Reproductive characteristics of the feral raccoon (Procyon lotor) in Hokkaido,Japan

Reproduction of feral raccoons (Procyon lotor) in Hokkaido, Japan, was examined during a 2-year period by analysis of placental scars or fetuses in the uterus. Of 242 collected females, 69 (29%) were juveniles, 71 (29%) yearlings, and 102 (42%) adults. The pregnancy rate averaged 66% in yearlings and was significantly lower than the 96% average observed in adults (p<0.01). Litter size ranged from 1 to 7 offspring per female, and averaged 3.6 in yearlings and 3.9 in adults. There was no significant difference in mean litter size between yearlings and adults. In Hokkaido, the raccoon mating season peaked in February and the majority of litters were born between March and May, similar to patterns described in North America, but some females mated in summer. The reproductive potential of feral raccoons in Hokkaido was similar to that reported in North America. The recent increase in raccoon numbers can be explained by their high productivity. Harvest data suggest that hunting pressure on juveniles is lower than that for older age classes when using box traps in summer. In order to reduce the feral raccoon population, alternative hunting methods that increase juvenile mortality rates are needed.     

iTRAQ-Based Proteomics Investigation of Critical Response Proteins in Embryo and Coleoptile During Rice Anaerobic Germination

Direct-seeding of rice has become popular in recent years due to its low cost and convenience, however, hypoxic condition limits seedling establishment. In this study, weedy rice WR04-6 with high germination ability under anaerobic conditions was used as a gene donor, and we successfully improved the seedling establishment rate of rice cultivar Qishanzhan (QSZ) based on selection of a new rice line R42 from the recombinant inbred line population. R42 inherited high anaerobic germination (AG) ability, and was used for isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative proteomic studies with QSZ to further explore the molecular mechanism of AG. A total of 719 differentially abundant proteins (DAPs) were shared by R42 and QSZ responded to AG, and thus defined as common response DAPs. A total of 300 DAPs that responded to AG were only identified from R42, which were defined as tolerance-specific DAPs. The common response and tolerance-specific DAPs had similar biochemical reaction processes and metabolic pathways in response to anoxic stress, however, they involved different proteins. The tolerance-specific DAPs were involved in amino acid metabolism, starch and sucrose metabolism, tricarboxylic acid cycle pathway, ethylene synthesis pathway, cell wall-associated proteins and activity of active oxygen scavenging enzyme. The in silico protein-protein interactions for the top 60 DAPs indicated that tolerance-specific DAPs had relatively independent protein interaction networks in response to an anoxic environment compared with common response DAPs. The results of physiological indicators showed that α-amylase and superoxide dismutase activities of R42 were significantly increased under anoxic conditions compared with aerobic conditions. Multiple lines of evidence from western blot, physiological analysis and quantitative real-time PCR jointly supported the reliability of proteomics data. In summary, our findings deepened the understanding of the molecular mechanism for the rice response to AG.     

Effects of alcohol extract of defatted soybean meal on growth performance and digestive physiology of yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

The effects of alcohol extract of soybean meal on growth performance, digestibility, and pancreatic enzyme and bile acid status of yellowtail Seriola quinqueradiata were studied. Fish were fed one of the following for 8 weeks: a fish meal (FM) control diet (FMD), a defatted soybean meal diet (SBMD), an alcohol-extracted SBM diet (ExSBMD), an ExSBMD with alcohol-extract diet (ExSBM + ExtD), and an FMD with alcohol-extract diet (FM + ExtD). Growth performance was significantly inferior in fish fed with SBM, ExSBM + ExtD, and FM + ExtD compared to FMD- or ExSBMD-fed fish. Total bile acid levels, and trypsin and lipase activities in the anterior intestine were significantly lower in fish fed with SBM, ExSBM + ExtD, and FM + ExtD than in fish fed with ExSBMD and FMD, despite similar values in gallbladder or pyloric caeca. Cholecystokinin mRNA levels of the former diet-fed fish were significantly lower than those of FMD- or ExSBMD-fed fish. Lipid and protein digestibility of fish fed with SBMD, ExSBM + ExtD, and FM + ExtD was significantly reduced in comparison with that of fish fed with FMD or ExSBMD. These findings indicate that the alcohol extract of soybean meal inhibited the secretion of bile acids and pancreatic enzymes by a decrease of cholecystokinin stimulation. This inhibition seemed to be responsible for a low growth performance through impairment of the assimilation of lipid and protein.     

Acoustic pressure sensitivities and effects of particle motion in red sea bream Pagrus major

The auditory pressure thresholds of red sea bream were examined using cardiac response in the field by placing fish subjects far from the sound source to prevent particle motion. Pressure and particle motion thresholds were also obtained using the auditory brainstem response (ABR) technique. The thresholds at 100 and 200 Hz were significantly higher when measured using the cardiac response in the far field than those obtained in previously conducted experiments in experimental tub. However, thresholds obtained using ABR from 200 to 500 Hz were not remarkably lower, although significantly different (0.01 < P < 0.05), compared with those obtained using cardiac response in the far field. Furthermore, calculated particle velocity thresholds indicated that fish probably detected particle motion within the frequency range of 50–200 Hz, even in fish with a deactivated lateral line. Although the ABR method is widely applied in fish auditory study, hearing thresholds are apparently affected by particle motion.     

Dental incremental lines in Sika deer (Cervus nippon); polarized light and fluorescence microscopy of ground sections

Periodic growth incremental lines are found universally in dental hard tissues. This periodicity theoretically allows for estimation of age, even in days, which would be useful in studies of wild animals. In the present study, enamel and dentin increments of the sika deer (Cervus nippon) were observed in ground sections with a polarized light microscope, and their periodicity was examined by the use of a chronological labeling method with fluorochromes. Enamel increments occurred at a mean interval of 10.6 (SD=1.5) microm, and mean spacing of dentin increments was 17.3 (SD=1.8) microm. Fluorochromic marking revealed that incremental lines form each day in enamel and almost every second day in dentin. The fluorescence-labeled lines suggest that enamel formation of the first molar is complete by the age of 5 months. Due to its longer interval of incremental lines and longer term of formation, we conclude that dentin is more suitable than enamel for day-age estimation in sika deer. Experimental confirmation of incremental growth periodicity in various species can improve the reliability of use of tooth increments for age estimation and life history reconstruction.     

Molecular ordering of organic molten salts triggered by single-walled carbon nanotubes

When mixed with imidazolium ion-based room-temperature ionic liquid, pristine single-walled carbon nanotubes formed gels after being ground. The heavily entangled nanotube bundles were found to untangle within the gel to form much finer bundles. Phase transition and rheological properties suggest that the gels are formed by physical cross-linking of the nanotube bundles, mediated by local molecular ordering of the ionic liquids rather than by entanglement of the nanotubes. The gels were thermally stable and did not shrivel, even under reduced pressure resulting from the nonvolatility of the ionic liquids, but they would readily undergo a gel-to-solid transition on absorbent materials. The use of a polymerizable ionic liquid as the gelling medium allows for the fabrication of a highly electroconductive polymer/nanotube composite material, which showed a substantial enhancement in dynamic hardness.     

Urinary free metanephrines measurement in dogs with adrenal gland diseases using a new simple liquid chromatography tandem mass spectrometry method

Measurement of urinary metanephrines in spot samples is used for the diagnosis of canine pheochromocytoma (PC). We describe a simple analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for measuring free metanephrine (MN) and normetanephrine (NMN) in spot urine samples. Using the developed method, we evaluated the stability of urinary free-MN and free-NMN at various storing conditions. In addition, we assessed the feasibility of urinary free-MN and -NMN measurement for diagnosing PC. Urine samples were mixed with stable isotope internal standards and thereafter purified by ultrafiltration. The purified samples were analyzed by LC-MS/MS in the multiple reaction monitoring mode after separation on a multimode octa decyl silyl column. The coefficient of variation of free-MN and -NMN measurement was 7.6% and 5.5%, respectively. The linearity range was 0.5–10 µg/l for both analytes. Degradation was less than 10% for both analytes under any of the storage conditions. The median free-NMN ratio to creatinine of 9 PC dogs (595, range 144–47,961) was significantly higher (P<0.05) than that of 13 dogs with hypercortisolism (125, range 52–224) or 15 healthy dogs (85, range 50–117). The developed method is simple and may not require acidification of spot urine. The results of this preliminary retrospective study suggest that the measurement of urinary free metanephrines is a promising tool for diagnosing canine PC.