Partial characterization of phylogeny,ecology and function of the fibrolytic bacterium Ruminococcus flavefaciens OS14, newly isolated from the rumen of swamp buffalo

The fibrolytic rumen bacterium Ruminococcus flavefaciensOS14 was isolated from swamp buffalo and its phylogenetic, ecological and digestive properties were partially characterized. Isolates from rumen contents of four swamp buffalo were screened for fibrolytic bacteria; one of the 40 isolates showed a distinctive feature of solubilizing cellulose powder in liquid culture and was identified as R. flavefaciens based on its 16S ribosomal DNA sequence. This isolate, OS14, was employed for detection and digestion studies, for which a quantitative PCR assay was developed and defined cultures were tested with representative forages in Thailand. OS14 was phylogenetically distant from other isolated and uncultured R. flavefaciens and showed limited distribution among Thai ruminants but was absent in Japanese cattle. OS14 digested rice straw and other tropical forage to a greater extent than the type strain C94 of R. flavefaciens. OS14 produced more lactate than C94, and digested para grass to produce propionate more extensively in co‐culture with lactate‐utilizing Selenomonas ruminantium S137 than a co‐culture of C94 with S137. These results indicate that phylogenetically distinct OS14 could digest Thai local forage more efficiently than the type strain, possibly forming a symbiotic cross‐feeding relationship with lactate‐utilizing bacteria. This strain might be useful for future animal and other industrial applications.     

Light-use properties in two sun-adapted shrubs with contrasting canopy structures

We investigated the impact of high solar irradiance and elevated temperature on carbon gain by two, co-occurring, sun-adapted, dwarf shrub species, Planchonella obovata var. dubia (Koidz.) Hatusima and Hibiscus glaber Matsumura, growing on sun-exposed ridges in the Bonin Islands, in the subtropical Pacific Ocean. Planchonella had steeply inclined, longer lived, sclerophyllous leaves, whereas Hibiscus has thinner, more horizontally oriented, and shorter lived leaves. We tested the hypothesis that leaf physiological tolerance to high light is lower in Planchonella than in Hibiscus. Under relatively high irradiances (photosynthetic photon flux density, PPFD, > 500 micromol m(-2) s(-1)), net photosynthetic rate (P(n)) was about 8.0 and 0.4 micromol m(-2) s(-1) in mature and young leaves of Planchonella, and about 12.4 and 10.3 micromol m(-2) s(-1) in mature and young leaves of Hibiscus, respectively. Both P(n) and photosystem II (PSII) quantum yield at a given PPFD were lower in Planchonella than in Hibiscus, whereas non-photochemical quenching (NPQ) at a given PPFD was higher in Planchonella. When leaf discs were exposed to high light (1900 micromol m(-2) s(-1) PPFD) at 37, 40 or 43 degrees C for 3 h, the recovery of PSII quantum yield (F(v)/F(m)) in the following 60-min dark period was slower in Planchonella than in Hibiscus, indicating that the ability of PSII to tolerate high light and high temperature was less in Planchonella than in Hibiscus. We postulate that there is a linkage between leaf display and leaf photochemical ability in sun-adapted shrub species.     

Leaf morphology and photosynthetic adjustments among deciduous broad-leaved trees within the vertical canopy profile

Photosynthetic acclimation of deciduous broad-leaved tree species was studied along a vertical gradient within the canopy of a multi-species deciduous forest in northern Japan. We investigated variations in (1) local light regime and CO2 concentration ([CO2]), and (2) morphological (area, thickness and area per mass), biochemical (nitrogen and chlorophyll concentrations) and physiological (light-saturated photosynthetic rate) attributes of leaves of seven major species on three occasions (June, August and October). We studied early successional species, alder (Alnus hirsuta (Spach) Rupr.) and birch (Betula platyphylla var. japonica (Miq.) Hara); gap phase species, walnut (Juglans ailanthifolia Carrière) and ash (Fraxinus mandshurica var. japonica Rupr.); mid-successional species, basswood (Tilia japonica (Miq.) Simonk.) and elm (Ulmus davidiana var. japonica (Rehd.) Nakai); and the late-successional species, maple (Acer mono Bunge). All but maple initiated leaf unfolding from the lower part of the crown. The [CO2] within the vertical profile ranged from 320-350 ppm in the upper canopy to 405-560 ppm near the ground. The lowest and highest ambient [CO2] occurred during the day and during the night, respectively. This trend was observed consistently during the summer, but not when trees were leafless. Chlorophyll concentration was positively related to maximum photosynthetic rate within, but not among, species. Leaf senescence started from the inner part of the crown in alder and birch, but started either in the outer or top portion of the canopy of ash, basswood and maple. Chlorophyll (Chl) to nitrogen ratio in leaves increased with decreasing photon flux density. However, Chl b concentration in all species remained stable until the beginning of leaf senescence. Maximum photosynthetic rates observed in sun leaves of early successional species, gap phase or mid-successional species, and late successional species were 12.5-14.8 micromol m(-2) s(-1), 4.1-7.8 micromol m(-2) s(-1) and 3.1 micromol m(-2) s(-1), respectively.     

Allocation of 14C-carbon in two species of larch seedlings infected with ectomycorrhizal fungi

The flow of labeled carbon in ectomycorrhizal and non-ectomycorrhizal seedlings of Japanese larch (Larix kaempferi Sarg.) and its F1 hybrid (Larix gmelinii Rupr. x L. kaempferi) was studied. Larch seedlings were grown in a greenhouse for 110 days with larch forest soil (FM) or Suillus grevillei (SM) inoculum, or in the absence of ectomycorrhizal fungi (NM). Shoots of colonized and NM seedlings were exposed to a pulse of 14CO2 for 1 h under natural light. Seedlings were harvested following 0, 6 and 24 h of exposure to 14CO2. At the final harvest, SM seedlings of Japanese larch and hybrid larch allocated 2.6 and 2.5% more 14C, respectively, to roots than NM seedlings. In contrast, FM seedlings of Japanese larch and hybrid larch allocated 6.5 and 18.0% more 14C, respectively, to the stem than NM seedlings. Of the total 14C detected in needle, stem and root fractions, FM and SM seedlings allocated a greater proportion than NM seedlings, perhaps because FM and SM seedlings had significantly (P < 0.05) higher photosynthetic rates than NM seedlings. As a result, FM and SM seedlings had greater dry masses than NM seedlings. Concentrations of nitrogen and phosphorus in FM and SM seedlings were significantly (P < 0.05) higher than in NM seedlings, as was stomatal conductance.     

Effect of collar insertion on soil respiration in a larch forest measured with a LI-6400 soil CO2 flux system

Little information is available on the effect of root cutting by the collar pre-insertion technique on soil respiration. In this study, we found that soil respiration rates decreased with increasing depth of collar insertion in both the with live roots intact and with live roots severed treatments, but the rate of decrease was substantially higher in the former. The cutting of roots, especially fine roots, may be responsible for this result.     

Ecological implications of mammal feces buried in snow through dung beetle activities

In early spring, snow-buried mammal feces simultaneously emerge on the ground with the melting of the snow across regions with heavy snowfall. Here, we evaluated the ecological implications of this phenomenon for the cool-temperate forest ecosystem in terms of the resource use of dung beetles (Coleoptera: Scarabaeidae), which play key roles in secondary seed dispersal and nutrient cycling. During May 2012 and 2013, we conducted cafeteria experiments in heavy snowfall regions with different types of forest cover in northern Japan by using pitfall traps baited with the snow-buried feces of Japanese macaques (Macaca fuscata), Japanese serows (Capricornis crispus), and Japanese hares (Lepus brachyurus angustidens). From the experiments, we identified 12 dung beetle species, indicating that the snow-buried feces could act as a valuable resource for vernal beetles. Most of the beetles had obvious fecal preferences. The snow-buried feces of serows were widely available resources for most vernal species, including dwellers and tunnelers, contributing to the biomass of those species. Although dung beetles using snow-buried feces in young broadleaf forests with rich feces supplies did not always exhibit the highest species richness, tunnelers frequently emerged in those forests, and Phelotrupes preferred macaque feces. This finding could have important implications for plant regeneration, as it relates to time-lagged mammal–beetle interactions. Endozoochorous seeds dispersed by macaques in the autumn have insufficient opportunities for secondary seed dispersal by beetles as the beetles are inactive in autumn, but those seeds are protected against predation by snow during winter and are safely preserved under soil by vernal tunnelers.     

Characteristics of the temperature coefficient, Q 10, for the respiration of non-photosynthetic organs and soils of forest ecosystems

The temperature coefficient, Q ₁₀ (fractional change in rate with a 10°C increase in temperature) describes the temperature sensitivity of soils, roots, and stems, as well as their possible performance in global warming processes. It is also a necessary parameter for the estimation of total CO₂ efflux from each element. A number of studies have focused on Q ₁₀ values to date; however, their conclusions are not universal and do not always agree. A review of these reported Q ₁₀ values therefore becomes necessary and important for a global understanding of the temperature sensitivity of different forest types and elements. The aims of our present paper are, first, to find the frequency distribution pattern of soils, roots, and stems (branches) and compare their temperature sensitivity; then, to find the Q ₁₀ differences between conifer and deciduous tree species and the effect of methodology on Q ₁₀ values; finally we want to give a perspective on future Q ₁₀-related studies. We found that most Q ₁₀ values of each element were concentrated in a relatively narrow range despite a total data distribution over quite a wide range. For soil respiration, the median Q ₁₀ value was 2.74 and the center of the frequency distribution was between 2.0 and 2.5 with a percentage of 23%. Most of the data (>80%) were within the range from 1.0 to 4.0. The median Q ₁₀ value for root respiration was 2.40 and the center of the frequency distribution was from 2.5 to 3.0 with a percentage of 33%. Most of the results (>80%) ranged from 1.0 to 3.0. For stem respiration, the median Q ₁₀ value was 1.91 and the frequency distribution was concentrated between 1.5 and 2.0. Over 90% of the data ranged from 1.0 to 3.0. Obvious differences in Q ₁₀ value were found between different elements, stem < root < soil including root < soil excluding root. The differences between woody organisms of stems, roots, and soils excluding roots were statistically significant (p<0.05), indicating that heterotrophic respiration from microorganism activity may be more sensitive to global warming. The duration of the period with leaves slightly affects the temperature sensitivity of woody organisms since the Q ₁₀ values for root and stem of coniferous evergreen trees did not differ significantly from deciduous trees (p>0.10). CO₂ analytical methods (soda lime absorption method, IRGA (Infra-read gas analysis), and chromatograph analysis) and root separation methods (excised root and trenched box) slightly affected the Q ₁₀ values of soil and root respiration (p>0.10), but an in vitro measurement of stem respiration yielded a significantly higher Q ₁₀ value than an in vivo method (p<0.05). In general, although the Q ₁₀ values of non-photosynthetic organisms stayed within a relatively conservative range, considerable variation between and within elements were still detectable. Accordingly, attention should be paid to the quantitative estimation of total CO₂ efflux by Q ₁₀-related models. In future studies, the biochemical factors and the environmental and biological factors controlling respiration should be emphasized for precise estimation of total CO₂ efflux. The difficulty is how to clarify the underlying mechanism for fluctuations of Q ₁₀ values for one specific habitat and element (e.g. temperature acclimation or adaptation of Q ₁₀ values) and then allow the Q ₁₀ values to be more conservative for representation of temperature sensitivity in global warming processes. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(4) [译自:植物生态学报, 2005, 29 (4)]     

Coordination of crown structure, leaf plasticity and carbon gain within the crowns of three winter-deciduous mature trees

We examined the vertical profiles of leaf characteristics within the crowns of two late-successional (Fagus crenata Blume and Fagus japonica Maxim.) and one early-successional tree species (Betula grossa Sieb. et Zucc.) in a Japanese forest. We also assessed the contributions of the leaves in each crown layer to whole-crown instantaneous carbon gain at midday. Carbon gain was estimated from the relationship between electron transport and photosynthetic rates. We hypothesized that more irradiance can penetrate into the middle of the crown if the upper crown layers have steep leaf inclination angles. We found that such a crown has a high whole-crown carbon gain, even if leaf traits do not change greatly with decreasing crown height. Leaf area indices (LAIs) of the two Fagus trees (5.26-5.52) were higher than the LAI of the B. grossa tree (4.50) and the leaves of the F. crenata tree were more concentrated in the top crown layers than were leaves of the other trees. Whole-crown carbon gain per unit ground area (micromol m(-2) ground s(-1)) at midday on fine days in summer was 16.3 for F. crenata, 11.0 for F. japonica, and 20.4 for B. grossa. In all study trees, leaf dry mass (LMA) and leaf nitrogen content (N) per unit area decreased with decreasing height in the crown, but leaf N per unit mass increased. Variations (plasticity) between the uppermost and lowermost crown layers in LMA, leaf N, the ratio of chlorophyll to N and the ratio of chlorophyll a to b were smaller for F. japonica and B. grossa than for F. crenata. The light extinction coefficients in the crowns were lower for the F. japonica and B. grossa trees than for the F. crenata tree. The leaf carbon isotope ratio (delta(13)C) was higher for F. japonica and B. grossa than for F. crenata, especially in the mid-crown. These results suggest that, in crowns with low leaf plasticity but steep leaf inclination angles, such as those of F. japonica and B. grossa trees, irradiance can penetrate into the middle of the crowns, thereby enhancing whole-crown carbon gain.     

Carbon dioxide exchange of larch (Larix gmelinii) cones during development

Larch (Larix gmelinii (Rupr.) Rupr.) cone scales are green, but little is known of their photosynthetic role in cone development or about how they differ in gas exchange characteristics from needle leaves. In contrast to leaf photosynthesis (Pleaf), we found that stomatal regulation of cone photosynthetic rate (Pcone) was marginal because the photosynthetic carbon came from internal recycling of respiratory carbon dioxide (CO2). Photosynthetic recycling of respired CO2 was confirmed by the finding that the intercellular CO2 concentration (Ci) in cone scales was much higher than ambient [CO2]; also, there was a positive correlation between Pcone and Ci, whereas Pleaf was almost constant as Ci varied. Low chlorophyll (Chl) concentration was a limiting factor for Pcone, but not for Pleaf, as indicated by the correlation between Pcone and chlorophyll concentration. Moreover, chlorophyll utilization efficiency (Psat/Chl a+b) for cone scales was lower than that for leaves. In both cones and leaves, nitrogen (N) was positively correlated with photosynthetic capacity (P), but the P/N value was much lower for cones than for leaves. For both organs, the ratio of respiration to N was broadly similar. Although mature cones have no photosynthetic capacity, Pcone of young cones was as high as 5.3 micromol m(-2) s(-1), about 1.26 times the value of Pleaf, and accounted for the refixation of 30-40% of the respiratory CO2 produced by cones, equivalent to the photosynthetic capacity of a bundle of short shoots near each cone. Thus, Pcone may be an important additional source of photosynthate for cones, given the weak assimilating capacity of leaves that are not fully expanded during cone development.     

Effects of prescription diet on dealing with stressful situations and performance of anxiety-related behaviors in privately owned anxious dogs

The objectives of this study were to evaluate the effects of a diet (CALM CANINE) supplemented with alpha-casozepine and L-tryptophan on the stress response of anxious dogs with a focus on anxiety-related behavioral parameters and physiological parameters. Forty-four privately owned dogs were first fed with the control diet, followed by the study diet, each for an 8-week period, with a transitional period of 1 week between the diets. After 7 weeks on a particular diet, owners reported their dogs' behavior by filling out a questionnaire. Using the Canine Behavioral Assessment and Research Questionnaire, the effect of the study diet was found to be significant for 4 anxiety-related behavioral parameters, possibly mirroring a placebo effect. After 7 weeks on each diet, the urine cortisol-to-creatinine ratio (UCCR) was measured to assess the stress response to a visit to a veterinary practice for toenail clipping. UCCR was measured to complement the interpretation of behavioral data with regard to the stress susceptibility of the dogs. A positive correlation was found between baseline UCCR, as measured in the home situations, and poststressor UCCR. The stressor-induced increase was significantly lower in the dogs when they were fed the study diet than earlier when they were fed the control diet. The supplementation of casozepine or the proportion of protein in a given diet could have been a determining factor. The study diet seems to improve the ability of an individual to cope with stress and may reduce anxiety-related behavior in anxious dogs.