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 | Acceso al texto completo restringido a Biblioteca INIA Las Brujas. Por información adicional contacte bibliolb@inia.org.uy. |
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha : |
13/07/2023 |
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Actualizado : |
13/07/2023 |
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Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
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Autor : |
DOS SANTOS-NETO, P.C.; CUADRO, F.; SOUZA-NEVES, M.; CRISPO, M.; MENCHACA, A. |
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Afiliación : |
P.C. DOS SANTOS-NETO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; F. CUADRO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; M. SOUZA-NEVES, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Uruguay; M. CRISPO, Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Uruguay; JOSE ALEJO MENCHACA BARBEITO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay. |
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Título : |
Refinements in embryo manipulation applied to CRISPR technology in livestock. |
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Fecha de publicación : |
2023 |
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Fuente / Imprenta : |
Theriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028 |
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ISSN : |
0093-691X |
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DOI : |
10.1016/j.theriogenology.2023.05.028 |
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Idioma : |
Inglés |
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Notas : |
Article history: Received 14 April 2023; Received in revised form 29 May 2023; Accepted 29 May 2023; Available online 9 June 2023. -- Correspondence author: Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; email:menchaca.alejo@gmail.com -- FUNDING: This study was financially supported by Fundación IRAUy, FOCEM (MERCOSUR Structural Convergence Fund), COF 03/11, and the Uruguayan National Research Agency (ANII, Uruguay). PCdSN received a scholarship from the National Council for Scientific Technological Development (CNPq, Brazil). AM, MC, and PCdSN are fellows of Sistema Nacional de Investigadores (SNI, ANII) of Uruguay and PEDECIBA. -- |
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Contenido : |
The implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), and subjected to vitrification/warming via the Cryotop method (n = 474), while a subset of embryos were left fresh as control group (n = 75). Embryos were transferred into the uterine horn of recipient females at prefixed time 8.5 days after the estrous synchronization treatment (i.e., approximately six days after ovulation). Pregnancy rate (30.8% vs. 48.0%), embryo survival rate (14.8% vs. 21.3%), and birth rate (85.7% vs. 75.0%) were not different (P[dbnd]NS) between vitrified and fresh embryos, respectively. In conclusion, the current study in sheep embryos reports (a) suitable developmental rate after CRISPR/Cas microinjection (i.e., 20%), even though it was lower than non-microinjected zygotes; (b) similar outcomes when Day 2-embryos were placed into the uterine horn instead of the oviduct, avoiding both time-consuming and invasive oviduct manipulation, and extended in vitro culture during one week; (c) promising pregnancy and birth rates obtained with vitrification of CRISPR/Cas microinjected embryos. This knowledge on in vitro embryo development, timing of embryo transfer, and cryopreservation of CRISPR/Cas microinjected zygotes have practical implications for the implementation of genome editing technology in large animals. © 2023 Elsevier Inc. MenosThe implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), ... Presentar Todo |
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Palabras claves : |
Early embryo transfer; Gene editing; In vitro embryo production; Minimum volume vitrification. |
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Asunto categoría : |
L10 Genética y mejoramiento animal |
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Marc : |
LEADER 04268naa a2200253 a 4500
001 1064251
005 2023-07-13
008 2023 bl uuuu u00u1 u #d
022 $a0093-691X
024 7 $a10.1016/j.theriogenology.2023.05.028$2DOI
100 1 $aDOS SANTOS-NETO, P.C.
245 $aRefinements in embryo manipulation applied to CRISPR technology in livestock.$h[electronic resource]
260 $c2023
500 $aArticle history: Received 14 April 2023; Received in revised form 29 May 2023; Accepted 29 May 2023; Available online 9 June 2023. -- Correspondence author: Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; email:menchaca.alejo@gmail.com -- FUNDING: This study was financially supported by Fundación IRAUy, FOCEM (MERCOSUR Structural Convergence Fund), COF 03/11, and the Uruguayan National Research Agency (ANII, Uruguay). PCdSN received a scholarship from the National Council for Scientific Technological Development (CNPq, Brazil). AM, MC, and PCdSN are fellows of Sistema Nacional de Investigadores (SNI, ANII) of Uruguay and PEDECIBA. --
520 $aThe implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), and subjected to vitrification/warming via the Cryotop method (n = 474), while a subset of embryos were left fresh as control group (n = 75). Embryos were transferred into the uterine horn of recipient females at prefixed time 8.5 days after the estrous synchronization treatment (i.e., approximately six days after ovulation). Pregnancy rate (30.8% vs. 48.0%), embryo survival rate (14.8% vs. 21.3%), and birth rate (85.7% vs. 75.0%) were not different (P[dbnd]NS) between vitrified and fresh embryos, respectively. In conclusion, the current study in sheep embryos reports (a) suitable developmental rate after CRISPR/Cas microinjection (i.e., 20%), even though it was lower than non-microinjected zygotes; (b) similar outcomes when Day 2-embryos were placed into the uterine horn instead of the oviduct, avoiding both time-consuming and invasive oviduct manipulation, and extended in vitro culture during one week; (c) promising pregnancy and birth rates obtained with vitrification of CRISPR/Cas microinjected embryos. This knowledge on in vitro embryo development, timing of embryo transfer, and cryopreservation of CRISPR/Cas microinjected zygotes have practical implications for the implementation of genome editing technology in large animals. © 2023 Elsevier Inc.
653 $aEarly embryo transfer
653 $aGene editing
653 $aIn vitro embryo production
653 $aMinimum volume vitrification
700 1 $aCUADRO, F.
700 1 $aSOUZA-NEVES, M.
700 1 $aCRISPO, M.
700 1 $aMENCHACA, A.
773 $tTheriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028
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INIA Las Brujas (LB) |
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Registro completo
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Biblioteca (s) : |
INIA La Estanzuela. |
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Fecha actual : |
20/01/2022 |
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Actualizado : |
18/03/2022 |
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Autor : |
CLOTHIER, K.A.; WATSON, K.D.; METE, A.; GIANNITTI, F.; ANDERSON, M.; MUNK, B.; MCMILLIN, S.; CLIFFORD, D.L.; RUDD, J.; SHIRKEY, N.; FAMINI, D.; WOODS, L. |
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Afiliación : |
KRISTIN A. CLOTHIER, California Animal Health and Food Safety Laboratory System, University of California-Davis, CA, USA.; KATHERINE D. WATSON, California Animal Health and Food Safety Laboratory System, University of California-Davis, CA, USA.; ASLI METE, California Animal Health and Food Safety Laboratory System, University of California-Davis, CA, USA.; FEDERICO GIANNITTI, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; California Department of Fish and Wildlife, Rancho Cordova, CA, USA.; DAN FAMINI, Sonoma County Wildlife Rescue, Petaluma, CA, USA.; LESLIE WOODS, California Animal Health and Food Safety Laboratory System, University of California-Davis, CA, USA. |
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Título : |
Generalized dermatophytosis caused by Trichophyton equinum in 8 juvenile black bears in California. |
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Fecha de publicación : |
2022 |
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Fuente / Imprenta : |
Journal of Veterinary Diagnostic Investigation, 2022, Volume 34, Issue 2, pp.279-283. doi: https://doi.org/10.1177/10406387211061143 |
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DOI : |
10.1177/10406387211061143 |
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Idioma : |
Inglés |
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Contenido : |
Abstract. From 2014?2019, 8 juvenile black bears (Ursus americanus) from different geographic regions were presented to the California Department of Fish and Wildlife because of emaciation, alopecia, and exfoliative dermatitis that resulted in death or euthanasia. Autopsy and histopathology revealed that all 8 bears had generalized hyperkeratotic dermatitis, folliculitis, and furunculosis. Skin structures were heavily colonized by fungal hyphae and arthrospores; fungal cultures of skin from 7 bears yielded Trichophyton equinum, a zoophilic dermatophyte reported only rarely in non-equid species. Additional skin conditions included mites (5), ticks (2), and coagulase-negative Staphylococcus sp. infections (2). No other causes of morbidity or mortality were identified. Molecular comparisons performed at the University of Texas Fungal Reference Laboratory determined that all isolates produced identical banding patterns, potentially representing a clonal population.
Dermatophytosis is commonly localized and limited to the stratum corneum of the epidermis and hair follicles. Generalized disease with dermal involvement is rare in immunocompetent individuals; illness, malnutrition, age, or immunosuppression may increase susceptibility. Underlying causes for the severe disease impact in these bears were not evident after physical or postmortem examination. The mechanism by which bears from different geographic locations had severe, T. equinum?associated dermatophytosis from a potentially clonal dermatophyte could not be explained and warrants further investigation. MenosAbstract. From 2014?2019, 8 juvenile black bears (Ursus americanus) from different geographic regions were presented to the California Department of Fish and Wildlife because of emaciation, alopecia, and exfoliative dermatitis that resulted in death or euthanasia. Autopsy and histopathology revealed that all 8 bears had generalized hyperkeratotic dermatitis, folliculitis, and furunculosis. Skin structures were heavily colonized by fungal hyphae and arthrospores; fungal cultures of skin from 7 bears yielded Trichophyton equinum, a zoophilic dermatophyte reported only rarely in non-equid species. Additional skin conditions included mites (5), ticks (2), and coagulase-negative Staphylococcus sp. infections (2). No other causes of morbidity or mortality were identified. Molecular comparisons performed at the University of Texas Fungal Reference Laboratory determined that all isolates produced identical banding patterns, potentially representing a clonal population.
Dermatophytosis is commonly localized and limited to the stratum corneum of the epidermis and hair follicles. Generalized disease with dermal involvement is rare in immunocompetent individuals; illness, malnutrition, age, or immunosuppression may increase susceptibility. Underlying causes for the severe disease impact in these bears were not evident after physical or postmortem examination. The mechanism by which bears from different geographic locations had severe, T. equinum?associated dermatophytosis from a potent... Presentar Todo |
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Palabras claves : |
Bears; Dermatophytosis; PLATAFORMA DE INVESTIGACIÓN EN SALUD ANIMAL; PLATAFORMA DE SALUD ANIMAL; Trichophyton equinum; Ursus americanus. |
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Asunto categoría : |
-- |
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Marc : |
LEADER 02641naa a2200337 a 4500
001 1062648
005 2022-03-18
008 2022 bl uuuu u00u1 u #d
024 7 $a10.1177/10406387211061143$2DOI
100 1 $aCLOTHIER, K.A.
245 $aGeneralized dermatophytosis caused by Trichophyton equinum in 8 juvenile black bears in California.$h[electronic resource]
260 $c2022
520 $aAbstract. From 2014?2019, 8 juvenile black bears (Ursus americanus) from different geographic regions were presented to the California Department of Fish and Wildlife because of emaciation, alopecia, and exfoliative dermatitis that resulted in death or euthanasia. Autopsy and histopathology revealed that all 8 bears had generalized hyperkeratotic dermatitis, folliculitis, and furunculosis. Skin structures were heavily colonized by fungal hyphae and arthrospores; fungal cultures of skin from 7 bears yielded Trichophyton equinum, a zoophilic dermatophyte reported only rarely in non-equid species. Additional skin conditions included mites (5), ticks (2), and coagulase-negative Staphylococcus sp. infections (2). No other causes of morbidity or mortality were identified. Molecular comparisons performed at the University of Texas Fungal Reference Laboratory determined that all isolates produced identical banding patterns, potentially representing a clonal population. Dermatophytosis is commonly localized and limited to the stratum corneum of the epidermis and hair follicles. Generalized disease with dermal involvement is rare in immunocompetent individuals; illness, malnutrition, age, or immunosuppression may increase susceptibility. Underlying causes for the severe disease impact in these bears were not evident after physical or postmortem examination. The mechanism by which bears from different geographic locations had severe, T. equinum?associated dermatophytosis from a potentially clonal dermatophyte could not be explained and warrants further investigation.
653 $aBears
653 $aDermatophytosis
653 $aPLATAFORMA DE INVESTIGACIÓN EN SALUD ANIMAL
653 $aPLATAFORMA DE SALUD ANIMAL
653 $aTrichophyton equinum
653 $aUrsus americanus
700 1 $aWATSON, K.D.
700 1 $aMETE, A.
700 1 $aGIANNITTI, F.
700 1 $aANDERSON, M.
700 1 $aMUNK, B.
700 1 $aMCMILLIN, S.
700 1 $aCLIFFORD, D.L.
700 1 $aRUDD, J.
700 1 $aSHIRKEY, N.
700 1 $aFAMINI, D.
700 1 $aWOODS, L.
773 $tJournal of Veterinary Diagnostic Investigation, 2022, Volume 34, Issue 2, pp.279-283. doi: https://doi.org/10.1177/10406387211061143
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