A selective medium to isolate airborne spores of Microdochium nivale,causing winter wheat scab

Winter wheat scab in Hokkaido, Japan is caused predominantly by Gibberella zeae and Microdochium nivale and can result in significant yield losses. A selective medium for isolation of G. zeae was previously developed, but not for M. nivale. The purpose of this study therefore was to develop a selective medium for isolation of airborne spores of M. nivale. Based on the basic composition of Komada’s Fusarium-selective medium, carbon and nitrogen sources and the most suitable vitamin B component for the basal composition were examined. Hyphal growth of M. nivale was promoted when galactose was replaced with lactose and combined with L-asparagine, while aerial hyphal formation increased with thiamine hydrochloride as the vitamin B source. In antimicrobial composition, colony formation of other filamentous fungi was greatly inhibited by spiroxamine. Thiophanate methyl, to which M. nivale shows resistance, selectively inhibited the growth of Fusarium spp. only. Spore trapping using the selective medium was subsequently performed in a wheat field. M. nivale formed characteristic pinkish colonies on the selective medium in the case of contamination with other filamentous fungi, making differentiation easy. Overall, the findings show that LATTS medium developed in this study is effective for isolation of airborne spores of M. nivale.     

Temperature controls temporal variation in soil CO2 efflux in a secondary beech forest in Appi Highlands, Japan

Forest soil is a huge reserve of carbon in the biosphere. Therefore to understand the carbon cycle in forest ecosystems, it is important to determine the dynamics of soil CO₂ efflux. This study was conducted to describe temporal variations in soil CO₂ efflux and identify the environmental factors that affect it. We measured soil CO₂ efflux continuously in a beech secondary forest in the Appi Highlands in Iwate Prefecture for two years (except when there was snow cover) using four dynamic closed chambers that automatically open after taking measurements. Temporal changes in soil temperature and volumetric soil water content were also measured at a depth of 5 cm. The soil CO₂ efflux ranged from 14 mg CO₂ m⁻² h⁻¹ to 2,329 mg CO₂ m⁻² h⁻¹, the peak occurring at the beginning of August. The relationship between soil temperature and soil CO₂ efflux was well represented by an exponential function. Most of temporal variation in soil CO₂ efflux was explained by soil temperature rather than volumetric soil water content. The Q ₁₀ values were 3.7 ± 0.8 and estimated annual carbon emissions were 837 ± 210 g C m⁻² year⁻¹. These results provide a foundation for further development of models for prediction of soil CO₂ efflux driven by environmental factors.     

Mechanism of macrophage activation by chitin derivatives

In order to analyze the detailed mechanisms responsible for macrophage activation by chitin derivatives, resident peritoneal macrophages were prepared and stimulated with chitin, chitosan and low-molecular weight chitosan. Our findings were as follows: (i) chitosan induced apoptosis of peritoneal macrophages, but this did not occur when chitin or water soluble low-molecular weight chitosan were used; (ii) chitosan treatment induced activation markers, such as the major histocompatibility complex (MHC) class I, class II, Fc receptors, transferrin receptor, mannose receptor, Fas, and macrophage inflammatory protein (MIP)-2, whereas chitin and low molecular weight soluble chitosan induced only the expression of MHC class I and II molecules; (iii) apoptosis induced by chitosan was mediated by the Fas signaling pathway, in response to phagocytosis via the mannose receptor. We conclude that since chitosan activates macrophages, this may be the mechanism by which it accelerates wound healing.     

Bovine paraoxonase 1 activities in serum and distribution in lipoproteins

Paraoxonase 1 (PON1) is an enzyme associated with high-density lipoprotein (HDL) and has a protective effect against oxidation of lipoproteins in mammals. We investigated PON1 enzyme activities in bovine serum and its distribution among bovine serum lipoproteins. Paraoxonase activity and arylesterase activity in serum (152 Holstein cows and 42 Japanese Blacks) were 275 +/- 55 U/ml and 130 +/- 27 U/ml (mean +/- SD), respectively. There was a high correlation (r=0.962) between the two enzyme activities, and the activity ratio of paraoxonase/arylesterase did not exhibit individual variation. More than 85% of both paraoxonase and arylesterase activities were detected in the HDL fraction separated by ultracentrifugation. The 43-kDa protein in the HDL fraction was identified as bovine PON1 by N-terminal amino acid sequence analysis. Bovine PON1 was purified by ultracentrifugation and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and an anti-bovine PON1 antiserum was developed. The concentration of PON1 protein determined by immunoblotting was closely correlated (r=0.976) with paraoxonase activity in serum. Bovine HDL was further fractionated into subpopulations, and the distribution of PON1 was examined. Paraoxonase activity and PON1 protein increased with decreasing HDL size and approximately 60% of total paraoxonase activity was distributed in the heavy HDL fraction. The different distributions of PON1 among HDL subpopulations might be concerned to the function and metabolism of bovine HDL.     

Relationship between growth hormone (GH) pulses in the peripheral circulation and GH-releasing hormone and somatostatin profiles in the cerebrospinal fluid of goats

Growth hormone (GH) is secreted in a pulsatile manner, but the underlying mechanisms of GH pulse generation remain to be resolved. In the present study, we investigated the relationship between GH pulses in the peripheral circulation and GH-releasing hormone (GHRH) and somatostatin (SRIF) profiles in the cerebrospinal fluid (CSF) of male goats. The effects of an intracerebroventricular (icv) injection of neuropeptide Y (NPY), galanin and ghrelin were also analyzed. Blood and CSF samples were collected every 15 min for 8 hr from the jugular vein and third ventricle, respectively. GH pulsatility in the goat was found to consist of distinct large pulses of 5 hr periodicity and small pulses of 1 hr periodicity. GHRH and SRIF in the CSF fluctuated in a pulsatile manner with 1 hr periodicity, and most of the descending phase of SRIF pulses were associated with the initiation of GH pulses. Icv injections of NPY, galanin and ghrelin stimulated GHRH release without affecting SRIF release. In addition, NPY suppressed, and galanin and ghrelin induced large GH pulses, although ghrelin was much more effective than galanin. These results suggest that an hourly fall in SRIF is involved in generating intrinsic circhoral rhythm of GH pulsatility. The mechanisms underlying the generation of large GH pulses of 5 hr periodicity remain unknown, while direct action of NPY and/or ghrelin on the pituitary might be involved.     

Experimental transmission of ovine herpesvirus-2 in sheep

Transmission of ovine herpesvirus-2 (OvHV-2) in sheep via natural contact and nasal secretions was examined. OvHV-2-free lambs were produced by separating newborn lambs from their mothers within 5 days of birth and raising them in an isolation facility. Transmission experiments via natural contact were conducted by keeping OvHV-2-free lambs with OvHV-2-infected sheep of different ages. Six of the infected ewes in this experiment were pregnant and gave birth during the experimental period. OvHV-2 was not transmitted from the adult sheep, though viral DNA was consistently detected in their peripheral blood leukocytes (PBL). On the other hand, OvHV-2 was transmitted from recently infected lambs to sheep at 10 or 12 weeks after the onset of contact. In addition, we attempted the experimental transmission of OvHV-2 via nasal secretions, by transferring nasal washings from infected sheep to the nostrils of uninfected sheep. Sheep receiving the nasal washings from infected adult sheep maintained their negative status for 15 months, whereas sheep receiving nasal washings from recently infected lambs acquired OvHV-2 by 8 months. The results of these experiments support that OvHV-2 is more easily transmitted to negative sheep by recently infected lambs than by adult sheep. Further, it is supposed that the nasal cavity is a portal for entry and shedding of infectious OvHV-2 in sheep.     

Relationship of disease progression and plasma histamine concentrations in 11 dogs with mast cell tumors

Plasma histamine concentrations (PHCs) were measured serially over 9 months or until death in 11 dogs with mast cell tumors (MCTs). Eight dogs had grossly visible disease and the other 3 dogs had microscopic disease. Initial PHCs in the dogs with gross disease were significantly higher than PHCs in healthy dogs (median, 0.73 ng/mL and 0.19 ng/mL respectively; P < .009), whereas initial PHCs in dogs with microscopic disease showed no difference from controls. Seven dogs subsequently had progressive increases in PHC, and developed hyperhistaminemia (median, 14.0 ng/mL; range, 5.11-30.1 ng/nL). These 7 dogs died from MCTs, and 1 had general weakness with rapid lysis of a large tumor burden after radiation therapy. PHCs of the other 4 dogs were less than 1 ng/mL during the study. These 4 dogs were still alive with adequate control of the tumor at the conclusion of the study. Four of the 11 dogs initially had gastrointestinal (G1) signs, which abated soon after administration of histamine-2 (H-2) blockers. No significant difference was found between PHCs in dogs with GI signs and those without GI signs (median, 0.86 ng/mL and 0.35 ng/mL. respectively). Thereafter, 7 dogs had serious GI complications for which H-2 blocker therapy was ineffective. PHCs in these 7 dogs were extremely high (median, 12.2 ng/mL; range, 3.42-30.1 ng/nL). Results of this study demonsrated that PHC was one factor related to disease progression, and indicated that marked hyperhistaminemia was associated with the GI signs refractory to H-2 blocker therapy in dogs with MCTs.