Molecular cloning and expression of the heat shock protein 70 gene in the Kumamoto oyster <Emphasis Type="Italic">Crassostrea sikamea</Emphasis>

Episodes of summer mortality of the Kumamoto oyster Crassostrea sikamea are a major problem for its cultivation. Expression of the heat shock protein 70 (HSP70) is induced by various environmental stresses, including heat. We cloned and sequenced hsp70 complementary DNA from C. sikamea to investigate the relationship between hsp70 expression and heat tolerance in this oyster. Quantitative real-time polymerase chain reaction was performed using gill tissue dissected from oysters before and after heat shock for 1 h. The results showed hsp70 expression was faster and greater in oysters cultured at 20–22 °C than at 10–12 °C, and survival was lower among oysters cultured at 20–22 °C than at 10–12 °C. Moreover, heat tolerance was investigated by a 1-h pre-heat treatment, followed by exposure to heat shock conditions 5 days later. Survival was higher and hsp70 expression was notably lower in oysters that received the pre-heat treatment compared with those that did not. We conclude that a pre-heat treatment of only 1 h may be useful for inducing heat tolerance in C. sikamea, and that a low level of hsp70 expression after heat shock is an important index in selecting for high heat tolerance in these oysters.     

Photosensitivity in mice caused by pyropheophorbide in the midgut gland of the scallop <Emphasis Type="Italic">Patinopecten yessoensis</Emphasis> observed in diarrhetic shellfish poisoning mouse bioassays

Photosensitivity was observed in the mice used in bioassays of diarrhetic shellfish poisoning. The mice were intraperitoneally injected with the extracts from the midgut glands of cultured scallops Patinopecten yessoensis gathered in early spring in Funka Bay, Hokkaido, Japan. The injections induced atypical symptoms in the mice within 24 h. The symptoms included piloerection, substantial eye discharge, swelling of the ears and head, and ear scratching. They were similar to those reported in studies of photosensitivity caused by pyropheophorbide in the midgut glands of abalones, which are herbivorous gastropods. The problematic scallops also accumulated a large amount of pyropheophorbide-a. The amount determined with high-performance liquid chromatography was 300–530 μg per 1 g of the homogenate of the midgut glands. The dose of pyropheophorbide sufficient to cause photosensitivity in a mouse is estimated to be 0.99–2.3 mg per mouse with 20-g body weight. Moreover, comparative tests in mice showed that the onset of the symptoms required light and a substantial amount of pyropheophorbide. Therefore, we determined that the observed symptoms were photosensitivity caused by pyropheophorbide. Scallops, which are suspension-feeding bivalves, can accumulate a large amount of pyropheophorbide in the midgut glands depending on the habitat environment.     

Feeding behavior under dark conditions in larvae of sutchi catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis>

Sutchi catfish Pangasianodon hypophthalmus hatch with morphologically immature sensory organs; however, sensory organs develop rapidly with larval growth. Two-day-old larvae commenced ingesting Artemia nauplii. The larvae displayed many taste buds on the barbels, the head surface, and in the buccal cavity. Other sense organs were also well developed at this stage. Feeding experiments revealed that 2-day-old larvae ingested Artemia under both light and dark conditions, moreover, the larvae could ingest frozen dead Artemia. The ingestion rates for 4- and 7-day-old larvae were significantly higher under dark conditions than under light conditions. The rates using frozen dead Artemia were mostly higher than the rates using live Artemia. Therefore, feeding behavior under dark conditions is most likely not mediated by visual or mechanical senses, but rather by chemosensory senses, such as taste buds. Larval fish are vulnerable to predators; thus, if they can search for and eat food at night, they can avoid diurnal predators. The behavior observed here appears to represent their survival strategy. Moreover, these results suggest a new possibility that sutchi catfish larvae can be reared under dark or dim light conditions in order to improve survival and growth rates as in the case of African catfish Clarias gariepinus.     

Morphological studies of peripheral blood cells of the Chinese sturgeon, <Emphasis Type="Italic">Acipenser sinensis</Emphasis>

The peripheral blood cells of one-year-old Chinese sturgeon (Acipenser sinensis) have been studied by light microscopy and transmission electron microscopy. The erythrocyte count was 84.86 × 10⁴ cell mm⁻³ in the peripheral blood of the fish and that of leukocytes was 2.24 × 10⁴ cell mm⁻³. The erythrocytes and four main types of leucocyte—thrombocytes, lymphocytes, granulocytes (including neutrophils and eosinophils), and monocytes, were identified in the peripheral blood. In addition to normal erythrocytes, reticulocytes and division of erythrocytes were observed. Thrombocytes were the most numerous among the leukocytes, and the number of neutrophils with lobated nuclei was larger than for other fish. The structures of the erythrocytes, lymphocytes, monocytes, granulocytes, and thrombocytes of the fish were studied. The erythrocytes were almost completely devoid of organelles, except for some mitochondria and granules. A large number of vacuoles and a few organelles were observed in cytoplasm of the monocytes. There were many microvilli on the membrane and pseudopodia-like cytoplasm bulge in the lymphocytes. The neutrophils were round or oval in shape with bilobed, trilobed, or multilobed nuclei whereas the eosinophils had big special granules, dark stained. There were many vesicles in some thrombocytes, which were related to its phagocytosis; some thrombocytes had almost no cytoplasm or organelles.     

Effects of mangrove deforestation on fish assemblage at Pak Phanang Bay,southern Thailand

Daytime sampling using a seine net was conducted at Pak Phanang Bay (Nakhon Si Thammarat Province, Thailand) in February and July 2006, to determine differences in fish assemblage structures between a mangrove site and an adjacent site completely cleared of mangroves. The overall numbers of fish species and individuals were significantly higher at the mangrove site than the cleared site in both months. Although benthic crustacean feeders showed more species and individual numbers at the mangrove site in both months, the opposite was found for zooplankton feeders. A cluster analysis, based on the abundance of each species, demonstrated that the fish assemblage structures were distinctly different between the two sites. In addition, significant differences in length frequency distributions for each of the four most abundant species were found between the sites in February and/or July. Small individuals of Scatophagus argus, Ambassis nalua, and Tetraodon nigroviridis were more common at the mangrove site, and of Chelon subviridis at the cleared site. These results suggest that mangrove deforestation exerts marked effects on fish assemblages.     

Parentage assignment of stocked black sea bream <Emphasis Type="Italic">Acanthopagrus schlegelii</Emphasis> in Hiroshima Bay using microsatellite DNA markers

The genetic contribution of 51 broodstock, comprising 29 females and 22 males, reared at Hiroshima City Marine Products Promotion Center for the production of stocked black sea bream was monitored during two consecutive years using seven microsatellite DNA loci. The high discrimination ability of these markers was reflected in the polymorphic identification content (PIC=0.831), the exclusion probability (Q≈1), and the low probability of identity index (I=3.635⁻¹⁰). The total number of breeders contributing to the mating process was estimated at 32 (62.7%) in 2000 and 30 (58.8%) in 2001. On pedigree reconstruction, 69.3% of the offspring were successfully assigned to a single broodstock pair. Loss of alleles accounted for 16.9% during seed production; nevertheless, 90.9% of males and 69.0% of females participated in the mating process. Based on microsatellite genetic tagging, 58.9% of the fish sampled during the two months after release were identified as hatchery stock, presenting no significant differences from wild conspecifics in either fork length or body weight.     

Relationship between soak time and number of enmeshed animals in experimentally lost gill nets

In order to determine the effects of ghost fishing by lost gill nets, the relationship was examined between soak time and number of enmeshed animals in experimentally lost gill nets by using diving observations. Two experimental gill nets were set at 13 m depth in Tateyama Bay, Chiba Prefecture, Japan for approximately 200 days. One gill net was deployed in a small trough surrounded by artificial reefs, and the other was deployed on an adjacent open sandy bed. Twelve species of crustaceans, six species of gastropods, and five species of bony fish were enmeshed in the experimental gill nets. The number of enmeshed animals in the artificial reef gill net was substantially larger than that in the sandy bed gill net. The number of enmeshed animals in the experimental gill nets increased rapidly within one month after deployment, and then declined gradually showing fluctuations caused by the decrease in newly enmeshed animals, and the drop off from gill nets caused by the decomposition of dead animals. The decrease in the number of enmeshed animals was expressed by logarithmic equations, and based on these equations, the duration of capture function for the lost gill nets was calculated to be 284–561 days in the artificial reef gill net and 200 days in the sandy bed gill net. The duration of capture function for the lost gill nets for non-commercial by-catch species such as small crustaceans and gastropods was longer than for commercial species such as Japanese spiny lobster and bony fish.     

Determination of the quality parameters of pike perch <Emphasis Type="Italic">Sander lucioperca</Emphasis> caught by gillnet,longline and harpoon in Turkey

ABSTRACT:   The effects of the different catching methods (gillnet, longline, harpoon) on sensory, chemical (pH, total volatile base nitrogen, K -value) and microbiological (total viable count [TVC]) changes in pike perch Sander lucioperca stored in ice were investigated. The same soaking time was used for both gillnet and longline fishing. The catching method had considerable influence on the freshness quality of pike perch. The acceptable shelf life was 15 days for pike perch caught by gillnet, and 22 days for longline and harpoon. The initial concentrations of inosine monophosphate (2.4 μmol/g) in pike perch caught by gillnet were significantly lower ( P  < 0.05) than longline (4.1 μmol/g), and especially by harpoon (16.7 μmol/g). However, the initial K -values for fish caught by harpoon were significantly ( P  < 0.05) lower (24.36%) than fish caught by longline and gillnet (57.69%, 64.41%, respectively). The average K , Ki, G and H -values at rejection day in terms of sensory assessment were approximately 90, 98, 156 and 40%, respectively, for all catching methods during ice storage. However, TVC reached 7.0 log cfu/g after approximately 11 days of storage for fish caught by gillnet, 19 days for fish caught by longline and 8 days for fish caught by harpoon. The result of this study suggests that the best catching method for preserving the freshness of pike perch is longline, based on the data obtained from the sensory and microbiological analysis.     

Carbon source and trophic position of pelagic fish in coastal waters of south-eastern Izu Peninsula,Japan, identified by stable isotope analysis

ABSTRACT:   It is important to clarify trophic dynamics in marine ecosystems for management of the fishing ground. Organic carbon sources and trophic position of pelagic fishes in the coastal waters of the south-eastern Izu Peninsula, Japan, were examined on the basis of carbon and nitrogen stable isotope distributions. The δ¹³C of the fishes was mostly distributed from −19 to −16‰ for nektonic fishes (13 species of adults and immatures) and planktonic fishes (10 species of larvae and juveniles), close to the δ¹³C values of particulate organic matter and planktonic decapods. These δ¹³C signatures for the inhabitants of the water column were in contrast with the high δ¹³C values (mainly −16 to −13‰) for demersal fishes of Scorpaeniformes and benthic polychaetes collected in the surf zone. These results indicate that nektonic and planktonic fishes depend on phytoplankton for carbon supply. The δ¹⁵N signatures suggest that the trophic position ranged 3.1–4.5 for the nektonic fishes and 2.9–3.7 for the planktonic fishes, premised on trophic level 3 for larval Japanese anchovy Engraulis japonicus . Thus, planktivorous fishes should be mainly assigned to trophic levels 3 and 4 in this area.     

Post-release movement and diel activity patterns of hatchery-reared and wild black-spot tuskfish <Emphasis Type="Italic">Choerodon schoenleinii</Emphasis> determined by ultrasonic telemetry

ABSTRACT:   Post-release movement and diel activity patterns of hatchery-reared and wild black-spot tuskfish were examined using ultrasonic telemetry. Five hatchery-reared and four wild fish were released in the sandy bottom of Urasoko Bay, Ishigaki Island, Okinawa, Japan, and monitored using automated monitoring receivers from November 2005 to February 2006. Both hatchery-reared and wild fish tended to stay near the release site for over two weeks, before leaving the release site. Both hatchery-reared and wild tuskfish showed diurnal rhythm intermittently; signals were recorded more frequently in the daytime and less frequently in the nighttime, suggesting that the fish of both origins were active during the day and inactive during the night. These findings indicate that the one-year-old hatchery-reared tuskfish have some consistent behavioral characteristics with those of the wild.