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Silvia Ortubay Víctor Cussac Miguel Battini Juan Barriga Juana Aigo Marcelo Alonso Patricio Macchi Mariana Reissig Joy Yoshioka Stanley Fox 《水产资源保护:海洋与淡水生态系统》2006,16(1):93-105
- 1. Laguna Blanca, in Laguna Blanca National Park, is a lake in Patagonia which has been designated as a Ramsar site since 1971 because of bird diversity and abundance and importance for nesting, particularly for the black neck swan, Cygnus melanocoryphus. It is also valued for its populations of endemic amphibians, Atelognathus patagonicus and Atelognathus praebasalticus.
- 2. Avian and amphibian populations have decreased dramatically in recent years. Percichthys colhuapiensis, Percichthys trucha (Pisces, Percichthyidae), Salmo trutta and Oncorhynchus mykiss (Pisces, Salmonidae) were introduced into Laguna Blanca in 1965. Since 1986, no Atelognathus frogs have been found. The abundance of swans and coots, which are strongly associated with macrophytes for nesting and feeding, has diminished drastically, whereas piscivorous birds have increased.
- 3. The fishless condition of some neighbouring small lakes with abundant pond weeds, aquatic birds and endemic amphibians, was assessed in order to compare the physical and chemical characteristics and the quantitative composition of the benthos among lakes. Fish presence at Laguna Blanca and its absence at El Burro, Antiñir and Jabón lakes, were confirmed. Compared with previous results, it seems that the abundance of Amphipoda (Hyalella), Copepoda and Cladocera at Laguna Blanca has decreased, while Acari has increased and Notostraca has disappeared. Water transparency has diminished in Laguna Blanca and now is lower than that of fishless lakes.
- 4. P. colhuapiensis were captured only in Laguna Blanca, with the highest captures in the littoral zone. The population shows rapid individual growth in the early years and an absence of fish older than 6 years. The preponderance of benthos and the presence of macrophytes in the gut contents of adult P. colhuapiensis appear to indicate that they are important consumers of these resources.
- 5. This paper concludes that fish introduction in Laguna Blanca led to a complex trophic cascade effect (fish predation on tadpoles, fish competition for avian food, bottom disturbance, zooplankton reduction) producing deleterious effects on the amphibian and bird populations.
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Fuminori TANIHARA Michiko NAKAI Hiroyuki KANEKO Junko NOGUCHI Takeshige OTOI Kazuhiro KIKUCHI 《The Journal of reproduction and development》2013,59(4):385-392
In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm
penetration or prevention of polyspermy is not well understood. In the present study, we
investigated the effects of the ZP on sperm penetration during in vitro
fertilization (IVF). We collected in vitro-matured oocytes with a first
polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two
treatments (pronase and mechanical pipetting), and the effects of these treatments on
sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per
oocyte) were evaluated. There was no evident difference in the parameters between the two
groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes
using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+
oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the
sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation).
The proportions of oocytes penetrated by sperm increased significantly with time in both
groups; however, the number of penetrated sperm per oocyte did not increase in ZP−
oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and
prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5
h group than in the control group. These results suggest that the ZP and oolemma are not
competent factors for prevention of polyspermy in our present porcine IVF system. However,
it appears that ZP removal is one of the possibilities for reducing polyspermic
penetration in vitro in pigs. 相似文献
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Quynh A. Le Maki Hirata Nhien T. Nguyen Koki Takebayashi Manita Wittayarat Yoko Sato Zhao Namula Masahiro Nii Fuminori Tanihara Takeshige Otoi 《Animal Science Journal》2020,91(1)
This study was conducted to investigate the effect of seven concentrations of Cas9 protein (0, 25, 50, 100, 200, 500, and 1,000 ng/µl) on the development and gene editing of porcine embryos. This included the target editing and off‐target effect of embryos developed from zygotes that were edited via electroporation of the Cas9 protein with guide RNA targeting Myostatin genes. We found that the development to blastocysts of electroporated zygotes was not affected by the concentration of Cas9 protein. Although the editing rate, which was defined as the ratio of edited blastocysts to total examined blastocysts, did not differ with Cas9 protein concentration, the editing efficiency, which was defined as the frequency of indel mutations in each edited blastocyst, was significantly decreased in the edited blastocysts from zygotes electroporated with 25 ng/µl of Cas9 protein compared with that of blastocysts from zygotes electroporated with higher Cas9 protein concentrations. Moreover the frequency of indel events at the two possible off‐target sites was not significantly different with different concentrations of Cas9 protein. These results indicate that the concentration of Cas9 protein affects gene editing efficiency in embryos but not the embryonic development, gene editing rate, and non‐specific cleavage of off‐target sites. 相似文献
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Fuminori Tanihara Maki Hirata Nhien T. Nguyen Quynh A. Le Takayuki Hirano Tatsuya Takemoto Michiko Nakai Dai‐ichiro Fuchimoto Takeshige Otoi 《Animal Science Journal》2019,90(1):55-61
Recently, we established the GEEP (“gene editing by electroporation of Cas9 protein”) method, in which the CRISPR/Cas9 system, consisting of a Cas9 protein and single guide RNA (sgRNA), is introduced into pig zygotes by electroporation and thus induces highly efficient targeted gene disruption. In this study, we examined the effects of sgRNA on the blastocyst formation of porcine embryos and evaluated their genome‐editing efficiency. To produce an animal model for diabetes, we targeted PDX‐1 (pancreas duodenum homeobox 1), a gene that is crucial for pancreas development during the fetal period and whose monoallelic disruption impairs insulin secretion. First, Cas9 protein with different sgRNAs that targeted distinct sites in the PDX‐1 exon 1 was introduced into in vitro‐fertilized zygotes by the GEEP method. Of the six sgRNAs tested, three sgRNAs (sgRNA1, 2, and 3) successfully modified PDX‐1 gene. The blastocyst formation rate of zygotes edited with sgRNA3 was significantly (p < 0.05) lower than that of control zygotes without the electroporation treatment. Our study indicates that the GEEP method can be successfully used to generate PDX‐1 mutant blastocysts, but the development and the efficiency of editing the genome of zygotes may be affected by the sgRNA used for CRISPR/Cas9 system. 相似文献
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Fuminori Tanihara Maki Hirata Satoru Iizuka Shinya Sairiki Masahiro Nii Nhien Thi Nguyen Quynh Anh Le Takayuki Hirano Takeshige Otoi 《Animal Science Journal》2019,90(6):712-718
The aim of this study was to investigate the ovarian follicular development, developmental competence of oocytes, and plasma anti‐Müllerian hormone (AMH) levels of Japanese wild boar crossbred (wild hybrid) gilts, whose litter size is inferior to that of European breeds. Ovary and plasma samples were collected from two different breeds of gilts (wild hybrid and Large White breeds). The ovaries from the wild hybrid gilts had a lower average numbers of secondary follicles and vesicular follicles in ovarian cross‐sections and of good quality oocytes collected from ovarian follicles as compared with those from Large White gilts (p < 0.05). The development rate to the blastocyst stage of good quality oocytes after in vitro maturation, fertilization and culture was also lower (p < 0.05) in wild hybrid gilts than in Large White gilts. Plasma AMH levels with >0.16 ng/ml were detected in 8.3% of the examined wild hybrid gilts and 33% of the Large White gilts. These results indicate that the low reproductive performance of wild hybrid breed may result in part from low numbers of vesicular follicles and good quality oocytes, and low developmental competence of oocytes. Moreover, plasma AMH levels may support low number of vesicular follicles in ovaries of wild hybrid gilts. 相似文献
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Thanh‐Van Nguyen Lanh Thi Kim Do Tamas Somfai Takeshige Otoi Masayasu Taniguchi Kazuhiro Kikuchi 《Animal Science Journal》2019,90(12):1530-1536
Chlorogenic acid (CGA) is known to protect oocytes from oxidative stress. Here we investigated the effects of CGA on porcine oocyte maturation under heat stress and subsequent embryonic development after parthenogenetic activation. For in vitro maturation (IVM) at 41.0°C (hyperthermic condition), supplementation of the maturation medium with 50 μM CGA significantly improved the percentage of matured oocytes and reduced the rate of apoptosis relative to oocytes matured without CGA (p < .05). CGA treatment of oocytes during IVM under hyperthermia tended to increase (p < .1) percentage of blastocyst formation after parthenogenesis and significantly increased (p < .05) the total cell number per blastocyst relative to oocytes matured without CGA. For IVM at 38.5°C (isothermic condition), CGA significantly improved the rate of blastocyst development compared with oocytes matured without CGA (p < .05), but did not affect oocyte maturation, apoptosis rate or the number of cells per embryo. Omission of all antioxidants from the IVM medium significantly reduced the rate of oocyte maturation, but the rate was restored upon addition of CGA. These results demonstrate that CGA is a potent antioxidant that protects porcine oocytes from the negative effects of heat stress, thus reducing the frequency of apoptosis and improving the quality of embryos. 相似文献
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Manita Wittayarat Maki Hirata Zhao Namula Yoko Sato Nhien T. Nguyen Quynh A. Le Qingyi Lin Koki Takebayashi Fuminori Tanihara Takeshige Otoi 《Animal Science Journal》2021,92(1):e13534
This study aimed to investigate the efficiency of KRAS gene editing via CRISPR/Cas9 delivery by electroporation and analyzed the effects of the non-homologous end-joining pathway inhibitor Scr7 and single-stranded oligodeoxynucleotide (ssODN) homology arm length on introducing a point mutation in KRAS. Various concentrations (0–2 µM) of Scr7 were evaluated; all concentrations of Scr7 including 0 µM resulted in the generation of blastocysts with a point mutation and the wild-type sequence or indels. No significant differences in the blastocyst formation rates of electroporated zygotes were observed among ssODN homology arm lengths, irrespective of the gRNA (gRNA1 and gRNA2). The proportion of blastocysts carrying a point mutation with or without the wild-type sequence and indels was significantly higher in the ssODN20 group (i.e., the group with a ssODN homology arm of 20 bp) than in the ssODN60 group (gRNA1: 25.7% vs. 5.4% and gRNA2: 45.5% vs. 5.9%, p < .05). In conclusion, the CRISPR/Cas9 delivery with ssODN via electroporation is feasible for the generation of point mutations in porcine embryos. Further studies are required to improve the efficiency and accuracy of the homology-directed repair. 相似文献