Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica

Repeated injections of salmon pituitary extract (20 mg per fish per week) induced vitellogenesis in feminized, cultivated Japanese eels (Anguilla japonica). Oocytes were attained at the migratory nucleus stage after 11 or 12 injections. Addition of 17,20-dihydroxy-4-pregnen-3-one (DHP) into the incubation medium induced germinal vesicle breakdown (GVBD) in the oocytes at the migratory nucleus stage. An injection of DHP (2 µg g^-1 BW), given 24h after an injection of salmon pituitary extract (20 mg fish^-1), succeeded in inducing maturation and ovulation in females which contained occytes at the migratory nucleus stage. Most fish ovulated 15–18h following the DHP injection. Eggs that were ovulated within 15h after the DHP injection showed high fertility and hatchability, but eggs ovulated 18 or 21h after the DHP injection, showed considerably lower fertility and hatchability. A delay between ovulation and stripping of the eggs rapidly decreased both the fertility and hatchability within 6–9h after ovulation, indicating that artificial fertilization must be carried out immediately after ovulation. Repeated injections of human chorionic gonadotropin (hCG) at a concentration of 1 IU g^-1 BW week^-1 induced spermatogenesis, spermiation, and the acquisition of potential for sperm motility in cultivated males. Most males spermiated after the fifth or sixth injection of hCG, and the milt weight gradually increased and remained constant (1–2 g) from the 11th to 31th injection. Sperm motility peaked 24h after each weekly injection, and decreased from the 3^rd day after the injection. Potassium ions are an essential constituent for the maintenance of motility in the eel spermatozoa. Artificial seminal plasma containing 15.2 mM KCl is applicable as a milt diluent. Using these techniques developed for female and male eels, we have succeeded in obtaining many fertilized eggs from cultivated eels.     

Bathymetric distribution of beni-zuwai crab <Emphasis Type="Italic">Chionoecetes japonicus</Emphasis> in the northern part of the Sea of Japan

Surveys of beni-zuwai crab were conducted to clarify the bathymetric distribution and fecundity of beni-zuwai crab populations without the effects of bias due to fisheries. The results were compared with previous ones obtained at the heavily exploited Yamato Bank. Two sites in the northern part of the Sea of Japan, off southwest Hokkaido (200–2500 m deep) and West-Tsugaru Basin (300–1900 m deep), both of which are experiencing low exploitation levels, were surveyed in August to September, 2007. There was an abundance of morphologically immature crabs of both sexes in the depth range 1800–2100 m, and a few occurred at depths >2200 m. Immature crabs of <40 mm carapace width were found to migrate downslope to depths of 1800–2100 m and then to migrate upslope during molting for males and after maturity molting for females. Although data were lacking for depths >2000 m at the West-Tsugaru Basin, these relationships were significant for the pooled data of both localities, and they were similar to the results at the Yamato Bank. Clutch fullness of ovigerous females was negatively correlated with depth, which indicates that the low clutch fullness was not induced by fisheries, although the similar phenomenon observed at the Yamato Bank had previously been considered to be due to fishery effects.     

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.     

Carbon source and trophic position of pelagic fish in coastal waters of south-eastern Izu Peninsula,Japan, identified by stable isotope analysis

ABSTRACT:   It is important to clarify trophic dynamics in marine ecosystems for management of the fishing ground. Organic carbon sources and trophic position of pelagic fishes in the coastal waters of the south-eastern Izu Peninsula, Japan, were examined on the basis of carbon and nitrogen stable isotope distributions. The δ¹³C of the fishes was mostly distributed from −19 to −16‰ for nektonic fishes (13 species of adults and immatures) and planktonic fishes (10 species of larvae and juveniles), close to the δ¹³C values of particulate organic matter and planktonic decapods. These δ¹³C signatures for the inhabitants of the water column were in contrast with the high δ¹³C values (mainly −16 to −13‰) for demersal fishes of Scorpaeniformes and benthic polychaetes collected in the surf zone. These results indicate that nektonic and planktonic fishes depend on phytoplankton for carbon supply. The δ¹⁵N signatures suggest that the trophic position ranged 3.1–4.5 for the nektonic fishes and 2.9–3.7 for the planktonic fishes, premised on trophic level 3 for larval Japanese anchovy Engraulis japonicus . Thus, planktivorous fishes should be mainly assigned to trophic levels 3 and 4 in this area.     

Linking fish ecomorphotypes to food categories and local-scale habitat uses along a Brazilian coastal stream

Fish ecomorphological analyses often deal with several traits simultaneously, making it challenging to interpret general patterns, and have not addressed the longitudinal component of streams in morphology-habitat relationships. We identified ecomorphotypes of fishes and correlated their morphological variation with food, structural/hydrological variables, and abundances to detect their links with diet and local-scale habitat categories. Eighteen morphometric traits were obtained from specimens collected by electrofishing in a coastal Atlantic stream in Brazil. To test for morphology-habitat associations, headwaters, middle, and mouth reaches were divided into shorter sampling sites, with the middle reach being classified into riffle, run, and pool mesohabitats. Multivariate analyses highlighted the morphological variations and associated the form categories of 18 fish species with food and habitat categories. It resulted in four combined trophic and habitat ecomorphotypes: (1) benthic AB/lithophilic, composed of smaller to longer loricariids, detritivores, of fast-water, and pebbly habitats; (2) benthic C/lithophilic of very fast-waters, algal and detritus scrapers, with longer intestines than the other benthic species, wider suctorial mouths, inhabitants of headwaters, and riffles habitats; (3) nektobenthic/lithophilic with streamlined bodies, invertebrate feeders, and dwellers of faster-water, cobbly habitats; and (4) nektonic/limnophilic composed mainly by characins, which had strong correlations with terrestrial insect consumption, and lentic/pool, sandy habitats. The morphology–environment correlations linked these ecomorphotypes to the broad habitat gradient arrayed in longitudinal and local scales. These results permit inferring the larger form patterns expected for local fish assemblages and emphasise the fish trait categorisation as a possible surrogate to reveal broader ecomorphological associations.     

Evaluation of three phytoplankton species as food for the pearl oyster <Emphasis Type="Italic">Pinctada fucata</Emphasis>

In the pearl cultivation farms of the Ehime Prefecture, Japan, mass mortalities of the pearl oyster Pinctada fucata have occurred since 1994. The occurrences of mass mortality roughly coincided with a shift of the dominant phytoplankton from Skeletonema and Chaetoceros to Chaetoceros and Nitzschia all of which belong to Bacillariophyceae. Hence, we evaluated Nitzschia, together with Chaetoceros and Isocrysis, as food for the oyster. Wet weights, lengths, widths, glycogen contents, and growth rates in terms of wet weight of the oysters in all the feeding treatments were significantly higher than those in the non-feeding treatment. The highest glycogen content (2.34%) and growth rate (2.21 g month⁻¹) were found in the Chaetoceros treatment. Growth rate in the Isocrysis treatment (1.63 g month⁻¹) was also high, although glycogen content in this treatment (0.41%) was low. In the Nitzschia treatment, growth rate of the oyster (0.94 g month⁻¹) was the lowest and glycogen content (0.83%) was also low relative to that in the Chaetoceros treatment. Chlorophyll a concentration in fecal pellets was lowest in the Nitzschia treatment (<2.7 μg mg⁻¹), suggesting more complete digestion of Nitzschia by the oyster. Thus, Nitzschia was edible and digestible but not assimilated by P. fucata. We propose the following scenario for the relationship between Nitzschia dominance and mass mortality. When Nitzschia dominates in a culture area, the physiological condition of P. fucata deteriorates due to low assimilation of Nitzschia by the oyster, followed by susceptibility of the oyster to infection by agents lethal to the oyster.     

The structure of iron in Earth's inner core

Earth's solid inner core is mainly composed of iron (Fe). Because the relevant ultrahigh pressure and temperature conditions are difficult to produce experimentally, the preferred crystal structure of Fe at the inner core remains uncertain. Static compression experiments showed that the hexagonal close-packed (hcp) structure of Fe is stable up to 377 gigapascals and 5700 kelvin, corresponding to inner core conditions. The observed weak temperature dependence of the c/a axial ratio suggests that hcp Fe is elastically anisotropic at core temperatures. Preferred orientation of the hcp phase may explain previously observed inner core seismic anisotropy.     

Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

Attractiveness and oviposition preference of Glycaspis brimblecombei Moore in Eucalyptus spp.

The red gum lerp psyllid Glycaspis brimblecombei Moore (Hemiptera: Psyllidae) was first recorded in Brazil in 2003 and since then has caused serious damage in Eucalyptus forests. The damage is mainly due to defoliation, sooty mold and dieback, thus requiring the development of efficient management strategies. The settling and ovipositional preference of G. brimblecombei by species and clones of Eucalyptus were evaluated in free- and no-choice tests. Preliminary tests were done to determine the density of psyllid couples that produced the greatest densities of eggs, leaf position selection for oviposition, and whether oviposition was affected by preimaginal conditioning. Apical leaves were used more frequently for oviposition by the psyllid, and the number of eggs decreased from the apex to the base. The densities of 10 and 15 psyllid couples per seedling produced the highest oviposition. There was no evidence of preimaginal conditioning. In the free-choice test, E. grandis, E. urophylla, VM-1, I-144, C-219 and H-13 were less attractive to G. brimblecombei adults, with H-13, E. grandis and E. urophylla having the least oviposition. In the no-choice test, E. urophylla, GG-100 and E. grandis also demonstrated a lower oviposition, but E. camaldulensis and 3025 were highly susceptible. The low preference for the genotypes E. grandis and E. urophylla suggests the occurrence of a non-preference type resistance against the red gum lerp psyllid. Our results can be utilized for the development of management programs for G. brimblecombei in Eucalyptus forests.     

Infection process of <Emphasis Type="Italic">Pseudocercospora musae</Emphasis> on banana leaf

Yellow Sigatoka that is caused by Pseudocercospora musae is an important banana disease. The aim of this study was to elucidate the infection process of P. musae in banana leaves by scanning electron microscopy. Leaf samples were inoculated on the abaxial surface with P. musae and then analysed at 12, 24, 36, 48, 72, 96, 120, 144, and 168 h post inoculation (hpi) and at 36 and 50 days post inoculation (dpi). The conidia were found to be germinated between 24 and 36 hpi and penetrated through the stomata between 96 and 120 hpi, or more generally from 144 hpi. P. musae colonized the spongy parenchyma at 36 dpi and the palisade parenchyma at 50 dpi. Sporulation occurred at 50 dpi on the adaxial surface of leaves through the emergence of conidia on conidiophores through the stomata. Considering the importance of yellow Sigatoka in banana production, our results provide a better understanding of the life cycle of the fungus for treatment processes.