Postnatal developmental changes in immunohistochemical localization of alpha-smooth muscle actin (SMA) and vimentin in bovine testes

The present study demonstrates the postnatal developmental changes in immunohistochemical localization of alpha-smooth muscle actin (SMA) and vimentin in the bovine testis. In the peritubular myoid cells of seminiferous tubules and the sub-epithelial and stromal cells of straight tubules and the rete testis, alpha-SMA starts appearing at around 4 months of age. Peritubular alpha-SMA attains the continuous mature pattern at around 5 months of age whereas sub-epithelial and stromal alpha-SMA increases with advancing age. Vimentin is localized in the perinuclear zone of Sertoli cells, peritubular and vascular wall cells, a few interstitial cells, and in the basal part of the epithelia of straight and rete tubules. Developmental changes are only evident in the Sertoli cell vimentin, which is basal and weak at birth and increases moderately until 4 months of age. From around 5 to 8 months of age when the Sertoli cells are under morphological transformation, vimentin intensity is considerably increased and the characteristic vimentin extensions connect the Sertoli nuclei to the basal membrane. These extensions get shorter at around 9 month of age as the Sertoli nuclei are positioned basally. The mature Sertoli cell perinuclear vimentin is strong and stable without infranuclear extension. In conclusion, the age of appearance of alpha-SMA coincides with the onset of postnatal division of spermatogonia, and vimentin may play a key role in stabilizing Sertoli cell nuclei during their transformation in bovine.     

Development of an SSR marker set for efficient selection for resistance to black spot disease in pear breeding

Black spot disease, which is caused by Alternaria alternata (Fries) Keissler Japanese pear pathotype, is one of the most harmful diseases in Japanese pear cultivation. Because of the potential harm of fungicides to consumers and the environment, resistant cultivars are desired. In this study, to enable efficient marker-assisted selection in pear breeding, we conducted comprehensive inoculation tests and genotyping with 207 pear cultivars. We identified a marker set (Mdo.chr11.27 and Mdo.chr11.34) suitable for selection for black spot resistance. In most susceptible cultivars, Mdo.chr11.27 amplified a 220-bp band and Mdo.chr11.34 amplified a 259-bp band. The genotype of Mdo.chr11.34 corresponds perfectly to the estimated genotype of Japanese pears susceptible to black spot disease. Using linkage analysis, we identified the positions of the gene for susceptibility to black spot disease in Chinese pear. Mdo.chr11.27 and Mdo.chr11.34 were tightly linked to susceptibility in Chinese pear, and the susceptibility gene was mapped at the top of linkage group 11, similar to that in Japanese pear. This marker set and the accumulation of phenotypic data will enable efficient marker-assisted breeding for black spot resistance in pear breeding.     

DNA profiling of pineapple cultivars in Japan discriminated by SSR markers

We developed 18 polymorphic simple sequence repeat (SSR) markers in pineapple (Ananas comosus) by using genomic libraries enriched for GA and CA motifs. The markers were used to genotype 31 pineapple accessions, including seven cultivars and 11 breeding lines from Okinawa Prefecture, 12 foreign accessions and one from a related species. These SSR loci were highly polymorphic: the 31 accessions contained three to seven alleles per locus, with an average of 4.1. The values of expected heterozygosity ranged from 0.09 to 0.76, with an average of 0.52. All 31 accessions could be successfully differentiated by the 18 SSR markers, with the exception of ‘N67-10’ and ‘Hawaiian Smooth Cayenne’. A single combination of three markers TsuAC004, TsuAC010 and TsuAC041, was enough to distinguish all accessions with one exception. A phenogram based on the SSR genotypes did not show any distinct groups, but it suggested that pineapples bred in Japan are genetically diversed. We reconfirmed the parentage of 14 pineapple accessions by comparing the SSR alleles at 17 SSR loci in each accession and its reported parents. The obtained information will contribute substantially to protecting plant breeders’ rights.     

Accumulation of Zinc and Copper in an Arable Field after Animal Manure Application

An experiment was conducted to examine the accumulation and mobility of heavy metals (Zn and Cu) at different depths in three types of arable soils (Brown Lowland soil, Andosol, and Brown Forest soil) amended with cattle and pig farmyard manures for 5 years. Nitric-perchloric acid digestion was performed for the determination of the total amounts of heavy metals, and 0.1 M hydrochloric acid extraction was performed for the determination of the amounts of soluble heavy metals. Results of the soil analysis indicated that pig farmyard manure application resulted in serious contamination of arable soils with Zn and potentially Cu. Especially, the Brown Forest soil displayed a high ability to accumulate heavy metals on the soil surface. Total-Zn concentration in surface soils was considerably affected by the holding capacity of soluble-Zn traction. Although the Andosol amended with pig farmyard manure showed higher concentrations of heavy metals related to the higher ability of retention on a weight basis, the soil did not contribute to high heavy metal accumulation because of its low bulk density. Heavy metals were easily leached in sandy soils such as Brown Lowland soil, and Cu was potentially stable compared with Zn. We suggest that long-term pig farmyard manure application to the Brown Lowland soil and Andosol with a light soil texture is associated with a higher risk of groundwater pollution than the application to the Brown Forest soil.     

Nitrate Decrease with Isotopic Fractionation In Riverside Sediment Column During Infiltration Experiment

This study determined isotopic fractionation of nitrate-nitrogen during denitrification in riparian aquifer sediments by sequential-column experiments at two different water flow rates. The purpose was to discuss the relationships of nitrogen isotopic enrichment factor of denitrification, denitrification rate, and infiltrating condition in sediment. Sandy sediment and groundwater were collected from floodplain in the middle reach of the Tama River where nitrogen isotopic enrichment factor for denitrification had been measured in a result of previous field observation. The denitrification rates and the isotopic enrichment factors of nitrate-nitrogen were estimated at 2.1 mgN dry-kg^?1 day^?1 and ?32.9‰ during the low water flow condition, and at 3.5 mgN dry-kg^?1 day^?1 and ?34.1‰ during the high water flow condition. The calculated water flow rates of the present column experiments were 300–400 times higher than that of the field observation. Because of the fast flow rate, nitrate was expected to continuously pass though active denitrifying sites in the small pore spaces of sediment, and nitrate supply to denitrifers by infiltration flow transport greatly exceeded the supply by diffusion. The observed denitrification rates was proportional to Peclet number as the ratio of solute nitrate transport to the diffusion, and apparent nitrogen isotopic enrichment factors during denitrification of our column experiments were lower than those obtained from the field observation. This study showed that denitrification rate and apparent nitrogen isotopic enrichment factor of denitrification in sediment depended on Peclet number.     

Real-time ultrasonographic evaluation of canine gastric motility in the postprandial state

Gastric motility is affected by several pathological conditions which may induce upper gastrointestinal clinical symptoms. The pathogenesis of canine gastric motility disorders is poorly understood because of methodological limitations. This study aimed at establishing a simple method for evaluating postprandial gastric motility in dogs. Gastric motility was ultrasonographically assessed in 7 healthy beagles using a technique previously described in humans. The motility index (MI), an indicator of gastric antral motility, was calculated by measuring the area of the gastric antrum in both a contracted and relaxed phase and by counting the number of contractions. The MI was measured every 30 min for 3 hr after feeding and compared with gastric emptying as assessed by a (13)C-octanoic acid breath test. The MI at 30 min had the lowest variability in the 7 dogs (mean SD, 9.77 ± 0.42; coefficient of variance, 4.25%), and a significant correlation was observed with gastric emptying coefficient (R(2)=0.8126, P=0.005) and half-emptying time (R(2)=0.654, P=0.027). When atropine was administered, a significant decrease in the MI at 30 min was observed compared with the control (9.77 ± 0.42 vs. 5.19 ± 0.22, P=0.0003). In conclusion, evaluation of the MI at 30 min is suitable for assessing gastric motility and enables us to assess gastric motility simply in a short time. By using this method, further studies for the pathogenesis of canine gastric motility disorders are warranted.     

Potato extract (Potein) suppresses food intake in rats through inhibition of luminal trypsin activity and direct stimulation of cholecystokinin secretion from enteroendocrine cells

Dietary proteins and trypsin inhibitors are known to stimulate the secretion of the satiety hormone cholecystokinin (CCK). A potato extract (Potein) contains 60% carbohydrate and 20% protein including trypsin inhibitory proteins. In this study, we examined whether Potein suppresses food intake in rats and whether it directly stimulates CCK secretion in enteroendocrine cells. In fasted rats, food consumption was measured up to 6 h after the oral administration of Potein or soybean trypsin inhibitor (SBTI). CCK-releasing activities of Potein and SBTI were examined in the murine CCK-producing cell line STC-1. Potein inhibited the trypsin activity in vitro with a potency 20-fold lower than that of SBTI. Oral administration of Potein dose-dependently suppressed food intake for 1-6 h. Potein, but not the SBTI, dose-dependently induced CCK secretion in STC-1 cells. These results suggest that Potein suppresses food intake through the CCK secretion induced by direct stimulation on enteroendocrine cells and through inhibition of luminal trypsin.