Quantitative mapping of kelp forests (Laminaria spp.) before and after harvest in coastal waters of the Shiretoko Peninsula,Hokkaido, Japan

In the Shiretoko Peninsula, a World Natural Heritage site, the sustainable management of kelp forests has drawn public attention because of the economic and ecological importance of kelp. We spatially estimated the distributions of kelp forests in the Shiretoko Peninsula before and after harvest. Field surveys were conducted in coastal waters (23.74 km²) at the ends of July and August 2008, immediately before and after harvest. Data on the presence or absence and thickness of the kelp forests were collected via acoustic observation. The data were interpolated using geostatistical methods. Before harvest, the kelp forests were continuously distributed over 5.64 km² (thickness 33–132 cm), especially near the north part of the study area. After harvest, they were sparsely distributed over 2.73 km² (thickness 35–105 cm). In the southern part of the study area, the influence of harvests was observed as declines in forest area. In addition, relatively thickly forested areas formed the majority of the part most likely to be harvested. Selective harvesting for area and size was confirmed though quantitative mapping of kelp forests. The quantitative mapping of both the distribution and harvest of kelp forests was successful.     

Development of microsatellite markers in cultivated and wild species of sections <Emphasis Type="Italic">Cepa</Emphasis> and <Emphasis Type="Italic">Phyllodolon</Emphasis> in <Emphasis Type="Italic">Allium</Emphasis>

The potential of microsatellite markers for use in genetic studies has been evaluated in Allium cultivated species (Allium cepa, A. fistulosum) and its allied species (A. altaicum, A. galanthum, A. roylei, A. vavilovii). A total of 77 polymerase chain reaction (PCR) primer pairs were employed, 76 of which amplified a single product or several products in either of the species. The 29 AMS primer pairs derived from A. cepa and 46 microsatellites primer pairs from A. fistulosum revealed a lot of polymorphic amplicons between seven Allium species. Some of the microsatellite markers were effective not only for identifying an intraspecific F₁ hybrid between shallot and bulb onion but also for applying to segregation analyses in its F₂ population. All of the microsatellite markers can be used for interspecific taxonomic analyses among two cultivated and four wild species of sections Cepa and Phyllodolon in Allium. Generally, our data support the results obtained from recently performed analyses using molecular and morphological markers. However, the phylogeny of A. roylei, a threatened species with several favorable genes, was still ambiguous due to its different positions in each dendrogram generated from the two primer sets originated from A. cepa and A. fistulosum.     

Comparison of soil depths between evergreen and deciduous forests as a determinant of their distribution, Northeast Thailand

In several areas in Northeast Thailand, evergreen and deciduous forests coexist under uniform terrain and climatic conditions. We compared depth and physical properties of soils between evergreen and deciduous forests in the Sakaerat Environmental Research Station to clarify what factor determines their distribution. The averaged soil depths were 79 ± 27 cm and 135 ± 58 cm in the deciduous and evergreen forests, respectively. The soils in the deciduous forests were relatively coarser in soil texture than those in the evergreen forests, particularly in the surface layers. The average available water capacity of the solum was lower in the deciduous forest soils (78 mm) than in the evergreen forest soils (123 mm). Compared with the evapotranspiration from the evergreen forest in the study area, the available water capacity of the evergreen forest soil was almost the same as the water deficit during the dry season (November–February), while that of the deciduous forest soil was lower and insufficient to maintain the evapotranspiration. These results suggest that the distribution of deciduous and evergreen forests in the study area was associated with soil water availability, which mainly depends on soil depth.     

Steroid injection therapy in a feline solitary bone cyst

A nine months old Japanese domestic cat with a solitary bone cyst, which is a benign tumor-like lesion and is particularly uncommon in feline practice was referred. Radiographic examination revealed an expansive radiolucency in the distal metaphysis of the right ulna and pathologic fracture. The histological examination demonstrated immature osteogenesis consisting of osteoblasts surrounded by connective tissue. We applied drainage and instillation of steroid solution into the cystic cavity. Clinical signs, including lameness and pain, disappeared three weeks after the therapy started. Fourteen months after the therapy, the cystic lesion of bone was remodeling successfully without re-developing. We conclude that our procedure was useful treatment in this case.     

Plasma concentration of estrone sulfate during pregnancy in different breeds of Japanese beef cattle

Plasma concentrations of estrone sulfate in different breeds of Japanese beef cattle and the relationship between those concentrations and feto-placental growth were examined in order to assess the possibility of monitoring abnormal growth of the fetus. Blood samples were obtained from cows from day 90 of gestation to parturition. The plasma concentration of estrone sulfate was measured by direct enzyme immunoassay. From day 180 of gestation, the mean concentration of estrone sulfate increased gradually and it was drastically elevated after day 240 of gestation with the maximum at day 285. Plasma concentrations of estrone sulfate on day 240 of gestation was significantly increased in F(1) cows (Holstein Friesian and Japanese Black) compared with those in other breeds of cow. From day 270 to 278 of gestation, estrone sulfate concentrations of Holstein Friesian cows inseminated by Holstein Friesian differed from those inseminated by Japanese Black. In the cow with retained placenta, the plasma concentration of estrone sulfate reached plateau at day 240 of gestation and did not increase thereafter. There was no significant relationship between estrone sulfate concentration and duration of gestation, calf birth weight, weight of placenta or viability of newborn calves. These results indicate that changes of plasma estrone sulfate concentration in Japanese beef cattle are very similar to those in Holstein dairy cattle. They also suggest that the plasma concentration of estrone sulfate is associated with the breed of pregnant cow and that its concentration is also affected by calf birth weight depending on the breed of bull. It seems possible to predict the incidence of retained placenta but not the calf birth weight and viability of newborn calves in Japanese beef cattle.     

Three-dimensional image analysis of a head of the giant panda by the cone-beam type CT

The cone-beam type CT (Computed Tomography) enabled us to collect the three-dimensional (3D) digitalized data directly from the animal carcass. In this study, we applied the techniques of the cone-beam type CT for a carcass head of the giant panda (Ailuropoda melanoleuca) to obtain the 3D images easily without reconstruction process, and could morphologically examine the sections from the 3D data by means of non-destructive observations. The important results of the study represent the two following points. 1) We could show the morphological relationships between the muscles of mastication and the mandible in non-destructive status from the 3D data. The exact position of the coronoid process could be recognized in the rostro-lateral space of the temporal fossa. 2) By the serial sections from the 3D data sets, the morphological characteristics in the nasal cavity were detailed with high resolution in this rare species. The nasal concha was well-developed in the nasal cavity. The ethmoidal labyrinth was encountered immediately caudal to the nasal cavity and close to the region of the olfactory bulb. The ethmoidal labyrinth consisted of the complicated osseous structure in this area. The data will be useful to discuss the olfactory function in the reproduction behavior of this species.     

A new preparation protocol for measurement of testicular sperm production

In studies of male reproductive toxicity, measuring daily sperm production is a quite important criterion. However, the accuracy of the values measured by the basic protocol is still controversial. In order to enhance the homogeneity of countable testicular sperm/spermatid heads, this report introduces a new enzymatic method with a subsequent detergent treatment. The testis of rat was firstly homogenized in phosphate-buffered saline. The homogenate (buffer mix) was then treated with collagenase and trypsin, and then sodium dodecyl sulfate (SDS) was added to produce detergent-resistant sperm/spermatid heads (detergent mix). After examination by hemocytometer, the coefficient of variation (CV) of the number of sperm/spermatid heads was compared with that obtained from the buffer mix. In addition, a MicroCell chamber was applied to the examination, and the CV was compared with other cases. In both examinations, homogeneity was improved by the detergent mix preparation. Counting with the hemocytometer showed an increased number of sperm/spermatid heads compared with that of the buffer mix (P<0.001), and the CV was decreased (P<0.05). In addition, when the MicroCell chamber was applied, the numbers increased about 3-hold compared with that of the buffer mix (P<0.001). The CV of the detergent mix was 23.7%, while that of the buffer mix was 38.9%. These results clearly demonstrate that the new preparation protocol generated in this study can provide more actual and accurate values when measuring daily sperm production.     

Effects of colostrum whey on immune function in the digestive tract of goats

The objective of the present study is to examine whether colostrum whey can have an effect on immune function in goats digestive tract. Two milliliters of colostrum whey (colostrum group) or water (control group) were administrated orally to goats every day for 3 weeks. Blood was collected twice a week for 3 weeks to measure immunoglobulin A (IgA), interleukin 8 (IL‐8), and IL‐10. At the end of the experimental period, the parotid glands, oral mucosa, lingua, esophagus, jejunum, and ileum were collected for immunohistochemical detection of IgA, cathelicidin‐7, and S100A8. The ratio of the length of IgA‐positive mucosal surface in the esophagus to the total esophageal length was significantly greater in the colostrum group than in the control group. The number of IgA‐positive cells in the labial gland and ileum in the colostrum group was significantly higher than that in the control group. There were no significant differences between the colostrum and control groups in the number of cathelicidin‐7‐positive cells in the jejunum and ileum and in the number of S100A8‐positive cells in the lingua, jejunum, and ileum. These results suggest that colostrum stimulates the recruitment of plasma cells into the labial gland, which then secrete more IgA into the saliva.     

Deficient proliferation and apoptosis in the granulosa and theca interna cells of the bovine cystic follicle

The aim of the present study was to examine the frequencies of cell proliferation and death of granulosa and theca interna layers during development of cystic follicles in order to understand the mechanisms of cystic follicle formation. Paraffin sections of cystic follicles were immunostained with antibodies against proliferating cell nuclear antigen (PCNA) and cleaved caspase-3 in order to observe proliferating and apoptotic cells, respectively. The concentrations of estradiol-17beta and progesterone in the follicular fluid of these follicles were measured by ELISA. The granulosa and theca interna layers contained both PCNA- and caspase-3-positive cells, although their numbers were limited. There was significant negative correlation between the estradiol-17beta and progesterone concentrations in the follicular fluid. Regression analysis revealed no significant correlation, except for that between the PCNA-positive cells in the theca interna and the caspase-3-positive cells in the granulosa layer. These results indicate that the granulosa and theca interna cells of the cystic follicle show weak proliferative activity and low apoptotic frequency; this implies that the cystic follicle grows slowly and then maintains a static condition without degeneration, which leads to long-term persistence of the follicle.