Effects of fish size and water temperature on the acute toxicity of boron to Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> and red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:     The acute toxicities of boron were examined for Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major in terms of fish size and water temperature. Japanese flounder of 0.1–70.0 g and red sea bream of 0.6–20.3 g were exposed to different concentrations of boron for 96 h at 20°C under semi-static conditions. In both fish species, the median lethal concentration (LC₅₀) for 96 h of boron increased linearly with increasing fish size, ranging from 108 to 252 mg B/L for the flounder, and from 97 to 172 mg B/L for the sea bream. The effect of water temperature on acute toxicity was examined for Japanese flounder of 0.6 and 1.5 g at 10, 15, 20 and 25°C, and for red sea bream of 0.6 and 2.4 g at 12, 15, 20 and 25°C. The toxicity of boron for the flounder increased linearly with increasing water temperature. The 96 h LC₅₀ values ranged from 299 to 108 mg B/L for the 0.6 g flounder and from 350 to 113 mg B/L for the 1.5 g flounder. A similar trend was shown for the 2.4 g red sea bream; however, the relationship for the 0.6 g red sea bream was not significant.     

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.     

Assessing the end-point temperature of heated fish and shellfish meats

ABSTRACT: Attempts have been made to assess the end-point temperature (EPT) of heated fish and shellfish meats by using the coagulation method together with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and enzyme activity determination. Unfrozen and frozen fish and shellfish meats were heat-treated at different selected temperatures with 0, 15 or 30 min holding times. Proteins were extracted with NaCl solution. The coagulation method was able to determine EPT of heated fish and shellfish meats between 60 and 67°C. SDS-PAGE patterns of the filtrates from heated meats were closely related to the results of the coagulation method and enzyme activity determination. Two proteins responsible for producing coagulum of fish meat extracts seem to be lactate dehydrogenase (LDH) and glyceraldehyde phosphate dehydrogenase (GAPDH). End-point temperatures determined by these methods were not significantly different between unfrozen and frozen samples. On the contrary, a highly thermostable protein with a molecular mass of 35 kDa was detected in heated shellfish meats up to 108°C. In scallop adductor muscle, this highly thermostable protein was found to be the tropomyosin subunit from its amino acid composition and their partial sequences. Tropomyosin could be used as an EPT indicator up to 108°C for heated shellfish meats.     

Myofibrillar proteolysis by myofibril-bound serine protease from white croaker <Emphasis Type="Italic">Argyrosomus argentatus</Emphasis>

ABSTRACT:   The modori phenomenon is defined as heat-induced myofibrillar degradation caused by endogenous serine protease(s) of fish muscle during Kamaboko fish meat gel production. This study was undertaken to analyze myofibrillar proteolysis of white croaker Argyrosomus argentatus muscle, which is an ingredient of high quality Kamaboko, by myofibril-bound serine protease (MBSP) under conditions corresponding to the modori phenomenon. White croaker MBSP was stable between pH 2–11 and below 65°C, and about 60% of its initial activity remained after incubation for 2 h under the conditions at 65°C and pH 7.5. About 60% of the enzyme activity was suppressed by 0.5 M NaCl. White croaker MBSP degraded various myofibrillar proteins between 40 and 70°C and pH 6.0–9.0, and preferentially degraded myosin heavy chain rather than other myofibrillar proteins. The enzyme degraded the myosin heavy chain most strongly at 55°C and pH 7.0, and a major part of the bands of myosin heavy chain and its degradation products disappeared for a period of 2 h. These degradation characteristics are very similar to those observed during the modori phenomenon, indicating that MBSP could be a modori-inducing protease involved in the modori phenomenon of white croaker Kamaboko production.     

Setting Ability of Threadfin Bream (Nemipterus japonicus) Meat as Affected by Freezing and Frozen Storage with Special Reference to Transglutaminase Enzyme Activity and Rheological Properties

The effect of freezing and frozen storage of threadfin bream fish (Nemipterus japonicus) meat on the setting and gel-forming ability has been evaluated. The dynamic viscoelastic properties of fish meat during setting and thermal gelation process were evaluated using Controlled Stress Rheometer under oscillatory mode. A sharp decrease in setting ability was recorded immediately after freezing as revealed by storage modulus (G?) values. The transglutaminase (TGase) enzyme activity of fish meat decreased from the initial value of 81.09 to 51.46 U/g meat/min at the end of 200 days of frozen storage. A decrease in setting ability of fish meat beyond 160 days of frozen storage is probably related to lower TGase enzyme activity. The gel-forming ability was related to setting ability during the frozen storage period. Although the protein solubility showed a decreasing trend during 200 days of frozen storage, the decrease was not significant. The effect of freezing and frozen storage on calcium-activated adenosine triphosphatase (Ca²⁺-ATPase) enzyme activity of fish mince was significant (P < 0.05). A reduction in protein solubility and Ca²⁺-ATPase enzyme activity is an indication of aggregation/denaturation of myofibrillar proteins.     

Experimental transmission of encephalopathy and retinopathy induced by nodavirus to sea bream, Sparus aurata L., using different infection models

The transmission of viral encephalopathy and retinopathy (VER) was investigated in juvenile sea bream, Sparus aurata L. Two different infection routes [intraperitoneal (i.p.) and intramuscular injection (i.m.)] were tested at two different temperatures (20 and 26 °C) using sea bream of mean weight 0.7, 2 and 4 g, as well as an immersion challenge performed at 26 °C with sea bream of 0.7 g. Successful transmission of the disease was only achieved by i.m. injection. Mortalities of 100% occurred in sea bream of 0.7 g at day 15 post-infection and 47% in sea bream of both 2 and 4 g at day 30 post-infection in all the experimental infections at 26 °C. No mortalities were ever observed with infections at 20 °C. When mortalities were observed, the virus was detected by immunoperoxidase staining in the SSN-1 cell line inoculated with tissues from infected fish. Histological examination of both normal and infected fish showed a vacuolization in the bipolar and granular layers of the retina of the infected sea bream. This is the first experimentally induced transmission of VER in sea bream. Differences were observed at the time of disease onset depending on water temperature, the route of infection and the age of the juvenile fish.     

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.     

Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder,Paralichthys olivaceus,and Red Sea Bream,Pagrus major

The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Self-feeding activity patterns in gilthead sea bream (Sparus aurata), red porgy (Pagrus pagrus) and their reciprocal hybrids

This study was conducted to monitor feeding activity of gilthead sea bream (Sa) and red porgy (Pp) and to investigate the hereditary effects on feeding in their hybrids (female Sa×male Pp, female Pp×male Sa), obtained after artificial crosses. All fish groups were held in 500-l tanks under ambient temperature (15–25°C) and light conditions (daylength from 9 h 30 min to 14 h 48 min). Fish (initial body weight 100–165 g) were fed by means of self-feeders, which were connected to a computer, recording fish feeding activity for a year. Results showed that there was a 24-h periodicity in all fish types over the experiment. In their daily feeding patterns, both parental species were diurnal: red porgies had a stable daily pattern over the experiment, while gilthead sea bream shifted their activity from a general daylight fluctuation in warm periods to an afternoon peak in cold periods. The Sa×Pp hybrids showed intermediate feeding patterns compared to their parents in warm periods and a pattern that was similar to the red porgy in cold periods, while the Pp×Sa hybrids had rather unchangeable daily feeding patterns, which were not related to the parental ones. The proportion of night feed demands increased in cold periods independent of fish group. The relationship between temperature and feed intake was species-dependent. Finally, this study indicated that feeding expression in hybrids was related to the combination of the parental genome, supporting our hypothesis for a hereditary influence on feeding activity in fish.     

Detection of cathepsin L in red cell membranes from fish blood

Cathepsin L, one of the cysteine proteases found in fish muscle, is considered to be the main cause of post-mortem autolysis of fish muscle. We have determined the presence of cathepsin L in the membranes of red blood cells of carp, amberjack, and red sea bream and measured its activity. Immunoblotting of an extract of the red blood cell membranes from these three fish species using human anti-cathepsin L antibody revealed the presence of cathepsin L of different molecular masses. The molecular masses of cathepsin L was estimated to be 120 and 85 kDa in the amberjack and 75 and 70 kDa in the carp. These proteins have higher molecular masses than the mature form of cathepsin L, suggesting that they are precursor forms. In contrast, the protein in the red sea bream was estimated to have a molecular mass of 30 kDa, suggesting that this cathepsin L is a mature form. The specific activity of cathepsin L was highest in the red blood cell membranes of the amberjack, followed by the carp and the red sea bream in descending order.     

Sensory Characteristics of Threadfin Bream (Nemipterus japonicus) Hydrolysate

ABSTRACT

Threadfin bream (Nemipterus japonicus) spray-dried hydro-lysate was prepared using Alcalase at predetermined optimum hydrolysis conditions consisting of pH 8.5, 60°C, 2% enzyme, and 2 h hydrolysis. The hydrolysate was evaluated for its preferred concentration in rice porridge. Its acceptability was compared with sauces and enzyme-treated rice and vegetable hydrolysate. The preferred concentration in rice porridge was found to be 3% (w/v) and bitterness increased with concentration of hydrolysate. Threadfin bream hydrolysate was preferred as much as soy sauce; however, it received a lower score for taste compared to fish sauce. In terms of color, threadfin bream hydrolysate received significantly (p < 0.05) lower score than Flavorzyme and Alcalase treated rice hydrolysate and Vegetable hydrolysate. Low preference to the hydrolysate could probably be due to its greenish yellow color in which the common color for flavorants are brownish. For overall acceptability, no significant difference was observed between threadfin bream hydrolysate and Alcalase-treated rice hydrolysate but was significantly lower than vegetable hydrolysate and Flavorzyme-treated rice hydrolysate.

The hydrolysis of threadfin bream with Alcalase produced hydrolysate having a combination of bitter, umami, salty, sweet and sour tastes with fishy flavor. Bitterness and umami were rated as having an intermediate intensity, salty as between weak and intermediate, and sour and sweet as weak. The three basic tastes for hydrolysate were bitter, umami and salty, with umami and bitterness being the determining tastes.     

Upper lethal and rearing temperatures for juvenile common carp, Cyprinus carpio L., and silver carp, Hypophthalmichthys molitrix (Valenciennes)

Abstract. The ultimate upper lethal temperature (UULT) was 43.0 or 46.0°C for common carp and 43.5 or 46.5°C for silver carp, depending on the method of calculation. The influence of fish age on lethal temperature values was low. Using the relationship between the UULT and the optimum growth temperature the latter was calculated as being 38°C for common carp and 39°C for silver carp. No significant differences were found in survival of fish reared at lower and higher temperatures when proper food was used.     

Study on Gel Properties of Silver Carp (Hypophthalmichthys molitrix) and White Croaker (Argyrosomus argentatus) Blended Surimi at Different Setting Conditions

Gel properties of blends of surimi from silver carp and white croaker set for a range of times and at different temperatures were evaluated for breaking force, breaking distance, and whiteness. Total protein content was 12% in all samples, and the concentration of white croaker surimi protein in the blends was designed at 0, 10, 20, 30, and 40%. The blended surimi had greater gel strength than either of the two types of surimi alone. Breaking force and breaking distance after setting were also significantly higher than that of the blended surimi cooked directly without setting. The blended surimi exhibited less modori than either of the two types of surimi alone heated to 50°C. The whiteness values of the blended surimi were significantly higher than that of white croaker alone. These results indicate there are advantages to be gained in utilizing surimi from freshwater silver carp in blends with surimi from marine captured white croaker which is becoming limited in supply.     

Ontogeny of tolerance to and avoidance of ultraviolet radiation in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli

ABSTRACT:   Ontogenetic changes of tolerance to, and avoidance of, ultraviolet-B radiation (UV-B) were examined in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli . In the tolerance experiment, larvae and juveniles (age 13–46 days) were put in beakers, and were exposed to one of five different levels of UV-B radiation (1.8, 1.1, 0.2, 0.1, and 0 W/m²) for one hour. Their survival rates were calculated either 12 or 24 h later. In the avoidance experiment, fish (age 3–49 days) were put in a long experimental tank, half of which was covered with UV-blocking film and placed under two levels of UV-B radiation (1.1 and 0.2 W/m²), and their avoidance indices were calculated. Black sea bream had significantly better survival compared to red sea bream for most ages. Only black sea bream of ages 37 and 49 days showed significant avoidance of UV radiation under the higher level of UV-B, whereas both species did not show avoidance on any days at the lower level. The present results suggest that black sea bream are significantly better adapted to habitats with high UV-B radiation, than red sea bream, reflecting that back sea bream live in shallower waters through their early life stages.     

Impact of aquatic acoustic noise on oxidative status and some immune parameters in gilthead sea bream Sparus aurata (Linnaeus, 1758) juveniles

This study evaluated the ambient noise effect of onshore and offshore aquaculture systems on oxidative status and on some immune parameters in gilthead sea bream (Sparus aurata) juveniles. The experiment was performed for 40 days, projecting two different playlists of acoustic stimuli: offshore aquaculture noise condition that created the typical acoustic field in proximity of an offshore sea cage for fish farming, using a random sequence of quite sea background and boat noises; and onshore aquaculture noise condition that represented the acoustic field inside an open concrete tank for fish farming. A third group without any acoustic projection was used as a control. After 40 days from the beginning of acoustic noise, fish exposed to the offshore aquaculture condition showed significantly higher levels of total oxidant status, lysozyme activity, antiprotease activity and white blood cells together with a lower albumin/globulin ratio in comparison to fish from the control group; whereas fish exposed to the onshore condition showed analytical data placed between the other groups, though never significantly different. These results demonstrate that the offshore aquaculture noise, and in particular the sea soundscape, adversely influences the oxidative status and the immune function of gilthead sea bream determining a mild stress condition that could affect the sea bream welfare.     

Supplemental effect of <Emphasis Type="Italic">Porphyra</Emphasis> spheroplasts on the growth and feed utilization of black sea bream

ABSTRACT:   A 56-day feeding trial was conducted to evaluate the effects of Porphyra spheroplasts (PS) as a feed additive on growth, carcass composition and feed utilization of black sea bream. Four experimental diets with or without PS (0, 1, 3, 5% inclusion level) were formulated to determine the proper incorporation level for best growth and utilization using white fishmeal as the principal protein source. Three replicate groups of fish averaging 1.28 ± 0.05 g (mean ± SD) were maintained in flow-through thermo-controlled sea water (32–34 psu, 25°C) aquaria (100 L) under the laboratory conditions (L12 : D12) and fed three times a day at 09:00, 13:00 and 17:00 hours by hand at approximately 3% body weight on each diet. The rearing trial revealed that growth performance (in terms of weight gain, specific growth rate) are almost the same level in all the dietary groups, nevertheless the survival, nutrient utilizations and retentions (feed efficiency, protein efficiency ratio, protein retention rate, lipid retention rate) were significantly higher ( P  < 0.05) in a group fed on the diet containing 3% spheroplasts. Further, the fish fed on the diet containing PS had significantly higher ( P  < 0.05) crude lipid level in their dorsal muscles and viscera. The fish fed a PS-based diet showed higher level of eicosapentaenoic acid and docosahexaenoic acid (DHA) in viscera and DHA in dorsal muscles. The results obtained in this feeding trial suggest that the optimum dietary PS supplementation level up to 3% for black sea bream can be considered as improved consequence on growth performance, nutrient utilization or body composition were noticed.     

Efficacy of taurine supplementation for preventing green liver syndrome and improving growth performance in yearling red sea bream Pagrus major fed low-fishmeal diet

ABSTRACT:   This study was performed to evaluate the efficacy of taurine supplementation for preventing green liver syndrome and improving growth performance in red sea bream Pagrus major fed a low-fishmeal (FM) diet. Yearling red sea bream were fed for 34 weeks on low-FM diets either supplemented with taurine, or without taurine, and the tissue taurine and bile pigment concentrations were measured. Compared to the fish fed the FM diet, fish fed the low-FM diet without taurine supplementation resulted in inferior feed performances and higher incidence of green liver related to the morphological transformation of the erythrocytes. In these fish, the hepatopancreatic taurine concentration was significantly lower and hepatopancreatic biliverdin concentration was high compared to the fish fed the FM diet. These parameters were markedly improved by taurine supplementation of the low-FM diet and were similar in levels to the fish fed the FM diet. These results indicate that green liver appearance and inferior feed performances of red sea bream fed the low-FM diet without taurine supplementation were caused by dietary taurine deficiency, and indicate the requirement of taurine supplementation to low-FM diets for red sea bream.     

条石鲷的耗氧率与窒息点研究

2008年4～8月,采用Winkler法研究了70日龄（平均全长37.68mm,平均体重1.07g）和105日龄（平均全长65.71mm,平均体重5.58g）的条石鲷在不同温度（15、20和25℃）、不同盐度（20、25和30）下的耗氧率、窒息点及耗氧率昼夜变化。结果表明,105日龄组的相对耗氧率明显低于70日龄组,但个体耗氧量明显高于70日龄组;耗氧率随幼鱼体重的增加而相对减少。盐度一定时,温度升高,相对耗氧率和个体耗氧量都随之增大。盐度对70日龄幼鱼组的耗氧率影响显著（P〈0.05）,对105日龄幼鱼组影响不显著（P〉0.05）。盐度25左右、温度20～25℃时是条石鲷苗种培育和生长的最适盐度和最适温度。条石鲷在温度15～25℃、盐度20～30下的窒息点为1.6021～2.9538mgO2/L,窒息点随着个体的增大而降低;温度、盐度对条石鲷窒息点的影响均不显著（P〉0.05）。条石鲷耗氧率的昼夜变化明显,白天的耗氧率平均为0.9459mgO2/g.h,夜间平均为0.2727mgO2/g.h,白天的耗氧率是夜间的3～5倍,条石鲷白天代谢旺盛并持续到夜间22∶00左右。     

Comparison of 5′-inosine monophosphate and p-nitrophenyl phosphate degrading activities among red,pink, and white muscle fibers of cultured carp

ABSTRACT:   As part of a study to clarify the differences in the temporal change in K -value among fish species, the temporal change in K -value and the 5'-inosine monophosphate (5'-IMP) and p-nitrophenol phosphate (p-NPP) degrading activities in the red, pink, and white muscle fibers in the dorsal muscle of the carp were compared. The temporal change in K -value was fastest in red, followed by pink, and white muscle fibers, at both 0°C and 32°C. Moreover, the 5'-IMP and p-NPP degrading activities were highest in red, followed by pink, and white muscle fibers at near optimum pH concentrations. The 5'-IMP degrading activity at pH 7.0 had a positive correlation with the increasing rate of K -value at 32°C for all types of muscle fibers. These results suggest that differences in increasing rates of K -values between red, pink, and white muscle fibers corresponded to the 5'-IMP degrading activities.