Effect of water exchange with mangrove enclosures based on nitrogen budget in <Emphasis Type="Italic">Penaeus monodon</Emphasis> aquaculture ponds

ABSTRACT:   A model experiment in which water was circulated between shrimp aquaculture ponds (Ponds 1, 2, and 5), and mangrove enclosures (Ponds 3 and 4), was carried out at the Samut Songkhram Coastal Aquatic Research Station, Thailand, from March through July 2003. Five ponds with a size of 40 m × 20 m and 1.5 m deep were used for this experiment. Ten thousand shrimp larvae Penaeus monodon at the PL-15 stage were stocked in Ponds 1 and 5, and 20 000 larvae were stocked in Pond 2. Mangrove Rhizophora mucronata (476 trees) had been planted in each of Ponds 3 and 4 in July 2002. Approximately 30% of the water in Ponds 2 and 5 was moved to the mangrove enclosures by siphon (Ponds 3 and 4, respectively) every Thursday, and the same amount was returned to the shrimp ponds by pump every Monday over the 136-day experiment. Shrimp incorporated 30.3, 25.6, and 33.7% of the input nitrogen as feed in Ponds 1, 2, and 5. Since 2.02 kg N in Pond 2 and 0.76 kg N in Pond 5 was transported to the mangrove ecosystem, and 4.26 kg N in Pond 2 and 3.98 kg N in Pond 5 was retained in water and mud, it was estimated that between 2.1 and 5.2 unit areas of mangroves is required to remove the nitrogen remaining in the aquaculture pond.     

Genetic and molecular analysis of the restoration of fertility ( Rf ) genes for Ogura male-sterility from a Japanese wild radish ( Raphanus sativus var. hortensis f. raphanistroides Makino)

Ogura male-sterile cytoplasm is one of the most extensively studied cytoplasms in Brassicaceae. In this study, in order to gain better understanding of the variation and evolution of the restoration of the fertility (Rf) gene for Ogura male-sterile cytoplasm, the nucleotide sequence of the orf687 homologue in the Japanese wild radish (Raphanus sativus var. hortensis f. raphanistroides Makino) was analyzed using an F₂ population made with a cross between a Japanese wild radish plant containing the Rf gene and ‘Uchiki-Gensuke’ (a maintainer of Ogura-male sterility). Segregation of male-fertile/-sterile plants in the F₂ generation suggested that another unidentified Rf gene unlinked to orf687 exists in the Japanese wild radish. The genotype of orf687 was determined for each F₂ plant by Southern hybridization with an orf687 gene probe, mismatch-specific endonuclease digestion of PCR products, and direct sequencing of a PCR product. Genotyping revealed that some fertility-restored plants are homozygotic for the ‘Uchiki-Gensuke’ type orf687 allele, supporting the idea that another gene different from orf687 also functions as an Rf gene for Ogura male-sterility. Protein analysis using an antibody raised against the Ogura-specific ORF138 protein suggests a mechanism of fertility restoration by the unidentified Rf similar to that by orf687. Sequence analysis of orf687 from a Japanese wild radish plant and ‘Uchiki-Gensuke’ revealed that both orf687 regions encode a mitochondrially-targeted protein consisting of 687 amino acids with 16 PPR motifs. Comparison of the deduced amino acid sequences with those of the known orf687 sequences from ‘Yuan hong’ and ‘Kosena’ containing Rf and recessive one (rf), respectively, showed that three unique amino acid replacements are present in ORF687 of the Japanese wild radish. Two of the three replacements, that from lysine to isoleucine at position 232 and from asparagine to asparate at position 240, confer negative charges to the protein. Since the Rf of ‘Yuan hong’ was reported to have a unique replacement that confers a negative charge to ORF687 (from asparagine to aspartate at position 170), it is proposed that the amino acid replacements conferring a negative charge to ORF687 are important for determining the status of the Rf/rf gene.     

Biochemical changes in oyster tissues and hemolymph during long-term air exposure

The Pacific oyster Crassostrea gigas is a sessile bivalve that inhabits the intertidal zone and therefore frequently exposed to air during the tidal cycle. It is highly adaptive to hypoxic conditions. We have studied the physiological state of oysters during long-term exposure to air. The oysters became hypoxic when exposed to air or hypoxic seawater. The 50% lethal time of oysters exposed to air at 4, 15 and 20°C was 47.8, 15.9 and 12.2 days, respectively. The hemolymph pH decreased by day 3; however, it showed a slight increase by day 5 at both 4 and 20°C. The adenylate energy charge (AEC) values decreased rapidly on the first day of air exposure in the adductor muscle, mantle, gill and body trunk, and these decreases were accompanied by decreases in ATP concentrations and increases in AMP concentrations. The AEC values in all of the tissues had fallen to below 30% by day 50 of air exposure at 4°C. These data suggest that the energy state of oysters deteriorates rapidly with air exposure. Consequently, AEC values may be useful indices of the physiological state of the oyster during long-term exposure to air.     

Phylogeography based on the nuclear ribosomal DNA internal transcribed spacer region of native Miscanthus sinensis (Poaceae) populations in Japan

Nucleotide sequence variations were investigated with respect to the geographical distribution patterns of Miscanthus sinensis populations that were sampled from 26 Japanese national parks and three populations of the Ryukyu Islands. Twelve homozygous sequences in the nuclear ribosomal DNA internal transcribed spacer region were detected. The populations of M. sinensis in mainland Japan mainly were composed of a monophyletic group with a symapomorphic character, whereas those in the Ryukyu Islands included a polyphyletic group. Only an internal transcribed spacer haplotype with a plesiomorphic character was found in both mainland Japan and the Ryukyu Islands. Thus, no clear geographical isolation was observed in this species. These facts might be caused by the ability of M. sinensis as a pioneer plant to have a high migration potential and high gene flow by outcrossing. On the basis of the results of this study (nuclear ribosomal DNA) and the previous study (chloroplast DNA), a phylogeographical history of M. sinensis was presumed where the ancestral and derived lineages were distributed in the southern and northern regions of Japan, respectively, without strict geographical isolation.     

A case of spontaneous rete testis adenoma in a Sprague–Dawley rat

A 104-week-old male CD (SD) rat exhibited enlargement of the left testis. Microscopically, this mass was demarcated from the testis by fibrous connective tissue and characterized by cystic dilatation with single-layered columnar cells and papillary proliferation connected to the solid growth area without clear boundaries. In the solid growth area, cells were dissected into irregular alveolar nests by scant fibrous tissue with small blood vessels. The nuclei of proliferating cells were variable in size and round- to oval-shaped, and their cytoplasm was pale or eosinophilic and sometimes contained vacuoles or eosinophilic granules. Immunohistochemically, the tumor cells were positive for vimentin and cytokeratin (CK) 7. Since CK7 was exclusively positive in the rete testis epithelium of the naïve rat, it was valuable to diagnose this tumor as rete testis-originated. Based on these results and the lack of apparent pleomorphism, mitotic figures, and metastasis, the present case was diagnosed as rete testis adenoma.     

Characterization and distribution of an arginine vasotocin receptor in mouse

A cDNA, which has a high homology with teleost Platichthys flesus [Arg8] vasotocin (AVT) receptor (GenBank: AK033957), was found in mouse genome database. Analyses of the deduced amino acid sequence revealed that a cDNA has several features of AVT receptor. We tentatively named it as a mouse vasotocin receptor (MVTR). A two-electrodes voltage clamp technique was applied to characterize the MVTR expressed in Xenopus laevis oocytes. AVT induced Ca2+-dependent Cl- currents in Xenopus oocytes injected with MVTR cRNA. On the other hand, [Arg8] vasopressin, oxytocin and isotocin did not induce such currents. RT-PCR showed that MVTR mRNA was specifically expressed in the brain. In situ hybridization analysis demonstrated significant expression of MVTR mRNA in suprachiasmatic nucleus, arcuate nucleus and medial habenular nucleus of mouse brain. These results suggest that MVTR may mediate a variety of physiological functions in mouse.     

Structure and function of the oxidation products of polyphenols and identification of potent lipoxygenase inhibitors from Fe-catalyzed oxidation of resveratrol

Polyphenols have recently attracted much attention as potent antioxidants and related bioactive substances. These potent antioxidative polyphenols are very oxidizable due to their chemical properties, and their oxidation products must accumulate in the oxidizing foods when they are contained as the active ingredients. In this investigation, 30 polyphenols and related phenolics were oxidized with oxygen in the presence of a catalytic amount of Fe ions. Piceatannol, catechin, epicatechin, hydroxytyrosol, carnosol, and carnosic acid were oxidized very quickly. Sinapic acid, caffeic acid, chlorogenic acid, rosmarinic acid, gallic acid, propyl gallate, α-tocopherol, quercetin, and nordihydroguaiaretic acid were moderately oxidized. Protocatechuic acid, syringic acid, taxifolin, resveratrol, gentisic acid, secoisolariciresinol, and ellagic acid were oxidized for 19-20 days; however, their oxidation was very slow and did not complete. The other phenolics were not oxidized. The obtained oxidation products were next subjected to a lipoxygenase inhibition assay and the results compared to those of the corresponding phenols. Very interestingly, the oxidation product from resveratrol showed a high inhibitory activity, whereas resveratrol itself had no activity and its oxidation efficiency was low. To clarify the inhibition principle of the oxidation product, an LC-MS analysis was carried out on the oxidation product. The analytical results showed that they are the oligomeric and degraded compounds of resveratrol. Among them, the structures of three dimeric compounds were successfully identified, and their activity data clarified that the closed ring dimers were potent lipoxygenase inhibitors, whereas the opened ring dimer was not. It should be noted that resveratrol had almost no lipoxygenase inhibitory activity, contrary to some researchers' findings.     

Isolation of Fusarium sp. from a claw of a dog with onychomycosis

An 8-year-old male Golden Retriever had lameness and claw abnormality in the second digit of the left forelimb. Radiography revealed osteomyelitis in the distal phalanx bone of the affected limb. Microscopic examination of the claw revealed numerous hyphae in the claw matrix. Fungal DNA fragments coding the ribosomal internal transcribed spacer region (ITS) were detected from the claw matrix as well as fungal colonies of the clinical isolates by PCR. Nucleotide sequencing revealed that the amplicons shared > 99% homology with Fusarium sp. Therapy including oral itraconazole resulted in regrowth of a new claw, in which no hyphae were detected. To the authors' knowledge, this is the first case report of canine onychomycosis in which Fusarium sp. was isolated from the affected claw.     

Early development of the silver catfish Rhamdia quelen (Quoy & Gaimard, 1824) (Pisces:Heptapteridae) from the São Francisco River Basin, Brazil

The silver catfish, Rhamdia quelen , is endemic to North, Central and South America with high aquaculture potential and wide acceptance in the market. Breeder fish were subjected to induced reproduction through hypophysation using a crude common carp pituitary extract. Egg characteristics, oocyte surface ultrastructure and histology of larval ontogenesis until whole yolk resorption were described for the first time for this species. Oocytes and semen were obtained by manual extrusion, and fertilization was conducted using the dry method. After fertilization, eggs were kept in incubators at 24 °C. The embryonic development was monitored using a stereomicroscope every 10 min until hatching. To analyse the larval development, larvae samples were collected from incubators daily until the fifth day, fixed in Bouin's fluid and subjected to routine histological techniques. The oocyte extrusion occurred 8 h after the second hormone dose at 26 °C. The oocytes were spherical, non-adhesive and yellow, with a diameter of 1471.75±47.63 μm. Scanning electron microscopy revealed a thin jelly coat covering the zona radiata in the animal pole around the micropyle. The blastopore closure occurred within 8 h after fertilization, and the fertilization rate was 79.9±5.2% at 24 °C. Embryonic development was completed within 25 h 30 min after fertilization. The complete resorption of the yolk and the formation of the digestive system organs and the mouth opening occurred on the fifth day, indicating a need for exogenous feeding. The results of this study provide information important for improvement in R. quelen culture and management.