Developmental Competence of Frozen-thawed Blastocysts from Fair-quality Bovine Embryos Cultured with β-Mercaptoethanol

Two-hundred-and-thirty-one fair-quality embryos at the compacted morula stage collected from 89 superovulated cows were cultured in TCM199 or Brinster's BMOC-3 medium with or without 100 microM beta-mercaptoethanol (beta-ME). After 24 h culture, a total of 142 fair-quality embryos developed to the blastocyst stage, of which 106 were subsequently frozen with 1.8 M ethylene glycol. The mean cell number and development rates of frozen-thawed blastocysts from the fair-quality embryos cultured in TCM199 containing beta-ME were higher than those of the fair-quality embryos directly frozen without culture. The pregnancy rates obtained with frozen blastocysts from fair-quality embryos tended to be lower than those of non-cultured fresh fair-quality embryos and cultured fresh blastocysts. These results indicate that the inclusion of beta-ME in pre-freezing culture media improve the development of frozen-thawed blastocysts from fair-quality embryos, but not the pregnancy rate.     

Factors that limit maintenance of recombinant rumen bacterium in sheep rumen

Factors limiting the maintenance of recombinant ruminal bacterium in the rumen were evaluated in vitro , using batch culture prepared from rumen fluid of sheep. Butyrivibrio fibrisolvens expressing a foreign xylanase gene ( B. fibrisolvens NO4) was used as a tested recombinant that was selectable on an erythromycin-containing agar medium. The recombinant tended to reduce its level slowly in the rumen fluid of sheep on a high hay diet, while its initial decrease was more apparent in the rumen fluid of sheep on a high concentrate diet. Incubation with cell-free ruminal fluid revealed a significant decrease of inoculated recombinant, suggesting the presence of antibacterial factors limiting maintenance of the recombinant. In particular, during the first 12 h of incubation this inhibition was more notable in culture prepared from rumen fluid of sheep given the high concentrate diet. Autoclaving the cell-free rumen fluid inactivated the inhibition. Numbers of the recombinant for inoculation did not influence the final level of survived recombinant, that is, the initial depression was larger as more recombinant was inoculated. Subculturing with xylan before inoculation and/or direct addition of xylan to the batch culture did not improve survival of the recombinant. From these results it is suggested that the level of survived recombinant is limited to 10^2–4/mL of in vitro batch culture with restricted energy supply and that initial depression of the recombinant is mainly caused by the heat-sensitive antibacterial factors not associating with microbial cells in the rumen.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

尼日利亚热带雨林地区的内陆山谷中土壤发育和肥力特征: 矿物成分和粒度分布

The particle-size distribution and mineralogical composition of the clay (< 2 μm) and fine-sand (0.25--0.10 mm) fractions in soils of two inland valleys in Abakaliki and Bende, Southeast Nigeria, were investigated to provide basic information on soil-forming processes and agricultural potentials. These soils were silty or clayey, deriving from Cretaceous or Tertiary shale materials. The particle-size distribution and its computation on a clay-free basis revealed relatively remarkable lithologic breaks in a couple of pedons. The effect of lithologic discontinuities on soil mineralogical composition was not, however, conspicuous. Petrographic investigation revealed that quartz predominantly comprised the fine-sand fraction in the soils at both study sites. Nevertheless, the clay mineralogical composition of the soils was a mixture of kaolinite, irregularly interstratified smectite-illite intergrades (S/I), hydroxyl-Al interlayered 2:1 type clays (HICs), vermiculite, smectite, halloysite and illite along with fine-sized quartz in Abakaliki. The soils of Bende predominantly contained smectite, which was partially interlayered with hydroxyl-Al and kaolinite. It is suggested that seasonal floodwater has slowed the disintegration of weatherable clay minerals inherited from the shale, while quartz originating from the sandstone is predominant in the fine-sand fraction. Additionally, a possible soil-forming process observed at the both study sites was ferrolysis, which was indicated by a clear decreasing pattern of HICs downward in the soil profiles. The entry of S/I and vertical distribution patterns for a couple of clay minerals in the pedon suggested that the soils in Abakaliki have developed under the significant influence of aeolian dust delivered by the harmattan. The findings might describe a site-specific deposition pattern of harmattan dusts as well as hydromorphic soil-forming processes in the wetlands of the inland valleys.     

Chemerin fragments show different effects on systemic blood pressure dependent on carboxyl-terminal cleavage site

Chemerin is an adipocytokine whose concentration in blood correlates positively with blood pressure (BP). We have recently revealed that acute intracerebroventricular (i.c.v.) injection of chemerin-9, an active fragment of human chemerin, increased systemic BP in normal Wistar rats, suggesting that chemerin is involved in the central nervous control of peripheral BP. After secreted as an inactive form as prochemerin, a mature form of active chemerin is produced through the cleavage of its carboxyl (C)-terminus by proteases. Although the activity of cleaved products of chemerin has been examined in vitro, in vivo effects remained to be elusive. In order to explore them, we performed acute i.c.v. injection of mouse chemerin-9 (mChemerin-9; 148F-156S), mouse chemerin-8 (mChemerin-8; 148F-155F), and mouse chemerin-7 (mChemerin-7; 148F-154A) into Wistar rats, and examined the effects on systemic BP. After chemerin fragment (1–30 nmol/head, i.c.v.) was cumulatively administered, systemic BP was measured by a cannulation method under an isoflurane anesthesia. mChemerin-9 but not mChemerin-8 and -7 induced a pressor response, which was concentration-dependent. In conclusion, we for the first time demonstrated that mChemerin-9 that corresponds to the C-terminal nine amino acids of active mouse chemerin156S increased systemic BP in rats, and also that chemerin fragments showed different effects on systemic BP dependent on how their C-terminus was cleaved.     

Epidermal hyperplasia of walleye, Stizostedion vitreum vitreum (Mitchill), associated with retrovirus-like type-C particles: prevalence, histologic and electron microscopic observations

Epidermal hyperplasia consisting of discrete translucent raised outgrowths of cells were observed on the skin of walleye, Stizostedion vitreum vitreum (Mitchill), during their spawning period in the spring. The cells constituting the hyperplastic growths were limited to the epidermal layer, and were associated with surface budded, 120-nm-diameter, retrovirus-like particles located in the expanded intercellular spaces. These tumour-like growths were distinct from the other virus-associated skin lesions of walleye including dermal sarcoma, lymphocystis disease and herpesvirus-associated hyperplasia. Lesions could be differentiated by careful observation in the field and comparison of portions of each growth by histologic and electron microscopic observations.     

Partial replacement of fish meal with plant protein sources using organic acids to practical diets for juvenile yellowtail,Seriola quinqueradiata

An experiment was conducted to investigate the effects of phosphorus (P), citric acid (CA) and formic acid (FA) supplementation on growth and loading of nitrogen (N) and phosphorus (P) in juvenile yellowtail fed fish meal (FM) and alternate plant protein (APP) diets. Six diets designated as F49 (490 g kg^?1 FM‐based), F49 + P (490 g kg^?1 FM with 5 g kg^?1 inorganic P), F31 + CA (310 g kg^?1 FM containing 200 g kg^?1 APP with 5 g kg^?1 CA), F31 + FA (310 g kg^?1 FM having 200 g kg^?1 APP with 4 g kg^?1 FA), F23 + CA (230 g kg^?1 FM containing 300 g kg^?1 APP with 5 g kg^?1 CA) and F23 + FA (230 g kg^?1 FM having 300 g kg^?1 APP with 4 g kg^?1 FA) were formulated. Yellowtails were fed one of the diets for 12 weeks under on‐site conditions at water temperature 22.0–27.0 °C. F49 + P gave the best growth, while F23 + CA the lowest. Specific growth rate and feed conversion ratio of F31 + CA were similar with control group. Addition of CA to APP diets significantly increased retention of P; hence, its excretion was lowered. The results of this study suggest that FM could be replaced up to 70% with the addition of 5 g kg^?1 CA to a low P‐containing plant protein sources diet without inorganic P supplementation in juvenile yellowtail diets, which in turn would enable a reduction in environmental pollution from aquafeeds.