Fisheries Science

We explored osmoregulatory ability and mechanisms of ion and water regulation in Japanese eel leptocephali. Tissue osmolality of leptocephali ranged from 360 to 540 mOsm/kg·H2O. Immunocytochemical observations revealed that Na+/K+-ATPase-immunoreactive mitochondrion-rich (MR) cells were distributed over the entire body surface of leptocephali. Using a fluorescent sodium indicator and the chloride test, we localized Na+ and Cl− secreting sites at the apical region of cutaneous MR cells. To further examine drinking behavior and water absorption in the intestine, leptocephali were exposed to seawater containing dextran labeled with Alexa Fluor. To calculate relative water absorption, fluorescent intensity was measured along the digestive tract. Whereas water was hardly absorbed in the stomach and intestine, water absorption predominantly took place in the rectum. Our findings indicate that Japanese eel exert hyposmoregulatory ability as early as during leptocephalus stages, secreting Na+ and Cl− through cutaneous MR cells and primarily absorbing water from ingested seawater in the rectum.

In the mid-1990s, Neoheterobothrium hirame suddenly appeared as a new species in olive flounder Paralichthys olivaceus in Japanese coastal waters. Anemia caused by the parasite has prevailed in wild and cultured populations of olive flounder since that time. In this study, to clarify the origin of N. hirame, two Neoheterobothrium species, namely unidentified Neoheterobothrium species (tentatively abbreviated as Neoheterobothrium sp. PL) and N. affine, were collected from Paralichthys lethostigma and Paralichthys dentatus, respectively, off the east coast of North America and compared with N. hirame collected in Japan. No substantial differences were detected in the morphology and DNA sequences of ITS1-5.8S rRNA-ITS2 and mitochondrial cytochrome oxidase subunit I (mt COI) regions between N. hirame and Neoheterobothrium sp. PL. On the other hand, the congeneric N. affine was clearly distinguished from both N. hirame and Neoheterobothrium sp. PL in its longer isthmus and the DNA sequences in ITS1 and mt COI. The absence of differences between N. hirame and Neoheterobothrium sp. PL and the clear difference between both of these and N. affine indicate that N. hirame is conspecific with Neoheterobothrium sp. PL infecting P. lethostigma and that N. hirame was recently introduced from North America to Japan.

Cellular and humoral effectors are imperative for the innate defense mechanisms of invertebrates, including the spotted babylon snail Babylonia areolata, a marine gastropod belonging to the family Buccinidae. In this study, we have investigated the influence of body size [5–8 g (small), 10–12 g (medium) and 15–17 g (large)], and culture waters of varied quality [natural seawater, low salinity, low alkalinity, high total ammonia nitrogen (TAN) and artificial seawater] on some hemato-immunological parameters [total hemocytic counts (THC), hemolymphatic glucose, total protein concentration, phenoloxidase (PO), agglutinating and lysozyme activity] of the snail. Growth performance measures and survival of test snails were also evaluated after rearing them in culture water of varied quality. Body size did not influence most parameters assessed, except that the lysozyme and PO activities of medium and large-size snails were higher than those of the small-size snails. Sub-optimal culture water quality affected hemato-immunological factors, growth and/or survival of the spotted babylon snail. Seawater with low salinity, low alkalinity and a high TAN level caused decreased THC, hemolymphatic glucose, and PO activity; increased lysozyme activity; and clearly retarded growth of the snail. The snails held in artificial seawater could not survive after 4 weeks, suggesting the lack of certain essential factors necessary for their survival.

Paralytic shellfish poisoning (PSP) toxins produced by Alexandrium isolates from Korea were analyzed by high-pressure liquid chromography. Species designation of the regional isolates was determined by morphological criteria and ribotyping inferred from sequences of the 28S rDNA D1–D2 region. Toxin analysis performed at the exponential growth phase, revealed that the two strains of A. fraterculus were non-toxic, while the strains of A. tamarense and A. catenella were toxic. Toxic isolates DPC7 and DPC8 of A. catenella produced GTX1, 2, 3, 4, 5, dcGTX2, 3, C1, 2, neo STX and STX with trace or non-detectable levels of C3 and C4, while isolates UL7, KDW981, SJW97043, SJW97046, KJC97111 and KJC97112 of A. tamarense produced GTX1, 2, 3, 4, dcGTX3, C1, 2, neoSTX with trace or non-detectable levels of C3, 4, dcSTX and STX, and no GTX5 and dcGTX2. The major toxins produced by A. catenella were C1+2, and those of A. tamarense were C1+2 and GTX4 in most of the isolates. A tamarense strains other than SJW97046 produced a relatively high proportion of carbamate toxins, reflecting the high toxicity scores of shellfish intoxication in sampled coastal areas. Two representative toxic isolates, A. tamarense SJW97043 and A. catenella DPC7, were cultured for 30 days in batch mode and subjected to toxin analysis at 5-day intervals. Comparison of toxin productivity in terms of total toxin content, toxin components, and their variations with culture age revealed marked differences between the two strains.

The elasmoid scales of fish represent a significant internal reservoir of calcium ions (Ca2+), but little is known about the contribution of these scales to the short-term regulation of Ca2+ homeostasis in the extracellular fluid (ECF). This gap in our knowledge is partly due to the technical challenges involved in measuring small Ca2+ fluxes around the scales of live fish in real time. Here, we describe a technique for exfoliating, mounting, and culturing intact living zebrafish Danio rerio scales, then subjecting them to examination using an extracellular, non-invasive, surface-scanning ion-selective electrode technique (SIET). In a Ca2+-sensitive configuration, the SIET can resolve Ca2+ flux values in the low-to-sub picomole/square centimeter/second range, with a spatial resolution of approximately 5 μm. We quantified the Ca2+ fluxes into and out of scales under different extracellular calcemic challenges set to mimic a variety of Ca2+ concentrations in the ECF and showed that the results were similar to those previously reported from isolated mouse metatarsal bone. Our new data extend our current understanding of the role played by fish scales in the short-term homeostatic regulation of Ca2+ concentration in the ECF. They also support the suggestion that scales might provide an inexpensive and complementary model for studying the fundamentals of bone-mediated homeostatic Ca2+ regulation and the diseases that result from its dysregulation.

Biwa salmon Oncorhynchus masou subsp. were tagged with ultrasonic transmitters and released in Lake Biwa, Japan. The temporal and spatial distributions were investigated by using two types of ultrasonic telemetry system, active tracking and passive monitoring. Biwa salmon cruised throughout the north basin horizontally and were distributed from the surface to the bottom vertically in the lake. An individual monitored for over 1 year showed the diurnal patterns of vertical distribution and the seasonal patterns of horizontal distribution. The vertical distribution of the fish during daytime in the stratified period was bimodal, suggesting a feeding strategy targeting both fish near the thermocline and the amphipods near the bottom. During the night, however, the fish maintained a fixed depth range. The fish were distributed over a wider horizontal area in the mixed period than in the stratified period. Three individuals swam in shallower water and experienced higher water temperatures during the spawning season than before moving between offshore and coastal areas.

We have developed immunological probes to quantify eggs and sperm of the Black-lip pearl (BLP) oyster Pinctada margaritifera. The western blot assay revealed that the polyclonal antibodies developed in this study specifically recognized only egg and sperm proteins. These polyclonal antibodies also showed high sensitivities to the antigens, detecting 0.31–10 μg/ml of the egg or sperm proteins in an indirect enzyme-linked immunosorbent assay (ELISA). Accordingly, we used an indirect ELISA to quantify the eggs and sperm in BLP oysters collected in December 2009 from Weno Island in Micronesia and in May 2010 from Tahiti. The gonad somatic index (GSI), a ratio of gonad weight to somatic tissue weight, of the females collected from Weno Island ranged from 3.6 to 18.4 %, while the GSI of the females collected from Tahiti ranged from 5.6 to 12.6 %. Similarly, the GSI of the male BLP oysters from Weno Island ranged from 0.8 to 8.5 %, while the GSI of the males from Tahiti ranged from 4.8 to 7.5 %. These results lead to the conclusion that the immunological probes developed in this study can be successfully applied to quantify BLP oyster gonadal tissues and that these probes can be used in studies of BLP oyster reproductive ecology based on their sensitivity, rapidity, and affordability.

Bluefin tuna maintain a higher body temperature than ambient sea water. Body heat is derived mainly from metabolic heat to elevate and maintain regional body temperature that is higher than the ambient, while heat loss is caused by heat transfer throughout the whole body surface and gills. Retention of high body temperature is thought to differ at each growth stage, so that a larger body mass maintains a higher body temperature. We evaluated the whole-body heat transfer coefficient, thermal difference between each tissue and water temperature, and metabolic heat in tissues during swimming of juvenile bluefin tuna as a function of fork length (FL) using a small thermometer and a treadmill-type flow tank. A system for maintaining high body temperature was well developed in fish with FL greater than 20.0 cm. Whole-body heat transfer coefficient was fitted to a −0.695 power of mass. Juvenile bluefin tuna showed a transition speed of 3.0 FL/s at which they switched from aerobic to anaerobic motion.