Plant species identification using fecal DNAs from red-eared slider and Reeves’ pond turtle in agricultural canals for rural ecosystem conservation

Fecal DNA samples from the red-eared slider and Reeves’ pond turtle, suspected pests of lotus root paddies, were used to identify the plant species eaten by these turtles in order to develop a strategy for rural ecosystem conservation. The fecal samples were obtained from young and adult individuals (mostly female) of both species living in agricultural canals surrounding lotus root paddies in Tokushima Prefecture, Japan. The samples were screened for the presence or absence of DNA from nine plant species using PCR and plant species-specific primers for the rbcL gene of chloroplast DNA. In the red-eared slider, our analysis identified seven plant species in the fecal DNA samples of adults and three plant species in those of young individuals. In Reeves’ pond turtle, our analysis identified two plant species from adult fecal samples and one species from those of young individuals. Thus, adult red-eared sliders consume a greater range of plants than young red-eared sliders or Reeves’ pond turtles. Both turtle species, independently of age, consumed lotus plants and were likely to cause feeding damage to lotus roots. Considering the plant species detected in adult red-eared sliders and these plant habitats, we suggest that this adult turtle is likely to travel between the agricultural canals and the lotus root paddies. These findings will help the development of strategies for preventing damage to lotus roots by these turtles; furthermore, they indicate that fecal DNA analysis will be applicable to investigation of the feeding habits of other animal species.     

Variable response of growth and arbuscular mycorrhizal colonization of maize plants to preceding crops in various types of soils

The effects of the preceding crops, sunflower (mycorrhizal host) and mustard (nonhost), on arbuscular mycorrhizal (AM) colonization and growth of succeeding maize were examined in 17 soils in an attempt to clarify the influence of soil characteristics on the effects of preceding crops. Shoot weight and P uptake of maize planted after sunflower were much higher than those after mustard in 14 soils, although the preceding crop had little effect on soil-P availability. AM colonization of maize after sunflower was much higher than that after mustard. The effect of the preceding crop was eliminated by soil sterilization. These results suggested that the differences in maize growth were caused by differences in the AM colonization. Correlation analysis of the effect of the preceding crop and soil properties showed that the difference in the effects on maize growth could not be explained by soil chemical properties, but only by the AM colonization of the preceding sunflower crop. In one of the 17 soils, however, the effect was not evident despite the higher AM colonization of sunflower. This soil was sterilized, and the effect of inoculation by AM fungi (AMF) on maize was examined. However, it was found that the inoculation increased AM colonization but did not improve maize growth at any P level, suggesting that the effect of AMF was unusually inhibited in this soil by unknown soil physicochemical properties. In most soils, however, the preceding mycorrhizal host crop, sunflower, improved the growth and AM colonization of maize depending on the AM colonization of sunflower.     

Age-related changes of genital systems in the female Crj:CD (SD) IGS rats during sexual maturation

The age-related changes of vaginal opening, body weight, the weights of the uterus and ovary, together with histological examination, serum 17beta-estradiol (E2) and progesterone levels were examined in intact female Crj:CD (SD) IGS rats between 21 and 36 days of age to understand the basic biological profile of changes of the female genital system during sexual maturation in the rat for female pubertal assays. With the beginning of the elevation of serum E2 level from 28 days of age, all parameters except body weight started to show drastic change until 31 days of age. The highest incidence of vaginal opening was recorded at 34 days of age. On macroscopic examinations, a number of rats showed uterine imbibition but vaginal opening. Immediately after the confirmation of the vaginal opening, the genital systems of three rats were observed microscopically. Both ovaries already had multiple corpora lutea, and degeneration of endometrial epithelial cells was observed. In conclusion, we obtained essential data on genital tract development of female Crj:CD (SD) IGS rats for in vivo screening assays that will contribute to detect potential endocrine active chemicals. In addition, it is assumed that the first ovulation precedes or occurs simultaneously with vaginal opening.     

Serum activities of tartrate-resistant acid phosphatase and bone specific alkaline phosphatase as indices of bone metabolism in the cow

The correlation between the serum hydroxyproline concentration and serum activity levels of TRAP and BALP was examined in 41 cows. The correlated coefficient (r) was 0.6391 for TRAP and 0.3147 for BALP, respectively. Judging from the significant correlation to the serum hydroxyproline concentration, serum TRAP activity is an index for bone metabolism in cows. Serum TRAP activity was therefore measured in 205 healthy cows (2-9 years old) in order to observe the changes in bone resorption with aging and milk production. TRAP levels differed slightly between group A (< or =4 yrs) and B (5 yrs< or =) at the same stage of lactation. The activity levels rose slightly at the height of lactation stage and during the dry stage.     

Temporal and spatial dynamics of an old-growth beech forest in western Japan

Dendrochronological approaches enable us to understand forest stand dynamics by estimation of disturbance history and age structure. The present study was conducted in an old-growth beech forest in a forest reserve in western Japan. Increment cores were taken for tree ring analysis from all canopy trees in a 50 m × 130 m study plot. Radial growth release criteria were developed to identify significant growth releases in each tree ring series and to characterize the disturbance history of the study site. The age structure of the forest was indicative of continuous establishment by Fagus crenata and simultaneous establishment by Magnolia obovata. A variety of low-intensity disturbances were identified in each decade, especially after the 1900s, but the occurrence of high-intensity catastrophic disturbance was rare, and likely played an important role in maintaining species diversity in the existing forest canopy. The results also suggest that F. crenata regenerates gradually before and after both large- and small-scale disturbances, whereas M. obovata and Betula grossa regenerate only after large-scale catastrophic disturbances.     

Leaf water relations in <Emphasis Type="Italic">Pinus densiflora</Emphasis> Sieb. et Zucc. on different soil moisture conditions

To elucidate the differences in the leaf water relations of Pinus densiflora Sieb. et Zucc. growing in different soil moisture conditions, we examined the pressure-volume curve and the diurnal changes in the stomatal conductance, the transpiration rate, and the leaf water potential. The leaf water relations were compared using field-grown 40-year-old pine trees growing on the upper and lower parts of a slope. We also compared the leaf water relations of potted 4-year-old saplings growing at pF 4.2 and pF 1.8 soil moisture levels for almost 1 year. The values of the ratio of symplasmic water at turgor loss point to symplasmic water at saturated point (V_p/V_o) and bulk modulus of elasticity () of both the adult trees on the upper part of the slope and the potted saplings growing on pF 4.2 soil moisture were higher than those values of both the adult trees on the lower part of the slope and the potted saplings growing on pF 1.8 soil moisture, respectively. The field-grown adult tree and the potted saplings growing under long-term water stress tended to reduce their stomatal conductance in response to the acute soil drying. It is suggested that P. densiflora growing under long-term water stress rapidly closed its stomata in response to soil drying and avoided losing water, and could also rapidly absorb water with reducing water loss because of the decrease in the leaf pressure potential derived from the high values.     

Comparison of stand dynamics after dieback caused by pine wilt disease among pine forests with different management regimes in western Japan

In pine forests damaged by pine wilt disease, in western Japan, the effect of protection regimes of pine trees on the stand dynamics were examined in the following four stands: (1) lightly damaged stand (age 30–40 years) with no procedure in operation for protecting pine trees; (2) severely damaged stand (age 30–40 years) with no procedure in place for protecting pine trees; (3) severely damaged stand (age 50 years) with a selective cutting of infected trees; (4) severely damaged stand (age 30–40 years) with a selective cutting of infected trees. All the stands had been abandoned before the pine wilt disease damage. The understory structure of the severely damaged stand with no protection procedure was similar to that of the lightly damaged stand. Frequent invasion by tree species and acceleration in the growth of understory trees occurred after the dieback in the selective cutting stand. These results suggest that a deficiency in the canopy layer caused by the dieback resulted in low disturbance intensity in the early stages after the dieback, but the selective cutting increased the intensity by the reduction in the understory as well as the canopy layer. The intensity of the disturbance in the selective cutting stands was larger in the younger stand because it had a higher density of selectively cut pine trees. The different stand structure of pine forests occurred after the dieback because the intensity of the disturbance varied as a result of the selective cutting operation and the stand age.     

Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for Fertilizing Oocytes

Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic reasons.     

Individual-level analysis of damage to residual trees after single-tree selection harvesting in northern Japanese mixedwood stands

Quantifying tree damage and mortality caused by single-tree selection harvesting is critical to understanding postharvest forest dynamics and management. In this study, we quantified the effects of tree size and species and the distance from residual trees to felled trees and skid trails on damage to residual trees and mortality in mixed coniferous–broadleaved stands of Hokkaido, northern Japan. Among the 4,961 trees that we studied, 373 (7.5 %) were damaged, and 148 of these trees (3.0 %) died during or immediately after logging. Hierarchical Bayesian modeling showed that the risk of damage to residual trees increased with increasing size of the felled trees and with increasing proximity to felled trees and skid trails. Smaller residual trees had the greatest risk of damage. Species differed in their susceptibility to damage; Abies sachalinensis (Fr. Schm.) Masters and Picea jezoensis (Sieb. et Zucc.) Carr. were the most susceptible species in our sample plots. Smaller damaged trees had the highest risk of mortality. The damaged trees that did not die at the time of logging had a higher risk of postharvest mortality than undamaged trees. Our results indicate that, to minimize logging-induced damage and mortality: (1) the spatial arrangement of skid trails should be optimized and fixed, (2) the risk of skidding damage should be considered before marking the trees to be harvested and choosing the felling direction, and (3) logging operators should be instructed to avoid damaging small trees, especially those of the species that are most susceptible to damage.     

Effects of cortisol and angiotensin II on the number and size of juxtaglomerular cells in masu salmon, Oncorhynchus masou

The authors previously reported that the number and size of juxtaglomerular cells (JGCs) in the kidney increased during smoltification in masu salmon, Oncorhynchus masou. In the present study, the effects of cortisol and/or angiotensin (Ang) II ([Asn¹, Val⁵]-Ang II) on the JGC number and size in masu salmon were examined to elucidate hormonal regulation of the changes in the JGC number and size during smoltification. These hormones were injected intraperitoneally every 2 days for a total of 6 injections. There was a significant increase in the JGC number and size with time following the start of the experiment in cortisol- and cortisol + Ang II-treated groups and no significant change in control and Ang II-treated groups. On both days 5 and 11, the JGC number and size in the cortisol-treated group were significantly large compared to those of control and Ang II-treated groups, respectively. The JGC number and size in the cortisol + Ang II-treated group were significantly large compared to those of control on both days 5 and 11, and those of the Ang II-treated group only on day 11, respectively. On the other hand, there was no significant difference in the JGC number and size between the Ang II-treated and control groups and between the cortisol- and cortisol + Ang II-treated groups during the experiment, respectively. The means of the JGC number and size in cortisol-treated group on day 11 were close to those previously reported in smolt. These results suggest that cortisol induces an increase in JGC number and size during smoltification in masu salmon. This revised version was published online in August 2006 with corrections to the Cover Date.