Anatomical and histological characterization of ileal and jejunal Peyer's patch in lesser mouse deer (Tragulus javanicus)

Tragulidae is a primitive ungulate family within the order Cetartiodactyla, suborder Ruminantia. Domestic ruminants such as cattle, sheep, and goat have two types of Peyer's patches (PP): jejunal and ileal PP, in which there are morphological and functional differences. In this study, lesser mouse deer (Tragulus javanicus) PP was studied by gross anatomical and histological procedures. At the fetal stage, both types of PP were formed in the small intestine. Ileal PP was observed as a single continuous aggregation of lymphatic follicles extending cranially from the ileo-caecal junction. However, jejunal PP was observed as multiple and discrete accumulations of lymphatic follicles. This study showed that the lesser mouse deer has two types of PP in the small intestine. In addition, the anatomical and histological characteristics of jejunal and ileal PP are quite similar to those of other ruminants' jejunal and ileal PP. Further studies are needed to analyze immune function of both PP in lesser mouse deer in order to determine the evolutionary process of Cetartiodactyla.     

A cytolethal distending toxin gene-based multiplex PCR assay for detection of Campylobacter spp. in stool specimens and comparison with culture method

In this study, we evaluated the applicability of cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR for the direct detection and identification of Campylobacter jejuni, C. coli and C. fetus from stool specimens of patients with gastroenteritis in comparison to culture methods. A total of 711 stool specimens were examined for the isolation or detection of campylobacters by using Skirrow's selective agar culture plates, a filtration method and the multiplex PCR assay. Forty-one and 36 C. jejuni strains were isolated by culture and filtration methods, respectively. In addition, 2 and 3 C. coli strains were isolated by Skirrow and the filtration methods, respectively. However, when the multiplex PCR was employed, the cdtB genes of C. jejuni and C. coli were detected in 45 and 4 stool samples, respectively, and 9 C. jejuni PCR-positive samples by multiplex PCR were negative by culture method. Sequence analysis of the PCR products obtained from 8 stool specimens from which campylobacters were not isolated by culture method but the sequences exactly matched with that of the cdtB gene of C. jejuni strain 81-176. None of the remaining stool samples which were culture negative for campylobacters produced any amplicon. Stool samples were defined as Campylobacter-positive if detected by any method. The sensitivity of the multiplex PCR was 83%, which was higher than Skirrow (74%) and filtration method (66%). These data indicate that cdtB gene-based multiplex PCR is a rapid and more sensitive method to identify the most important species of Campylobacter for human diseases. (248).     

Comparative studies on the distribution and population of immunocompetent cells in bovine hemal node, lymph node and spleen

The distribution and population of immunocompetent cells in bovine hemal node, mesenteric lymph node and spleen were analyzed comparatively by immunohistochemistry and flow cytometry. Many CD8(+) cells, CD172a(+) cells and γδ T cells were found in the lymphatic cord along the sinus of the hemal node and the splenic red pulp. A few CD8(+) cells and γδ T cells were distributed diffusely in the paracortex and medullary cord of the mesenteric lymph node. Many germinal centers were recognized in the lymphatic regions such as the cortex and white pulp of these lymphoid organs. The populations of CD8(+) cells and γδ T cells in the hemal node and the spleen were higher than those of the mesenteric lymph node. In addition, the populations of CD21(+) cells and MHC class II(+) cells in the hemal node and the mesenteric lymph node were higher than those of the spleen. The results suggest that the hemal node has an important role in both cellular and humoral immunity as well as the lymph node and the spleen in cattle.     

Bilateral segmental aplasia with unilateral uterine horn torsion in a pomeranian bitch

Bilateral segmental aplasia of the uterine horns with unilateral pyometra and uterine horn torsion were diagnosed in a Pomeranian bitch that presented with chronic abdominal distension and an acute onset of anorexia and lethargy. Because radiographic and ultrasonographic findings revealed the presence of markedly enlarged bilateral uterine horns filled with fluid in the caudal abdomen, a tentative diagnosis of either pyometra or hydrometra with uterine horn torsion was made. Exploratory laparotomy showed bilateral, segmentally distended uterine horns with unilateral uterine horn torsion. Ovariohysterectomy was performed, and bilateral segmental aplasia of the uterine horns with the development of unilateral uterine horn torsion was diagnosed histopathologically. To the authors' knowledge, this is the first report of uterine horn torsion in conjunction with segmental aplasia of the uterine horn in a bitch.     

Relation of reproductive performances and rectal palpation for luteum function of heifers 7 days after estrus

Diagnosis of corpus luteum (CL) function by rectal palpation (RP) has been widely used for recipient selection of embryo transfer (ET), a technology essential for genetic improvements in cattle. To examine the accuracy of RP diagnosis method, the relationship between RP‐based CL function and reproductive performance was compared in this study. In Experiment 1, CL of Holstein heifers on day 7 after estrus was classified into functional or hypoplastic by RP, and the results were compared with ultrasonographic (US) images and plasma progesterone (P4) levels. As a result, heifers with functional CL judged by RP had a mean maximum CL diameter of 20.1 ± 3.1 mm on US and a mean P4 concentration of 8.1 ± 2.3 ng/mL. These values were significantly greater than those of heifers with hypoplastic CL (12.4 ± 5.4 mm, 4.0 ± 2.8 ng/mL) (P < 0.001). In Experiment 2, the length of the estrus cycle was examined between functional CL and hypoplastic CL. The rate of heifers with a normal estrus cycle length with 18–25 days was significantly lower with hypoplastic CL than with functional CL (16/24 vs. 43/46, P < 0.01). In Experiment 3, 543 inseminated heifers were similarly classified by CL function by RP 7 days after estrus. The heifers with functional CL showed higher pregnancy rate compared with the heifers with hypoplastic CL (75.2 vs. 47.9%, P < 0.0001). Finally, the CL function of 66 heifers was examined by RP on day 7 post‐estrus, and ET was performed in 49 (74.2%) heifers with functional CL. As a result, 27 (55.1%) of them became pregnant. Taken together, these results reconfirm that RP on day 7 after estrus is useful for selection of heifers with functional CL.     

Effects of transplantation sites on tumour growth,pulmonary metastasis and ezrin expression of canine osteosarcoma cell lines in nude mice

To determine the influence of the transplantation site of canine osteosarcoma (OS) cell lines on tumour growth and pulmonary metastasis, three OS cell lines were transplanted into nude mice via subcutaneous (SC), intratibial (IT) or intravenous (IV) injection. IT‐xenografts exhibited greater potential for developing primary masses and pulmonary metastasis than SC‐xenografts. In IT and IV xenografts, lung micrometastases along with phosphorylated ezrin–radixin–moesin (p‐ERM) overexpression were found in mice xenografted with HMPOS and OOS cells after 1 week and metastasis was found with decreased p‐ERM expression at later time points. The expression of ezrin and p‐ERM in the primary tumours of IT‐xenografted mice was higher than those in SC‐xenografted mice with HMPOS and OOS cells. The results suggest that the orthotopic transplantation site plays an important role in the spontaneous metastasis of canine OS and that ezrin phosphorylation may be involved in the early metastatic mechanism of canine OS cells.     

Effects of thinning intensity on species diversity and timber production in a conifer (Cryptomeria japonica) plantation in Japan

In conifer plantations, enhancement of species diversity has become an important management goal. Although thinning is a useful method to enhance diversity, determining optimum thinning intensities may be rather complicated because of potential trade-offs among a broad array of management goals (e.g., recovery of biodiversity, increasing individual tree sizes, increasing net primary production, saving management costs). To evaluate the optimum thinning intensity by analyzing these relationships, we conducted a thinning experiment with three different thinning intensities—unthinned, 33% thinned, and 67% thinned—in a Cryptomeria japonica plantation in 2003, and investigated the number, diameter at breast height (DBH), and diversity of hardwoods (height?>?1.5?m) in 2008, and the growth of conifers over five?years. In hardwoods, the number of individuals, number of species, mean DBH, and total basal area were greatest in the 67% thinned treatment, irrespective of successional status. However, Shannon’s diversity index did not differ among the three treatments due to a disproportionate increase with thinning intensity in the abundance of a mid-successional species, Cornus controversa. Diameter growth of conifers was also highest in the 67% thinned and lowest in the unthinned treatment, whereas the reverse was true for stand volume increment. These results suggest that intensive thinning is a reliable method to convert conifer plantations into conifer–hardwood mixed forests at canopy level much more quickly and consistently than weak thinning, although primary production is to some extent reduced. If forest managers prefer sustainable timber production of conifers rather than full recovery of diversity, weak thinning may be suitable.     

Risk assessment of ozone impact on <Emphasis Type="Italic">Fagus crenata</Emphasis> in Japan: consideration of atmospheric nitrogen deposition

Tropospheric ozone (O₃) is considered to be the air pollutant relating to the decline of Fagus crenata forest in Japan. In the present study, we assessed a risk of O₃ impact on the growth of F. crenata in Japan, giving consideration to the effects associated with atmospheric nitrogen (N) deposition based on the experimental study, national monitoring data for oxidant concentration and atmospheric N deposition, and a national vegetation survey. The average and maximum O₃-induced relative growth reduction (RG_red) of F. crenata across Japan were estimated to be 3.2 and 9.7%, respectively. Current levels of atmospheric N deposition were found to significantly affect the sensitivity of F. crenata to O₃. When the N deposition was assumed as zero, the estimated average and maximum RG_red were 2.3% and 5.7%, respectively. The inclusion of atmospheric N deposition data thus increased the estimated values for average and maximum RG_red (by 38% and 71%, respectively). Our results demonstrate that a change in the sensitivity to O₃ associated with atmospheric N deposition is an important consideration in the risk assessment of O₃ impact on the growth of F. crenata in Japan.     

Detection and verification of QTLs associated with heat-induced quality decline of rice (Oryza sativa L.) using recombinant inbred lines and near-isogenic lines

Decline in the apparent quality of rice (Oryza sativa L.) grain due to high temperatures during ripening recently became a major concern in many areas in Japan. The occurrence of white-back kernels (WBK) is one of the main problems of heat-induced quality decline. We identified QTLs associated with the occurrence of WBK using recombinant inbred lines (RILs) and verified their effects using near-isogenic lines (NILs). The QTL analysis used F₇ and F₈ RILs derived from ‘Hana-echizen’ (HE), which is tolerant to high temperature, × ‘Niigata-wase’ (NW), which is sensitive to high temperature. Four QTLs were identified on chromosomes 3, 4, 6, and 9 (qWB3, qWB4, qWB6 and qWB9). To verify the effects of qWB6 and qWB9, we developed two NILs in which qWB6 or both were introduced from HE into the NW background. The HE allele at qWB6 significantly decreased WBK under multiple environments. The combination of qWB6 and qWB9 in an F₂ population derived from a cross between a NIL and NW showed that the NW allele at qWB9 significantly decreased WBK if the qWB6 allele was HE. These results will be of value in marker-assisted selection for the breeding of rice with tolerance to heat-induced quality decline.     

Effect of various drought stresses and subsequent recovery on proline,total soluble sugar and starch metabolisms in Rice (Oryza sativa L.) varieties

ABSTRACT

Rice (Oryza sativa L.) is one of the most important staple foods in the world, however most improved rice varieties are susceptible to drought stress. A two-year study was conducted to explore the effects of various drought stresses and subsequent recovery on the accumulation and degradation of proline, total soluble sugar and starch in different rice varieties at vegetative stage. The results showed that relative water content in the leaves and sheaths of rice varieties significantly decreased under drought stresses, but not at the same rate. Under control and drought conditions, the water content in sheaths was higher than that in leaves. Interestingly, under severe drought stress in 2015, the leaf water content was higher than the sheath water content. The water distribution between leaves and sheaths might be a response of plants to protect leaf system from devastation by drought. Proline was highly accumulated under drought stress but rapidly decreased after re-watering. The drought tolerant variety DA8 expressed higher ability in accumulation of proline than susceptible varieties. In general, total soluble sugar and starch contents in leaves and sheaths of varieties decreased under drought stress conditions. Total soluble sugar and starch content of DA8 were less affected than other varieties under drought conditions. Our study indicated that metabolisms of total soluble sugar and starch in rice were affected by both environmental conditions and characteristics of varieties. Proline accumulation ability of varieties can be used as a useful indicator for drought tolerant potential in rice breeding for water-limited environments.