A genetic linkage map of the Japanese flounder, Paralichthys olivaceus

We report the first genetic linkage map of the Japanese flounder (Paralichthys olivaceus) constructed with 111 microsatellite markers and 352 AFLP fragments. The parental male linkage map consisted of 25 linkage groups while the female map consisted of 27 groups, with an average resolution of 8 and 6.6 cM, respectively. We have identified linkage among 96% of the markers and the total map length was estimated to be around 1000–1200 cM. This study reports the first low-density linkage map for the Japanese flounder and describes differences in sex recombination. Recombination rates were higher in male flounder compared to the female (7.4:1), a rare condition among vertebrates. This map is a starting point for the mapping of single loci and quantitative traits in flatfish species.     

Comparison of calpain and calpastatin activities in skeletal muscle of broiler and layer chickens

1. The objective of this study was to estimate the difference between broiler and layer chicks in the activities of calpain and calpastatin (inhibitor of calpain) in breast muscle. Differences between broilers and layers in body weight, daily gain at 3 weeks of age and fractional growth rate (FGR) during 2 and 3 weeks of age were statistically significant (P < 0.01).

2. Calpain and calpastatin activities were measured at three weeks of age with alkali‐denatured casein as a substrate. The m‐calpain (calpain activated by millimolar calcium concentration) activities in units/g muscle and units/ mg extractable muscle protein were 0.779 and 0.353 for broilers, and 1.042 and 0.440 for layers, respectively. The calpastatin activities in units/g muscle and units/mg extractable muscle protein were 0.332 and 0.153 for broilers, and 0.262 and 0.112 for layers, respectively.

3. Broilers with high FGR showed low m‐calpain and high calpastatin activities. In contrast, layers with low FGR showed high m‐calpain and low calpastatin activities.

4. These results suggest that m‐calpain and calpastatin activities in skeletal muscle vary between breeds which have different rates of muscle production.     

Classification of Southeast Asian mints (Mentha spp.) based on simple sequence repeat markers

Mentha is a complex genus encompassing many species as a consequence of their interspecific hybridization and polyploidy. Southeast Asian mints have been poorly distinguished though they are widely used for culinary and medical purposes. In this study, we have analyzed Southeast Asian mints and known varieties as well as a related Lamiaceae species (Nepeta sp.) using simple sequence repeat (SSR) markers and leaf morphology. Two types of mints were clearly distinguished based on their venation pattern and leaf shape index. We developed 12 SSR markers that allowed good amplification in the Mentha and another Lamiaceae species. In the SSR-based phylogram, the Mentha lines could be delimited into groups I–VI. The Southeast Asian mints divided into groups I and II, and the phylogram separated most of the available species, with groups I and II containing the known species M. × cordifolia and M. arvensis, respectively. The separation of the two groups was supported by a population structure analysis. The SSR markers developed in this study enabled the simultaneous classification of mints and will help improve our understanding of the genetic composition of known mint varieties and as yet unclassified Southeast Asian mints.     

Inhibitory effects of (-)-epigallocatechin gallate on the mutation,DNA strand cleavage,and DNA adduct formation by heterocyclic amines

Green tea is known to be a potential chemopreventive agent against cancer. In this study, we investigated the inhibitory activities of tea extracts, and in particular the polyphenolic component (-)-epigallocatechin gallate (EGCG), against heterocyclic amine-induced genotoxicity. The tea extracts displayed inhibition of 2-hydroxyamino-6-methyldipyrido[1,2-a,3',2'-d]imidazole (Glu-P-1(NHOH))-induced mutagenicity. This inhibition can be accounted for by the presence of EGCG in the extracts. The mutagenic effect of Glu-P-1(NHOH), which induces single-strand cleavage in supercoiled circular DNA under neutral conditions, was inhibited by EGCG. Using the Drosophila repair test, a test for gross DNA damage, and DNA adduct detection by (32)P-postlabeling, we showed that EGCG prevented 2-amino-3,8-dimethylimidazo[4,5-f]quinoline-induced DNA damage and adduct formation in insect DNA. EGCG was found to accelerate the degradation of Glu-P-1(NHOH) in vitro. This observation suggested that the inhibition by EGCG is associated with an accelerated degradation of metabolically activated heterocyclic amines.     

Differences in free amino acid composition between testis and ovary of sea urchin <Emphasis Type="Italic">Anthocidaris crassispina</Emphasis> during gonadal development

ABSTRACT:   For the purpose of clarifying the differences of taste components between the testis and ovary of the sea urchin Anthocidaris crassispina , variations in free amino acid composition in gonadal extract during gonadal development were investigated using chemical and histopathological methods. Sea urchins were collected at Iki Island, Nagasaki, Japan between 30 April and 5 June 2003, and maturation stages and the free amino acid compositions were determined. The sweet amino acids threonine (Thr), glycine (Gly), and alanine (Ala), the umami amino acid glutamic acid (Glu), and the bitter amino acids lysine (Lys) and arginine (Arg) were the main constituents accounting for more than 80% of total free amino acids. In the testis and ovary, Gly levels were the highest among them and increased with gonadal development. The other free amino acids consisted mainly of bitter amino acids and either showed no variation or decreased with gonadal development. These variations in free amino acid composition were observed more clearly in the ovary than the testis. These findings indicate the possibility of differences in the flavor of sea urchin gonad by sex and gonadal development stages.     

Food habits of small fishes in seagrass habitats in Trang,southern Thailand

To clarify the feeding habits of seagrass fishes, we examined the gut contents from 42 fish species collected in seagrass habitats in Trang. Thirteen species showed ontogenetic and/or seasonal changes in food-use patterns. Smaller individuals generally preyed on small planktonic items (e.g., copepod larvae) or small benthic/epiphytic crustaceans (e.g., harpacticoid copepods), with subsequent changes to other prey items (e.g., shrimps, crabs, detritus and filamentous algae) with growth. The most important dietary items for the seagrass fish assemblages comprised benthic/epiphytic crustaceans, detritus, and planktonic copepods. Cluster analysis based on dietary overlaps showed that the seagrass fishes comprised eight feeding guilds (large benthic/epiphytic crustacean, detritus, planktonic animal, small benthic/epiphytic crustacean, mollusc, invertebrate egg, polychaete, and fish feeders). Of these, the first three guilds were the most abundantly represented, whereas the last three were each represented by only a single species.     

Effect of Fucoidan Extracted from Mozuku on Experimental Cartilaginous Tissue Injury

We investigated the effect of fucoidan, a sulfated polysaccharide, on acceleration of healing of experimental cartilage injury in a rabbit model. An injured cartilage model was surgically created by introduction of three holes, one in the articular cartilage of the medial trochlea and two in the trochlear sulcus of the distal femur. Rabbits in three experimental groups (F groups) were orally administered fucoidan of seven different molecular weights (8, 50, 146, 239, 330, 400, or 1000 kD) for 3 weeks by screening. Control (C group) rabbits were provided water ad libitum. After the experimental period, macroscopic examination showed that the degree of filling in the fucoidan group was higher than that in the C group. Histologically, the holes were filled by collagen fiber and fibroblasts in the C group, and by chondroblasts and fibroblasts in the F groups. Image analysis of Alcian blue- and safranin O-stained F-group specimens showed increased production of glycosaminoglycans (GAGs) and proteoglycans (PGs), respectively. Some injured holes were well repaired both macroscopically and microscopically and were filled with cartilage tissues; cartilage matrices such as PGs and GAGs were produced in groups F 50, F 146, and F 239. Thus, fucoidan administration enhanced morphologically healing of cartilage injury.     

Patterns of aquaporin expression in the canine eye

Aquaporins (AQPs) function as water channels in many types of cells involved in fluid transport. More than 10 isoforms have been identified, and these are differentially expressed in many types of mammalian cells in the body. Six AQPs (AQP0, AQP1, AQP3, AQP4, AQP5, and AQP9) have been identified in the eyes of humans and/or rodents. The unique permeability characteristics and distribution of AQPs indicate their diverse roles in the regulation of water homeostasis in the eye. The aim of this study was to investigate the localisation of AQPs in normal canine eyes, with AQP0 protein expressed in the crystalline lens and retina. Although AQP1 mRNA was detected in various areas of the canine eye, its protein expression was limited to the cornea, iris and ciliary body. AQP4 was identified in the iris, retina and optic nerve. AQP3 and AQP5 were found in the cornea and conjunctiva, and their expression was particularly high in the limbus. AQP3 and AQP5 were present in the nictitating membrane indicating that they play a role in water transport within the membrane. The observations suggested that several subtypes of the AQP family are involved in the regulation of water homeostasis in the canine eye.     

DIFFERENCES IN ACQUISITION OF ORGANIC NITROGEN IN SOILS BETWEEN BOK CHOY AND TOMATO

Seedlings of bok choy and tomato were grown in soils with different nitrogen (N) sources [no N (?N), ammonium sulfate (AS), and cattle farmyard manure (CM)]. Comparison between soils treated with ?N and CM indicated that the growth and N accumulation in bok choy were significantly enhanced by CM treatment, whereas no difference was found in tomato. In the rhizosphere soils, the highest protease activity was detected in CM treatment irrespective of species. Correlation analysis between rhizospheric protease activity and total N accumulation of plant treated with –N and CM showed a significant positive correlation only for bok choy. The determination of amino acid absorption rate in excised roots indicated that glycine was taken up at a significantly higher rate in bok choy than tomato. This study suggested that at least two possible factors affected the acquisition of organic N: rhizospheric protease activity and ability to absorb amino acids in roots.