulfate showed the best results for A. tonsa and C. weissflogii cultures. No differences were observed for A. improvisus between the treatments. A. tonsa survival rates showed no differences from the control at nystatin concentrations up to 0.005 g L^?1 in combination with the antibiotics. The biofilm bacterial density decreased up to 94% under this treatment, and fungal growth was prevented. Species of other planktonic groups should also be tested to improve our understanding of the effectiveness of the treatments proposed.

     

Effect of different total suspended solids levels on a Litopenaeus vannamei (Boone, 1931) BFT culture system during biofloc formation

In a Biofloc Technology System (BFT), there is constant biofloc formation and suspended solids accumulation, leading to effects on water quality parameters that may affect the growth performance of cultured shrimp. This study aimed to analyse during biofloc formation the effect of different total suspended solids (TSS) levels on water quality and the growth performance of Litopenaeus vannamei shrimp in a BFT system. A 42‐day trial was conducted with treatments of three ranges of TSS: 100–300 mg L^?1 as low (TL), 300–600 as medium (TM) and 600–1000 as high (TH). The initial concentrations of 100 (TL), 300 (TM) and 600 mg L^?1 (TH) were achieved by fertilization before starting the experiment. Litopenaeus  vannamei juveniles with an average weight of 4.54 ± 1.19 g were stocked at a density of 372 shrimp m^?3. Physical and chemical water parameters and shrimp growth performance were analysed. After 6 weeks, TSS mean concentrations were 306.37, 532.43 and 745.2 mg L^?1 for, respectively, TL, TM and TH treatments. Significant differences (P < 0.05) were observed in TSS, settleable solids, pH, alkalinity and nitrite, especially between the TL and TH treatments. Similarly, differences (P < 0.05) were observed in the growth performance parameters, specifically final weight, survival, feed conversion and productivity. The water quality parameters at lower range of total suspended solids concentration (TL) treatment resulted in a better performance of L. vannamei in the BFT system. The maintenance at range of 100–300 mg L^?1 TSS is thus important to the success of shrimp culture.     

Use of Ox‐Aquaculture© for disinfection of live prey and meagre larvae,Argyrosomus regius (Asso, 1801)

Live prey used for marine larval fish (rotifers and Artemia) as well as intensive larval rearing conditions are susceptible to the proliferation of bacteria that are the cause for reduced growth and larval mortality. Hydrogen peroxide has been recently proved a good disinfectant in aquaculture, either for eggs, larvae or live prey. In this study the effects of a hydrogen peroxide‐based product, Ox‐Aquaculture^©, on live prey (rotifers and Artemia) and meagre larvae bacterial load, composition and final status have been tested. A 34.6% reduction of total heterotrophic bacteria and 59.7% of Vibrionaceae were obtained when rotifers were exposed for 15 min to 40 mg L^?1 of the product. A 34.3% reduction of total heterotrophic bacteria and 37.7% of Vibrionaceae were obtained when Artemia were exposed for 5 min to 8000 mg L^?1 of the product. More than 95% reduction of total heterotrophic bacteria and 75% of Vibrionaceae were obtained when meagre larvae were exposed for 1 h to 20 mg L^?1 of the product. Furthermore, disinfection of enriched live prey with the product did not change the fatty acid composition and survival of the live prey and improved final larval survival.     

Bacterial load reduction of live prey for fish larval feeding using Ox-Aquaculture©

Reduction of the bacterial load in live prey is paramount to achieving efficient microbiological control during fish larval rearing. Several methods have been tested using physical and chemical disinfection procedures. Nevertheless, chemicals are difficult to deal with because of the frequent problems encountered when disposing of their residual products and because of the sensitivity of the live prey. Hydrogen peroxide is a good disinfectant that has been used for a long time in other fields of research and decomposes in non‐toxic products. However, it has not been applied for disinfection in aquaculture until recent years. The effects of a hydrogen peroxide‐based product, Ox‐Aquaculture^©, on rotifers and Artemia nauplii survival and on their associated microbial population have been tested in the present study, a disinfection protocol is proposed, and its effects on live prey fatty acids composition after enrichment were checked. More than 80% survival and a 90% reduction of total heterotrophic bacteria and Vibrionaceae were obtained in rotifers exposed for 15 min to 40 mg L^?1 of the product. In the case of Artemia nauplii, 90% survival and 94.5% reduction of heterotrophic bacteria were obtained after 5 min exposure to 8000 mg L^?1 with a further 82.8% reduction of Vibrionaceae. No differences in the fatty acid composition of the live prey were detected after disinfection with hydrogen peroxide. No oxidation of polyunsaturated fatty acids was observed. A short time exposure and easy removal of the product from the treated live prey make hydrogen peroxide an interesting chemical for industrial application.     

Use of formalin and copper to control ichthyophthiriosis in the Australian freshwater fish silver perch (Bidyanus bidyanus Mitchell)

Infestations of the protozoan parasite, Ichthyophthirius multifiliis, cause the serious disease ichthyophthiriosis in freshwater fish throughout the world. Formalin is a recommended treatment for ichthyophthiriosis in the Australian fish silver perch (Bidyanus bidyanus Mitchell), but the disease is difficult to control in ponds, particularly at low water temperatures. Experiments were carried out to develop an improved treatment regime for formalin and to evaluate copper as a therapeutant. Silver perch fingerlings infested with I. multifiliis were stocked into 55 L aquaria at temperatures of 14.8–17.6 °C and alkalinities of 70–110 mg L^?1. Formalin (34–38% formaldehyde) or copper (24.5% copper sulphate) were added to the aquaria and then monitored and readjusted to nominal concentrations daily. A concentration of 30 mg L^?1 formalin controlled ichthyophthiriosis, but fish treated with 20 mg L^?1 remained infested with theronts and trophonts on day 17; survival at both concentrations was 100%. A concentration of 10 mg L^?1 formalin did not control ichthyophthiriosis and all fish were dead from the infestation by day 17. Fish treated with 0.1 or 0.2 mg L^?1 copper were free of theronts and trophonts by days 17 and 14, respectively, and survival was 100%. Survival at 0.05 mg L^?1 copper was 100%, but fish remained infested. At 0.25 mg L^?1 copper, survival was 82.5% and there were no theronts or trophonts on gill and skin tissues of fingerlings by day 14. There was total mortality of fish treated with 0.5 or 1.0 mg L^?1 copper suggesting these concentrations are toxic to silver perch. All fish in infested‐control treatments died. In earthen ponds containing silver perch, 0.2 mg L^?1 copper was depleted to below 0.1 mg L^?1 within 24 h, and concentrations of 25–38 mg L^?1 formalin were depleted to below 15 mg L^?1 within 48 h. Treatment regimes involving daily applications of formalin or copper controlled ichthyophthiriosis in silver perch in earthen ponds at costs of $US466.37 and $US65.58 hectare^?1 day^?1 respectively. This study has developed a new formalin‐treatment regime for the control of ichthyophthiriosis, and demonstrated that copper sulphate is a potential therapeutant for this serious disease of silver perch.     

Effect of calcium and magnesium hardness on acute copper toxicity to Indian major carp, Labeo rohita (Hamilton) and catfish, Channa punctatus (Bloch)

Four experiments were conducted to evaluate the effects of calcium and magnesium hardness on the acute toxicity of copper sulphate to Indian major carp, rohu (Labeo rohita, Hamilton) fingerlings and juvenile catfish (Channa punctatus, Bloch) in medium alkalinity experiments. A preliminary bioassay determined the 96 h LC₅₀ of copper sulphate to be 0.56 mg L^?1 for L. rohita fingerlings and 11.78 mg L^?1 for juvenile C. punctatus placed in water with calcium hardness and total alkalinity set at 100 mg L^?1 CaCO₃. In the first experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 350 mg L^?1 CaCO₃ and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced rohu mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 7% at 350 mg L^?1 CaCO₃. In the second experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 350 mg L^?1 CaCO₃ with total alkalinity set at 100 mg L^?1 CaCO₃. Hundred percent mortality was observed in magnesium‐based hardness treatments. In the third experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 400 mg L^?1 and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced catfish mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 4% at 400 mg L^?1 CaCO₃. In the fourth experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 400 mg L^?1 CaCO₃, with total alkalinity set at 100 mg L^?1 CaCO₃. In this case, 100% mortality was also observed in magnesium‐based treatments. Mortality rates in magnesium hardness treatments were consistent with those in the second experiment. These data suggest a calcium‐specific mechanism with respect to acute copper toxicity both in rohu and catfish.     

Alternative protein blends as a replacement for fish meal in diets for white seabass,Atractoscion nobilis

An 8‐week feeding trial was conducted to determine the effectiveness of replacing fish meal (FM) with blends of alternative proteins in diets for white seabass (WSB, Atractoscion nobilis) at a starting weight of 5.6 g. Five diets were formulated with 400–440g kg^?1 crude protein (380g kg^?1 digestible). These included a high 520g kg^?1 FM control diet, a series of three diets with a sequential replacement of FM containing 410g kg^?1, 510g kg^?1 and 630g kg^?1 of a soy‐based protein blend (SPC) and 200g kg^?1, 100g kg^?1 and 0g kg^?1 FM, respectively and a fifth diet containing 550g kg^?1 of a corn‐based protein blend (CGM) and 100g kg^?1 FM. Survival was highest in the FM control group at 99% but all other performance measures (weight gain, feed conversion ratio, specific growth rate and protein retention efficiency) were worse than the other treatment groups. Weight gain reached a maximum of 595% in the SPC 200g kg^?1 FM treatment group. Performance decreased as inclusion of the soy‐based protein blend increased. The CGM treatment performed comparably to the SPC 100g kg^?1 FM treatment among all measures, except for survival, which was higher in the CGM 100g kg^?1 FM treatment. With nutrient levels and alternative protein blends used in this study, FM can be reduced to 100g kg^?1 of the diet for WSB without reductions in performance.     

Effects of compost manure on the body burden of cadmium and growth of common carp, Cyprinus carpio (L.)

One-hundred-and-eighty-day bioassays were made in outdoor cement tanks (volume 300 L) with common carp, Cyprinus carpio (L.), exposed to eight different treatments, each with three replicates. Four treatments contained 2.5 mg L^?1 cadmium (Cd): Cd alone; Cd with 6.7 g L^?1 of compost manure made from the freshwater macrophyte Pistia stratiotes (L.); Cd with 1.0 g L^?1 of the same manure; and Cd with a cumulative treatment of the manure (1.0 g L^?1 month^?1). Four treatments contained no Cd: the control (without manure); and treatments with only manure (6.7 g L^?1,1.0 g L^?1 and 1.0 g L^?1 month^?1). The concentration of dissolved (filterable) Cd in water ranged from 0.74 to 0.89 mg L^?1 after 24 h (one day) of treatment. The addition of compost manure significantly reduced the Cd concentration of the water and induced a quicker removal of Cd from the water. The quickest removal of Cd from water (60 days) was found when a high dose of compost manure (6.7 g L^?1) was added. The cumulative (monthly) treatment and the one-time treatment of the manure at a low dose (1.0 g L^?1) required 75 and 90 days, respectively, for removal of Cd, while the metal persisted for up to 120 days in water when no manure was added. All doses of compost manure significantly reduced the bioaccumulation of Cd in plankton, but maximum reduction was found under cumulative treatment at a low dose of the manure. A single treatment at a high dose or cumulative treatment at a low dose of the manure helped in reducing the total body burden of Cd. A dose of 2.5 mg L^?1 Cd did not show any adverse effect on the growth of fish, but compost manure, irrespective of its dose and combination with Cd, markedly increased the growth of fish and primary productivity of water. On the other hand, a high dose of manure resulted in much higher levels of NH₃-N, N0₂-N and P0₄-P in the water in comparison with other treatments     

Evaluating bioencapsulation of florfenicol in rotifers (Brachionus plicatilis)

The rotifer Brachionus plicatilis is one of the most important live‐feed organisms used in the larval culture of marine fish. Bioencapsulation of florfenicol in rotifers was investigated by delivering it directly to the organisms as particles and the doses ranged from 50 to 1000 mg L^?1. Analysis of the florfenicol concentrations in rotifers was performed using high‐performance liquid chromatography. The uptake increased with the dose and the enrichment time, and the highest concentration achieved in this study, 1.25 ng rotifer^?1, was obtained using a dose of 1000 mg L^?1 and 150 min of enrichment. The optimal time for satisfactory enrichment of rotifers with florfenicol was found to be dose dependent and ranged from 30 to 120 min for doses between 400 and 800 mg L^?1. The parameters affecting the concentrations achieved in rotifers were the size of the florfenicol particles, enrichment time and dose.     

Effects of crushed conspecifics on growth and survival of Penaeus monodon Fabricius post larvae

In the present study, the hypothesis tested was that Penaeus monodon post‐larvae (PL) experience lower growth when exposed to crushed conspecifics, which was achieved by exposing individual P. monodon PL with abundant food for 4 weeks to a gradient from 0 to 100 crushed conspecific PL L^?1. Both dry weight (48.5±7.2 mg) and body size (28.0±1.3 mm) of animals exposed to 1 macerated PL L^?1 were significantly (P≤0.011) higher than those of animals in treatments with 0, 5 and 10 crushed PL L^?1 (average over treatments: 25.6±3.2 mg; 23.4±0.5 mm). All animals died within 1 week when exposed to 70 and 100 crushed PL L^?1, and within 3–4 weeks when exposed to 50 and 30 crushed PL L^?1. Exposure time affected mortality and it appeared that LC₅₀ values decreased from 60 to 13 crushed PL L^?1 from 1 to 4 weeks' exposure. Survival of P. monodon PL was negatively correlated to pH, biological oxygen demand, ammonia and nitrate. In conclusion, low dose of crushed conspecifics has a stimulatory effect on P. monodon PL, as larvae were heavier and larger, while high doses cause high mortality.     

Alcalase treatment for elimination of stickiness in pikeperch (Sander lucioperca L.) eggs under controlled conditions

Elimination of egg stickiness is an important factor in artificial reproduction of pikeperch (Sander lucioperca L.). This study was conducted to evaluate the effectiveness of Alcalase enzyme to remove the adhesive layer of pikeperch eggs. The eggs were treated with Alcalase at 0.5, 1.0, 1.5, 2.0 and 5.0 mL L^?1 or a milk/talc solution 2 min post insemination. Duration of exposure was 2 min in Alcalase and 60 min in milk/talc. The highest, albeit not significant, hatching rate (85.4%) was found with 1.5 mL L^?1 of Alcalase, but hatching rates were similar in 0.5, 1.0 and 2.0 mL L^?1. Hatching rates were significantly lower groups treated with 5.0 mL L^?1 Alcalase enzyme (56.4%) compared to groups treated with milk powder and talc (61.3%). Nominally complete removal of adhesiveness was observed in 1.5 and 2 mL L^?1. All Alcalase treatments led to significantly lower incubation duration compared with the traditional milk/talc treatment. The application of Alcalase successfully eliminated pikeperch egg stickiness in less time than with traditional milk/clay/talc methods.     

Effects of egg stocking density on egg hatchability,larval quality and water quality for pinfish,Lagodon rhomboides,and pigfish,Orthopristis chrysoptera

Pinfish (Lagodon rhomboides Linnaeus) and pigfish (Orthopristis chrysoptera L.) are relatively new, cultured species commonly used as marine baitfish in the south‐eastern United States and currently have no defined protocols for egg incubation. Thus, experiments were conducted to determine efficient egg stocking densities during static and flow‐through incubation that yielded higher quality larvae. Eggs of each species were incubated statically at 250, 500, 1000 and 2000 eggs L^?1, and after incubation, egg hatching success, larval survival to first feeding, larval morphometrics and water quality were assessed. Stocking densities above 250 eggs L^?1 led to significant reductions in pinfish hatching success and water quality degradation, evident from decreased dissolved oxygen and pH levels and increased nitrogenous wastes. Increased stocking densities for pigfish also resulted in significant water quality degradation, although hatching success was unaffected up to a density of 1000 eggs L^?1. A high flow‐through water exchange rate of 2000% daily resulted in significant reductions in nitrogenous wastes and greater stability in dissolved oxygen and pH levels during incubation when compared to static treatments of the same egg density. Additionally, the high exchange rate of 2000% was critical in maintaining high hatching success and larval survival to first feeding at stocking densities of 1000 eggs L^?1 for pinfish and up to 4000 eggs L^?1 for pigfish. No clear patterns in larval morphometrics were observed among stocking densities. Static incubation densities of 250 and 1000 eggs L^?1 are recommended for pinfish and pigfish, respectively.     

Administration of hot‐water extract of Padina boergesenii via immersion route to enhance haemolymph‐immune responses of Fenneropenaeus indicus (Edwards)

Various polysaccharides extract from marine algae to increase the non‐specific immune system in crustacean. In this study, effects of hot‐water extract on total haemocyte count, total plasma protein, Phagocytic activity, bacterial clearance efficiency and bactericidal activity while the shrimp Fenneropenaeus indicus (10.12±2.18 g) were immersed in seawater (40 g L^?1 and 25±0.8 °C) containing hot‐water extract of brown algae Padina boergesenii at 100, 300 and 500 mg L^?1 for 1–4 h, were investigated. These parameters increased significantly (P<0.05) when the shrimp were immersed in seawater containing hot‐water extract at 100 mg L^?1 after 3 h and 300 and 500 mg L^?1 after 2 h. Fenneropenaeus indicus that were immersed in hot‐water extract at 300 and 500 mg L^?1 had significantly increased phagocytic activity and increased clearance efficiency to Vibrio harveyi after 2 h. But bactericidal activity increased after 1 h immersed in 500 mg L^?1 concentration.