Diluted isoflurane as a suitable alternative for diethyl ether for rat anaesthesia in regular toxicology studies

Despite its explosive properties and toxicity to both animals and humans, diethyl ether is an agent long used in Japan in the anaesthesia jar method of rat anaesthetises. However, in response to a recent report from the Science Council of Japan condemning diethyl ether as acceptable practice, we searched for an alternative rat anaesthesia method that provided data continuous with pre-existing regular toxicology studies already conducted under diethyl ether anaesthesia. For this, we examined two candidates; 30% isoflurane diluted with propylene glycol and pentobarbitone. Whereas isoflurane is considered to be one of the representatives of modern volatile anaesthetics, the method of propylene glycol-diluted 30% isoflurane used in this study was our modification of a recently reported method revealed to have several advantages as an inhalation anaesthesia. Intraperitoneal pentobarbitone has long been accepted as a humane method in laboratory animal anaesthesiology. These 2 modalities were scrutinized in terms of consistency of haematology and blood chemistry with previous results using ether. We found that pentobarbitone required a much longer induction time than diethyl ether, which is suspected to be the cause of fluctuations in several haematological and blood chemical results. Conversely, only calcium ion concentration showed a slight difference from traditional results in the case of 30% isoflurane. Additionally, serum prolactin and corticosterone levels indicated that 30% isoflurane induced less stress than ether, confirming that 30% isoflurane can both provide results consistent with diethyl ether, while at the same time remove its disadvantages. As such 30% isoflurane appears to be a strong alternative anaesthetic agent for future regular toxicology studies in Japan.     

Isolation of Listeria monocytogenes from the skin of slaughtered beef cattle

We attempted to isolate Listeria monocytogenes from skin, contents of large intestines and carcasses of cattle introduced to a slaughterhouse in order to identify source of contamination for this pathogen. Sixty skin samples, 60 samples of the contents of large intestines and 30 carcass samples were colleted in June, August and November 2003 for use in this study. Listeria spp. and L. monocytogenes were isolated from 30 (50%) and 3 (5%) of the cattle skin samples, respectively. However, no Listeria spp., including L. monocytogenes, were isolated from intestinal contents or carcasses. Seven isolates were obtained, of which five and two strains were serotypes 1/2a and 1/2b, respectively. Genetic analysis suggested that there was persistent inhabitation of the pathogen around the area investigated in this study.     

Isolation and identification of F-spondin in the boar testis and its production during testis growth

F-spondin/vascular smooth muscle cell growth-promoting factor (VSGP), purified from the follicular fluid of adult bovine ovaries, has been identified as a promoter of neuronal differentiation and vascular smooth muscle growth. The objectives of the present study were (1) to clarify whether F-spondin is also produced in the testis, which is ontogenically equivalent to the ovary, and (2) to examine whether production of this protein changes with testicular growth. To isolate F-spondin from the testis, testicular homogenates obtained from 8-week-old boars were sequentially subjected to heparin-Sepharose chromatography, diethylaminoethyl (DEAE)-Sepharose chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). The isolated protein had a molecular mass of approximately 110 kDa and was cross-reactive with anti-F-spondin antibody by Western blotting. The purified protein was further characterized by amino acid sequence analysis of its internal peptide. The sequence obtained was GEQCNIVPDN VD, and a homology search indicated that the purified protein is a homologue of rat, human, and bovine F-spondin. By fractionation of the same amounts of testis tissue obtained from 1-, 8-, 16-, and 40-week-old boars, we analyzed age-related production of F-spondin in the testis. Western blotting of the fractions obtained from RP-HPLC revealed the presence of a band at approximately 110 kDa, corresponding to F-spondin, in the testes obtained from boars between 1 and 16 weeks old, but this band was not detected at 40 weeks. These results clearly indicate that (1) the porcine testis produces F-spondin and that (2) production of this protein is evident in the immature porcine testis, but not the adult testis.     

Development of Centrocestus armatus in different final hosts

In this study, Centrocestus armatus metacercariae were fed orally to hamsters, albino rats, mice, and chicks. Animals were sacrificed and dissected at 1, 3, 5, 7, and 14 days post-infection to determine the development and recovery rate of worms. Results indicated that the average worm recovery rate in hamsters was 25% on the first day post-infection and recovery continued until the 14th day with a gradual decrease in the percentage. Worms were also recovered from mice and albino rats from the first until the third day post-infection, but no worms were recovered thereafter. In chicks, worms were not observed on first day but recovery was positive at 12 h post-infection. Among the four animal hosts, feces from hamsters were the only ones positive for eggs; these were initially observed from the third day and recovery continued until 14 days post-infection. In our study, hamsters are the animal model most suitable for the study of C. armatus when compared to rats, mice, and chicks.     

Clinical and molecular analysis of GM2 gangliosidosis in two apparent littermate kittens of the Japanese domestic cat

This case report documents clinical and molecular findings in two littermate kittens of the Japanese domestic cat with GM2 gangliosidosis variant 0. Analysis included detailed physical, magnetic resonance imaging, biochemical, pathological and genetic examinations. At first, these littermate kittens showed typical cerebellar signs at approximately 2 months of age. About 2 months later, they progressively showed other neurological signs and subsequently died at about 7 months of age. Magnetic resonance imaging just before the death showed an enlarged ventricular system, T1 hyperintensity in the internal capsule, and T2 hyperintensity in the white matter of the whole brain. Histological findings suggested a type of lysosomal storage disease. Biochemical studies demonstrated that the kittens were affected with GM2 gangliosidosis variant 0, and a DNA assay finally demonstrated that these animals were homozygous for the mutation, which the authors had identified in a different family of the Japanese domestic cat. The findings in the present cases provide useful information about GM2 gangliosidosis variant 0 in Japanese domestic cats.     

Variation of indole acetic acid (IAA) amounts in cambial-region tissues in 7- and 24-year-old sugi (Cryptomeria japonica) trees

Recently, breeding programs have attempted to produce high growth rates for shorter rotation cycles in plantation trees. In these trees, the ratio of juvenile wood increases; thus, the juvenile wood properties should be improved for structural use. To this end, it is important to understand the influences on juvenile wood properties precisely. In this study, we report on the indole acetic acid (IAA) amounts of juvenile sugi (Cryptomeria japonica) trees in September and compare the IAA amounts to those in mature trees. The IAA amounts at the lower trunks in juvenile trees were significantly larger than those in mature trees and the IAA amounts decreased with tree height. In each stand, except a mature tree stand, there is no significant effect of IAA amounts on latewood width and MFA. However, put together all samples, the latewood width and MFA increased with IAA amounts in samples with IAA <200 ng/cm². The samples at lower trunk in juvenile trees had significantly larger IAA amounts, larger MFA and larger latewood width than the samples in mature trees (p < 0.01). The very large IAA amounts may have a certain relation with juvenile wood properties.     

Relationship between sea-surface temperature and catch fluctuations in the Pacific stock of walleye pollock in Japan

ABSTRACT:   This paper investigates the relationship between sea-surface temperature (SST) and catch fluctuations in the Pacific stock of walleye pollock Theragra chalcogramma in Japan. Incorporating time lags between years of birth and harvest, the correlation coefficients between the catch and SST in two regions off the east coast of Hokkaido were calculated. The catch in year t had a high negative correlation with the SST during January–April and November–December of the years t- 2 and t- 3 in the spawning area. These results coincided well with the correlation observed in the northern 'Sea of Japan' stock. Both analyses suggested that the long-term catch fluctuations of the two stocks could be explained by the same mechanism, that is, the fluctuations would be explained by the SST in their spawning area during the spawning season using 2–3 or 3–5 years time lags, which corresponded to the dominant age of the catch within these two stocks.     

Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.