Accumulation of organotin compounds and marine birnavirus detection in Korean ascidians

ABSTRACT:   Recently, a serious disease spread extensively in aquaculture sites of the ascidian Halocynthia roretzi in Korea. To understand circumstances of ascidians in Korean aquaculture sites, residue levels of organotin compounds were analyzed, and detection of a marine birnavirus (MABV) in tissues of H. roretzi was attempted. Korean H. roretzi showed high concentrations of butyltins (mono, di, and tributyltins), especially in the gill, hepatopancreas, and digestive tract. However, there was no significant difference in the residues of butyltins in the hepatopancreas between diseased and non-diseased ascidians. The positive rate of MABV detection was high in the hepatopancreas, but also no significant difference was observed between diseased and non-diseased individuals. These observations suggest that an accumulation of tributyltin and a latency of MABV in H. roretzi tissues does not directly relate to the occurrence of the disease.     

Identification of ommastrephid squid paralarvae collected in northern Hawaiian waters and phylogenetic implications for the family Ommastrephidae using mtDNA analysis

ABSTRACT:   A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu  I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.     

Moisture distribution in rice grains used for sake brewing analyzed by magnetic resonance imaging

Water absorption in individual white-core and non-white-core grains of rice varieties used in sake brewing was observed during soaking by magnetic resonance imaging (MRI). Water was absorbed faster in white-core grains than in non-white-core grains and was pooled in the core. Moisture distributions were compared using the line profiles of nuclear magnetic resonance signal intensities (SI profiles) generated from magnetic resonance images of grains soaked for 2 h. SI profiles of white-core and non-white-core grains overlapped despite slight differences in the central and intermediate regions of the grains. A white-core-like structure with loosely packed starch granules was found to exist in the endosperm of non-white-core grains. SI profiles of grains polished to 70% of the yield weight of brown rice were similar in shape to those of grains polished to 90% of the yield, but their overall moisture content was higher after the removal of the hydrophobic grain periphery. A varietal difference in the thickness of the intermediate region, which may affect fissuring resistance in the polishing process, was detected in SI profiles. Differences in water absorbability between parents and their offspring were also apparent from their SI profiles. MRI enabled the detection of variety-specific water-absorption properties in grains.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

Changes of lymphocyte subpopulations and natural killer cells in mice sensitized with Toxoplasma lysate antigen before and after Babesia infection

When 8-week-old BALB/c mice were sensitized with two intramuscular injections of Toxoplasma lysate antigen (TLA) at 2 week interval, the numbers of sIg(+), Thy-1,2(+), Lyt-1,2(+) Lyt-2,2(+), and Asialo GM1(ASGM1)(+) cells in the spleen, liver and peripheral blood increased by 2 to 4 times over those found in unsensitized mice of the same age. When TLA-sensitized and unsensitized mice were infected with Babesia, 4 of 10 (40%) of the TLA-sensitized mice survived infection, while none of the unsensitized control mice lived longer than 14 days after Babesia infection. By contrast, sensitization of nude mice with TLA had no effect on survival, and mice did not live more than 12 days. The number of thymic Thy-1,2(+) cells decreased in TLA-sensitized and unsensitized BALB/c mice by almost 80% within 10 days after infection (AI). During the same time, the numbers of B cells, T cells, and NK cells increased in the spleen, liver and peripheral blood of both sensitized and unsensitized mice. Especially notable were increases in numbers of Lyt-2,2(+) cells in the spleen and blood and increases in numbers of NK cells in the spleen, liver and blood in both TLA-sensitized and unsensitized mice. When spleen cells from TLA-sensitized and unsensitized mice were cultured in the presence or absence of TLA for 6 days, assays for cytotoxicity using NK-insensitive P-815 target cells and NK-sensitive YAC-1 target cells demonstrated higher rates of cytotoxicity in cultures of TLA-sensitized spleen cells.     

Narthecium asiaticum Maxim. Poisoning of grazing cattle: observations on spontaneous and experimental cases

A total of 39 Holstein cattle were grazed in tracts of wild grassland on account of shortage in pasture grass. Twenty-nine cattle were affected and 26 of them died during a 21-day period. The main signs were depression, anorexia, ascites, and oliguria. There was elevated serum urea nitrogen and sugar and protein in the urine. Pathological examination revealed turbid swelling of the kidney, an increase in the amount of fluid in the body cavity, edema in the perirenal adipose tissue and hemorrhage in various visceral organs and tissues. Histologically, acute tubular necrosis in the kidney, hypoplasia of the erythroblast series in the bone marrow, atrophy and degeneration of the lymphatic tissue and focal necrosis of the liver were observed in many of the cattle. Among cows experimentally fed Narthecium asiaticum Maxim., Polygonum sachalinense Fr. Schum., and Vitis coignetiae Pulliat which were presumed to have been ingested in large amounts by grazing cattle in the field, the cows fed N. asiaticum revealed the clinical, biochemical and pathological changes similar to those noticed in naturally affected cattle. Cows fed P. sachalinense and V. coignetiae showed no distinct systemic symptoms except transient anorexia and hypothermia.