Lysozyme in the ovary of tilapia (Oreochromis mossambicus): its purification and some biological properties

Lysozyme was purified from the ovary of tilapia, Oreochromis mossambicus, with two steps, chitin coated-cellulose and Sephadex G-100, and its biological properties were investigated. Purified lysozyme had a molecular mass of 15kDa on SDS-PAGE under reducing condition. Analyses with antibody (a-EL) against the purified lysozyme revealed that serum and egg extract reacted with a-EL and the precipitin lines fused completely. The enzyme activities in serum and egg extract were inhibited by adding serially diluted a-EL. Therefore, egg extract and serum lysozyme was immunologically identical. Immunohistochemically, lysozyme was observed in the ooplasm of the oocytes laden with yolk but not in the follicle layers, egg envelope or immature oocytes (the peri-nucleolus stage). In addition, the enzyme activity in the large oocytes was higher than that in the small ones. These results suggest that lysozyme detected in the oocytes is derived from extra-ovarian tissue and transfers from the maternal circulation. Lysozyme activity in the serum of female tilapia increased with oocyte development, suggesting that the change in the enzyme level may be partially related to the reproductive events (especially vitellogenesis) of the female fish.     

Potential impacts of non‐native channel catfish on commercially important species in a Japanese lake,as inferred from long‐term monitoring data

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Effect of injection on bed shear stress and turbulence characteristics in a closed conduit flow

Bed shear stress and turbulence quantities are important parameters to understand sediment erosion, transport, and hydraulic processes in most hydraulic studies. An experimental investigation was conducted to understand the effect of injection on the bed shear stress and turbulence characteristics of flows during low sediment transport rate in a closed conduit, which is similar in construction to the erosion function apparatus. In particular, the effect of injection on the bed shear stress, mean velocity profiles, turbulence intensities, Reynolds shear stress, and higher-order moments of the closed conduit flows in the seepage zone as well as at the upstream edge of the seepage zone was examined. The instantaneous velocities were measured using two-dimensional particle image velocimetry (PIV) to evaluate the turbulence structure in both the seepage zone and at the upstream edge of the seepage zone. The bed shear stress estimated by the Reynolds shear stress approach was found to be more appropriate than that estimated by the usual logarithmic law approach. However, a spatial fluctuation in the bed shear stress was noticed as the injection intensity was increased. Injection was found to decrease the velocity near the bed and to increase the velocity near the center of the conduit in comparison to the no-seepage condition in both zones. The injection resulted in more of a decrease in bed stability in the seepage zone in comparison to the upstream edge of the seepage zone as the injection intensities were increased. The introduction of injection increased the magnitudes of the various turbulence parameters in comparison to the no-seepage condition in the seepage zone. The effect of injection was not only visible in the near-bed region, but in both zones as the water depth (measured from top of the sediment surface) increased.

     

Delineation of Japanese soil temperature regime map

The soil temperature regime map provides for utilitarian classification that can be superimposed on soil classification to permit more precise interpretations and assessments of land use. The objects of this study are (1) to clarify the relationship between soil temperature and meteo-geographical factors, and then (2) to delineate detailed soil temperature regime map (1?km grid) as Japanese land resources inventory. There was a parallel relationship between mean annual soil temperature (MAST) and mean annual air temperature (MAAT), but this relationship was affected to some extent by the mean annual wind speed and mean annual global irradiation in this study. Furthermore, the difference between MAST and MAAT [Diff(MAST–MAAT)] showed the highest correlation with elevation. The map of RK_Diff(MAST–MAAT) was computed using this meteo-geographical relationship with the regression-kriging approach, and then the map of MAST and the soil temperature regime map were delineated using the map of MAAT and the RK_Diff(MAST–MAAT). The root mean square error of this delineation procedure was 0.47°C. It was clear that the majority of the Japanese soils had “mesic” soil temperature regime, and Japanese agricultural land was mainly distributed at “mesic” area and followed by “thermic”, “frigid”, and “hyperthermic” area. For promoting this land resource inventory, the soil temperature regime map will be uploaded on “Soil Information Web Viewer (http://agrimesh.dc.affrc.go.jp/soil_db/)”, which is provided by the National Institute for Agro-Environmental Sciences.     

Dual inoculation of peanut with Glomus sp. and Bradyrhizobium sp. enhanced the symbiotic nitrogen fixation as assessed by 15N-Technique

Abstract

The response of peanut (Arachis hypogaea L.) to inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi (Glomus etunicatum) and Bradyrhizobiurn sp. was studied in pots by the acetylene reduction activity (ARA) and ‘A-value’ methods. The soil used was a Light-coloured Andosol and the treatments consisted of the inoculation of VAM fungi only, inoculation of Bradyrhizobium only, dual inoculation of VAM fungi and Bradyrhizobium and control, under non-sterilized and sterilized soil conditions.

In the non-sterilized soil the ARA and nitrogen fixation determined by the ‘A-value’ method increased significantly only by dual inoculation of VAM fungi and Bradyrhizobium at 100 days after planting (DAP), but no significant difference was observed at 70 DAP. In the case of dual inoculation, 75% of the nitrogen of the plant was derived from fixation whereas the plants inoculated only with Bradyrhizobium derived 68% of their nitrogen from fixation and the control plants, 64%. Amount of P in plant increased significantly only by dual inoculation with VAM fungi and Bradyrhizobium.

In the sterilized soil a highly significant increase in the ARA was observed of the dual inoculation at all the sampling times. Nitrogen fixation determined by the A-value technique and N and P contents in plant also increased significantly by dual inoculation. Results obtained by the A-value method showed that plants with dual inoculation derived 68% of their nitrogen from fixation while the plants inoculated only with Bradyrhizobium, 38%.

From our this study we conclude that nitrogen fixation as well as N and P contents in peanut increased significantly only by dual inoculation with VAM fungi and Bradyrhizobium.     

Effect of water flow on the mobility of the root-knot nematode Meloidogyne incognita in columns filled with glass beads, sand or andisol

In the present study, the migration of nematodes was studied in columns filled with three materials of different textures and chemical properties. The role of soil pores that enable root-knot nematode (Meloidogyne incognita) second stage juveniles (J2) to escape rapid water flow in soil was demonstrated using columns filled with glass beads, sand or andisol that maintained a constant water flow. Under a constant flow flux of 36 cm h⁻¹, living J2, dead J2 or anion bromine tracer (Br⁻) was injected in the middle of the column and then drainage water equivalent to two pore volumes (PV) was collected. The passive transport of the anion tracer in water flow could be explained by a convection dispersion equation. The dead J2 showed a pattern similar to that of Br⁻. However, the living J2 resisted movement in the water flow and remained in the column even at the highest water flow rate of 93.3 cm h⁻¹ in glass beads. The mobility of living J2 was affected by the filling materials; the number of J2 passing through the column was much lower in the andisol-filled column than in the other two columns but the total number of J2 in drainage water was 5% or less of the number injected for all columns. We suggest that J2 were affected not only by soil water flow but also by soil pore structure and have the ability of withstanding or avoiding movement in soil water flow.     

Nebulin and N-WASP cooperate to cause IGF-1-induced sarcomeric actin filament formation

Insulin-like growth factor 1 (IGF-1) induces skeletal muscle maturation and enlargement (hypertrophy). These responses require protein synthesis and myofibril formation (myofibrillogenesis). However, the signaling mechanisms of myofibrillogenesis remain obscure. We found that IGF-1-induced phosphatidylinositol 3-kinase-Akt signaling formed a complex of nebulin and N-WASP at the Z bands of myofibrils by interfering with glycogen synthase kinase-3β in mice. Although N-WASP is known to be an activator of the Arp2/3 complex to form branched actin filaments, the nebulin-N-WASP complex caused actin nucleation for unbranched actin filament formation from the Z bands without the Arp2/3 complex. Furthermore, N-WASP was required for IGF-1-induced muscle hypertrophy. These findings present the mechanisms of IGF-1-induced actin filament formation in myofibrillogenesis required for muscle maturation and hypertrophy and a mechanism of actin nucleation.     

Molecular mapping of a gene conferring resistance to Aphanomyces root rot (black root) in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Caused by Aphanomyces cochlioides Drechsler, Aphanomyces root rot is a serious disease of sugar beet (Beta vulgaris L.), for which sources of resistance are scarce. To identify the segregation pattern of the rare resistance trait found in Japanese sugar beet line ‘NK-310mm-O’, F₁ and BC₁F₂ seedings, drawn from a cross between ‘NK-310mm-O’ and susceptible line ‘NK-184mm-O’, were inoculated with zoospores and their survival evaluated in the greenhouse. Resistance segregation followed was that of a single dominant gene, which was designated Acr1 (Aphanomyces cochlioides resistance 1). Molecular markers tightly linked to Acr1 were identified by bulked segregant analysis of two BC₁F₂ populations. Fourteen AFLP markers linked to Acr1 were identified, the closest located within ±3.3 cM. Three F₅ lines and two BC₂F₁ lines, selected on the basis of their Acr1-AFLP markers, were tested for their resistance to Aphanomyces root rot in a highly infested field. Results indicated that Acr1 conferred significant resistance to Aphanomyces root rot at the field level. Based on its linkage with CAPS marker tk, a representative marker for chromosome III, Acr1 was located on this chromosome. The clear linkage between tk and Rhizomania resistance trait Rz1, suggests the clustering of major disease resistance genes on chromosome III.     

Monoclonal antibodies preventing invasion of Neospora caninum tachyzoites into host cells

Twelve monoclonal antibodies (MAbs) against Neospora caninum tachyzoites were produced to specify the antigens related to the invasion of tachyzoites into host cells. In the assay to evaluate the inhibition activity, all these MAbs prevented the cultured Vero cells from the invading by the tachyzoites. These MAbs recognized approximately a 73 kDa antigen in Western blot analysis. Immunofluorescence assay and immune electron microscopy revealed that this 73 kDa antigen is a part of the surface antigens of N. caninum tachyzoite, and that the tachyzoite antigen identified plays an important role for invasion of host cells.     

The first record of sucking louse, Neohaematopinus callosciuri, infesting pallas squirrels in Japan

A species of sucking louse, Neohaematopinus callosciuri, was found for the first time in Japan. The species was found on an invasive species of squirrel, Pallas squirrel, Callosciurus erythraeus, in the Kamakura district, Kanagawa Prefecture, Japan. A total of 52 lice were obtained from 22 of 104 squirrels captured. The lice were about three times more prevalent in male squirrels than in females and were detected most frequently in the winter. As N. callosciuri has never been reported on wild animals in Japan, this species probably was introduced into Japan along with their host, Pallas squirrels.