TMAOase, trimethylamine-N-oxide demethylase, isa thermostable and active enzyme at 80°C

ABSTRACT:   Purified trimethylamine- N -oxide demethylase (TMAOase)from walleye pollack muscle is a thermostable protein that was notinactivated after heating at 80°C for 30 min.The heated enzyme was electrophoresed in the same manner as fornative enzyme. Circular dichroism (CD) spectra for purified enzymechanged reversibly in the temperature range of 10–80°C.As the enzyme was still active at 80°C, the CD spectralchange did not directly relate to enzyme activity. TMAOase activity inthe myofibrillar fraction decreased sharply above 30°C,but was extracted and recovered from the heated myofibrillar fraction,suggesting that the activity seemed to be interrupted and apparently inactivateddue to the thermal alteration of myofibrillar proteins or some unknownfactors. The complicated profile found in dimethylamine (DMA) formationfrom trimethylamine- N -oxide (TMAO) in walleye pollack muscleduring heating consisted of both enzymic and non-enzymic processes.Most DMA was produced enzymatically below 40°C and interruptedabove 40°C. Therefore, DMA and trimethylamine was formednon-enzymatically at high temperatures regardless of the presenceof native enzyme. A new, simple and easy purification method wasproposed based on the thermostable nature of the enzyme.     

Total activity of transglutaminase at various temperatures in several fish meats

ABSTRACT: Transglutaminase seems to be related to the setting phenomenon of fish meat paste that occurs at temperatures below 40°C. In many reports on the relationship between transglutaminase and setting phenomenon, the enzyme activity has been measured at 25°C. However, it is known that the setting phenomenon is complicated and the effect of calcium and the 3-D structure of myofibrils, which are sensitive to temperature, play important roles in the reaction. In the present study, total activities of transglutaminase of threadfin bream, white croaker, red sea bream and carp meats were measured at various temperatures. Total transglutaminase activities at 25°C of tested fish meats are arranged in order as follows; white croaker, red sea bream, carp, threadfin bream. In contrast, optimal temperature of carp meat is 30°C, red sea bream 40°C, threadfin bream 50°C and white croaker 50–55°C. In carp meat, the enzyme activity at optimal temperature (30°C) became 8.5-fold higher than at 25°C. The data of the total activity at various temperatures are useful in order to comprehend the details of the reaction.     

Preventive method of color deterioration of yellowtail dark muscle during frozen storage and post thawing

ABSTRACT:   The aim of this study was to develop a practical preventive method for color deterioration of sliced or filleted yellowtail muscle, especially of dark muscle, during frozen storage and post thawing. When the sliced meats were packaged in a vacuum with low oxygen permeable flexible films and then stored frozen below −40°C, no significant discoloration or browning of dark muscle was observed for 9 months or more. For higher temperature storage at −20°C or −30°C, nitrogen gas substituted packaging was a useful practical method for storing sliced meats for 6 weeks. In order to prevent color deterioration of sliced meats after thawing and subsequent storage at 0°C, the efficacy of materials of packaging was investigated. The most desirable result was obtained by using a film with oxygen permeability of 50–90 cm³/m² per 24 h.     

Chemical, physical and sensory changes of small abalone meat during cooking

ABSTRACT:   Small abalone meats were heated at 80°C and 98°C for 0–120 min and the differences in chemical, physical and sensory changes of the cooked meats were investigated. The decrease in moisture and weight and the increase in browning and Hunter's b -value were relatively higher for cooking at 98°C than at 80°C. After cooking for 20–120 min, the total amount of adenosine triphosphate and its related compounds on a dry weight basis decreased by 17–27% at 80°C and by 30–39% at 98°C; the total amount of free amino acids on a dry weight basis changed insignificantly at 80°C but decreased by 22–35% after cooking at 98°C. The meats cooked at 80°C were higher in cutting force whereas the levels in the samples cooked at 98°C did not decrease until samples had been cooked for 60 min. The hydroxyproline content showed little change during cooking except for in samples cooked at 98°C for 120 min, in which the content was found to be low. The extended cooking at 80°C improved the acceptability of small abalone meat, whereas only the acceptability score of aroma increased significantly for cooking at 98°C.     

Heat treatment affects protein quality and protease activity in decapsulated cysts of Artemia when used as starter food for larvae of African catfish Clarias gariepinus (Burchell)

Decapsulated cysts of Artemia subjected to different heat treatments (40, 60, 80 and 96 °C) were fed to African catfish Clarias gariepinus larvae. Heated cysts, untreated cysts and live Artemia nauplii as control constituted the experimental diets. Protein denaturation and solubility, total alkaline protease and specific trypsin activities in the cyst diets were evaluated. The growth of catfish larvae and the proteolytic activity of larval samples during development were also determined. Heat treatment of cysts increased protein denaturation and decreased protein solubility. The protease activity in the cyst diets decreased with higher heating temperatures. The growth of catfish larvae differed according to the diet; higher fish growth was achieved with nauplii and cysts heated at 40 °C. The digestive enzyme activity in larval samples remained similar in all dietary treatments during larval development. The quality of food protein and the way this protein is processed might be more important for successful larval growth than exogenous enzyme supply.     

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.     

Effects of temperature on disease progression and swimming stamina in Ichthyophonus-infected rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss , were infected with Ichthyophonus sp. and held at 10 °C, 15 °C and 20 °C for 28 days to monitor mortality and disease progression. Infected fish demonstrated more rapid onset of disease, higher parasite load, more severe host tissue reaction and reduced mean-day-to-death at higher temperature. In a second experiment, Ichthyophonus -infected fish were reared at 15 °C for 16 weeks then subjected to forced swimming at 10 °C, 15 °C and 20 °C. Stamina improved significantly with increased temperature in uninfected fish; however, this was not observed for infected fish. The difference in performance between infected and uninfected fish became significant at 15 °C ( P  =   0.02) and highly significant at 20 °C ( P  =   0.005). These results have implications for changes in the ecology of fish diseases in the face of global warming and demonstrate the effects of higher temperature on the progression and severity of ichthyophoniasis as well as on swimming stamina, a critical fitness trait of salmonids. This study helps explain field observations showing the recent emergence of clinical ichthyophoniasis in Yukon River Chinook salmon later in their spawning migration when water temperatures were high, as well as the apparent failure of a substantial percentage of infected fish to successfully reach their natal spawning areas.     

Quality improvement by vacuum treatment of boiled skipjack loins for the production of dried skipjack

ABSTRACT: This study attempted to reduce the lipid and water contents in boiled skipjack loins by vacuum treatment and the quality of vacuumed boiled meat was then evaluated during the chilled and frozen storage. The boiled meats were vacuumed at a pressure of 400 Pa for 30 min using a vacuum freeze dryer as an experimental convenience in order to degrease, cool and dehydrate rapidly. The vacuumed boiled meats were stored at 0.0 ± 0.2°C for 10 days and at −20 ± 0.5°C for 90 days. The unvacuumed boiled meats were cooled by air-cooling and stored as the control. Lipid content on a wet basis (w.b.) was reduced as much as 0.5% in the ventral part and 0.4% in the dorsal part of the boiled meats after vacuum treatment. Water content (w.b.) was reduced as much as 2.5% in the ventral part and 3.0% in the dorsal part. The thiobarbituric acid and volatile basic nitrogen values in vacuumed boiled meats were significantly lower ( P < 0.05) than that in unvacuumed boiled meats during storage. Inosine monophosphate (IMP) was only detected as a major nucleotide (about 70%) in boiled meats, and the decomposition of IMP in vacuumed boiled meats was smaller than that in unvacuumed boiled meats during storage. It was concluded that the quality of boiled skipjack loins could be improved by vacuum treatment as a preprocessing procedure in the production of dried skipjack such as katsuobushi .     

Two iridovirus-susceptible cell lines established from kidney and liver of grouper, Epinephelus awoara (Temminck & Schlegel), and partial characterization of grouper iridovirus

Two iridovirus-susceptible cell lines were established and characterized from grouper Epinephelus awoara kidney and liver tissues. These cell lines have been designated GK and GL, respectively. The cells multiplied well in Leibovitz's L-15 medium, supplemented with 10% foetal bovine serum, at temperatures between 20 and 32 °C, and have been subcultured more than 120 times, becoming continuous cell lines. The cell lines consist of a heterogeneous mixture of fibroblastic and epithelial cells. The viability of cells, stored frozen in liquid nitrogen (−196 °C), was 95% after 1 year. Chromosome morphologies of GK and GL cells were homogeneous. Both cell lines were susceptible to grouper iridovirus, and yielded high titres of up to 108 TCID50 mL⁻¹. In addition, both cell lines effectively replicated the virus, which could be purified to homogeneity by cesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 170±10 nm in diameter, and were hexagonal in shape. Virus-infected cells showed an abundance of virus particles inside the cytoplasm. These results show that the GK and GL cell lines effectively replicate grouper iridovirus, and can be used as a tool for studying fish iridoviruses.     

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.