Effect of temperature and cell density on ethanol fermentation by a thermotolerant aquatic yeast strain isolated from a hot spring environment

ABSTRACT: A thermotolerant, fermentative yeast strain named RND13 from a hot spring drainage was evaluated for its ethanol-producing ability at elevated temperatures at a high substrate concentration [15% (w/v) glucose] close to the level reflecting industrial practice. The RND13 was capable of utilizing glucose almost completely at 40°C with increasing inoculum size, producing ethanol up to 6.6% (w/v), which is comparable to levels (7.0–7.2%) at 30°C. The maximum rate of ethanol production by the RND13 was found to be 9.0 g/L per h at 40°C in an inoculum sized 5% (w/v). At 43°C, however, the RND13 could not utilize glucose to completion and showed a slight drop in the extent of produced ethanol [6.0% (w/v)]. Thus, the culture at 40°C with a 5% cell inoculum was considered to be the optimal condition for ethanol production at higher temperatures in terms of batch fermentation. In the phylogenetic analysis based on the small-subunit rDNA sequence, the strain was grouped together with both Candida glabrata and Kluyveromyces delphensis , which are relatively close to Saccharomyces cerevisiae .     

Synergetic effects of cell immobilization in polyurethane foam and use of thermotolerant strain on degradation of mixed hydrocarbon substrate by Prototheca zopfii

ABSTRACT:   The synergetic effects of an immobilized cell system in an oleophilic polyurethane foam (PUF) and use of a thermotolerant strain on degradation of hydrocarbons by colorless green microalga Prototheca zopfii are reported. Two strains of P. zopfii , i.e. thermotolerant RND16 and non-thermotolerant ATCC30253, were immobilized in PUF to compare their abilities to biodegrade a mixed hydrocarbon substrate (MHS) containing aliphatic and polycyclic aromatic hydrocarbons (PAHs), as a function of temperature. The thermotolerant strain RND16 degraded MHS at 35°C, while ATCC30253 did not degrade hydrocarbons at temperatures higher than 30°C. Immobilization of P. zopfii in PUF resulted in shortened lag for growth-associated biodegradation of n -alkanes in MHS, the effect of which was most significant in cultures of RND16 at 25°C. Nevertheless, the decrease in the amount of degraded PAHs was caused by PUF immobilization and the level of this decrease was marked in the cultures of RND16, in which rate and extent of n -alkane degradation were higher than for ATCC30253.     

Thermal tolerance of a rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> strain selected by high-temperature breeding

ABSTRACT:   Thermal tolerance was studied in a rainbow trout strain successively selected through high temperature breeding at 20–27°C since 1966 in Miyazaki Prefecture, Japan. The hatching rate and fry mortality at high temperatures were examined in the selected strain along with normal strains cultured at water temperature of 9–17°C. The hatching rate of embryos fertilized at either 10 or 14°C and subsequently subjected to high temperatures in the blastula or neurula stage of the selected strain, was marginally higher than that of the normal strain counterparts. The upper 50% lethal temperatures (LT₅₀) for embryos in the early segmentation, blastula and neurula stages of the selected strain were also higher than those of the normal strain counterparts. Death temperatures and LT₅₀ of fries acclimated to 20°C of the selected strain were significantly higher than those of the normal strains. However, no difference in the critical thermal maximum was detected between the different strains. These results suggest that the selected strain of rainbow trout established by selecting successively for many generations at high temperatures acquired a degree of thermal tolerance.     

Influence of decreasing water temperature and shortening of the light phase on macronutrient self-selection by rainbow trout Oncorhynchus mykiss and common carp Cyprinus carpio

SUMMARY: Macronutrient self-selection patterns in rainbow trout and common carp, which were maintained in groups, were examined when both water temperature and duration of light phase gradually decreased. Three semipurified macronutrient diets composed mainly of protein (casein and gelatin; crude protein (CP) = 65%), fat (pollock oil and soybean oil; crude fat (CF) = 38%), or digestible carbohydrate (dextrin and gelatinized starch; crude starch (CS) = 57%) were offered to five groups of trout (20 fish/group, 51 g/fish) and carp (15 fish/group, 36 g/fish) using three self-feeders. A standard diet (ST) containing the three macronutrients at an appropriate ratio (CP = 47%, CF = 14%, CS = 25%) was also offered to three groups each by a single self-feeder. After an initial 3 weeks of experiencing a long photoperiod (14 h light : 10 h dark cycle) and high water temperature (trout, 17°C; carp, 25°C), the light phase and temperature were gradually decreased in the following 3 weeks to 10 h light : 14 h dark, and 8°C for trout and 13°C for carp, and feeding was continued for a further 3 weeks under low temperature and short photoperiod conditions. Growth and protein utilization parameters between the ST diet self-fed groups and macronutrient diet self-selection groups were not different ( P > 0.05) for both trout and carp. Trout preferred the high-protein diet (HP) to the high-fat and the high-carbohydrate diets ( P < 0.05) regardless of the changes in water temperature and photoperiod. Carp also preferred the HP diet ( P < 0.01) under the high temperature and long photoperiod conditions; however, this preference gradually disappeared with lower temperatures and shorter photoperiods. These results suggest that the most appropriate macronutrient ratios in carp diets change with environmental conditions.     

Short-term salinity tolerance of northern pike, Esox lucius, fry, related to temperature and size

Abstract  The short-term tolerances of northern pike, Esox lucius L., fry reared in a freshwater hatchery, to salinity were examined in the laboratory. Survival of two size groups of pike fry (mean length 21 ± 2 mm SD and 37 ± 4 mm SD) was examined over 72- to 96-h periods at 9–14 ppt salinity in combination with temperatures of 10, 14 and 18 °C. A parametric survival model found a significant correlation between survival of pike fry and temperature and salinity, respectively. L _C50 values after 72 h were between 11.2 and 12.2 ppt, being lowest at 10 °C. Pike fry did not survive more than 13 ppt. Mortality at 12 ppt was significantly faster at 18 °C than 10 or 14 °C. Moreover, mortality was higher and faster for large than for small pike fry at 12 ppt and 14 °C. These results imply that pike raised in fresh water can survive stocking into brackish waters below 11 ppt at least for a short time.     

Effects of temperature on hatchability, development and growth of eggs and yolksac fry of Oreochromis mossambicus (Peters) under artificial incubation

Abstract. The eggs and yolk sac fry of the mouth brooding tilapia. Oreochromis mossambicus (Peters), were artificially reared in an experimental hatchery system at different temperatures. The hatchability, embryonic development of eggs and growth of yolk sac fry were studied under controlled conditions. It was shown that the upper lethal temperature for O. mossambicus eggs was above 40°C and the lower lethal temperature between 11°C and 17°C. For yolk sac fry these values lay between 34 and 40°C and 17 and 20°C respectively. Fry survival between 24·3 and 34·0°C was near to 100% while al 20°C it was less than 60%. The somatic growth rates of fry at temperature between 20 and 34·5°C up to 10 days post hatch are presented and were found to differ significantly. At 34·6°C a negative SGR was recorded during the period 6–9 days post hatching, but, during the first 6 days, the SGR at this temperature was more than four times higher than at 20°C. At the elevated temperatures, the utilization of yolk is faster and the loss of weight observed was due to starvation. This should be taken into account when incubating under artificial rearing condition so that initiation of exogenous feeding is timed most appropriately.     

Histopathology of snakehead, Channa striatus (Bloch), experimentally infected with the specific Aphanomyces fungus associated with epizootic ulcerative syndrome (EUS) at different temperatures

Abstract Snakeheads, Channa striatus (Bloch), were inoculated with a spore suspension of the specific pathogenic Aphanomyces , isolated from fish affected by epizootic ulcerative syndrome (EUS), in South East Asia. Fish were held at three different temperatures: 19, 26 and 31 °C. Histological changes induced by the infection are described. In the early stages of the disease, degenerative changes were observed in all samples, but inflammatory infiltrate was much more marked in fish kept at 26 and 31 °C. By 8 days post-injection, extensive mycotic granulomatosis was observed in the samples kept at 26 and 31 °C. The fish kept at 19 °C developed a severe invasive myonecrosis with limited macrophage response. From 14 to 28 days post-injection, healing became well established at 26 and 31 °C and surviving fish kept at these temperatures recovered completely by 28 days. The lesion was still progressing at 21 days post-injection in fish kept at 19 °C, and all such fish succumbed by this time. Thus, mortalities in the fish kept at 19 °C were considerably higher than in the groups of fish kept at 26 and 31 °C. The findings help to explain why mortalities from EUS occur when water temperatures are low.     

Effect of temperature and feeding on conditioning of Ostrea chilensis Philippi, 1845 reproductors

Abstract. Adult oysters of Ostrea chilensis Philippi 1845, collected from the Quempilén River estuary, Ancud, Chile, were subjected to different temperature (14,17 and 20°C) and feeding treatments (daily rations of dry weight algae equivalent to 0.75% and 1.5% mean dry flesh wt) to measure their influence on gonad maturation. Males spawned under all these experimental conditions, after 25-27 conditioning days, while females spawned only at the same time at the higher temperatures (17 and 20°C). At high food ration (1.5%), the incubation period lasted for 29 days at 20°C, and 44 days at 17°C. At half food ration (0.75%), however, the incubation period was 24 days at 20°C and lasted for more than 54 days at 17°C, longer than under the natural estuary conditions. Larger larvae and better settlement were obtained from oysters kept at 17°C and high food ration (1.5%).     

Characteristics of burnt meat in cultured yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

ABSTRACT:   In order to understand the characteristics of burnt meat in cultured yellowtail Seriola quinqueradiata , fish were kept at two different temperatures (13 and 30°C) and slaughtered by either spinal cord destruction (SCD) or suffocation in air (SA). Early postmortem changes during storage at 32°C were analyzed by rheological, biochemical, and histological methods. The burnt meat (with lightness parameter, L* ≥ 55) was observed at 1-h storage in the SA 30°C group, at 2 h in SCD 30°C, and at 4 h in SA 13°C; meat was normal for the SCD 13°C group until 6 h of storage. Breaking strength scores were higher for the normal meat (200 g/cm²) than burnt meat (70 g/cm²) at 4 h of storage. Expressible water content was higher for the burnt meat than for the normal meat. Adenosine triphosphate concentrations for the SCD groups were higher than for the SA counterparts. Moreover, pH decrease was much faster in the 30°C groups, showing pH 5.6 at 2 h of storage. A negative correlation between the pH and lactic acid contents in muscle ( P  < 0.001) was found. Histological analysis evidenced a larger pericellular area (40%) in the burnt samples than in the normal samples (16%). It was confirmed that a higher fish-keeping water temperature and a stressful slaughter method (faster glycolytic process) were determinative factors that influence the occurrence of burnt muscle in yellowtail, and that the effect of the former is larger than the latter.     

Use of marine yeasts as an available diet for mass cultures of Moina macrocopa

A 4‐week feeding trial was conducted to test the use of marine yeasts purified from seawater and sediments as a dietary source in cultivating a cladoceran, Moina macrocopa, a potential alternative live food for fish larvae. Optimal growth conditions of two yeast strains were obtained for NaCl concentration, pH and temperature. Moina macrocopa was cultivated using different diets: marine yeasts (Debaryomyces hansenii Yeast‐14 and Candida austromarina Yeast‐16) and a commercial diet (Erythrobacter sp. Sπ‐I). The essential amino acids of both the yeast strains were nearly as great as those in M. microcopa. Further, the yeast‐fed M. macrocopa had essential amino acid profiles similar to the documented values for rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. Sπ‐I lacked n‐3 polyunsaturated fatty acids, 20:5n‐3 and 22:6n‐3, which were also low but detected in both yeasts. An increase in the 20:5n‐3 [eicosapentaenoic acid (EPA)] and 22:6n‐3 [docosahexaenoic acid (DHA)] levels, compared with the levels in yeast strains themselves, was more pronounced in the 22:6n‐3 level of Moina fed the C. austromarina, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their δ¹³C values shifted gradually towards the values of the switched diets. Diet switch from Erythrobacter sp. Sπ‐I to C. austromarina Yeast‐16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the δ¹³C values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the δ¹³C values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly C. austromarina Yeast‐16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietary values than the commercial diet (Erythrobacter sp. Sπ‐I).     

Effects of temperature on biochemical reactions and drug resistance of virulent and a virulent Aeromonas salmonicida

Abstract. Incubation temperatures of 11°, 18° and 28° did not substantially affect biochemical reactions of either virulent or avirulent forms of Aeromonas salmonicida subspecies salmonicida. The only change observed, amygdalin fermentation, was positive at 11° and 18° but negative at 28°C. Several isolates utilized sucrose, a characteristic not normally recognized for A. salmonicida subspecies salmonicida. Antimicrobial susceptibility screening indicated resistance to novobiocin increased at the higher incubation temperatures. Standardized drug sensitivity testing procedures and precise zone diameter interpretive standards for bacterial fish pathogens are needed.     

Salinity-induced quiescence in eggs of the calanoid copepod Acartia tonsa (Dana): a simple method for egg storage

We report the effect of salinity and temperature on the viability of stored culture-based subitaneous eggs of the calanoid copepod Acartia tonsa for use of copepods in fish larvae culture. Quiescence induction was recorded at 17 and 25 °C, in salinities from 0 to 30. Quiescence was strongly induced at 0 salinity and partially at 5 in both temperatures. Eggs incubated at 0 salinity for up to 12 days at both temperatures showed a decline in the fraction able to be induced into quiescence by abrupt salinity changes. The hatching success of eggs that were able to enter quiescence stabilized after a 1-day incubation and remained ∼25% viable for 12 days in 17 °C. On the contrary, the 25 °C trial showed a gradual decline in viability until stabilizing ∼10% at day 7 and onwards. Longterm 17 °C incubation for 35 days showed that eggs remained quiescent with a viability of ∼14%. Hence, we recommend salinity storage of A. tonsa subitaneous eggs as a relevant shortterm technique, and a suitable alternative to the recently proposed cold storage of eggs when eggs are to be shipped from the copepod producer to a given fish larvae hatchery.     

Suppression of fish meat softening by strict control of storage temperature

ABSTRACT:   Changes in meat firmness and structure of cultured yellowtail at several storage temperatures, from −1.5°C to 10°C, were compared. Firmness decreased less at −1.5°C and 10°C than at the other temperatures. During storage at temperatures from −0.5 to 4.0°C, expansion of intercellular space, which indicates weakening of the connecting force between muscle cells, began to increase after 8 h storage, and the ratio of space to the whole area increased by 3.4–4.9 times after 24 h of storage. However, at −1.5°C and 10°C, the increase of intercellular space was suppressed, and the ratio was 2.4–2.6 times, even after 24 h storage. Collagen fibrils connecting muscle cells disintegrated after 24 h storage at all storage temperatures. Conversely, in 10°C storage, the fine structure of muscle, except for collagen fibrils, was maintained better than at other storage temperatures. These findings indicate that −1.5 and 10°C storage could maintain fish-meat quality for longer periods than storage at −0.5, 1.0, or 4.0°C, the normal refrigeration temperature.     

Qualitative and quantitative changes of F<Subscript>o</Subscript>F<Subscript>1</Subscript>-ATPase in Japanese flounder and red sea bream associated with rearing temperatures

ABSTRACT:   Fast skeletal muscles of Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major were examined for quantitative and qualitative changes of mitochondrial ATP synthase (F_oF₁-ATPase) in association with rearing temperatures. The specific activities of F_oF₁-ATPase from Japanese flounder reared at 10°C, 15°C and 25°C for 4 weeks were determined to be 81 ± 11, 74 ± 13 and 83 ± 11 nmol/min·mg mitochondrial protein, respectively. The corresponding activity from red sea bream reared at 8°C for 5 weeks was determined to be 65 ± 9 nmol/min·mg mitochondrial protein, which was higher than 33 ± 9 nmol/min·mg mitochondrial protein in fish reared at 23°C. The contents of α- and β-F₁-ATPase in total mitochondrial proteins were not significantly different between fish reared at different temperatures for the two fish species. However, the contents of β-F₁-ATPase in the total fast skeletal muscle extracts, prepared from Japanese flounder reared at 10°C, were 2.1- and 2.9-fold higher than those for fish reared at 15°C and 25°C, respectively. The corresponding content from red seabream reared at 8°C was 2.2-fold higher than that for fish reared at 23°C. Therefore, the changes in F_oF₁-ATPase depending on rearing temperatures were species-specific.     

Effect of temperature on growth,digestion and immunity of green,white and purple sea cucumbers,Apostichopus japonicus (Selenka)

Three color morphs (white, green and purple strains) of Apostichopus japonicus (Selenka) were cultured in artificial seawater, for approximately 90 days, in three temperature ranges: 27–22°C (high), 22–17°C (mid) and 17–12°C (low). All strains grew in all temperature ranges. Temperature significantly affected growth rate, digestibility, digestive enzymes and immune‐related enzymes. Highest specific growth rates were exhibited in 4‐month‐old sea cucumbers at mid and high temperatures, and in 16‐month‐old sea cucumbers at mid and low temperatures. Specific growth rates of green and purple strains were not significantly different, but were significantly higher than that of the white strain at mid temperatures. The digestibility of each strain was significantly higher at 27°C, 22°C and 17°C than at 12°C. Green‐strain digestibility was higher than that of purple and white strains at specific temperatures. Protease and amylase activities of all strains followed bell‐shaped temperature curves with maximum digestive enzyme activity at 17°C. The activities of alkaline and acid phosphatases were higher in the guts of the green strain than in the white or purple strains at the same temperature. Superoxide dismutase activity was higher in the purple strain than in white and green strains.     

Differences in self-feeding activity between thermally selected and normal strains of rainbow trout Oncorhynchus mykiss at high temperatures

ABSTRACT:   Feeding activity was examined at high temperatures by using a demand feeder for thermally selected and normal rainbow trout Oncorhynchus mykiss strains. When the water temperature was raised in experiment 1 from 17.5 to 25.7°C at 0.3°C/day in 21 days, the average daily food consumption rate in the thermally selected strain rose to 7.1%, which was significantly higher than that of the normal strain (4.1%, P  < 0.05). The corresponding rate was also significantly higher in the thermally selected (0.8%) than in the normal strain (0.2%, P  < 0.05) of fish in experiment 2 where water temperature was raised to 24.1°C in 0.5°C/day increments. When water temperature was raised rapidly in experiment 3 from 16.7 to 21.7°C in one day and gradually to 24.4°C in 28 days at 0.1°C/day, the average daily food consumption rates were 1.0 and 0.1% for the thermally selected and normal strains, respectively, with significant difference ( P  < 0.01). These results suggest that the thermally selected strain has acquired thermal tolerance as a result of artificial selection.     

Effects of Temperature on Egg Hatch,Larval Growth and Metamorphosis for Hatchery-Cultured Southern Flounder,Paralichthys lethostigma

ABSTRACT

Egg hatch, larval growth, and metamorphosis of southern flounder, Paralichthys lethostigma, were examined at 13, 17, 21, and 25°C in laboratory experiments. The experiments were separated into four developmental phases: (1) from fertilization until hatch; (2) from hatch until mouth opening; (3) from first feeding until the onset of metamorphosis; (4) from the onset until the completion of metamorphosis. Time to egg hatch were 109, 58, 39, 30 hours at 13, 17, 21, and 25°C, respectively. Size at hatch varied little among temperatures, while percent hatch was significantly higher at 17°C (P < 0.05). There was no significant difference in total length at mouth opening among temperatures (P > 0.05). However, those larvae raised at higher temperatures reached mouth opening earlier, as it took on average 7.4, 3.9, 2.8, and 2.2 days from hatch at 13, 17, 21, and 25°C, respectively. During phase three, the growth rate was optimal at 17°C, as growth rates were 0.046, 0.110, 0.106 and 0.096 mm/day at 13, 17, 21, and 25°C, respectively. Larvae in phase four completed metamorphosis sooner at higher temperatures than at lower temperatures. It took 26.4, 15.7, and 13.1 days to complete metamorphosis at 17, 21, and 25°C, respectively, while only 16% of those at 13 progressed to stage “G” within the 27 days that phase four encompassed. No difference in total length was found among treatments (P > 0.05) at the completion of metamorphosis, but fish in the 21 °C treatment had a significantly higher survival (P < 0.05). It can be concluded that southern flounder could be produced in the highest quantity and at the largest size by raising them at 17°C from fertilization until the onset of metamorphosis, and at 21 °C during metamorphosis.     

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.     

Effects of temperature on vitellogenesis in Japanese eel Anguilla japonica

ABSTRACT:   Cultured immature female Japanese eels acclimated to sea water at either 10 or 20°C were treated weekly with salmon gonadotropin fraction (sGTH) in order to investigate the effects of water temperature on artificial induction of ovarian maturation. In eels maintained at 20°C, ovulation was induced in 11 of 18 fish during the experimental period of 13 weeks, whereas at 10°C all fish showed a low gonadosomatic index (GSI) at the end of the experiment. Plasma vitellogenin levels were higher in eels kept at 20°C than in eels at 10°C throughout the experiment. However, no significant differences were observed in the plasma testosterone and estradiol-17β levels between groups. Eels pretreated with sGTHs at 10°C for 13 weeks were separated to two different temperature (10 and 20°C) groups, and received the same weekly sGTH injections. In eels transferred to 20°C, plasma vitellogenin levels, GSI and oocyte diameter were increased, but these values were maintained at low levels in eels that remained at 10°C. These results clearly indicate that water temperature is an important factor regulating vitellogenesis in the Japanese eel.     

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.