Strategies for Development of Rotifers as Larval Fish Food in Ponds

Strategies to sustain rotifer peak biomass, distribution of rotifer resting eggs in the sediment, and relationship between rotifers and larval fish growth were studied in a series of pond experiments. After the ponds were filled with water, herbivorous rotifers (e.g., Brachionus calyciflorus ) developed first, but were gradually replaced by predatory rotifers (e.g., Asplanchna ). Subsequently, herbivorous cladocerans (e.g., Moina sp) eventually replaced rotifers and dominated the zooplankton community. The occurrence of Asplanchna and Moina indicated the decline of B. calyciflorus . Peak rotifer biomass developed 8–10 d after the ponds were filled with water at 20–25 C, 10–15 d at 17–20 C, 15–20 d at 15–17 C, 20–30 d at 10–15 C, and >30 d at < 10 C. The abundance of resting eggs in the top 5-cm sediment varied from 6 to 83/cm². About 25% of resting eggs were buried in the top 5-cm sediment but the number of resting eggs decreased with increased sediment depth. Optimum rotifer biomass for silver carp Hypophthalmichthys molitrix larvae stocked at 1,500,000/ha was 20–40 m/gL. High rotifer biomass (>20 mg/L) usually lasted 3–5 d, but could be prolonged by pond fertilization or cladoceran controls. A weekly application of dipterex at 0.05 mg/L reduced cladoceran biomass but enhanced rotifer biomass. Our results indicate with a careful management plan it is possible to synchronize the rotifer development with larval fish stocking.     

Effect of gamma-aminobutyric acid and porcine growth hormone on survival of the euryhaline rotifers <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> sp. complex preserved at low temperature

Gamma-aminobutyric acid (GABA) at 50 μg/ml and porcine growth hormone (GH) at 0.025 IU/ml were tested to see whether these chemicals would reduce the stress experienced by euryhaline rotifers Brachionus plicatilis species complex (L-, S- and SS-morphotypes) during low temperature (4–12°C) storage. Rotifers cultured at 25°C were transferred to 4–12°C for 10–30 days and transferred back to 25°C for recovery. GABA or GH were added to the rotifers at three different time points: 6 h before transfer from 25°C to low temperature (6h−), on day 7 after preservation at low temperature (7d+) and on the first day of recovery. For L-type rotifers, the GH treatment before the transfer to 4°C for 30 days was effective for better survival, while the GABA treatment was most effective for the S-type preserved at 10°C for 14 days. For the SS-type, the chemical treatments were not effective when the rotifers were preserved at 12°C for 14 days. After low temperature preservation, GABA treatments with the S- and SS-type rotifers just after their transfer to 25°C induced a relatively faster recovery of the rotifer population.     

Effect of combination feeding of <Emphasis Type="Italic">Nannochloropsis</Emphasis> and freshwater <Emphasis Type="Italic">Chlorella</Emphasis> on the fatty acid composition of rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> in a continuous culture

ABSTRACT:   A continuous culture of rotifer was conducted to investigate the effect of combination feeding of both a high density of Nannochloropsis oculata (N) and condensed freshwater Chlorella (FC) on the fatty acid composition of L-type rotifer Brachionus plicatilis in a continuous culture system. The algal feeding of the rotifers was carried out in three successive steps: N-feeding → N+FC-feeding → FC-feeding. The culture was conducted at 24°C and 25–27 psu in a 2000 mL bottle with 50% of water exchanged daily. The combination N+FC-feeding was effective in increasing rotifer density. The rotifers fed on N+FC (N+FC-R) had more non-polar lipids than polar ones, similar to those on N (N-R), opposite to the rotifers fed on FC (FC-R). N+FC-R contained higher levels of 16:2, 18:2n-6 (linoleic acid [LA]) and 20:2n-6, but lower levels of 18:1, 20:4n-6 (arachidonic acid), 20:5n-3 (eicosapentaenoic acid [EPA]) and 22:5n-3 (docosapentaenoic acid [DPA]) compared with N-R. Whereas N+FC-R contained higher levels of 16:1n-7, EPA and DPA, but lower levels of 16:2 and LA compared with FC-R. N+FC-R had more DPA in polar lipids than in non-polar ones. The Σn-6/Σn-3 ratio in N+FC-R was 0.9–1.0, significantly different from those in N-R (0.4) and FC-R (6.6–8.4). Therefore, it is inferred that the fatty acid profile of the N+FC-R cultured in a continuous culture system was affected by both N and FC. Also, the combination N+FC-feeding may be effective in manipulating the Σn-6/Σn-3 ratio in continuously cultured rotifers.     

维生素B12对圆型臂尾轮虫种群增长、繁殖和个体大小的影响

为了解维生素B 12 对海水圆型臂尾轮虫( Brachionus rotundiformis )种群增长、繁殖和个体大小的影响,采用四种不同浓度的维生素B 12 (0、0.3、0.6、1.2 μg/mL)强化培养轮虫7 d。每天测定轮虫的种群密度和怀卵率,每隔一天测定轮虫背甲长和宽。试验结果显示:0.3 μg/mL维生素B 12 组轮虫的种群增长、怀卵率显著高于其他组( P<0.05);0.3 μg/mL维生素B 12 组轮虫的背甲长和宽均显著小于其他三组( P>0.05),而其他三组差异不显著。试验结论:在本试验条件下,维生素B 12 对圆型臂尾轮虫种群增长、繁殖和个体大小影响的最适浓度为0.3 μg/mL。     

Ontogeny of the digestive tract of larval percula clownfish, Amphiprion percula (Lacépède 1802): a histological perspective

An understanding of the development of the digestive system of marine fish larvae is of critical importance in determining optimal feeding regimes for their culture. The present study provides information on the histomorphological development of the digestive system of clown fish, Amphiprion percula , larvae during the first month of life. Before hatching, clownfish larvae possess an alimentary tract, liver and pancreas with absorptive and digestive capabilities. The yolk sac is completely consumed within 5–7 days at 25 °C. Clownfish larvae readily accept rotifers after hatching and a complete dietary shift from rotifer to Artemia can be accomplished at 10 days after hatch (DAH). Gastric glands in the stomach first develop 11 DAH and proliferate by 15 DAH. Both non-staining vacuoles (NSV) and supranuclear inclusion vesicles (SIV) appear at 11 DAH in the midgut and hindgut respectively. Pinocytosis and extracellular digestion coexist for about 2 weeks after hatching. While SIV disappeared completely at 25 DAH, NSV continued to be a prominent feature of the midgut during the first month.     

Effect of water temperature and diet on growth and mortality of Neptunea arthritica juveniles

The growth and mortality of Neptunea arthritica juveniles hatched from different egg masses (controlled conditions and field) reared at 10 and 15 °C, and provided with sardine (D1) and polychaete (D2) as foods were evaluated. Results showed that diet type and water temperature significantly affect the growth and mortality. Interaction between these factors reflected robust relationship among them, but restricted exclusively to growth. Through the study period, juveniles treated at 10 °C showed maximum increments of 2.5 mm (shell length), 2.3 mm (shell width) and 0.25 g (body weight) from their initial size. While maximum values of those treated at 15 °C for the above-mentioned parameters were 7.6 mm, 5.5 mm and 1 g respectively. In both cases, these increments were recorded for individuals fed with sardine. Mortality was significantly affected by diets and water temperature, which significantly increased under D2 [10 °C (20–67%), 15 °C (70–87%)] or 15 °C [D1 (30–83%), D2 (70–87%)]. According to the results, juveniles of N. arthritica could be maintained under controlled conditions, which allows growth improvement. Mortality appears to be the limiting factor, but this could be improved by the implementation of efficient culture method, basically oriented to increase the water quality.     

Natural spawning, early development and first feeding of the semicircle angelfish [Pomacanthus semicirculatus (Cuvier, 1831)] in captivity

Successful natural spawning of Pomacanthus semicirculatus in captivity from 11 September to 18 October, 2006 is described for the first time. Each female laid an average of 230 000 eggs during the spawning period. Fertilized eggs were spherical, transparent and buoyant and had a mean diameter of 0.61 ± 0.03 mm (mean ± SD). Embryonic development lasted 18–21 h at 28.5 °C. Newly hatched larvae were 1.35 ± 0.02 mm in total length (TL) with 27 (12+15) myomeres and had an oil globule in the ventroposterior area of the yolk sac. Larvae completed yolk absorption within 3 days post hatching at 2.37 ± 0.05 mm TL. Larvae were fed either 100% microalgae ( Nannochloropsis sp.), 100% s-type rotifers ( Brachionus rotundiformis ), 100% dinoflagellates ( Gonyaulax sp.) or different combinations of the three (50%:50%:0%, 30%:35%:35%) to determine the effect of live feed on the survival rate. The survival was significantly ( P <0.001) better in larvae fed a combination of diets (30%: 35%: 35%) than others. These results indicate that P. semicirculatus is a potential species for captive-breeding programmes and the use of a combination of diets (microalgae plus s-type rotifers and dinoflagellates) may be a suitable first food for fish larvae.     

Optimum temperature and salinity conditions for growth of green algae <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis> and <Emphasis Type="Italic">Nannochloris oculata</Emphasis>

ABSTRACT:   The effects of temperature and salinity on growth of green algae Chlorella ellipsoidea and Nannochloris oculata were determined to compare the optimum culture conditions. A four-temperature (15, 20, 25, and 30°C) × three-salinity (10, 20, and 30) factorial design with triplicates was applied. Specific growth rate (SGR), maximum density, and duration to reach maximum density of C. ellipsoidea were significantly affected by both temperature and salinity. The highest SGR was observed in C. ellipsoidea at 25°C and salinity 10, but the maximum density was very low. The highest maximum density was achieved in C. ellipsoidea at 15°C and 10. The slope constant of the linear relationship between semilogarithmic growth of C. ellipsoidea and day of culture was highest at 15°C and 10. The SGR and duration to reach maximum density of N. oculata were significantly affected by both temperature and salinity. However, maximum density of N. oculata was significantly affected by temperature, but not salinity. The highest maximum density was achieved in N. oculata at 25°C and 30, but SGR was significantly lower than that of N. oculata at 25°C and 10. The slope constant of the linear relationship between semilogarithmic growth of N. oculata and day of culture was highest at 25°C and 30. Based on these results, the condition of 15°C and salinity 10 seemed to be optimal for maximum density of C. ellipsoidea , and the condition of 25°C and 10 and 30 for SGR and maximum density for N. oculata , respectively.     

Temperature and salinity tolerances of yellowfin sea bream, Acanthopagrus latus, at different salinity and temperature levels

Abstract Yellowfin sea bream Acanthopagrus latus (Houttuyn) (0.98 ± 0.27 g) were examined for temperature and salinity tolerances at nine different environmental regimes (0.3, 15 and 33 ppt combined with 10, 25 and 32 °C), in which the fish were subjected to gradual and sudden changes in temperature and salinity respectively. The 50% CTMax (critical thermal maximum) and the UILT (upper incipient lethal temperature) were in the range of 34.8–38.2 °C and 32.8–36.4 °C respectively. The 50% CTMin (critical thermal minimum) and the LILT (lower incipient lethal temperature) were in the ranges of 4.9–9.4 °C and 7.1–17.6 °C respectively. The 50% CSMax (critical salinity maximum) and the UILS (upper incipient lethal salinity) were in the ranges of 54–69‰ and 9–44‰, respectively. The fish at lower temperature (10 °C) and salinity of 33‰ tolerated temperature as low as 6 °C when the temperature was decreased gradually. The fish at 33 and 15‰ and lower temperature (10 °C) tolerated salinity as high as 66–67‰, whereas those at freshwater salinity level (0.3‰) and 32 °C tolerated salinity to 50‰ when salinity was increased gradually. The relationships among UILT, LILT, 50% CTMax, 50% CTMin, UILS, 50% CSMax, salinity and temperature are given.     

Improved performance of an intensive rotifer culture system by using a nitrifying inoculum (ABIL)

Abstract A dense nitrifying culture (ABIL) has been examined for its capacity to stimulate rotifer growth in a labscale culture system. The nitrifiers were applied in different ways. When ABIL was added directly to rotifer batch cultures, it gave rise to significantly higher population densities (factor 1.5–2.5 higher, P  < 0.05). The nitrifiers were subsequently examined for their capacity to enhance the start-up of bioreactors, commonly installed in aquaculture rearing tanks. Of the different carrier materials used in these bioreactors, i.e. CaCO₃, gravel and a PVC matrix (Bionet), CaCO₃ gave by far the best results. In a third set of experiments, effectively nitrifying bioreactor systems were connected to rotifer culture tanks and operated over a period of up to 10 days. It was demonstrated that the ABIL inoculated CaCO₃-based bioreactor allowed excellent rotifer growth reaching rotifer densities up to 5500 rotifers per mL. Moreover, a new system in which the ABIL culture was recirculated through hollow fibres was developed and demonstrated to be effective for supporting rotifer growth up to 3500 rotifers per mL. Overall, the use of the dense nitrifying culture either in seed batch cultures, conventional bioreactors or hollow fibre bioreactor systems in support of rotifer cultures was demonstrated to be effective for improving the water quality and the rotifer growth.     

Determination of the optimal water temperature for the culture of juvenile dusky kob Argyrosomus japonicus Temminck and Schlegel 1843

The effects of temperature on growth, food-conversion ratio (FCR) and feeding intensity of juvenile dusky kob Argyrosomus japonicus (23.7±2.6 g fish⁻¹) were assessed over the temperature range 17.5–28.5 °C in a 42-day growth trial. Growth increased with increasing temperature up to an optimum after which it declined. Specific growth rates were 2.05% and 1.2% day⁻¹ for the fastest (25.3 °C) and the slowest (17.5 °C) treatments respectively. Food-conversion ratio peaked at a lower temperature than growth. Optimal (0.72 kg kg gain⁻¹) and least efficient (1.40 kg kg gain⁻¹) FCR were found at 21.7 and 17.5 °C respectively. Feeding intensity was linearly related to temperature within the range of 17.5–28.5 °C. These results corresponded to the thermal preference (25–26.4 °C) and natural temperature distribution (12–28 °C) of South African dusky kob. Determination of the temperature range that does not limit growth is a prerequisite to assess the relationship between growth and environmental variables such as light intensity, feeding regime and stocking density. Consequently, experiments to determine the effects of these environmental variables on growth and aquaculture potential of dusky kob should be conducted at 24–26 °C.     

Effects of water temperature on early development of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   To determine an optimal temperature range for efficient production of healthy eel larvae Anguilla japonica , the effect of water temperature on hatching, survival, and deformity rates was examined. The early ontogeny of morphological features in this species by incubating eggs at five different temperatures (19, 22, 25, 28 and 31°C) was examined. Hatching occurred at 24, 28, 46 and 58 h after fertilization when incubated at 28, 25, 22 and 19°C, respectively; no eggs hatched at 31°C. The growth rate of prefeeding larvae increased as water temperature was elevated and acquisition of feeding ability was also accelerated at higher temperature. Significantly high hatching rates (76–86%, P  < 0.05) and survival rates (61–86%, P  < 0.05) were observed at relatively high temperatures (22–28°C). The deformities were 'open lower jaw', 'pericardial edema', and 'notochordal bending', in which the mouth was deformed with a downward projecting lower jaw, the pericardial cavity was swollen, and the notochord was bent or twisted to various degrees, respectively. Open lower jaw and pericardial edema were especially predominant at 19°C, with rates of 68 and 92%, respectively, compared with 31 and 10% at 25 and 28°C, respectively. The occurrence of notochordal bending was not affected by temperature. The optimal temperature for incubation and rearing A. japonica eggs and prefeeding larvae is approximately 25–28°C.     

Isolation of ice-nucleating active bacterium from mackerel and its properties

ABSTRACT: An ice-nucleating bacterium, designated MACK-4, was isolated from ice-stored mackerel ( Scomber australasicus ) and identified as a Pseudomonas fluorescens . The optimal temperature and pH for its growth in nutrient broth with 2.5% glycerol (NB-G) were 15°C and 6.5, respectively. The maximal ice-nucleating activity (INA) was obtained after 54 h incubation at 15°C. However, the INA was almost completely lost after 48 h incubation at 25°C or higher. The growth and INA decreased with increase of NaCl added in NB-G within 0.0–4.0%. The INA of MACK-4 was very stable at 5–25°C, pH 4.0–9.5, while that of isolated ice-nucleating matter from MACK-4 was stable at 5–25°C, pH 5.5–9.0.     

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.     

Production, disinfection and evaluation for aquaculture applications of rotifer resting eggs from Bohai Bay, P.R. of China

The dynamics of resting egg production of the rotifer Brachionus plicatilis originating from a wild population in the Tanggu Saltworks (P.R. China) was investigated. In the natural environment as well as in semi-controlled rearing conditions an increased resting egg production was noticed with declining food availability. Processed resting eggs had a hatching efficiency of 3 x 10 6 rotifers per gramme irrespective of their origin. Hatching started 22 h after the initiation of incubation and was completed after 36 h. Rotifers obtained from resting eggs could be further cultured on Culture Selco® and enriched with Super Selco®. The fatty acid profile of these rotifers were not divergent from reference rotifers (originating from the Laboratory of Aquaculture and Artemia Reference Centre) demonstrating that this strain was not catabolizing essential fatty acids and could be used for enrichment purposes. Storage of resting eggs at 4°C resulted in a 50% lower hatching after 1 year but remained stable during the next 2 years. The resting eggs used for storage could easily be disinfected without affecting their hatching characteristics. These results indicate that this material could be used as inocula for mass cultures of live food for commercial hatcheries.     

Effects of hormone treatments and temperature on sex-reversal of Nigorobuna Carassius carassius grandoculis

ABSTRACT: Functional sex reversal of all-female nigorobuna Carassius carassius grandoculis to phenotypic males was examined by immersion exposure of fry to 17-methyltestosterone (17-MT) and controlled water temperature during early development. Fry were reared in water containing different concentrations of 17-MT at 24 and 30°C for 80 days starting 20 days after hatching. Although the fish exposed to 0.1 and 1.0 μg/L 17-MT at 24°C were all male, treatment with 10.0 μg/L 17-MT resulted in 43% females. Twenty-two percent males appeared in the control treatment at 30°C but the control at 24°C was entirely female. The proportion of males in treatments exposed to 0.01– 1.0 μg/L 17-MT at 30°C was slightly lower than in the respective treatments at 24°C. These results indicate that the phenotypic expression of sex in nigorobuna is thermolabile and that sex determination is under the control of genetic factors and temperature. Also, control of temperature during early development has been shown to be important for the production of all-female offspring for use as breeding stock for pond culture of fish suitable for preparation as 'funazushi'.     

Recirculation rate of rearing water affects growth of Japanese smelt Hypomesus nipponensis larvae

Larvae of Japanese smelt Hypomesus nipponensis have unique salinity adaptability and a small gape size; consequently, no system has been developed to feed them for stable high growth and survival. Therefore, suitable conditions for larval growth of Japanese smelt remain to be studied. In an earlier study, we developed a Japanese smelt rearing method using a closed recirculating system. Using that system for this study, we examined recirculation rate effects on Japanese smelt larval growth because the recirculation rate can affect larval feeding opportunities through regulation of the rotifer retention time in the rearing tanks. Using 0.2–0.3% salinity and water temperatures of less than 20 °C, we fed Japanese smelt larvae with SS-type rotifer and commercial feed. Results indicated that larval growth with a recirculation rate of less than 648% per day (LR) is superior to that achieved at a rate of over 2160% per day (HR). Moreover, rotifer density under the LR condition declines more slowly than under the HR condition. These results demonstrated that the recirculation rate affects their growth, perhaps by affecting the opportunities for larvae to ingest the rotifers. Furthermore, SS-type rotifers are available for feeding teleosts in a wide range of water temperatures and salinities.

     

Growth-independent effects of a fluctuating thermal regime on the life-history traits of the Japanese medaka (Oryzias latipes)

Abstract –  Temperature fluctuation has been shown to affect somatic growth rates and metabolic functions of fish, but little is known about the effects of fluctuating temperatures on life-history traits. We examined these effects on female Japanese medaka ( Oryzias latipes ) reared from hatch to maturity at two mean temperatures (27 and 30 °C) with diel temperatures either held constant or allowed to fluctuate ±5 °C. Feeding regimes were manipulated to equilibrate medaka growth rates in all treatments. Age and length at maturity decreased as a result of thermal fluctuation, whereas there was no effect of temperature fluctuation on the proportion of fish attaining maturity or on the number of eggs produced by a mature female. We hypothesise that short-term high temperatures associated with temperature fluctuation decreases development time, whereas reduced energy conversion efficiency at higher temperatures does not allow for increased egg production.     

The effect of different rotifer feeding regimes on the growth and survival of yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae

First feeding success is critical to larval marine finfish and optimization of live feed densities is important for larval performance and the economics of commercial hatchery production. This study investigated various rotifer feeding regimes on the prey consumption, growth and survival of yellowtail kingfish Seriola lalandi larvae over the first 12 days post hatch (dph). The common practice of maintaining high densities of rotifers (10–30 ind. mL^?1) in the rearing tank was compared to a low density feeding technique, where 5–8 ind. mL^?1 of rotifers were offered. A ‘hybrid’ feeding regime offered rotifers at the high density treatment until 5 dph and the lower feeding densities thereafter. There was no significant difference in larval survival (hybrid: 28.9 ± 7%, low density: 17.3 ± 5% and high density: 17.2 ± 9%) or growth (hybrid: 6.12 ± 0.18 mm, low density: 6.03 ± 0.10 mm and high density: 6.11 ± 0.23 mm) between treatments. Rotifer ingestion was independent of rotifer density throughout the trial and increased with larval age, with larvae at 4 dph ingesting 22 ± 1.5 rotifers larvae^?1 h^?1 and by 11 dph ingesting 59 ± 1.6 rotifers larvae^?1 h^?1. These data demonstrate that from first feeding, yellowtail kingfish larvae are efficient at capturing prey at the densities presented here and consequently significant savings in rotifer production costs as well as other potential benefits such as facilitation of early weaning and improved rotifer nutritional value may be obtained by utilizing lower density rotifer feeding regimes.     

Effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder <Emphasis Type="Italic">paralichthys olivaceus</Emphasis>

ABSTRACT:   The effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder Paralichthys olivaceus which were acclimated to 3, 14 or 20°C in advance were investigated. The oxygen consumption rate (OCR) of individuals (total length >28 cm) that were acclimated to 3°C, maintained a relatively low value of 6.9 ± 2.3 mL (mean ± SD) O₂/kg ww (wet weight)/h till 5.8°C, which is considered a 'minimum' rate. Thereafter, the OCR gradually increased up to 18.7 mL O₂/kg ww/h at the temperature of 10.6°C. The OCR of individuals acclimated to 14°C stabilized with 124.2 ± 24.1 mL O₂/kg ww/h, when tested at the relatively constant temperature of 14.4 ± 0.17°C. Maximum entropy spectral analysis revealed that the peak OCR occurred at 23.9 h intervals, which could be a circadian rhythm. The mean OCR showed a peak at 06.00–08.00 hours. For the temperature above 25°C in which the fish were acclimated to 20°C, the rhythmic patterns of OCR were observed until the experimental temperature reached 26.4°C. For the temperature above 28.4°C, the metabolic activity rhythm of the OCR was dampened and then the OCR abruptly rose again. It is assumed that the oscillator of the endogenous rhythm lost the governing of normal metabolic activity in Japanese flounders above 28.4°C. These results suggest that the temperatures of around 5.8, 26.4 and 28.4°C are the 'lower limit temperature', 'upper incipient lethal temperature', and the 'critical thermal maximum', respectively, for the endogenous rhythm of oxygen consumption in the Japanese flounder.