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Registros recuperados: 68
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Advantages and Applications of Cryopreservation in Fisheries Science BABT
Tsai,S.; Lin,C..
Cryopreservation is a long-term storage technique to preserve the biological material without deterioration for extended period of time at least several thousands of years. The ability to preserve and store both maternal and paternal gametes provides a reliable source of fish genetic material for scientific and aquaculture purposes as well as for conservation of biodiversity. Successful cryopreservation of fish sperm have been achieved for more than 200 fish species and many fish species have been adequated for the purpose of cryobanking. Cryopreservation of fish embryo is not viable, mainly because of the same limitations as in fish oocytes, i.e., high chilling sensitivity and low membrane permeability. However, cryopreservation of isolated embryonic...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cryopreservation; Sperm; Embryo; Oocyte; Blastomere.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000300014
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Association of recombinant bovine somatotropin (rBST) with equine chorionic gonadotropin (eCG) on antral follicle count and oocyte production in Holstein and Tabapuã heifers R. Bras. Zootec.
Morais,Hévea de; Spuri,Renata; Gonçalves,Tarcísio de Moraes; Carvalho,Rafaela Rodrigues de; Andrade,Renato Campos; Pinto,Tássia Louregiani Carvalho; Souza,José Camisão de.
The objective of this study was to investigate whether the use of rbST and eCG prior to ultrasound-guided follicular aspiration (OPU) improves oocyte yield and quality in Tabapuã and Holstein heifers. The study was conducted in two phases, 20 days apart, in a change-over design. The dominant follicle was ablated two days (D-2) before two treatments: stimulation (6 Holstein and 8 Tabapuã), 500 mg of rbST (Boostin®) on D0 and 500 IU of eCG (Folligon) on D2; and control (6 Holstein and 8 Tabapuã), in which heifers received injections of the excipient. Heifers were aspirated on D4. Oocytes were subjected to a well established commercial in vitro embryo production protocol (Vitrogen®) and embryos were evaluated seven days after fertilization. There was an...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryo; Hormonal stimulation; Ovary.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982012001200004
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Bovine conceptus of Bos indicus produced by somatic cell nuclear transfer and parthenogenesis present morphological variations since the blastocyst stage Arq. Bras. Med. Vet. Zootec.
Oliveira,F.D.; Sangalli,J.R.; Meirelles,F.V.; Perecin,F.; Silva Filho,P.P.C.; Watanabe,Y.F.; Miglino,M.A.; Assis Neto,A.C..
In cattle, embryo development is characterized by the appearance of two distinct cell layers, the trophectoderm and the inner cell mass. The latter will undergo differentiation to form the embryonic disc consisting of the epiblast and hypoblast. The aim of this study was to ultrastructurally characterize the bovine embryo from different in vitro production techniques, with emphasis on trophectoderm and inner cell mass cells. Bovine embryos on day 7 (conception = D1) of pregnancy, derived via in vitro production techniques, were fixed for light and transmission electron microscopy processing. Results suggested that embryos produced by nuclear transfer of somatic cells and parthenogenesis showed significant changes in macroscopic and microscopic structure....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryo; Blastocyst; In vitro production; Ultrastructure.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352015000601483
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Bovine embryo induces an anti-inflammatory response in uterine epithelial cells and immune cells in vitro: Possible involvement of interferon tau as an intermediator OAK
Talukder, Anup K.; Yousef, Mohamed S.; Rashid, Mohammad B.; Awai, Kensuke; Acosta, Tomas J.; Shimizu, Takashi; Okuda, Kiyoshi; Shimada, Masayuki; Imakawa, Kazuhiko; Miyamoto, Akio.
Recent observations suggest that the bovine uterus starts to react to the early embryo immediately after its arrival from the oviduct. The present study aimed to investigate the effect of the early developing embryo on the immune-related gene profile in bovine uterine epithelial cells (BUECs) in vitro, and to further examine the impact of conditioned media (CM), either from embryo-BUEC co-culture or embryo culture alone, on gene expression in peripheral blood mononuclear cells (PBMCs). First, BUECs were co-cultured with morulae (n = 10) for D5-D9 (D0 = IVF), and gene expression in BUECs was analyzed. Subsequently, PBMCs were cultured in CM from embryo-BUEC co-culture or D5-D9 embryo culture, and gene expression was evaluated. In BUECs, the embryo induced...
Palavras-chave: Bovine; Embryo; Immunity; Interferon tau (IFNT); Uterus.
Ano: 2017 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4532
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Bovine Oocytes and Early Embryos Express mRNA Encoding Glycerol Kinase but Addition of Glycerol to the Culture Media Interferes with Oocyte Maturation OAK
OKAWARA, Sumika; HAMANO, Seizo; TETSUKA, Masafumi; 手塚, 雅文.
Glycerol plays multi-functional roles in cellular physiology. Other than forming the backbone molecule for glycerophospholipid and triglyceride (TG), glycerol acts as an energy substrate for glycolysis. Spermatozoa are known to utilize glycerol for energy production, but there are no reports of this in oocytes. In this study, the value of glycerol as an energy substrate for bovine oocyte maturation (Exp. 1) and the gene expression of glycerol kinase (GK), an enzyme crucial for cellular glycerol utilization, in bovine oocytes and early embryos (Exp. 2) were examined. In Exp. 1, in vitro maturation (IVM) was conducted using synthetic oviduct fluid supplemented with/without glucose (1.5 mM) and/or glycerol (1.0 mM), and maturation rate, degree of cumulus...
Palavras-chave: Bovine; Embryo; Glycerol; Glycerol kinase (GK); Oocyte maturation.
Ano: 2009 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2829
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CHEMICAL COMPOSITION AND TETRAZOLIUM TEST OF Syagrus romanzoffiana (Cham.) Glassman SEEDS Rev. Bras. Frutic.
IOSSI,EMERSON; MORO,FABIOLA VITTI; VIEIRA,BRUNO GUILHERME TORRES LICURSI; BARBOSA,RAFAEL MARANI; VIEIRA,ROBERVAL DAITON.
ABSTRACT Queen palm is a tropical species of ecological importance and economic potential in various uses, including food. However, the knowledge about the nutritional value of fruits and the estimation of seed viability are still scarce. The aims of this work were to study the biometrics and chemical composition of fruits and seeds; describe the anatomy of embryos and assess the viability of the tetrazolium test. Mature fruits were collected, processed and submitted to biometry, water content, pulp and seeds chemical composition, germination and emergence in the laboratory evaluations. For the tetrazolium test, whole embryos were immersed in solution at 40 °C for one, two, four and six hours and compared with germination. The fruit pulp is rich in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Queen palm; Embryo; Germination; Palm; Pyrene.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452016000402001
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Co-culture of Mouse Two-cell Embryos on Human Endometrial Stromal Cells in Proliferative and Secretory Phases International Journal of Morphology
Fakheri,Taravat; Nankali,Anisodowleh; Khazaei,Mozafar; Ghanbari,Ali; Abedini,Mehrandokht; Saeidiborojeni,Hamidreza.
There were no significant differences in the distribution of embryos reaching to 2- cells, 4- cells, morula or blastocysts culturing on human endometrial stromal cells (Secretory or proliferative phases). The percent of morula in stage A (without fragmentation), stage B (<25% fragmentation), stage C (25-50% fragmentation) and stage D (&gt;50% fragmentation) and did not showed significant differences between two coculture groups. Thus, the phase that the endometrial stromal cells were in thereby did not affect on the quality of embryos.
Tipo: Journal article Palavras-chave: Co-culture; Embryo; Endometrium; Mice; Proliferative phase; Secretory phase.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022012000400014
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Comparison of Some Relative Gene Expressions in the Yolk Sac Membrane and Small Intestine of the Embryos in the Layer Hybrid and Pure Breeds Rev. Bras. Ciênc. Avic.
Onbaşilar,EE; Erdoğan,M; Erol,S; Kahraman,M; Güngör,ÖF.
ABSTRACT Pure breeds are important due to the fact that they adapt well to poor husbandry conditions. However, gene expression of digestive enzymes and nutrient transporters of pure breeds and layer hybrids have not been studied yet. Therefore, this study was directed to detect the gene expression of digestive enzymes and nutrient transporters in the yolk sac membrane and small intestine of the embryos in layer hybrid and pure breeds. A total of 354 hatching eggs were collected from 29 wk old Denizli, Gerze and Lohman White (LW) breeder flocks. Twelve eggs from each genotype were selected at embryonic day 19 (E19) and day 21 (E21). APN, SI, PepT1, EAAT3, CAT1, B0AT, SGLT1 and GLUT5 mRNA expressions were evaluated in the yolk sac membrane (YSM) and small...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryo; Gene expression; Layer hybrid; Pure breed.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2018000300435
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Composición y remoción de nutrimentos y calidad del fruto de mango (Mangifera indica L.) en diferentes ambientes. Colegio de Postgraduados
Mellado Vázquez, Adriana.
Se estudió la influencia de las condiciones de cultivo (clima, riego o temporal y tipo de suelo) en la composición nutrimental de los tejidos del fruto (epicarpio, pulpa, testa y embrión) y en la remoción de nutrimentos por tejido y por tonelada de fruto fresco de los cvs. Ataulfo, Kent, Keitt y Tommy Atkins. Asimismo, en ‘Ataulfo’ y ‘Tommy Atkins’ se estudió la relación entre el contenido nutrimental en suelo, hojas, epicarpio y pulpa (materia seca) y pulpa fresca, con la calidad postcosecha. Se trabajó con árboles altamente productivos (producción superior a los 150 kg·árbol-1), seleccionados en el 2008 de los que se cosecharon frutos en madurez fisiológica. En la materia seca de cada tejido se determinó la concentración de: N, P, K, Ca, Mg, S, Fe, Cu,...
Palavras-chave: Mangifera indica; Epicarpio; Pulpa; Testa; Embrión; Calidad postcosecha; Skin; Pulp; Seed coat; Embryo; Post-harvest quality; Doctorado; Fruticultura.
Ano: 2012 URL: http://hdl.handle.net/10521/736
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Cryopreservation of mouse morulae in glycerol, sucrose and honeybee royal jelly Braz. J. Vet. Res. Anim. Sci.
VISINTIN,José Antônio; GARCIA,José Fernando; PANTANO,Thais; D’ÁVILA ASSUMPÇÃO,Mayra Elena Ortiz.
Compacted mouse morulae were frozen at 0.3ºC/min. or 0.5ºC/min. from -6ºC to -24ºC or -32ºC in 10% of glycerol plus different sucrose concentrations with or without 0.1% of honeybee royal jelly. Embryos were thawed in water bath at 22ºC for 20 seconds and cryoprotectant dilution was done in three steps. Embryos were cultured in Whitten’s medium for 24, 48 and 72 hours at 37ºC, 5% of CO2 and 100% of humidity. The in vitro development ranged from 56.6% to 100% after 72 hours. Expanded blastocysts were transferred to pseudopregnant recipients on the third day of the estrous cycle. Viable fetuses rates for embryos frozen to -24 or -32ºC at 0.3ºC/minute in 10% glycerol + 10% sucrose, 10% glycerol + 10% sucrose + 0.1% honeybee royal jelly, 10% glycerol + 0.1%...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mice; Embryo; Cryopreservation; Sucrose; Royal jelly.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962000000400009
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Cryopreservation of mouse morulae through different methods: slow-freezing, vitrification and quick-freezing Braz. J. Vet. Res. Anim. Sci.
MELLO,Marco Roberto Bourg de; QUEIROZ,Vinícius Seixas; LIMA,Alessandra Sobreira de; TAVARES,Liliam Mara Trevisam; ASSUMPÇÃO,Mayra Elena Ortiz D'Avila; WHEELER,Mathew B.; VISINTIN,José Antonio.
The in vitro and in vivo development of mouse morulae after cryopreservation through different methods was examined. The slow-freezing involved an equilibration in 1.5M ethylene glycol (EG) and cooled at 0.5; 0.7; 1.0 or 1.2ºC/minute. The vitrification involved a 3 minutes equilibration in 20% EG and 60 seconds in solution containing 40% EG, 18% ficoll and 10.26% sucrose. The quick-freezing involved an equilibration in 3M EG + 0.3M sucrose for 5 minutes and 2 minutes in nitrogen vapor. In all three methods the straws were thawed in air for 10 seconds and in water at 25ºC for 20 seconds and the embryos cultured in vitro for 72 hours to estimate blastocyst rate. To assess viability in vivo, frozen morulae as well as fresh embryos were transferred into...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cryopreservation; Embryo; Mouse.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962001000400003
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Detection of equine herpesvirus 1 DNA in a single embryo and in horse semen by polymerase chain reaction Arq. Bras. Med. Vet. Zootec.
Carvalho,R.; Passos,L.M.F.; Oliveira,A.M.; Henry,M.; Martins,A.S..
The genome of one equine embryo and three equine semen specimens collected from a Brazilian farm were tested by polymerase chain reaction (PCR) for the presence of EHV-1 and EHV-4-specific timidine kinase (TK) sequences. The PCR detected specific EHV-1 TK gene sequences in all samples tested. The peripheral blood leukocytes (PBL) of the embryo donor mare also was amplified by EHV-1 TK primers. Infectious virus was not recovered from any specimens. The animals did not show any clinical signal of EHV-1 or EHV-4 infections. EHV-4 was not detected in the studied specimens. The results indicate that PCR was more sensitive than virus isolation in cell culture for detecting EHV-1 in semen of carrier horses.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Equine; Semen; Embryo; EHV-1; EHV-4; PCR.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352000000400002
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Differential bioaccumulation behaviour of Ag and Cd during the early development of the cuttlefish Sepia officinalis ArchiMer
Lacoue Labarthe, T; Warnau, A; Oberhansli, F; Teyssie, J; Koueta, Noussithe; Bustamante, Paco.
Cuttlefish eggs were exposed to background concentrations of dissolved Ag and Cd, using the radiotracers Ag-110m and Cd-109. At different time of the embryonic development (50 days), some eggs were placed in non-contaminating conditions. During the experiment, the uptake and depuration kinetics, and distribution of these metals among the egg compartments (i.e. eggshell, vitellus, peri-vitelline fluid and embryo) were assessed. In parallel, experiments were conducted with sub-lethal concentrations of stable Ag and Cd (2 and 1 mu g l(-1), respectively) to compare the metal behaviour at higher concentrations. From the spawning date up to I month of development, both metals were taken up efficiently by the eggs, reaching load/concentration ratio (LCR) of 1059...
Tipo: Text Palavras-chave: Cephalopod; Permeability; Eggshell; Embryo; Biokinetics; Silver; Cadmium.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-3882.pdf
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Does supplemental LH changes rate and time to ovulation and embryo yield in Santa Ines ewes treated for superovulation with FSH plus eCG? Ciência Rural
Oliveira,Maria Emilia Franco; Cordeiro,Mabel Freitas; Ferreira,Roberta Machado; Souza,Samuel Figueirêdo; Pieroni,Juliana Souza Pinto; Rodrigues,Luiz Fernando de Souza; Fonseca,Jeferson Ferreira da; Vicente,Wilter Ricardo Russiano.
The objective was to evaluate if supplemental LH given at the end of FSH treatment would synchronize the time of ovulation and increase the ovulation rate and embryo yield in Santa Ines ewes. Twenty superovulatory (SOV) programs were accomplished in cross-over design (60d interval). On D0, a CIDR device was inserted, and the device was replaced with a new one 7 days later, when 37.5µg of d-cloprostenol was administered. On D12, we started the SOV treatment, administering 256mg of pFSH 8 times, 12h apart. On D14, the CIDR was removed, and 200IU of eCG and 37.5µg of d-cloprostenol were administered. On D15, the ewes were allocated into one of two groups, a Control group (n=10) that received no supplemental LH and a LH group (n=10) treated with 7.5mg of LH...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Superovulation; LH; Ovulation; Embryo; Sheep.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782012000600021
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Effect of breeder age on eggshell thickness, surface temperature, hatchability and chick weigh Rev. Bras. Ciênc. Avic.
Gualhanone,A; Furlan,RL; Fernandez-Alarcon,MF; Macari,M.
Two experiments were carried out to study the effect of breeder age on incubation parameters (hatchability, eggshell thickness, egg surface temperature and chick weight). In Exp. 1, fertile eggs (30- and 60-wk-old breeders) were incubated at three different temperatures (36.8, 37.8 and 38.8 ºC). Eggshell surface temperature was measured by attaching a thermocouple to the shell and data were collected in a datalogger every ten minutes. This study was conducted according to a 3 x 2 factorial design (three temperatures and two breeder ages). Data revealed that eggshell surface temperature changed according to incubation temperature, with the main increase occurring between 10 and 13 days of incubation, and that the maximum increase in eggshell surface...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Broiler breeder; Development; Embryo; Temperature; Incubation.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2012000100002
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Effect of broiler breeder age on pancreas enzymes activity and digestive tract weight of embryos and chicks Rev. Bras. Ciênc. Avic.
Maiorka,A; Santin,E; Silva,AVF; Routman,KS; Pizauro Jr.,JM; Macari,M.
A study was carried out to evaluate the effect of broiler breeder age on the development of the digestive tract of embryos and chicks. Fertilized eggs Cobb from 30 and 60 week-old broiler breeder was utilized in this experiment. The results showed that eggs from older (60 weeks of age) broiler breeders were heavier (p = 0.001) than those from younger (30 weeks of age) broiler breeder. In addition, older broiler breeder had larger (p = 0.001) embryos showing a higher yolk sac (p = 0.001) and higher gastrointestinal tract relative weight (p = 0.007) than those from younger broiler breeder. The activities of pancreatic lipase and trypsin enzymes were also higher in embryos from older broiler breeder than those from younger broiler breeder (p = 0.001 and p =...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Broiler breeder age; Embryo; Enzymatic activity; Gastrointestinal tract.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2004000100003
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Effect of Broiler Breeder Age on the Intestinal Mucosa Development of the Embryos at 20 Days of Incubation Rev. Bras. Ciênc. Avic.
Maiorka,A; Silva,AVF; Santin,E; Bruno,LDG; Boleli,IC; Macari,M.
ABSTRACT This study investigated the effect of broiler breeder age on the morphological development of the small intestine broiler embryos (villus height, crypt depth, microvillus height, and villus density) at 20 day of incubation. Eggs obtained from 30- and 60-wk-old broiler breeders were used. The results showed that embryos from older broiler breeders presented longer villi in the duodenum, jejunum, and ileum compared with younger broiler breeders. In addition, embryos from older broiler breeders presented deeper crypts in the jejunum and ileum, longer microvilli in jejunal enterocytes, and lower villus density (microvillus number/mm2) in the duodenum and ileum than younger breeders. These results suggest that breeder age influences the gut mucosa...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Broiler breeder age; Embryo; Microvillus; Small intestine; Villus.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2016000600079
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Effect of Fibroblast Growth Factor 2 (FGF2) and Insulin Transferrin Selenium (ITS) on In Vitro Maturation, Fertilization and Embryo Development in Sheep BABT
Mondal,Sukanta; Mor,Avantika; Reddy,Ippala Janardana; Nandi,Sumanta; Parameswaragupta,PS.
The present study evaluated the effect of fibroblast growth factor 2 (FGF2) and insulin-transferrin-selenium (ITS) to the in vitro maturation and embryo culture media on ovine oocyte maturation, cleavage and embryo development. Oocytes having more than five layers of unexpanded cumulus cells and granular homogenous ooplasm were cultured into 50 μL droplets of eight different culture systems: (i) TCM-199 (Tissue Culture Medium-199); (ii) TCM-199+10 ng/mL FGF2; (iii) TCM-199+20 ng/mL FGF2; (iv) TCM-199+30 ng/mL FGF2; (v) TCM-199+10 ng/mL ITS; (vi) TCM-199+20 ng/mL ITS; (vii) TCM-199+30 ng/mL ITS and (viii) TCM-199+20 ng/mL ITS+20 ng/mL FGF2 in a CO2 incubator at 38.50C for 24 h. All the oocyte culture media were supplemented with 10% FBS, FSH (10 μg/mL) and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Fibroblast growth factor 2; Insulin transferrin selenium; Oocyte; Embryo; Sheep.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000400521
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Effect of Incubator Type and Broiler Breeder Age on Hatchability and Chick Quality Rev. Bras. Ciênc. Avic.
Araújo,ICS; Leandro,NSM; Mesquita,MA; Café,MB; Mello,HHC; Gonzales,E.
ABSTRACT Two experiments were conducted to evaluate the effect of broiler breeder age and incubator type on hatching parameters, hatch window, embryo diagnosis results, and hatchling physical quality. The treatments consisted of a combination of three broiler breeder ages (29, 35 and 59 weeks of age) and two incubator types (single stage, SS; or and multiple stage, MS). A completely randomized design in a 3x2 factorial arrangement was applied. In Experiment I, 1,896 eggs were used and 360 eggs in Experiment II. There was an interaction between breeder age and incubator type only for hatchling physical quality score. Independently of incubator type, hatchability rate, late embryo mortality, and egg contamination were higher in the eggs laid by older...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryo; Hatch window; Multiple-stage and single-stage incubation.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2016000600017
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Effect of Preconception Gamma Irradiation on Morphometric Assessment of Adult Female Mice and Embryo International Journal of Morphology
Edagha,Innocent A; Shokunbi,Matthew T; Adenipekun,Adeniyi A; Bassey,Rosemary B; Ekong,Moses B.
The aim was to study the effect of preconception gamma irradiation on the gross morphometry of the adult female mice and its embryo. Twenty-seven mice; 18 females and 9 males: subdivided into 3 groups namely (Control, Non-Irradiation and Radiation) containing 6 females and 3 male mice each in 2:1 ratio. A gamma irradiation dose of 1Gy/min was delivered to each batch of mice exposed by a Cobalt 60, Theratron 780c model, by Atomic Energy of Canada Limited (AECL) at the Radiotherapy department of the University College Hospital, Ibadan. All the animals were mated 1 week post irradiation. Vaginal plugs were confirmed, and the pregnant females were sacrificed on day 14 of gestation by chloroform inhalation. The gross morphology of the female mice and their...
Tipo: Journal article Palavras-chave: Preconception; Irradiation; Gamma ray; Spinal cord; Embryo.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022013000200039
Registros recuperados: 68
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