Comparison of cell populations derived from canine olfactory bulb and olfactory mucosal cultures

OBJECTIVE: To evaluate the numbers and proportions of olfactory ensheathing cells (OECs) in cell cultures derived from the olfactory bulb (OB) and olfactory mucosa of dogs. ANIMALS: 7 dogs. PROCEDURES: OB tissue and olfactory mucosa from the nasal cavity and frontal sinus were obtained from euthanatized dogs and prepared for cell culture. At 7, 14, and 21 days of culture in vitro, numbers and proportions of OECs, astrocytes, and fibroblasts were determined via immunocytochemistry. Antibody against the low-affinity nerve growth factor receptor p75 was used to identify OECs, antibody against glial fibrillary acidic protein was used to identify astrocytes, and antibody against fibronectin was used to identify fibroblasts. RESULTS: Cultured OECs derived from the olfactory mucosa of the nasal cavity and frontal sinus had similar characteristics. However, whereas OECs in the OB cell cultures constituted approximately 50% of the cells at 7 days and approximately 75% at 21 days the proportion of OECs in cultures derived from both mucosal types was much lower, with approximately 40% OECs at 7 days and approximately 25% at 21 days. Analysis of OEC numbers revealed that these changes were accompanied by corresponding decreases and increases in the population of cells with fibronectin receptors. CONCLUSIONS AND CLINICAL RELEVANCE: Although olfactory mucosal cell cultures yielded a sufficient number of OECs for spinal cord transplantation procedures in dogs, modification of culture conditions would be required to ensure that the derived cell population contained a sufficient proportion of OECs.     

Anthocyanin composition and antioxidant activity of the Crowberry (Empetrum nigrum) and other berries

The anthocyanin composition and antioxidant activity of the crowberry ( Empetrum nigrum) were studied. High-performance liquid chromatography (HPLC) combined with a diode array detector and electrospray ionization mass spectrometry were used for identification and quantification of individual anthocyanins. Freeze-dried crowberry powder was extracted with 80% methanol containing 0.5% acetic acid and subjected to HPLC. Thirteen kinds of anthocyanins were identified. The major anthocyanins were cyanidin-3-galactoside and delphinidin-3-galactoside, at 8.04 and 8.62 mg/g extract, respectively. The HPLC profile of crowberry extract was similar to bilberry and blueberry. The total content of anthocyanins in crowberry was 41.8 mg/g extract, higher than the other nine major berry species (2.5-38.8 mg/g extract). The antioxidant activity was also evaluated using the 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis(3-ethybenzothiazoline-6-sulfonic acid) radical quenching assays and the ferric reducing activity power assy. Crowberry extract exerted the strongest antioxidant activity. In conclusion, individual anthocyanins in crowberry were identified and then quantified in this study. Additionally, crowberry is suggested to be associated with a reduction in the risk of developing chronic diseases because of its strong antioxidant activity.     

Validation and standardization of virus neutralizing test using indirect immunoperoxidase technique for the quantification of antibodies to rabies virus

A virus neutralizing test using an indirect immunoperoxidase technique (VNT-IIP) for rabies has been developed for the titration of dog and cat serum samples in Japan. The VNT-IIP has the advantage that results obtained can be viewed by the naked eye. The purpose of this study was to validate the VNT-IIP and compare it with one of the international standard methods, the fluorescent antibody virus neutralization test (FAVNT). The VNT-IIP showed satisfactory repeatability, high analytical specificity and good accuracy. Regarding the comparison between the VNT-IIP and the FAVNT, the VNT-IIP showed good agreement (91.9%), high sensitivity (92.8%) as well as specificity (87.0%) and good correlation (r = 0.92). As described above, the validation of the VNT-IIP was satisfactory and the performances of the test proved to be equivalent to those of an international standard method.     

Effects of elevated CO2 concentration on the respiratory behavior of the branches of field-grown hinoki cypress (Chamaecyparis obtusa)

The effects of elevated atmospheric CO₂ concentrations on the nighttime respiration were examined for two sample branches of a hinoki cypress tree (Chamaecyparis obtusa) growing in the field with an open gas exchange system for a one-year period from July 1994 to June 1995. The branches were of a similar size and located at a similar position within the crown. One branch was subjected to an elevated CO₂ concentration of 800 μmol mol⁻¹ and the other was subjected to ambient air which had a CO₂ concentration of about 370 μmol mol⁻¹. Nighttime respiration rate was higher in elevated CO₂ level than in ambient CO₂ level. The relationship between nighttime respiration and the corresponding nighttime air temperature was fitted by the exponential function in every month of the year. The segregation of regression lines between the two CO₂ treatments increased gradually as the seasons progressed during the treatment period. TheQ ₁₀ values for nighttime respiration were lower in elevated CO₂ (1.9 ≤Q ₁₀ ≤ 3.7) than in ambient CO₂ (2.4 ≤Q ₁₀ ≤ 4.5) in every month of the year. TheQ ₁₀ was inversely related to the monthly mean nighttime air temperature in both elevated and ambient CO₂. The estimated daily nighttime respiration rate under both CO₂ treatments had a similar seasonal pattern, which almost synchronized with the temperature change. The respiration ratio of elevated CO₂ to ambient CO₂ increased gradually from 1.1 to 1.6 until the end of the experiment. Our results indicate that the CO₂ level and the temperature have a strong interactive effect on respiration and suggest that a potential increase in respiration of branches will occur when ambient CO₂ increases.     

Quantitation of flavonoid constituents in citrus fruits.

Twenty-four flavonoids have been determined in 66 Citrus species and near-citrus relatives, grown in the same field and year, by means of reversed phase high-performance liquid chromatography analysis. Statistical methods have been applied to find relations among the species. The F ratios of 21 flavonoids obtained by applying ANOVA analysis are significant, indicating that a classification of the species using these variables is reasonable to pursue. Principal component analysis revealed that the distributions of Citrus species belonging to different classes were largely in accordance with Tanaka's classification system.     

Fishery biology of mud crabs <Emphasis Type="Italic">Scylla</Emphasis> spp. at Iriomote Island,Japan: species composition,catch, growth and size at sexual maturity

The fishery biology of mud crabs Scylla spp. was examined using baited traps and gill nets from September 2001 to August 2005 at Iriomote Island, Japan. To elucidate the growth of the crabs, artificially produced S. serrata juveniles were released and recaptured at the study site. The sizes at which 50% of females and males of S. serrata reached sexual maturity (SM₅₀) were estimated as an external carapace width (ECW) based on the morphology of the abdomen and the chela respectively. Two species, S. serrata and S. olivacea, were identified in the area with S. serrata being the dominant species (>95% of the catch). Changes in the mean ECW and the results of the release and recapture experiments suggested that the recruitment of young crabs to the fishery occurred from December/January to April/May. The SM₅₀ of females and males occurred at 132.4 and 150.7 mm ECW respectively. The body size composition of S. serrata revealed that immature crabs comprised approximately 40 and 65% of the catch for females and males respectively. To maintain a sustainable fishery for S. serrata, a minimum landing size based on the SM₅₀ estimates should be implemented as a fishing regulation.     

A transdisciplinary research of coastal fisheries co-management: the case of the hairtail <Emphasis Type="Italic">Trichiurus japonicus</Emphasis> trolling line fishery around the Bungo Channel,Japan

Under the coastal fisheries co-management regime in Japan, local fishers play important roles in deciding on and implementing local management measures. Therefore, using the case study of a hairtail trolling line fishery, we conducted a transdisciplinary research with fishers, processors, government officers, etc. Taking social and biological factors into account, we defined management criteria and targets (resource, economic, human community, and local food culture) in collaboration with local fishers. We chartered local fishing vessels to determine parameters that were easily understood by local fishers. We established a local consultative committee comprising fishers, processors, and distributors, and developed a fish distribution strategy to nurture the local hairtail food culture. The integrated effects of various management scenarios were evaluated by the operating model. We found that co-ordinated management by both the trolling line fishery and purse seiners in the neighbouring prefecture was required to achieve targets. Two realistic scenarios are recommended: a conservative scenario that could achieve targets even with few strong year classes, and an adaptive scenario that protects strong year classes, but relies on a higher frequency of these. We shared these results with the local fishers and government officers, which led to actual improvements in management measures.     

Conservation of small hairtail <Emphasis Type="Italic">Trichiurus japonicas</Emphasis> by using hooks with large artificial bait: effect on the trolling line fishery

Overfishing has reduced the stock of hairtail Trichiurus japonicus around the Bungo Channel, Japan. To determine whether using larger bait in the trolling line fishery could avoid catching small/undersize hairtail, we developed and trialled a new large artificial bait (a soft plastic bait, 6 inches long). A traditional lure with natural bait (type-1), a new lure of the new artificial bait hook without natural bait (type-2), and a new lure with natural bait (type-3) were tested in fishing operations around the Bungo Channel. Compared with type-1, type-2 and type-3 caught fewer undersized and immature female individuals. The number of fish caught, yield, and composition of commercial size grades per recruitment were calculated from field data for each lure and compared. The catch sizes (number of fish per recruitment) were smaller for type-2 and type-3 than for type-1, but yield per recruitment was higher for type-3 than for type-1. Compared with type-1, type-2 and type-3 caught more large individuals, which are more valuable. The newly developed artificial bait conserves hairtail stocks by targeting larger fish, which is economically beneficial for the hairtail trolling line fishery.     

Comparison of morphological and DNA-based techniques for stomach content analyses in juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>: a case study on diet richness of juvenile fishes

To identify the stomach contents of marine organisms, morphological observations are commonly conducted. However, the results obtained by this traditional method are frequently biased, as it is difficult to detect partially digested, soft-bodied organisms. To resolve this, recent studies have used DNA-based (DNA barcoding) analyses to examine the diet breadth of marine organisms. Here, we compared the prey richness of juvenile fishes using morphological observations and DNA-based analyses, with a focus on juvenile chum salmon Oncorhynchus keta. A higher number of prey taxa were successfully identified using DNA-based analyses than morphological observations. However, we also noticed several shortcomings of the DNA-based analyses, as reported in other diet-analysis studies that used molecular techniques. For example, the degree of digestion among prey taxa might have resulted in differential sensitivity to DNA detection. Additionally, several prey taxa could not be precisely identified, as the sequence data for some of the targeted organismal groups are unavailable in public gene databases. Remarkably, it is also possible that DNA-based analyses detected secondary prey, and therefore, the richness of prey taxa was likely overestimated. Thus, dietary analyses of juvenile fishes need to be carefully conducted, considering the respective advantages and disadvantages of DNA-based and morphological techniques.