Purification and characterization of feline ghrelin and its possible role

Ghrelin, a novel 28-amino acid peptide with an n-octanoyl modification at Ser3, has been isolated from rat and human stomach as an endogenous ligand for the growth hormone secretagogue receptor. Here, we purified feline ghrelin and examined its possible physiological role in cats. The major active form of feline ghrelin is a 28-amino acid peptide octanoylated (C8:0) at Ser3; except for one amino acid residue replacement, this structure is identical to those of rat and human ghrelins. However, much structural divergence in peptide length and fatty acid modification was observed in feline ghrelin: peptides consisting of 27 or 26 amino acids lacking Gln14 and/or Arg28 were found, and the third serine residue was modified by octanoic acid (C8:0), decanoic acid (10:0), or unsaturated fatty acids (C8:1, C10:1 and C10:2). In agreement with the structural divergence, two kinds of cDNA with different lengths were isolated. Administration of synthetic rat ghrelin increased plasma growth hormone levels in cats, with a potency similar to that in rat or human. Plasma levels of ghrelin in cats increased approximately 2.5-fold after fasting. The present study indicates the existence of structural divergence in feline ghrelin and suggests that, as in other animals, ghrelin may play important roles in GH release and feeding in cats.     

Anaphylaxis after vaccination for cats in Japan

Severe adverse reactions in cats after vaccination were examined from 316 cases reported to the Ministry of Agriculture, Forestry and Fisheries (MAFF) in Japan during 15-year period from April 2004 to March 2019. We found that 130 (41%) showed anaphylaxis, and 99 (76%) of the 130 cases of anaphylaxis resulted in death. Veterinarians should be well prepared to deal with vaccine-associated anaphylaxis in cats. Bovine serum albumin (BSA) as indicator of purification was detected at high levels in commercially available feline vaccines. BSA might derive from fetal calf serum in culture media. This study provides useful information about anaphylaxis including critical details of the potential clinical signs associated with adverse events to feline vaccination.     

Temporal shifts in diversity and quantity of nirS and nirK in a rice paddy field soil

Denitrification is an important part of the nitrogen cycle in the environment, and diverse bacteria, archaea, and fungi are known to have denitrifying ability. Rice paddy field soils have been known to have strong denitrifying activity, but the microbes responsible for denitrification in rice paddy field soils are not well known. Present study analyzed the diversity and quantity of the nitrite reductase genes (nirS and nirK) in a rice paddy field soil, sampled four times in one rice-growing season. Clone library analyses suggested that the denitrifier community composition varied over sampling time. Although many clones were distantly related to the known NirS or NirK, some clones were related to the NirS from Burkholderiales and Rhodocyclales bacteria, and some were related to the NirK from Rhizobiales bacteria. These denitrifiers may play an important role in denitrification in the rice paddy field soil. The quantitative PCR results showed that nirK was more abundant than nirS in all soil samples, but the nirK/nirS ratio decreased after water logging. These results suggest that both diversity and quantity changed over time in the rice paddy field soil, in response to the soil condition.     

Establishment of a potency test by ELISA for a rabies vaccine for animal use in Japan

The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.     

Survey on wild rodents for endoparasites in Iwate Prefecture, Japan

Wild rodents (58 Apodemus speciosus, 29 A. argenteus and 7 Microtus montebelli) were surveyed for endoparasites in Iwate Prefecture, Japan, from October to December 1995 and from April to October 1996. Two trematodes (Echinostoma macrorchis, Plagiorchis muris), 4 or more cestodes (Hymenolepis diminuta, Raillietina coreensis, Cladothyridium spp., Cysticercus fasciolaris), 12 nematodes (Carolinensis minutus, Eucoleus sp., Heligmosomoides kurilensis, H. protobullosus, H. speciosus, Heterakis spumosa, Rhabditis (Pelodera) orbitalis, Rictularia cristata, Syphacia emileromani, S. frederici, S. montana, Trichuris sp.) and 3 protozoans (Giardia sp., Trichomonas sp., Trypanosoma sp.) were identified. The two species of Apodemus were similar to each other, but they were extremely different from M. montebelli in parasite fauna.     

Induction of unseasonable hibernation and involvement of serotonin in entrance into and maintenance of its hibernation of chipmunks T. asiaticus

Chipmunks that had been housed at 22 degrees C under a light-dark cycle of 14L:10D for at least one year were exposed to a short photoperiod (10L:14D) and low temperature to induce unseasonable hibernation. We were able to induce hibernation at any time of year and there was no significant difference in the duration of the hibernation bout, the duration of interbout euthermia and duration of bouts of torpor throughout the year; however entrance into hibernation took about 60 days in summer but only about 30 days in any other seasons. In addition, interbout euthermia predominantly occurred during the light phase in winter, whereas in spring interbout euthermia occurred equally in the light and dark phases. These results suggest that both the circadian and circannual systems are linked to hibernation in chipmunks. Subcutaneous infusion of a serotonin antagonist, para-chlorophenylalanine (PCPA), facilitated entrance into and interrupted hibernation in aroused and hibernating chipmunks in summer, respectively. On the other hand, opioid antagonist, naloxone, did not affect hibernation, but extended the period of interbout euthermia. These results suggest that the role of serotonin in entrance into and maintenance of hibernation in chipmunks is independent of the circannual system, and that opioid system may not be involved in hibernation in chipmunks.     

Cross-sectional cutting of bamboo with a pair of shearing blades for bamboo cube production

As a new use of bamboo, bamboo cubes may be useful as a shot-blast material for surface treatments. However, a suitable processing technique for bamboo cubes has not been established. In this study, to obtain basic knowledge regarding the processing technology for bamboo cubes, we installed shearing blades (upper and lower blades) in a universal testing machine to test cross-sectional shearing of bamboo. The shearing force generated in this shearing was composed of a vertical component (F _v), a forward component (F _f), and a side component (F _s). This shearing force (F _v, F _f, and F _s) and the machining accuracy were investigated under various processing conditions. The shearing force became larger as the thickness of bamboo increased. In particular, F _v showed a tendency to increase rapidly. Changes in the shear angle of the upper blade had a remarkable influence on F _v. This result suggests that the shearing force could be greatly decreased by adjusting the shear angle. The shearing force within bamboo of 3mm in thickness was almost unaffected by the blade angle, and the change of shearing force with increasing clearance was almost indiscernible. It was clearly demonstrated that a large number of bamboo cubes could be made when the shear angle was large and cross-sectional shearing was performed from the bark side. However, a consequence of a large shear angle is that burr area increases. Part of this article was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, March 2003     

First report of trunk rot caused by <Emphasis Type="Italic">Fomitiporia torreyae</Emphasis> in Kyoto prefecture on cultivars of Japanese cedar with no relatedness to ‘Sanbu-sugi’

Trunk rot, caused by Fomitiporia torreyae, is one of the most economically important sap rot diseases on Cryptomeria japonica, especially on a cutting cultivar ‘Sanbu-sugi’. This disease had been reported only from Chiba and Ibaraki prefectures in Japan; however, a similar trunk rot on Japanese cedars was found recently in Kyoto prefecture. We identified the causal agent of the trunk rot on Japanese cedars in Kyoto prefecture, clarified the genotype of ‘Sanbu-sugi’ (sensu stricto) using microsatellite markers, and then investigated the relatedness of these infected cedars with ‘Sanbu-sugi’. The pathogen was identified as F. torreyae by its DNA sequences and morphological characteristics of the fruiting body. The 25 ramets of ‘Sanbu-sugi’ had only one genotype based on eight microsatellite loci. Infected Japanese cedars in Kyoto were comprised of different genotypes and showed no close relatedness to ‘Sanbu-sugi’. This is the first report of trunk rot caused by F. torreyae on cultivars of Japanese cedar with no relatedness to ‘Sanbu-sugi’ outside of Chiba and Ibaraki prefectures and showed the potential for expansion of this disease to Japanese cedar plantations in other areas.     

Effect of particle shape on linear expansion of particleboard

Hinoki (Japanese cypress) strand with 0.6mm thickness was hammer-milled to prepare three types of particles that had the same thickness but different lengths and widths. Screen analysis and image analysis were conducted to evaluate the shape and distribution of the particle dimensions. Laboratory-scale particleboards were fabricated at three density levels using these particles. To determine the true effect of particle shape on the linear expansion (LE) of the board, these boards exhibited almost the same temperature behavior during pressing, the same density profile, and the same bending properties at each board density level. LE at 40°C and 90% relative humidity of the board was found to be affected by the particle shape. The board composed of small particles showed a larger LE at the same density level. It was considered that the out-of-plane orientation angle of the particles affected the LE of the boards. Thickness swelling and internal bond strength were also affected by the particle shape.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000