Toxicity of a synthetic phenolic antioxidant,butyl hydroxytoluene (BHT), in vertebrate model zebrafish embryo (Danio rerio)

The synthetic antioxidant 3,5‐di‐tert‐butyl‐4‐hydroxytoluene (BHT) is widely used as an additive in the food, cosmetic and plastic industries to increase the tenability of food and plastic for the past 70 years. BHT is degraded to 3,5‐di‐test‐butyl‐4‐hydroxybenzaldehyde (BHT‐CHO) in mammals, as well as in the natural environment such as in river and water. The average daily intake of BHT for human being is estimated to be 0.3 mg/kg body weight. Even though it is considered safe for human at authorized level, but its ubiquitous presence in the aquatic environment and the controversial toxicological data are of great concern for human as well as aquatic life. The experimental findings of zebrafish embryo toxicity test (ZFET) showed that the acute toxicity of 96‐hr (LC₅₀) exposure during the embryogenic stage was found to be 4.388 mg/L and the effective concentration (EC₅₀) was 1.375 mg/L. The reduce heart rate from the sublethal concentrations indicates the chemical to be cardiotoxic but a further review is to be needed. The Teratogenic Index (TI) calculated to be 3.19, which implies the compound may be a potential teratogen in aquatic life. The findings obtained in this study will stretch more evidence regarding developmental toxicity of BHT, which will be of much importance in further risk assessment of ecotoxicological studies.     

Allozyme variation of littleneck clam Ruditapes philippinarum and genetic mixture analysis of foreign clams in Ariake Sea and Shiranui Sea off Kyushu Island, Japan

ABSTRACT:   Allozyme variation of the littleneck clam Ruditapes philippinarum was evaluated in four samples from Nameishi and Matsuo in the Ariake Sea, Ryugatake and Ushibuka in the Shiranui Sea off Kyushu Island, Japan, and in one sample from Jinzhou, China, in the Bohai Sea. A Ruditapes bruguieri sample imported from the Korean Bay off Nampo, North Korea was also studied. Among the R. philippinarum samples, heterozygosity varied from 0.265 to 0.301 and F _IS estimates indicated significant homozygosity excess in 15 of 40 loci analyzed. Deviations from Hardy–Weinberg equilibrium were significant in all samples ( P  < 0.05). Pairwise F _ST estimates indicate that genetic differences between the Chinese and Japanese samples were very low, but significantly different from zero. Mixture proportions with 95% confidence intervals of Chinese R. philippinarum in Nameishi and Matsuo were estimated at 0.4098 [0.2512, 0.5705] and 0.4899 [0.3262, 0.6540], respectively. However, genetic invasion of stocked Chinese R. philippinarum into wild populations in the Ariake Sea remains uncertain due to the low precision of the estimates caused by the high similarity of allele frequencies between Jinzhou and the Ariake Sea.     

Detection of the nonsense mutation of OPA3 gene in Holstein Friesian cattle with dilated cardiomyopathy in Japan

Bovine dilated cardiomyopathy (DCM) is an autosomal recessive genetic disorder causing congestive heart failure and subsequent death. Recently, a nonsense mutation c.343C>T in the bovine optic atrophy 3 (OPA3) gene had been reported to cause the DCM in Holstein cattle in Switzerland. However, the mutation has not been confirmed in bovine DCM outside Switzerland. Nine Holstein Friesian cows that were macroscopically and histologically diagnosed with or suspected of DCM and 12 control cows kept in Japan were tested for the mutation. The mutation surrounding OPA3 DNA fragment was amplified by PCR and subjected to direct sequences. The homogeneous c.343C>T mutation was proved to occur in all the affected cows and not in the control cows. The present study is the first report of the mutation in the DCM affected cows outside Switzerland.     

A network of control mediated by regulator of calcium/calmodulin-dependent signaling

Calmodulin (CaM) is a major effector for the intracellular actions of Ca2+ in nearly all cell types. We identified a CaM-binding protein, designated regulator of calmodulin signaling (RCS). G protein-coupled receptor (GPCR)-dependent activation of protein kinase A (PKA) led to phosphorylation of RCS at Ser55 and increased its binding to CaM. Phospho-RCS acted as a competitive inhibitor of CaM-dependent enzymes, including protein phosphatase 2B (PP2B, also called calcineurin). Increasing RCS phosphorylation blocked GPCR- and PP2B-mediated suppression of L-type Ca2+ currents in striatal neurons. Conversely, genetic deletion of RCS significantly increased this modulation. Through a molecular mechanism that amplifies GPCR- and PKA-mediated signaling and attenuates GPCR- and PP2B-mediated signaling, RCS synergistically increases the phosphorylation of key proteins whose phosphorylation is regulated by PKA and PP2B.     

Orbital malignant schwannoma and embryonal rhabdomyosarcoma in rats caused by leakage of locally injected nickel subsulfide to the orbit

Nickel subsulfide (Ni₃S₂) is known to induce intraocular neoplasms when injected intravitreally into the eyes of rats. Here, we found two extraocular orbital neoplasms in two different rat strains, presumably due to the leakage of locally injected Ni₃S₂ to the extraocular orbital tissues. In the F344/DuCrlCrlj rat, an orbital mass arose at 30 weeks after injection, and invaded into the cranium. Histologically, the orbital mass was composed of areas arranged in parallel bundles formed by densely packed elongated or spindle-shaped cells with indistinct cytoplasmic borders, and of areas of hypocellular arrangement consisting of round cells in eosinophilic myxoid-like substances. Metastases were observed in the right submandibular and cervical lymph nodes. The neoplastic cells were immunopositive for S-100 protein and vimentin. Transmission electron microscopy revealed that the neoplastic cells had cellular processes and pericytoplasmic basal laminae. In the RccHan^TM:WIST rat, an orbital mass arose at 36 weeks after injection. Histologically, the mass consisted of rhabdoid-like large round cells with proliferation of small round-to-polygonal cells, and these neoplastic cells infiltrated into the extraocular muscles. Immunohistochemically, the neoplastic cells were positive for desmin and vimentin. Transmission electron microscopy detected immature myofibrils with Z-band structures in the cytoplasm of these neoplastic cells. Consequently, the tumors were diagnosed as an orbital malignant schwannoma in an F344/DuCrlCrlj rat and an orbital embryonal rhabdomyosarcoma in a RccHan^TM:WIST rat. The results of this case report suggest that leakage of Ni₃S₂ to the orbit caused the induction of orbital malignant schwannoma or rhabdomyosarcoma in rats.     

Distribution,body length,and abundance of blue shark and shortfin mako offshore of northeastern Japan,as determined from observed pelagic longline data, 2000–2014

Longline surveys have been conducted in the Northwest Pacific Ocean from 2000 to 2014 using chartered commercial longline vessels. Each year, two cruises were conducted offshore of northeastern Japan from mid‐April to mid‐June. For each longline set during the surveys, onboard scientists collected detailed biological information about the species caught, such as the size and sex, and recorded the catch numbers for all species. Blue shark (Prionace glauca) and shortfin mako (Isurus oxyrinchus) have eurythermal distributions, but the application of a generalized additive model (GAM) showed that the sea surface temperatures (SSTs) at catch sites positive for shortfin mako were warmer than those for blue shark. On the basis of the GAM, the probabilities of occurrence of both sharks differed by size category: small sharks had a narrower SST range than that of large sharks. Most catches of both sharks were juveniles, and the nominal catch rate of blue shark was more than 10 times that of shortfin mako. The standardized catch per unit effort (CPUE) for both species was calculated using a generalized linear model (GLM) with negative binomial errors, or a delta‐lognormal GLM. The standardized CPUE for blue shark in the second quarter of the year peaked in the mid‐2000s and then decreased, but it has been increasing since 2012. The CPUE for shortfin mako in the second quarter generally increased, with fluctuations.     

Glomus tumor of the liver in a cow

An 11-year-old Holstein-Friesian cow exhibited anorexia and jaundice. A large mass was found in the liver during necropsy. Macroscopically, the mass was composed of dark red multilobular tissue and a centrally located abscess, which was connected to the hepatic duct. Histologically, the mass consisted of proliferation of small neoplastic cells and was demarcated from the hepatic parenchyma by a thick region of granulation tissue. The neoplastic cells were predominantly arranged in solid sheets, but they also formed blood-filled cancellous structures, and proliferating foci were seen around blood vessels. Periodic acid-Schiff reaction demonstrated that a fine basement membrane-like structure surrounded the neoplastic cells. Immunohistochemically, the neoplastic cells were positive for vimentin and alpha smooth muscle actin and negative for cytokeratin, factor VIII-related antigen, chromogranin and desmin. Based on its histopathological features, the hepatic neoplasm was diagnosed as a primary glomus tumor. This is the first report about a primary glomus tumor of the liver in a cow.     

Effect of polymorphism in egg white lysozyme on muramidase and antibacterial activities as well as hatchability in the Japanese quail (Coturnix japonica)

Lysozyme is one of the best characterized antimicrobial proteins in egg white. Three phenotypes of egg white lysozyme in Japanese quail, Coturnix japonica, (namely fast; slow; and the combination, FS) were observed by acid polyacrylamide gel electrophoresis. The fast phenotype showed faster mobility on Acid-PAGE than the slow phenotype. Comparison of the coding sequences for lysozyme derived from the slow and fast phenotypes revealed a nonsynonymous SNP at nucleotide position 115 from the translation initiation site, which alters AA sequence of lysozyme. This nonsynonymous SNP converted glutamine (Q) in the slow phenotype to lysine (K) in the fast phenotype at AA residue 21 of mature lysozyme (Q21K). Here, we investigated the effect of these phenotypes on muramidase activity, antibacterial activity, and hatchability. Muramidase activity toward isolated cell walls of Micrococcus lysodeikticus was in the order: fast allozyme > slow allozyme > chicken (Gallus gallus), but no significant difference was found among the 3 (P > 0.05). Antibacterial activity against live Staphylococcus aureus cells was significantly greater for the fast allozyme than the slow allozyme from 20 h after incubation (P < 0.05). For the antibacterial effects against live Escherichia coli cells, the activity of fast was significantly higher than that of slow at 16 h after incubation (P < 0.05). Hatchability was estimated for reciprocal crosses of Japanese quail with the FF (fast) and SS (slow) genotypes. Hatchability was 92.5% in FF male × SS female crosses and 87.2% in SS male × FF female crosses. A Cochran-Mantel-Haenszel test revealed a significant difference between the crosses (P < 0.05) and indicated that the female-derived slow phenotype led to improved rates of hatching. Our results suggest that the nonsynonymous SNP in Japanese quail lysozyme influences the electrophoretic migration, muramidase activity, and antibacterial activity of the protein, in addition to the hatchability of the eggs. These results demonstrate, for the first time, a significant difference in antibacterial activity and hatchability between 2 lysozyme phenotypes in Japanese quail.