The existence of aspolin and its trimethylamine-<Emphasis Type="Italic">N</Emphasis>-oxide demethylating activity in the muscle of freshwater fish

ABSTRACT:   Aspolin is a polyaspartic acid-like protein, which is originally isolated from walleye pollack Theragra chalcogramma muscle as trimethylamine- N -oxide (TMAO) demethylase. Although carp Cyprinus carpio muscle contains a trace amount of the enzyme substrate, TMAO, aspolin can be extracted and purified by acid treatment, successive chromatographies and polyacrylamide gel electrophoresis, and has twice the amount of that in walleye pollack muscle. Carp aspolin showed a low enzymatic activity in the presence of Fe²⁺ and reductants, and its Km value (100 mM) to TMAO was extremely high. It was a thermostable protein and had an unfolded conformation. The amino acid sequence of carp aspolin 1 deduced from cDNA revealed that it contained a long Asp polymer, an uninterrupted stretch of 138 Asp residues, followed by four amino acid residues, His-Glu-Glu-Leu, in C-terminus. The chain length was shorter by 42 Asp residues than that of its walleye pollack counterpart.     

Discovering the dominance of the non-native European eel in the upper reaches of the Tone River system,Japan

To investigate the presence of non-native anguillid eels in Japanese waters, 141 eels were collected from seven sampling sites throughout the Tone River system. Genetic species identification showed an extraordinary dominance of the non-native European eel Anguilla anguilla in the uppermost site of the study area. Estimation of age from otoliths suggested that the European eels were introduced into the river in the 1990s, corresponding to previous reports from other Japanese water systems. Comparison of the von Bertalanffy growth curve parameters indicated that the European eels caught in the Tone River system appeared to have a similar or even higher growth rate than the same species in the original habitats in Europe. The long-term inhabitation and the normal development of European eels in Japanese waters suggest that regulations prohibiting the release of non-native eels and safeguards against accidental escape from culture ponds must be strictly maintained in order to ensure the conservation of the native Japanese eel.     

Transport and survival of larvae of pelagic fishes in Kuroshio system region estimated with Lagrangian drifters

ABSTRACT:   Transport and survival of larvae of pelagic fishes in the Kuroshio system region were studied using Lagrangian drifter data recorded from 1990 to 2003. A large portion of the drifters from the Kuroshio area south of Japan spread around the Kuroshio Extension up to 170°E, while some moved south to the offshore area of the Kuroshio because of a recirculation gyre. The monthly mean eastward movement from areas south of Japan was approximately 800–900 km, which was smaller than previous numerical estimates. The results of a survival model assuming optimal temperatures for larvae suggest that surface waters during the observation period were too warm for larval Japanese sardine, which has an optimal temperature of 16°C, and the adult abundances did not increase during the observation period. In contrast, the spawning ground temperatures and transport conditions from an area south-west of Japan in April–June matched the requirement for the larval Japanese anchovy, which has an optimal temperature of 22°C. The combined effects of temperature variations due to seasonality and water mass mixing are suggested to play an important role in determining the environmental temperatures that occur in an area.     

Changes to vertical thermoregulatory movements of juvenile bigeye tuna (Thunnus obesus) in the northwestern Pacific Ocean with time of day,seasonal ocean vertical thermal structure,and body size

Vertical movements related to the thermoregulation were investigated in 12 juvenile bigeye tuna (Thunnus obesus) in Japanese waters using archival tag data. Movements changed with time of day, season, and body size. During daytime, bigeye tuna descended to greater depths, presumably to feed in the deep scattering layer (DSL). Thereafter, they repeatedly ascended to shallower layers, suggesting attempts at behavioral thermoregulation, although the beginning of vertical thermoregulatory ascents might reflect a shift in DSL depth. By the end of such movement, the whole‐body heat‐transfer coefficient might decrease because, although the depth and ambient temperature of the upper layers did not change, the body temperature gradually decreased significantly just after ascent for thermoregulation. Seasonal patterns indicated that the vertical thermal structure of the ocean might influence this ascent behavior. For example, from January to May, bigeye tuna made fewer ascents to less shallow waters, suggesting that they respond to increasing depths of the mixed surface layer by reducing energy expenditure during vertical migration. In addition, as body size increased, fewer thermoregulatory ascents were required to maintain body temperature, and fish remained deeper for longer periods. Thus, vertical thermoregulatory movements might change with body size as bigeye tuna develop better endothermic and thermoregulatory abilities. We hypothesize that bigeye might also increase cold tolerance as they grow, possibly due to ontogenetic shifts in cardiac function.     

Vertical behavior of juvenile yellowfin tuna Thunnus albacares in the southwestern part of Japan based on archival tagging

The behavior of juvenile yellowfin tuna Thunnus albacares in southwestern Japan was investigated using archival tag data from five fish (fork length 52.5–92 cm, days at liberty 26–280 days) released near the Nansei Islands (24–29°N, 122–130°E). Vertical behavior was classified into three patterns: “shallow” (≥50 % of daytime hours at depth of <50 m), “deep” (≥50 % of daytime hours at ≥100 m), and “intermediate” (other than “shallow” or “deep”). The pooled proportion of the number of days of each behavior was 29, 25 and 46 %, respectively. The proportion of “shallow” behavior increased with fish size. The proportion of time spent near the surface at nighttime increased in the colder season, when the thermal gradient was relatively small. Surface-oriented behavior (fish remained at a depth of <10 m for more than 10 min) occurred mainly during nighttime and between November and January. Dives exceeding 500 m were occasionally observed (0.02 day^?1), and one fish dived to 1230 m. The results of our study show that yellowfin tuna were typically distributed in the mixed layer or upper thermocline where the water temperature was close to the sea surface temperature and that the vertical behavior was variable.     

Vertical Changes in Bacterial Communities in Percolating Water of a Japanese Paddy Field as Revealed by PCR-DGGE

Percolating water was sampled from the plow layer and subsoil layer in a Japanese paddy field, and the bacterial communities were compared together with floodwater by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) targeting a partial 16S rRNA gene and subsequent sequencing. The number of DGGE bands ranged from 16 to 28 with no significant differences among the sampling sites and times. Only 2 bands were common for the three sources of water samples. DGGE bands specific for the floodwater samples and percolating water samples from the plow layer were identified, while percolating water samples from the subsoil layer did not show specific bands but displayed common bands to those of the floodwater samples (7 bands) and percolating water samples from the plow layer (1 band). Cluster analysis of the DGGE banding patterns showed a distinct clustering in the samples of percolating water from the plow layer and a closer relationship between the others. These results suggest that the bacterial communities in percolating water changed during downward movement through the plow layer and subsoil layer. Sequences of the DGGE bands specific for the samples of percolating water from the plow layer showed a close relationship with anaerobic bacteria such as iron-reducers or uncultured bacterial DNA isolated from environments that are considered to be less oxic. On the other hand, the sequences of the bands specific for the samples of floodwater and percolating water from the subsoil layer showed a close relationship with uncultured bacterial DNA isolated from freshwater environments.     

Links between de novo fatty acid synthesis and leptin secretion in bovine adipocytes

Leptin secretion by adipose tissue is involved in many physiological control systems, including those that determine growth, development, body composition, milk production, and reproductive function. In the adipocyte of monogastric animals, malonyl CoA (coenzyme A) seems to link the flux of energy substrates to the control of leptin production. In this study, we tested this for ruminants by examining the effect of cerulenin, an inhibitor of de novo fatty acid synthesis at the step from malonyl CoA to palmitate, on leptin production by cultured bovine adipocytes derived from intermuscular fat. Purified preadipocytes were obtained by the ceiling culture method, and adipogenic media were used to induce their differentiation into adipocytes. We found that leptin concentrations increased significantly with time in culture, and with increases in glucose concentration. Addition of 2-deoxy-D-glucose to the medium, a competitive inhibitor of glucose transport and metabolism, suppressed leptin secretion. In media with high glucose concentrations, cerulenin enhanced leptin secretion. We conclude that, as in monogastrics, malonyl CoA may play a key role in the control of leptin secretion in ruminants.     

Isolation of Mycobacterium spp. from milking buffaloes and cattle in Nepal

In Nepal, mycobacterial isolates obtained from the milk and feces of buffaloes and cattle that were positive for the single intradermal cervical tuberculin (SICT) tests were genetically identified. A total of 36 mycobacterial strains were isolated from 39% of the buffaloes (14 of 36) and 34% of the cattle (11 of 32). Of the 36 strains, 13 were identified as M. bovis, and these strains were isolated from 17% of the buffaloes (6 of 36) and 16% of the cattle (5 of 32). M. bovis was isolated from both the milk and feces of one buffalo and one cattle, the milk alone of three buffaloes and three cattle, and the feces alone of two buffaloes and one cattle. These results suggest that milking buffaloes and cattle infected with M. bovis exist in Nepal. The remaining 23 strains were atypical mycobacteria. A program for the elimination of bovine tuberculosis should be implemented as soon as possible, and the public health education and proper hygienic practices may be required.     

Brain lesions and transmission of experimental equine herpesvirus type 9 in pigs

We demonstrated that pigs are susceptible to acute infection by equine herpesvirus type 9 (EHV-9). Six 8-week-old SPF pigs were inoculated intranasally and four were inoculated orally with different doses of EHV-9, and observed for 6 days. Although neurological signs did not develop in any of the infected pigs, the six intranasally infected pigs and one of the orally infected pigs developed lesions of encephalitis consisting of neuronal necrosis, neuronophagia, and intranuclear inclusion bodies, distributed mainly in the rhinencephalon. EHV-9 antigen was localized in the necrotic neuronal cells and was closely associated with the presence of inclusion bodies. These findings clearly demonstrate that pigs are fully susceptible to EHV-9 infection following intranasal inoculation (but less so following oral inoculation), and that EHV-9 in pigs has a highly neurotropic nature.