Changes in metabolite concentrations in the abdominal muscle of the kuruma shrimp Marsupenaeus japonicus in response to different salinities

Invertebrates change the metabolite concentrations in their bodies to adapt to environmental salinity. The kuruma shrimp Marsupenaeus japonicus is one such invertebrate. It lives in coastal areas and is also known to change free amino acid concentrations depending on environmental salinity. To examine the relationship between the concentrations of metabolites, including amino acids, in the kuruma shrimp with environmental salinity, metabolome analysis was performed on the abdominal muscle of shrimps acclimated at 17, 34 and 40‰ salinity for 24 h. Principal component analysis revealed that the accumulation patterns of metabolites using the 111 metabolites detected in the shrimps exposed to different salinities were depicted in a salinity-dependent manner. The concentrations of alanine and glutamine were increased following exposure to increasing levels of salinity, suggesting that these free amino acids function in intracellular osmoregulation of the kuruma shrimp. Furthermore, the concentration of glycolytic metabolites was significantly decreased at high salinity. The concentrations of most of the metabolites related to the tricarboxylic acid (TCA) cycle also tended to decrease at high salinity. Considering the levels of expression of the genes related to various metabolic pathways, it seems that glycolysis is accelerated at high salinity.

     

Changes in rheological properties of grass carp fast skeletal myosin induced by seasonal acclimatization

To elucidate the effects of seasonal temperature acclimatization on thermal gelation of grass carp myosin, myosins from fish in different seasons were prepared and investigated for the changes in dynamic viscoelastic parameters including storage modulus (G′), loss modulus (G″) and damping factor (tan δ) upon heating. Myosins from fish in spring and summer had a temperature region of 38–44°C for the first marked increase of G′ higher than that of myosins from fish in autumn and winter (28–33°C). The measurement temperature-dependent changes in dynamic viscoelastic parameters such as G″ and tan δ were also different among the four myosins. While gel formation was observed with the spring and summer myosins, apparently in two steps, three steps were found in the autumn myosin. Furthermore, the winter myosin exhibited more than three steps for gel formation. These differences in rheological properties among the four myosins were considered to be attributed to the differences in thermodynamic and structural properties of these myosins previously reported.     

Differential expression of heat-shock proteins in F2 offspring from F1 hybrids produced between thermally selected and normal rainbow trout strains

Rainbow trout Oncorhynchus mykiss is a cold-water aquaculture species, and a thermally selected strain has been developed by multigenerational high-temperature breeding in Japan. We examined the expression of heat-shock proteins as candidates responsible for thermotolerance in rainbow trout using F2 offspring from F1 hybrids produced between thermally selected and normal strains. From F2 offspring, two groups were selected for western blot analysis, namely, low- and high-thermotolerance groups (times to loss of equilibrium were <30 and ??60?min, respectively). We demonstrated that the expression levels of Hsp70, Hsp60, and Hsp40 in tail fin tissues were significantly higher in the individuals with high thermotolerance than in those with low thermotolerance under non-heat-shock conditions. In particular, Hsp70 was expressed only in the individuals with high thermotolerance. These results suggest that Hsp70 is a major protein responsible for conferring thermotolerance in rainbow trout.     

Population model of Rana japonica crossing agricultural concrete canals: evaluating population conservation by improving the migration routes of frogs

Agricultural canals with deep concrete walls are commonly installed in paddy field areas following land consolidation projects in Japan. Such canals create a critical migration barrier for frogs, as escape is not possible after falling in. Hence, countermeasures that allow migrating frogs to cross agricultural canals, such as lid structures to prevent frogs from falling in or the creation of partial slopes to allow escape, have been developed to conserve endangered frog populations. The objective of this study was to design a method for evaluating the population viability of the endangered Japanese Brown Frog, Rana japonica, following the implementation of countermeasures. Two types of model simulations were examined: (1) a ??basic model,?? based on an age-structured model and (2) a ??crossing model,?? to evaluate migrating frogs crossing canals. Parameters, such as survival rate and fecundity, were estimated from field observation data of age-dependent population fluctuations. Comparison of a basic model simulation with the field observation data suggests model validation. The crossing model simulation, which had an initial population of 1,000 individuals, showed that a crossing proportion of greater than 30?C40% would achieve 95% population viability after 5?years. The simulation also showed that a crossing proportion of over 80% would also achieve 95% population viability after 40?years. These results indicate that frog survival during the migration period is insured, as long as no additional countermeasures are necessary after installation.     

Peripheral lymphocyte subsets as a prognostic indicator of mortality and morbidity in healthy dogs

To evaluate the relationship among immune status and increased morbidity and mortality, peripheral blood lymphocytes (CD3(+), CD4(+), CD8(+) and CD21(+) cells) from 32 healthy dogs over 8 years of age were analyzed. Twenty-five of the 32 dogs were followed-up for 3 years after the analysis; and 14 dogs were found to be diseased, and nine dogs died. There was no notable difference between the ages of the dogs that died compared with the ones that survived. The relative percentage of CD4(+) and the CD4(+):CD8(+) ratio decreased notably in dogs falling ill compared with healthy dogs. The relative percentage of CD3(+) lymphocytes showed a notable decrease in dogs that died within 3 years in comparison with dogs that survived. In a discriminant analysis of morbidity and mortality, most patients were correctly classified as diseased or not and surviving or dead, respectively. These results indicate that the immunophenotypes of peripheral blood lymphocytes in older dogs offer promise as parameters for evaluating mortality and morbidity.     

Exploration of the antibacterial proteins in pearl oyster <Emphasis Type="Italic">Pinctada fucata</Emphasis> induced by bacterial inoculation

The aim of this research was to characterize immune-related antibacterial substances from pearl oyster Pinctada fucata induced by bacterial invasion. Bacteria inoculation was performed by injecting 0.1 ml of 1.0 × 10¹² colony-forming units/ml Vibrio parahaemolyticus into adductor muscle. Acidic extracts were prepared with 0.1% trifluoroacetic acid from different tissues after 8 h of injection, and antibacterial activity against V. parahaemolyticus was determined via the microdilution broth method. The acidic extracts from gills of inoculated oysters (AEg) showed stronger antibacterial activity than those from non-inoculated ones. Based on this result, antibacterial proteins were purified from AEg via two-step gel filtration chromatography, followed by high-performance liquid chromatography using a TSkgel G3000 column. Protein components were analyzed by both sodium dodecyl sulfate and native polyacrylamide gel electrophoresis. As a result, two antibacterial proteins, APg-1 (with a molecular mass of approximately 210 kDa) and APg-2 (of approximately 30 kDa), were obtained from AEg. Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and partial amino acid sequences revealed that these proteins might be novel antibacterial proteins. These results indicate that antibacterial proteins are potentially upregulated in the gill of pearl oysters or released therefrom for defense against bacterial invasion.     

Tissue distribution of tetrodotoxin and its analogs in Lagocephalus pufferfish collected in Vietnam

Fisheries Science - Pufferfish belonging to Lagocephalus are composed of several species, some of which have been recognized to be non-toxic. Although fish belonging to this genus inhabit a wide...