N -ethyl- N -nitrosourea induces retinal photoreceptor damage in adult rats

Seven-week-old male Lewis rats received a single intraperitoneal injection of N-ethyl-N-nitrosourea (ENU) (100, 200, 400 or 600 mg/kg), and retinal damage was evaluated 7 days after the treatment. Sequential morphological features of the retina and retinal DNA damage, as determined by a TUNEL assay and phospho-histone H2A.X (γ-H2AX), were analyzed 3, 6, 12, 24 and 72 hr, 7 days, and/or 30 days after 400 mg/kg ENU treatment. Activation of the nuclear enzyme poly (ADP-ribose) polymerase (PARP) was analyzed immunohistochemically by poly (ADP-ribose) (PAR) expression in response to DNA damage of the retina. All rats that received ≥ 400 mg/kg of ENU developed retinal degeneration characterized by the loss of photoreceptor cells in both the central and peripheral retina within 7 days. In the 400 mg/kg ENU-treated rats, TUNEL-positive signals were only located in the photoreceptor cells and peaked 24 hr after ENU treatment. The γ-H2AX signals in inner retinal cells appeared at 24 hr and peaked at 72 hr after ENU treatment, and the PAR signals selectively located in the photoreceptor cell nuclei appeared at 12 hr and peaked at 24 hr after ENU treatment. However, degeneration was restricted to photoreceptor cells, and no degenerative changes in inner retinal cells were seen at any time points. Retinal thickness and the photoreceptor cell ratio in the central and peripheral retina were significantly decreased, and the retinal damage ratio was significantly increased 7 days after ENU treatment. In conclusion, ENU induced retinal degeneration in adult rats that was characterized by photoreceptor cell apoptosis through PARP activity.     

Growth of larval and early juvenile Japanese anchovy, Engraulis japonicus, in the Kuroshio-Oyashio transition region

Larvae, juveniles, and adults of Japanese anchovy, Engraulis japonicus , were distributed throughout the Kuroshio-Oyashio transition region off northern Japan as far offshore as 170°E in 1996 and 1997. The growth trajectories of individual larvae and early juveniles were backcalculated using the biological intercept method based on the allometric relationship between otolith radius and somatic length. Mean larval growth rates ranged from 0.49 to 0.71 mm day^–1 in the transition region, and were comparable to those reported from the Pacific coastal waters of central Japan, which is the principal distribution range of E. japonicus . In terms of growth, the Kuroshio-Oyashio transition region seemed to be a favourable nursery area for larval E. japonicus . Larval growth tended to decline from the inshore to the offshore waters in the transition region. Thermal conditions did not show an inshore–offshore trend in the survey area and did not explain the longitudinal trend in growth rates.     

Bioremediation of organically enriched sediment deposited below fish farms with artificially mass-cultured colonies of a deposit-feeding polychaete <Emphasis Type="Italic">Capitella</Emphasis> sp. I

ABSTRACT:   For bioremediation of organically enriched sediment deposited below fish farms, the extremely high potential for population growth of a deposit-feeding polychaete, Capitella sp. I, in the organically enriched sediment, and the effect on decomposition of organic matter in the sediment, were examined. A mass-culturing technique was conducted for this species. Bioremediation experiments were conducted on the organically enriched sediment in a fish farm in Kusuura Bay, Japan in 2003–2006. Approximately 1.7 million individuals of the worms were placed on the sediment below one net pen in December 2003, 9.3 million individuals in November 2004, and 2.2 million individuals in November 2005. After the worms were spread on the sediment, they rapidly increased in number and reached the highest densities of approximately 134 000 inds/m² in February 2004, 527 000 inds/m² in March 2005 and 103 000 inds/m² in January 2006. In the process of rapid population growth, the decomposition of the organic matter of the sediment was enhanced markedly. Our results demonstrate that the promotion of population growth by spreading cultured colonies of Capitella can enhance the decomposition rate of organic matter markedly in organically enriched sediment below fish farms. This method is promising for minimization of the negative effects of fish farms.     

Thioacetamide-induced Hepatocellular Necrosis Is Attenuated in Diet-induced Obese Mice

To assess modification of thioacetamide-induced hepatotoxicity in mice fed a high-fat diet, male C57BL/6J mice were fed a normal rodent diet or a high-fat diet for 8 weeks and then treated once intraperitoneally with thioacetamide at 50 mg/kg body weight. At 24 and 48 hours after administration, massive centrilobular hepatocellular necrosis was observed in mice fed the normal rodent diet, while the necrosis was less severe in mice fed the high-fat diet. In contrast, severe swelling of hepatocytes was observed in mice fed the high-fat diet. In addition, mice fed the high-fat diet displayed more than a 4-fold higher number of BrdU-positive hepatocytes compared with mice fed the normal rodent diet at 48 hours after thioacetamide treatment. To clarify the mechanisms by which the hepatic necrosis was attenuated, we investigated exposure to thioacetamide and one of its metabolites, the expression of CYP2E1, which converts thioacetamide to reactive metabolites, and the content of glutathione S-transferases in the liver. However, the reduced hepatocellular necrosis noted in mice fed the high-fat diet could not be explained by the differences in exposure to thioacetamide or thioacetamide sulfoxide or by differences in the expression of drug-metabolizing enzymes. On the other hand, at 8 hours after thioacetamide administration, hepatic total glutathione in mice fed the high-fat diet was significantly lower than that in mice fed the normal diet. Hence, decreased hepatic glutathione amount is a candidate for the mechanism of the attenuated necrosis. In conclusion, this study revealed that thioacetamide-induced hepatic necrosis was attenuated in mice fed the high-fat diet.     

Susceptibility to N-methyl-N-nitrosourea-induced retinal degeneration in different rat strains

To evaluate the potential role of genetic background in the susceptibility to retinal degeneration induced by N-methyl-N-nitrosourea (MNU), female rats of the Sprague-Dawley (SD), Long-Evans (LE) and Copenhagen (CH) strains were administered 50 mg/kg MNU or saline at 7 weeks of age. Retina morphology and morphometric analysis of all rats was performed 7 days after MNU administration. Atrophy of both the peripheral and central outer retina occurred in all rat strains exposed to MNU. Decreased photoreceptor cell ratio and increased retinal damage ratio were observed. The severities of the retinal atrophy were similar among all three rat strains. In conclusion, MNU-induced photoreceptor degeneration developed consistently in all three strains regardless of the absence (SD rats) or presence (LE and CH rats) of melanin in the retina, suggesting that genetic and melanin factors did not affect photoreceptor cell death after MNU.     

Fertilisation of bovine oocytes by the injection of immobilised, killed spermatozoa

Immobilised (killed) bovine spermatozoa were microinjected into bovine oocytes matured in vitro and cultured for six to nine days in vitro. A co-culture system with cumulus cells was used for the embryonic development. After one to two, three to four, five to six and seven to eight days the proportions of the oocytes which had developed to the two to four-cell, six to 12-cell, morula and blastocyst stages were 12.0 per cent (61 of 507), 9.3 per cent (47 of 507), 5.9 per cent (30 of 507) and 7.8 per cent (nine of 115), respectively. In contrast, none of the sham-operated group developed beyond the six-cell stage. This is the first report to show that bovine oocytes matured in vitro can undergo cleavage to the blastocyst stage after the injection of sperm in vitro. In addition, normal calves were obtained from the transfers of some of the embryos to recipient cows.     

Genotyping of Giardia intestinalis from domestic and wild animals in Japan using glutamete dehydrogenase gene sequencing

To determine the genotypes of Giardia intestinalis from domestic and wild animals in Japan, Giardia isolates obtained from feces of 24 dogs kept in households and breeding kennels, three companion cats, five dairy calves and three wild monkeys, Macaca fuscata, were genotyped using the 177 bp sequence of the glutamete dehydrogenase gene (gdh). The genotypes were assemblages A, C, D or A/D for dog isolates, Assemblage F for cat isolates, assemblages A or E for calf isolates and assemblage B for monkey isolates. This is the first report on the genotypes of Giardia isolates from cats, calves and wild monkeys in Japan.     

Geographical variation in skull morphology of gray-bellied squirrel Callosciurus caniceps

The geographical variation of the gray-bellied squirrel (Callosciurus caniceps) was examined using osteometry of skull in Southeast Asia. From the principal component analysis (PCA), the plots of the northern localities from Nan to Kanchanaburi and those of the southern localities from Narathiwat to Kuala Lumpur in male were completely separated. In female, the plots of the locality from Uttradit to Kanchanaburi and those of the locality from Pattani to Negri Sembilan were completely separated. We called these northern localities and southern localities which are distinguished by the PCA as N group and S group. The size and shape of the skulls of these squirrels indicated the differences between N group and S group from t-test and U-test. These results may be influenced by the two transitions of the phytogeography around the southernmost locality in N group and the northernmost locality in S group in the peninsular Thailand and Malay Peninsula. Localities which are located between N and S groups were called the Middle (M) group. From the PCA among N, S groups and each locality of M group, the plots of localities such as Prachuap Khiri Khan, Chumphon, Krabi, Nakhon Si Thammarat and Trang in both sexes of M group could not be separated from those of N and S groups. We suggest that the sympatric distribution of N and S groups and the hybrid of N and S populations may be seen in these localities of M group.