Seasonal differences in rumen bacterial flora of wild Hokkaido sika deer and partial characterization of an unknown bacterial group possibly involved in fiber digestion in winter

Rumen digesta was obtained from wild Hokkaido sika deer to compare bacterial flora between summer and winter. Bacterial flora was characterized with molecular‐based approaches and enrichment cultivation. Bacteroidetes was shown as a major phylum followed by Firmicutes, with similar proportions in both seasons. However, two phylogenetically unique groups in Bacteroidetes were found in each season: unknown group A in winter and unknown group B in summer. The ruminal abundance of unknown group A was the highest followed by Ruminococcus flavefaciens in winter. Moreover, the abundance of these two was higher in winter than in summer. In contrast, the abundance of unknown group B was higher in summer than in winter. In addition, this group showed the highest abundance in summer among the bacteria quantified. Unknown group A was successfully enriched by cultivating with oak bark and sterilized rumen fluid, particularly that from deer. Bacteria of this group were distributed in association with the solid rather than the liquid rumen fraction, and were detected as small cocci. Accordingly, unknown group A is assumed to be involved in degradation of fibrous materials. These results suggest that wild Hokkaido sika deer develop a rumen bacterial flora in response to changes in dietary conditions.     

Growth of rumen papillae in weaned calves is associated with lower expression of insulin‐like growth factor‐binding proteins 2, 3, and 6

This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin‐like growth factor‐binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real‐time PCR, and the presence of insulin‐like growth factors I and II (IGF‐I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk‐continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk‐continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk‐continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF‐I and IGF‐II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.     

Relationship between inbreeding coefficients and plant height of 1-year-old seedlings in crosses among Japanese pear (Pyrus pyrifolia Nakai) cultivars/selections

Plant height, a vigor trait, in 1-year-old seedlings made from Japanese pear (Pyrus pyrifolia) cultivars/selections was measured using 994 individuals from 29 families. The family mean of plant height was negatively correlated (r = −0.72**) to the inbreeding coefficients (F). The regression of the family mean (Fm) on the F value (Fm = 130 − 104F) showed that inbreeding depressions were 8%, 20%, and 40% for F = 0.1, 0.25, and 0.5, respectively. According to the regression, the family mean at F = 0 was estimated at 130 cm. These results showed that the vigor was greatly influenced by inbreeding in Japanese pear. Within-family variances, the genetic segregation of offspring in a family, differed according to family. The proportions of offspring with plant height above 130 cm (estimated Fm for F = 0) were extremely low, i.e., 0–17% for 0.5 ≤ F < 0.60 and 0–8% for F = 0.75.     

Arrangement of cortical microtubules in compression wood tracheids of Taxus cuspidata visualized by confocal laser microscopy

Cortical microtubules (MTs) in differentiating compression wood tracheids of Taxus cuspidata stems were visualized by confocal laser microscopy. They were oriented obliquely at an angle of about 45° to the tracheid axis during formation of the secondary wall. Artificial inclination altered the pattern of alignment of MTs. Banding MTs were helically oriented late during the formation of the secondary walls. These results indicate that MTs might control the orientation and localized deposition of cellulose microfibrils in the secondary walls of compression wood tracheids.Part of this report was presented at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 1996     

Photosensitivity in mice caused by pyropheophorbide in the midgut gland of the scallop <Emphasis Type="Italic">Patinopecten yessoensis</Emphasis> observed in diarrhetic shellfish poisoning mouse bioassays

Photosensitivity was observed in the mice used in bioassays of diarrhetic shellfish poisoning. The mice were intraperitoneally injected with the extracts from the midgut glands of cultured scallops Patinopecten yessoensis gathered in early spring in Funka Bay, Hokkaido, Japan. The injections induced atypical symptoms in the mice within 24 h. The symptoms included piloerection, substantial eye discharge, swelling of the ears and head, and ear scratching. They were similar to those reported in studies of photosensitivity caused by pyropheophorbide in the midgut glands of abalones, which are herbivorous gastropods. The problematic scallops also accumulated a large amount of pyropheophorbide-a. The amount determined with high-performance liquid chromatography was 300–530 μg per 1 g of the homogenate of the midgut glands. The dose of pyropheophorbide sufficient to cause photosensitivity in a mouse is estimated to be 0.99–2.3 mg per mouse with 20-g body weight. Moreover, comparative tests in mice showed that the onset of the symptoms required light and a substantial amount of pyropheophorbide. Therefore, we determined that the observed symptoms were photosensitivity caused by pyropheophorbide. Scallops, which are suspension-feeding bivalves, can accumulate a large amount of pyropheophorbide in the midgut glands depending on the habitat environment.     

Thermal-softening properties and cooling set of water-saturated bamboo within proportional limit

Thermal-softening properties and cooling set of water-saturated bamboo were investigated using stressrelaxation measurements in heating and cooling processes, followed by residual deflection measurement. In the heating process, an obvious decrease in relative relaxation modulus due to thermal-softening of lignin was found at around 60°C. On the other hand, no clear change in the relative relaxation modulus was recognized in the cooling process. After the cooling process, about 65% and 75% of residual set was measured when the specimen was loaded on the epidermis and endodermis side, respectively. Also, residual set depended on the maximum temperature reached in the heating process and the unloaded temperature in the cooling process. From these results, it was deduced that the glass transition of lignin from the rubbery to glassy state is important to fix the deformation. Comparing thermal-softening behavior between bamboo and wood, the relative relaxation modulus of wood decreased steeply at higher temperatures than for bamboo. On the other hand, while about 75% of residual set was also found for wood, almost the same as for bamboo, the recovery of deformation with time was larger for wood than for bamboo. Part of this article was presented at the 53rd Annual Meeting of the Japan Zairyou Society, Okayama, May 2004     

Liquid-chromatographic hybrid triple–quadrupole linear-ion-trap MS/MS analysis of fatty-acid esters of dinophysistoxin-1 in bivalves and toxic dinoflagellates in Japan

Fatty-acid esters of dinophysistoxin-1 (DTX1) in scallops Patinopecten yessoensis, mussels Mytilus coruscus, and toxic dinoflagellate Dinophysis species, collected from Japanese seawater, were analyzed by liquid-chromatography mass spectrometry (LC–MS). Precursor ion monitoring, multiple reaction monitoring for 18 fatty-acid esters of DTX1, and full-scan MS/MS spectra obtained with a hybrid triple–quadrupole linear-ion-trap mass spectrometer showed that 14:0, 16:0, and 16:1 esters were the most abundant 7-O-acyl-DTX1 analogues in bivalves. Fatty-acids esters formed by conjugations at hydroxyl positions other than the 7-position of DTX1 were not detected in the bivalves. DTX1 and okadaic acid-16:0 fatty-acid esters have been reported as the most abundant ester in bivalves in several previous studies; however, we found that 7-O-16:1-DTX1 was the most abundant ester in some mussels in which 16:1 was more dominant than 16:0 in the free fatty-acid profile. Comparison between 7-O-acyl-DTX1 and free fatty-acid profiles in the same bivalve samples suggests that polyunsaturated fatty acids are selectively excluded in enzymatic acylation of DTX1. No 7-O-acyl-DTX1 was detected in any single-cell isolates of D. fortii, D. acuminata, D. mitra, D. norvegica, D. tripos, D. infundibulus, and D. rotundata.     

Growth and water quality for growing-out of juvenile spotted Babylon, Babylonia areolata, at different water-exchange regimes in a large-scale operation of earthen ponds

Growth and water quality for growing-out of juvenile spotted Babylon, Babylonia areolata, were determined at three water-exchange regimes of 7-, 15- and 30-day intervals in large-scale operation of earthen ponds over a 5-month period. Results showed that growth was significantly different among the water-exchange treatments (P < 0.05). The higher body weight gains and shell length increments were observed in snails held at water exchange of 7- and 15-day intervals when compared with those held at water exchange of 30-day intervals. At the end of the experiment, average body weight gains were 4.22, 3.73 and 2.77 g for snails held in water-exchange treatments of 7-, 15- and 30-day intervals, respectively, and 16.63, 15.04 and 13.78 mm for those of shell length increments, respectively, The average final survivals were 83.60, 80.90 and 74.20% for snails held in water-exchange treatments of 7-, 15- and 30-day intervals, respectively. Seawater temperature, salinity, pH, dissolved oxygen, nitrite–nitrogen and ammonia–nitrogen had gradually changed throughout the culture period for all water-exchange treatments. With one exception, the total alkalinity showed the greatest changes during the culture period for all water-exchange treatments ranging from 58.67–97.50 mg/l.     

Amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis> interleukin-10 negatively suppresses host cell-mediated immunity

Interleukin-10 (IL-10) is an anti-inflammatory cytokine and negatively regulates cell-mediated immunity (CMI) induction by inhibiting cytokine production in type 1 T helper cells. IL-10 genes have been isolated from several fish, and inflammatory cytokine inhibition by IL-10 has been well examined. However, a CMI regulator of IL-10 in fish has not yet been identified. In this study, we cloned the IL-10 gene in amberjack Seriola dumerili and analyzed its function using its recombinant protein (rIL-10). In an in vitro culture experiment, gene expression of inflammatory cytokines was suppressed in leukocytes incubated with rIL-10 compared with cells that only received Nocardia seriolae stimulation. This result suggests amberjack IL-10 has conserved function as an inflammatory cytokine inhibitor. Bactericidal activity of amberjack cells against intracellular pathogen stimulation was decreased in a rIL-10 dose-dependent manner. Furthermore, a significant reduction in the T-bet/GATA-3 ratio was observed in N. seriolae living cell (LC)?+?rIL-10-injected fish. Taken together, these results suggest amberjack rIL-10 suppresses CMI induction both in vitro and in vivo. In addition, the number of IgM⁺ cells among spleen leukocytes in N. seriolae?+?rIL-10-injected fish was higher than in only N. seriolae LC, suggesting that Th2-dominant immunity was induced by adding rIL-10.     

Long‐term changes in recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) and environmental conditions around Japan

Recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) from 1952 to 2014 was examined by a sequential regime shift detection method. The regime shifts in recruitment were detected in 1957, 1972, 1980, 1994 and 2009. The durations of regime shift ranged from 8–15 years and averaged 13.0 years. In both the total (1952–2014) and data rich (1980–2014) periods, negative relationships were found between recruitment and the Pacific Decadal Oscillation in autumn, and positive relationships were found between recruitment and sea surface temperature (SST) anomalies in the northern part of the East China Sea, in the southwestern part of the Sea of Japan, and in the waters off Shikoku and Tokai in summer and autumn. The 1994 and 2009 regime shifts in recruitment occurred in the same years as shifts in SST anomalies in the northern part of the East China Sea in summer. These results suggest that the ocean conditions in the northern part of the East China Sea are closely related to recruitment of Pacific bluefin tuna, and that the warmer conditions result in higher recruitment of the species.