Forage and migration habitat of loggerhead (Caretta caretta) and olive ridley (Lepidochelys olivacea) sea turtles in the central North Pacific Ocean

Satellite telemetry from 26 loggerhead (Caretta caretta) and 10 olive ridley (Lepidochelys olivacea) sea turtles captured and released from pelagic longline fishing gear provided information on the turtles’ position and movement in the central North Pacific. These data together with environmental data from satellite remote sensing are used to describe the oceanic habitat used by these turtles. The results indicate that loggerheads travel westward, move seasonally north and south primarily through the region 28–40°N, and occupy sea surface temperatures (SST) of 15–25°C. Their dive depth distribution indicated that they spend 40% of their time at the surface and 90% of their time at depths <40 m. Loggerheads are found in association with fronts, eddies, and geostrophic currents. Specifically, the Transition Zone Chlorophyll Front (TZCF) and the southern edge of the Kuroshio Extension Current (KEC) appear to be important forage and migration habitats for loggerheads. In contrast, olive ridleys were found primarily south of loggerhead habitat in the region 8–31°N latitude, occupying warmer water with SSTs of 23–28°C. They have a deeper dive pattern than loggerheads, spending only 20% of their time at the surface and 60% shallower than 40 m. However, the three olive ridleys identified from genetics to be of western Pacific origin spent some time associated with major ocean currents, specifically the southern edge of the KEC, the North Equatorial Current (NEC), and the Equatorial Counter Current (ECC). These habitats were not used by any olive ridleys of eastern Pacific origin suggesting that olive ridleys from different populations may occupy different oceanic habitats.     

Growth, haematological parameters and tissue lipid peroxidation of soft-shelled turtles, Pelodiscus sinensis, fed diets supplemented with different levels of ferrous sulphate

Soft-shelled turtles, Pelodiscus sinensis , with an average weight of 5.55 g, were fed diets supplemented with eight levels of ferrous sulphate for 8 weeks. The analysed iron content ranged from 50.8 to 482.9 mg kg⁻¹. Growth rate of turtles fed the control diet with no iron supplementation was the lowest among all dietary groups. Haematological parameters including red blood cell, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration of the turtles fed the control diet were also significantly ( P  < 0.05) lower relative to the other groups. Thus, dietary iron at 50.8 mg kg⁻¹ (no supplemented iron) was deemed deficient for growth and ineffective at preventing anaemia in juvenile soft-shelled turtle. Whereas, a supplementation of 50 mg kg⁻¹ ferrous sulphate (a total dietary iron of 91.8 mg kg⁻¹) was enough to normalize the haematological values of soft-shelled turtles to the level similar to other iron supplement-fed groups. Within the tested dietary iron range, liver iron content curve-linearly ( r ² = 0.99) increased with increasing dietary iron level. Furthermore, thiobarbituric acid-reactive substances in liver tissues of the turtles have also increased when liver iron content increased. The dietary iron requirement of soft-shelled turtle is 120–198 mg kg⁻¹ when ferrous sulphate is used as the source of iron.     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

Relationship between spatial distribution of the Patagonian stock of Argentine anchovy, Engraulis anchoita, and sea temperatures during late spring to early summer

The relative abundance of adult Argentine anchovies ( Engraulis anchoita ) and the thermal structure of the sea between 41° and 45°30'S during four research surveys were compared. Acoustic data were collected while making parallel transects along which CTD (conductivity, temperature, depth) stations were regularly distributed. Anchovy abundance was related to both the sea surface temperature and the stratification of the water column, as classified according to the φ parameter of stability ( Simpson, 1981 ). Regarding the whole water column, the sea temperatures where adult anchovies were recorded ranged from 8.5 to 16.5°C, but anchovy echo traces from waters above 14°C were obtained mainly at night. The sea surface temperature bounds for anchovy distribution were 11 and 17°C, with a preferential range between 12.5 and 16°C, but no absolute value of sea surface temperature was found to be most favourable for the species. The highest fish abundances were related to sharp thermal gradients, either horizontally recorded in frontal zones (≈ 0.02°C or more per km) or vertically associated to the occurrence of a thermocline (stability > 10 J m^–3). Sea fronts seemed to be the axes for the distribution of anchovy shoals, and the annual changes in the positions of the fronts seemed to be followed by analogous changes in the locations of the main anchovy concentrations. Within thermally stratified areas and during the daytime, anchovies apparently preferred the thermocline layer or the layer immediately above that, although a few fish shoals were located below the thermocline.     

Biomass and production of the euphausiid Euphausia pacifica along the coastal waters off north-eastern Japan

ABSTRACT:   Biomass and production of Euphausia pacifica off south-eastern Hokkaido (41–43°N), Sanriku (38–41°N), and Joban (36–38°N) were investigated every two months from March 1997 to February 1998. High biomass was found in summer–autumn off south-eastern Hokkaido, and in the late winter–early summer off Sanriku and Joban. Annual mean biomass was 381, 314 and 258 mg C/m² off south-eastern Hokkaido, Sanriku, and Joban, respectively. The total production (sum of flesh, moults and eggs) during the survey period off south-eastern Hokkaido (3829 mg C/m²) was comparable with that off Sanriku (3872 mg C/m²); both were much higher than that off Joban (2243 mg C/m²). Somatic production during the survey period contributed the highest proportion (51.5–70.9%) to the total production in each coastal area. The somatic production–biomass ratio in each coastal area (5.2–6.9) was an intermediate value among 25 populations of 16 euphausiid species previously reported. This effect resulted from the negative factor of stagnated growth of adults during summer–winter, coupled with positive factors including continuous occurrence of larvae which show high growth rate throughout the year off Sanriku and Joban, and the numerous larvae occurring in October off south-eastern Hokkaido.     

The influence of different levels of dietary vitamin E on sea bass Dicentrarchus labrax flesh quality

Sea bass Dicentrarchus labrax fillet quality was investigated after feeding with four diets (A, B, C or D) containing different levels of dietary vitamin E (139 mg kg^–1, 254 mg kg^–1, 493 mg kg^–1 and 942 mg kg^–1, respectively). Six-hundred and eighty fish (mean initial weight 208 g) were equally divided into four 20 m³ tanks and fed for 87 days. Filtered seawater with a temperature ranging from 18.2 to 26.3 °C was supplied continuously. At the end of the experiment, fish were stored at 1 °C for 12 days. At one, three, six, nine and 12 days, 20 fish per group were processed for proximate composition, vitamin E and induced thiobarbituric acid reactive substances (TBARs) analyses. No significant differences in proximate composition were registered between groups. The flesh lipid content ranged from 88.0 g kg^–1 (group B) to 96.8 g kg^–1 (group A). Vitamin E fillet content was significantly different between groups, reaching levels of 98.0, 150.7, 225.2 and 302.0 μg g^–1 lipids for group A, B, C and D, respectively. Induced TBARs values were statistically different only for group A compared with the other groups. No significant variations were registered in relation to preservation time. Because of the positive influence of vitamin E on seafood quality and the correlation between its dietary level and flesh deposition, the α-tocopherol content of the diet should be well above fish minimum requirements.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Evaluation of lipid spray beads for the delivery of water-soluble materials to a marine suspension-feeder, the Manila clam Tapes philippinarum (Deshayes 1853)

We describe the development and evaluation of a new microparticle for delivering low-molecular weight, water-soluble materials to suspension feeders. Spray beads successfully incorporated materials dissolved in an aqueous phase or as dry particulate, within a triacylglyceride bead composed of tripalmitin, 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ triolein, or 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil.
Riboflavin was successfully incorporated (up to 44 mg g⁻¹ lipid) and retained (up to 98% over 24 h in seawater) as dry particles in all three mixtures of lipid. Aqueous oxytetracycline hydrochloride or polymeric dye were incorporated (45.6 mg g⁻¹ lipid and 18.1 mg g⁻¹ lipid, respectively) and retained best (99% and 94%, respectively) in spray beads composed of tripalmitin. The addition of triolein or fish oil to the lipid bead reduced incorporation and retention efficiencies for aqueous core materials by up to 75%.
Manila clam seed readily ingested and digested lipid microparticles, spray beads and lipid-walled microcapsules. Microparticles composed of tripalmitin were excreted with their payloads intact. Intact microparticles composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were largely absent in faecal strands suggesting successful release and delivery of microparticle contents to clams.
Spray beads composed of tripalmitin softened with 400 mg g⁻¹ fish oil represent an effective microparticle type for delivering low-molecular weight, water-soluble materials to aquatic suspension feeders.     

Enhancing the TurtleWatch product for leatherback sea turtles,a dynamic habitat model for ecosystem‐based management

Fishery management measures to reduce interactions between fisheries and endangered or threatened species have typically relied on static time‐area closures. While these efforts have reduced interactions, they can be costly and inefficient for managing highly migratory species such as sea turtles. The NOAA TurtleWatch product was created in 2006 as a tool to reduce the rates of interactions of loggerhead sea turtles with shallow‐set longline gear deployed by the Hawaii‐based pelagic longline fishery targeting swordfish. TurtleWatch provides information on loggerhead habitat and can be used by managers and industry to make dynamic management decisions to potentially reduce incidentally capturing turtles during fishing operations. TurtleWatch is expanded here to include information on endangered leatherback turtles to help reduce incidental capture rates in the central North Pacific. Fishery‐dependent data were combined with fishing effort, bycatch and satellite tracking data of leatherbacks to characterize sea surface temperature (SST) relationships that identify habitat or interaction ‘hotspots’. Analysis of SST identified two zones, centered at 17.2° and 22.9°C, occupied by leatherbacks on fishing grounds of the Hawaii‐based swordfish fishery. This new information was used to expand the TurtleWatch product to provide managers and industry near real‐time habitat information for both loggerheads and leatherbacks. The updated TurtleWatch product provides a tool for dynamic management of the Hawaii‐based shallow‐set fishery to aid in the bycatch reduction of both species. Updating the management strategy to dynamically adapt to shifts in multi‐species habitat use through time is a step towards an ecosystem‐based approach to fisheries management in pelagic ecosystems.     

Acute LD50 and kidney histopathology following injection of erythromycin (Erythro-200) and its carrier in spring chinook salmon, Oncorhynchus tshawytscha (Walbaum)

We evaluated the toxicity of commercial injectable erythromycin (Erythro-200) and the injectable drug carrier alone (PEG-400, ethyl alcohol and ethyl acetate) in sub-adult spring chinook salmon, Oncorhynchus tshawytscha (Walbaum). Both compounds were toxic to salmon when administered in high dosages. The 50% lethal dose (LD₅₀) of the erythromycin within its carrier (Erythro-200) at 12 °C was estimated at 429 mg erythromycin kg^–1 or 2.145 mL kg^–1 when measured by volume. The LD₅₀ of the drug carrier alone was 2.95 mL kg^–1 by volume or equivalent to 3.21 g kg^–1 of the carrier. We evaluated and scored the histopathology in kidney sections removed from fish 96 h after injection with Erythro-200, the drug carrier alone, or sterile phosphate buffered saline (PBS) that served as a reference control. Kidneys from fish injected with saline had lowest average pathology scores; those injected with the carrier had moderate scores; and the pathology score was highest in tissues from erythromycin-injected fish. In tests at 12 °C, vacuoles were more prominent in proximal and distal tubules of fish injected with higher dosages of 403 and 545 mg erythromycin kg^–1 than in tissues from fish injected with 299 mg kg^–1.     

Ecological variations and El Niño effects off the southern coast of the Korean Peninsula during the last three decades

To explore ecosystem response to environmental changes, we investigated interannual and decadal changes in the marine ecosystem using data collected from the southern coast of the Korean Peninsula during the 1960s–1990s. Water properties such as sea surface temperature (SST) and salinity in April did not show large variation during the 1970s, but a relatively cool water mass appeared from the early to mid 1980s and a warm water mass replaced it after 1989. Chl a concentration in April, which was converted from Secchi disc information, was low (mean=1.18 mg m⁻³) during the period 1968–1980 excluding 1974, but there was a period of greater biomass for 7 years from 1981 (mean=2.78 mg m⁻³). A negative correlation ( r =−0.387, P  < 0.05) between SST and chl a in April matched high chl a during the low-SST period in the early and mid 1980s. Annual zooplankton biomass ranged from 37 to 132 mg m⁻³, but abundance was frequently high after 1984 and low during 1972–1982. The most representative pelagic fishes in the South Sea seemed to respond to the increase in planktonic organisms. Anchovy, mackerel and sardine increased in abundance since the mid 1970s. High catches of anchovy and mackerel lasted until the 1990s, whereas sardine decreased after the early 1990s. From the matrix of simple correlation coefficient, the Southern Oscillation Index (SOI) exhibited a high correlation with SST in December in the study area ( P  < 0.05). Catches of anchovy and mackerel were positively correlated with chl a and zooplankton during their early life periods, as well as with SST in December.     

Effects of dietary protein and lipid level, and water temperature, on the post-feeding oxygen consumption of Atlantic cod and haddock

Tank respirometry was used to study the interactive effects of protein:lipid level (55%:11% vs. 42%:16%; both diets isoenergetic) and temperature (11, 6 and 2 °C) on the magnitude and duration of specific dynamic action (SDA) in juvenile Atlantic cod and haddock. The protein:lipid level did not affect any measured variable. However, numerous temperature and species effects were observed. For example, although the maximum post-feeding oxygen consumption (30–50% above routine metabolic rate; RMR) and SDA duration (∼55–85 h; SDA_DUR) were not affected by temperature, SDA_DUR g⁻¹ of food increased from 11 to 2 °C (from ∼3 to 12 h g food⁻¹). While absolute SDA (mg O₂) decreased by ∼60–65% in cod and ∼75% in haddock from 11 to 2 °C, due to a concomitant decrease in food consumption from ∼2.0% to 0.6% body mass, SDA comprised between 3.3% and 5.2% of the dietary energy content at all temperatures. Finally, RMR at 11 and 2 °C and SDA_DUR at 2 °C were 25–35% and 25% greater in cod, respectively, as compared with haddock. These results suggest that feeding reduced protein diets at low water temperatures is unlikely to improve the growth of these species.     

Interaction between dietary levels of vitamins C and E on growth and immune responses in channel catfish, Ictalurus punctatus (Rafinesque)

This study was conducted to evaluate the effect of dietary levels of vitamins C (0, 100 and 2000 mg kg⁻¹), E (0, 50 and 500 mg kg⁻¹) and their interaction on the growth performance, liver contents of ascorbic acid and α-tocopherol, haematology and immune response of channel catfish, Ictalurus punctatus . Each diet was fed to catfish in triplicate aquaria to apparent satiation twice daily for 12 weeks. The results indicate that the amount of vitamin E contained in the basal diet (23.1 mg kg⁻¹) was sufficient to promote good growth, feed efficiency and survival, but its supplementation was needed to maintain high haematological values and liver vitamin E. Supplementation of vitamin C (100 mg kg⁻¹) to the basal diet containing 10.5 mg kg⁻¹ was required for good growth, feed efficiency, survival and prevention of vertebral deformity and optimum haematological indices. Liver storage of ascorbic acid and α-tocopherol increased with increasing dietary levels of each vitamin. Dietary vitamin E levels had no effect on liver ascorbic acid content, but increasing dietary vitamin C increased liver α-tocopherol. Some measured immune parameters (serum protein and superoxide anion production) were enhanced by supplementation of vitamin C or E. Chemotaxis ratio and phagocytosis were not affected by treatments.     

Efficacy of tricaine methanesulphonate and clove oil as anaesthetics for juvenile cobia Rachycentron canadum

Six experiments were designed to determine the optimal anaesthetic dosage of tricaine methanesulphonate (TMS) and clove oil that could be used safely on juvenile cobia Rachycentron canadum of two sizes [G1=4.9±0.8 g; G2=13.9±3.1 g]. We documented the stage of anaesthesia and the acute toxicity as 96 h LC₅₀ (lethal concentration 50% population) at various exposure times of the two anaesthetics. At 10 min induction time, the TMS 96 h LC₅₀ was 93.9 mg L⁻¹ in G1 and 97.0 mg L⁻¹ in G2. Compared with clove oil, the 96 h LC₅₀ was 60.0 mg L⁻¹ in G1 and 69.8 mg L⁻¹ in G2. The difference between the two groups (G1, G2) did not influence anaesthesia safety ( P >0.05). Rachycentron canadum achieved stage 3 anaesthesia more rapidly at a lower clove oil concentration level (40 mg L⁻¹, 10 min) than TMS (60 mg L⁻¹, 10 min), but the recovery period of clove oil, was significantly longer. Clove oil was the most effective in reducing the short-term stress induced by routine biometry (20 mg L⁻¹, 10 min) and also by transporting (1 mg L⁻¹, 8 h). Whereas, for long-term exposure, 40 mg L⁻¹ TMS was found to be safe.     

Comparison of lipid-walled microcapsules and lipid spray beads for the delivery of water-soluble, low-molecular-weight materials to aquatic animals

Microparticles (< 40 μm diameter) composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were used to encapsulate the low-molecular-weight (mol. wt 460) antibiotic oxytetracycline in the form of either oxytetracycline hydrochloride (OTC.HCl) or oxytetracycline hemicalcium salt (OTC.HEM). Dry, finely ground particles of core material were encapsulated in spray beads. Dissolved core material was encapsulated in lipid-walled microcapsules.
Oxytetracycline (OTC) was most efficiently delivered (≈ 46.5 mg g⁻¹ lipid after 24 h suspension in seawater) as a hemicalcium salt in spray beads. Lipid-walled microcapsules were most efficient for delivering OTC (≈ 8.7 mg g⁻¹ lipid) as OTC.HCl dissolved in 0.2  M HCl at a concentration of 300 mg mL⁻¹.
Spray beads containing OTC.HEM were very stable over 1 month in storage. Lipid-walled microcapsules containing aqueous OTC.HCl lost ≈ 30% of their core material during storage. Freeze-drying of both microparticle types did not improve storage of spray beads, but showed promise for reducing leakage from lipid-walled microcapsules during storage and delivery to suspension feeders.     

Environmental preferences of longlining for yellowfin tuna (Thunnus albacares) in the tropical high seas of the Indian Ocean

A survey of yellowfin tuna, Thunnus albacares , fishing ground was carried out on board of the Chinese longliners from September 15 to December 12, 2005 in the tropical high seas of the Indian Ocean. The depth at which each yellowfin tuna was hooked was estimated using a stepwise regression analysis of theoretical hook depth and observed average hook depth measured using a temperature depth recorder. Water temperature, salinity, chlorophyll  a , dissolved oxygen and thermocline, which are important variables influencing yellowfin tuna habitats, were measured in the survey. Catch rates of yellowfin tuna were then analyzed with respect to depth, temperature, salinity, chlorophyll  a , dissolved oxygen and thermocline. We suggest that the optimum ranges of swimming depth, water temperature, chlorophyll  a and dissolved oxygen concentration for yellowfin tuna are 100.0–179.9 m, 15.0–17.9°C, 0.090–0.099  μ g L⁻¹, 2.50–2.99 mg L⁻¹, respectively; that salinity has less influence on the vertical distribution of adult yellowfin tuna; and that yellowfin tuna are mainly distributed within the thermocline in the high seas of the Indian Ocean. Our results match the yellowfin tuna's vulnerability to deep longline fishing gear well.     

Nutrient values of dietary ascorbic acid (l-ascorbyl-2-polyphosphate) on growth, survival and stress tolerance of larval shrimp, Litopenaeus vannamei

l -ascorbyl-2-polyphosphate (ApP) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the larval white shrimp, Litopenaeus vannamei . Five isoenergetic and isonitrogenous fish meal-fish protein hydrolysate-based diets with five levels of ApP, AsA equivalent to 91.8, 188, 271, 360 and 436 mg kg⁻¹ diet were fed to triplicate groups of L. vannamei (mean initial wet weight 1 mg) for 32 days. The diet with AsA 91.8 mg kg⁻¹ showed high cumulative mortality after 10 days of feeding. After the 32-day trial, the shrimp that fed the diet had significantly lower survival and weight gain (WG, %) than those that fed 188, 271, 360 and 436 mg AsA kg⁻¹ diets. Specific growth rate (SGR, % day⁻¹) and final body wet weight (FBW, mg) showed the same pattern as WG (%). There were no significant differences in growth performance (FBW, WG and SGR) among the groups that fed 188, 271, 360 and 436 mg kg⁻¹ of AsA at the termination of feeding trial. Broken-line regression analysis on WG indicated that 191 mg AsA kg⁻¹ in the diet was the optimum for larval L. vannamei . On the contrary, dietary level of more than 360 mg AsA kg⁻¹ was needed to ensure high resistance to stressful conditions such as low dissolved oxygen stressors.     

Haematological response and growth performance of Nile tilapia (Oreochromis niloticus L.) fed diets containing folic acid

Haematological response and growth performance over 150 days, and resistance to a low-temperature stress of Nile tilapia fed diets with increasing folic acid (FA) levels were evaluated. The experiment was conducted in a completely randomized design with eight FA levels (0.0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mg kg⁻¹ feed) supplemented in purified diets (32.0% CP and 13 398 kj DE kg⁻¹). One hundred and ninety-two fingerlings were randomly assigned to 32 net cages distributed in eight 1000 L aquaria with a physical and biological filter and a temperature control system (26.0 ± 1.0 °C). For cold-induced stress, fish were transferred to 24 30 L-aquaria with individual biofilters and aeration. The water temperature was gradually reduced until it reached 13 °C. Haematological parameters evaluated before and after cold stress were total erythrocytes and leucocytes count, differential leucocyte, haemoglobin, haematocrit, total plasmatic protein and haematometric indices. Growth performance parameters were mean weight gain, feed conversion ratio and survival. Dietary FA supplementation did not influence erythropoiesis under normal temperature conditions; cold stress impaired erythropoiesis, causing hypochromic microcytic anaemia and leucopoiesis, and also neutrophilia. Growth performance is influenced by folate and supplementation between 0.5 and 1.0 mg FA kg⁻¹ diet, which makes up for nutritional demands, guaranteeing production and health under appropriate temperature conditions.     

Effect of combined photoperiod, water calcium concentration and pH on survival, growth, and moulting of juvenile crayfish (Procambarus clarkii ) cultured under laboratory conditions

The red swamp crayfish, Procambarus clarkii (Girard), is one of the most commonly farmed freshwater species in inland China due to its high market value and consumer demand. The aim of this study was to determine the optimum combinations of photoperiod, water calcium concentration and pH for juvenile survival, growth and moulting. In our orthogonal experiment, the three environmental factors were varied at three levels (photoperiod: 16L:8D, 12L:12D and 8L:16D; calcium concentration: 45.5, 65.5 and 85.5 mg L⁻¹; and pH: 6.8, 7.8 and 8.8). Range analysis showed that the maximum survival of juvenile crayfish occurred at photoperiods of 16L:8D or 8L:16D, water calcium concentration of 45.5 mg L⁻¹ and pH of 7.8; maximum weight gain at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; maximum length increase at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; and the highest moult frequency at photoperiod 12L:12D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8. Analysis of variance indicated that photoperiod, water calcium concentration and pH significantly influenced only the weight gain of juvenile crayfish ( P <0.05). Taking growth into consideration, we suggest that a photoperiod of 16L:8D, calcium concentration of 65.5 mg L⁻¹ and pH 7.8 might be optimal conditions for rearing juvenile P. clarkii .     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.