Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus     

Toru Kobayashi  Ryo Ishibashi  Shinji Yamamoto  Satoshi Otani  Koichi Ueno  Osamu Murata 《Aquaculture Research》2011,42(2):230-239

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.  相似文献   

Targeted Knock-in of a Fluorescent Protein Gene into the Chicken Vasa Homolog Locus of Chicken Primordial Germ Cells using CRIS-PITCh Method     

Ryo Ezaki  Kennosuke Ichikawa  Mei Matsuzaki  Hiroyuki Horiuchi 《The Journal of Poultry Science》2022,59(2):182

In chickens, primordial germ cells (PGCs) are effective targets for advanced genome editing, including gene knock-in. Although a long-term culture system has been established for chicken PGCs, it is necessary to select a gene-editing tool that is efficient and precise for editing the PGC genome while maintaining its ability to contribute to the reproductive system. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) and CRISPR-mediated precise integration into the target chromosome (CRIS-PITCh) methods are superior as the donor vector is easier to construct, has high genome editing efficiency, and does not select target cells, compared to the homologous recombination method, which has been conventionally used to generate knock-in chickens. In this study, we engineered knock-in chicken PGCs by integrating a fluorescent protein gene cassette as a fusion protein into the chicken vasa homolog (CVH) locus of chicken PGCs using the CRIS-PITCh method. The knock-in PGCs expressed the fluorescent protein in vitro and in vivo, facilitating the tracking of PGCs. Furthermore, we characterized the efficiency of engineering double knock-in cell lines. Knock-in cell clones were obtained by limiting dilution, and the efficiency of engineering double knock-in cell lines was confirmed by genotyping. We found that 82% of the analyzed clones were successfully knocked-in into both alleles. We suggest that the production of model chicken from the knock-in PGCs can contribute to various studies, such as the elucidation of the fate of germ cells and sex determination in chicken.  相似文献   

Oocyte transport: Developmental competence of bovine oocytes arrested at germinal vesicle stage by cycloheximide under air     

Hashimoto S  Kimura K  Iwata H  Takakura R 《The Journal of reproduction and development》2003,49(1):61-66

The effects of the medium (TCM 199 or SOFaa) and temperature (20 or 39 C) during meiotic arrest by cycloheximide (CHX) under air on the developmental competence of bovine oocytes after in vitro maturation (IVM) and fertilization (IVF) were investigated. Oocytes were maintained in meiotic arrest by 10 microg/ml CHX in a 50-microl droplet of 25-mM HEPES-buffered TCM 199 (H199) at 39 C or synthetic oviduct fluid (HSOFaa) at 20 or 39 C in air for 24 h. After release from the arrest, the oocytes was matured and fertilized in vitro and their developmental competence was examined. The developmental rate of oocytes arrested in HSOFaa at 20 C to the blastocyst stage was similar to that of non-arrested oocytes but was significantly higher (P<0.05) than that of oocytes arrested at 39 C in H199 or in HSOFaa. In consideration of oocyte transport conditions, we also investigated the meiotic arrest of oocytes maintained in a 0.25-ml straw by CHX individually with 10 microl HSOFaa or as a group (40-50 oocytes) with 170-200 microl HSOFaa at 20 C in air for 24 h. After release from meiotic arrest, the developmental competence of these oocytes was assessed similarly. The developmental rate of oocytes treated with CHX individually was similar to that of those treated with CHX in 50-microl droplet of HSOFaa at 20 C. However, the developmental rate of oocytes treated with CHX as a group was lower than that of oocytes treated with CHX in a 50-microl droplet. Five blastocysts developed from oocytes maintained in meiotic arrest in a plastic straw were transferred to five recipient heifers. Consequently, three recipients became pregnant and 2 calves were delivered. The results of the present study indicate that bovine oocytes treated with CHX in HSOFaa at 20 C under air retain the same developmental competence as non-arrested oocytes.  相似文献   

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence     

Wakita Y  Kawano J  Shimizu A  Hajek V  Tomisaka E  Yasuda R  Matsuo E 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(7):603-605

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.  相似文献   

Distribution of mecA-harboring staphylococci in healthy mares     

Yasuda R  Kawano J  Matsuo E  Masuda T  Shimizu A  Anzai T  Hashikura S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(9):821-827

The prevalence of staphylococci that harbor the mecA gene responsible for methicillin resistance was examined in healthy breeding mares. Staphylococci often cause diseases of horses such as metritis, keratitis, and abscess. Methicillin-resistant staphylococci would make antibiotic treatments ineffective, so it may be significant to know the distribution of mecA-harboring staphylococci in mares. Isolation of mecA-harboring staphylococci was achieved from nares and pasterns of 100 mares in Hokkaido, Japan. From 13% of the mares, mecA-harboring staphylococci, including 15 isolates of Staphylococcus sciuri and 3 of Staphylococcus lentus, were isolated. Isolates of S. sciuri were found to be genetically polyclonal by pulsed-field gel electrophoresis. These isolates produced no PCase and showed low or no resistance to beta-lactam and other classes of antibiotics. Distribution of staphylococcal species and levels of antibiotic resistance were found to be different between isolates from the present mares and those previously reported from riding-horses. Antibiotic pressure may lead to these differences. In addition, it appears that mecA-harboring S. sciuri may be native to horses.  相似文献   

Quantitative structure-activity relationships of herbicidal N′-substituted phenyl-N-methoxy-N-methylureas     

Ichiki Takemoto  Ryo Yoshida  Seizo Sumida  Katsuzo Kamoshita 《Pesticide biochemistry and physiology》1985,23(3):341-348

Relationships between three types of herbicidal activity of N′-substituted phenyl-N-methoxy-N-methylureas and substitution at the benzene ring were analyzed by the Hansch-Fujita method. First, the Hill inhibitory activity was correlated with electronic (σ) as well as hydrophobic (π) substituent constants. The existence of an optimum value of hydrophobicity for substituents was suggested to reach the target site of action. Second, bliaching activity observed for the 3-substituted but not for 4-substituted compounds was correlated with π, σ, and steric substituent constant, E_s. Third, the postemergent herbicidal activity was shown to correlate linearly with the Hill inhibitory activity, pI₅₀, and hydrophobic parameter, π.  相似文献   

Variations in the morphology of Rhizomucor pusillus in granulomatous lesions of a Magellanic penguin (Spheniscus magellanicus)     

Fumiko SUZUTA  Kumiko KIMURA  Ryo URAKAWA  Yukio KUSUDA  Shogo TANAKA  Yasuko HANAFUSA  Makoto HARITANI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(8):1029-1031

This report presents a new case of mucormycosis encountered in penguin characterized by morphological variation of hyphae and presence of sporangia with numerous sporangiospores. A 4.5-year-old Magellanic penguin (Spheniscus magellanicus) died after exhibiting anorexia, poor nutritional condition and dyspnea. Multiple nodular lesions were observed in the thoracic and abdominal regions. Histopathologically, hyphae of various sizes were seen in the lungs, air sac and nodular lesions. Myriad sporangiospores and several sporangia were observed in/around the bronchi or parabronchi. The very narrow and short hyphae in the nodules were not consistent with the characteristics of Mucorales. However, for most hyphae, including those in the nodules, sporangiospores and sporangia, immunohistochemistry revealed Mucorales-positive reactions. In addition, these fungi were identified as Rhizomucor pusillus by gene analysis.  相似文献   

Interactions of knowledge systems in shiitake mushroom production: a case study on the Noto Peninsula,Japan     

Ryo Kohsaka  Mitsuyuki Tomiyoshi  Osamu Saito  Shizuka Hashimoto  Leah Mohammend 《Journal of Forest Research》2015,20(5):453-463

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Estimating harvest costs and projecting quantities of logging residues for small-scale forestry in Nasushiobara,Tochigi Prefecture,Japan     

Chikara Nakahata  Ryo Uemura  Masashi Saito  Kanae Kanetsuki  Kazuhiro Aruga 《林业研究》2014,25(4):965-974

We used GIS on a regional scale to estimate and compare supply potentials and costs of small-scale logging systems, a mini-forwarder and a 4-ton truck operated by private logging contractors, and manual logging and a light truck operated by individual forest own- ers, with the mechanized operational system of the Forest Owners＇ Asso- ciation. Total potential yields of timber and logging residues were esti- mated as 418,895 m3 and 254,962 m3, respectively. The economic bal- ances were estimated and available amounts were projected as supply potentials from profitable sub-compartments. As a result, available amounts of timber and logging residues were estimated at 375,466 m3 （89.9%） and 203,850 m3 （80.0%）, respectively. Because their transport expenses were lower than for other systems the most profitable sub-compartments were operated by private logging contractors who sold logging residues at a plant. The profitable sub-compartments operated by individual forest owners were few because the extracting distances were usually greater than 20 m. Raising logging residue prices from 3,000 yen·m-3 to 4,080 yen·m-3 or 6,800 yen·m-3, and establishing forest roads, which reduced some extracting distances to less than 20 m, increased the number and area of profitable sub＊compartments, and increased available amounts of logging residues.  相似文献   

Examining the Optimal Method to Extract Logging Residues from Small-Scale Forestry in the Nasunogahara Area,Tochigi Prefecture,Japan     

Chikara Nakahata  Kazuhiro Aruga  Ryo Uemura  Masashi Saito  Kanae Kanetsuki 《Small-Scale Forestry》2014,13(2):251-266

This study examines the optimal method to extract logging residues from small, fragmented and scattered forests separately from the mechanized operational systems used on aggregated forestry operation sites. First, small-scale logging systems operated by a private logging contractor and an individual forest owner were investigated. Regression equations for estimating felling and processing cycle times were established as a function of stem and log volumes, respectively. Equations to estimate the forwarding and transporting cycle times were established as a function of the forwarding and transporting distances using the forwarding and transporting velocities, respectively. Then, equations to estimate productivities and costs were established with the cycle times, volumes and hourly operational expenses consisting of labour and machinery expenses. Finally, costs of the small-scale systems—mini-forwarder and 4 t truck operated by a private logging contractor, and manual logging and light truck operated by an individual forest owner—were estimated and compared with the mechanized operation system operated by the Forest Owners’ Association. The mechanized operation system was found to have the highest cost and could not compete for small forestry operational sites and small stem volumes. The small-scale systems could be effective for harvesting small areas. This was especially true when conducting extracting operations without machines, as done by the individual forest owner, and transporting operations by a private logging contractor, in which the hourly income of the individual forest owner was higher than the hourly labour expenses.  相似文献