Browsing by Author "Argudo Garzón, Daniel Ernesto"

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Ajuste del tiempo de inseminación con semen sexado en vacas holstein en producción superovuladas
(2018) Argudo Garzón, Daniel Ernesto; Soria Parra, Carlos Alonso; Durán López, Juana Catalina; Bravo Mosquera, Sandra Elizabeth; Alvarado Molina, Juan Pedro; Alberio, Ricardo Horacio
The embryo production results after use of sex-sorted sperm to inseminate superovulated cows (SOV) have not been very encouraging so far, due to the low quantity of sperms used as well as their short half- life. In a previous study conducted in Brazil in Holstein cows, it was shown that using the protocol of SOV P36/Lh60 and inseminating with sexsorted sperm at 18 and 30h (interval of 12hs between the two inseminations) after application of the inductor of ovulation it was obtained a similar quantity of transferable structures than inseminating at the 12 and 24h with non-sorted sperm. The aim of this investigation was to adjust the moment of the artificial insemination (AI) in superovulated and inseminated cows with sex-sorted sperm to achieve better synchrony with ovulations and by this way to increase the quantity of transferable embryos obtained. For this, we evaluated 2 moments of AI in which the interval between both were of 6hs. We used 30 lactating Holstein cows treated with the protocol P36/LH60 and, at insemination, were divided at random in three groups: 1-group AI18/30 (control group; n=10), the AI were carried out at the 18 and 30h after application of the inductor of the ovulation (GnRH); 2- group AI18/24 (n=10) the AI inseminations were made at 18 and 24 h after GnRH and 3- group AI24/30(n=10) the AI were made at 24 and 30h. In all cases, each insemination was doing with 2.1 x 106 sex-sorted sperm. The results between groups were no different in the quantity of transferable embryos (1,9±0,6;4,1±1,5 and 1,3±0,4respectively; p>0,05) nor in the proportion of transferable from total structures (38,8%; 60,1% y 34,1% respectively). In conclusion, the low number of cows in each group and the high variability of responses do not permit to conclude on significant differences in the alternatives evaluated. However, the numerical results lead to propose new studies to increase the number of animals treated and so, reevaluate these results.
Ajuste del tiempo de inseminación con semen sexado en vacas superovuladas
(2016) Argudo Garzón, Daniel Ernesto; Soria Parra, Carlos Alonso
The results of the combination of sex-sorted sperm and superovulation (SOV) have not been very encouraging so far, due to the low quantity of sperms as well as their short life. In a previous study conducted in Brazil in Holstein cows, it was shown that using the protocol of SOV P36/Lh60 and inseminating with sex-sorted sperm at the 18 and 30 hours of application of the inductor of the ovulation (interval of 12 hs between the two inseminations) it is obtained an equal quantity of transferable structures than inseminating at the 12 and 24 hours with non-sorted sperm. The aim of this investigation was to adjust the hours of the artificial insemination (AI) in superovulated and inseminated cows with sex-sorted sperm to achieve better synchrony with ovulations, so in this way to increase the quantity of transferable embryos. This adjustment evaluated 2 moments of AI in which the interval diminished to 6 hs. For this, 30 superovulated lactating Holstein cows were used with the protocol P36/LH60 and were divided in three groups in an aleatory form; in the group IA18/30 (control group; n=10) the inseminations were carried out at the 18 and 30 hs of application of the inductor of the ovulation (GnRh); in the group IA18/24(n=10) was inseminated at the 18 and 24 hs of the GnRH and the group IA24/30(n=10) was inseminated at the 24 and 30 hs (it was used 2.1 x 106 sex-sorted sperm/insemination). It was not found statistical difference in the three groups; however, the group IA18/24 showed numeric advantage over the group IA24/30 and the Control (4,1±1,5 vs 1,3±0,4 and 1,9±0,6 respectively) in the quantity of transferable embryos, concluding that this adjustment in the hours of AI with sexed semen in superovulated lactating Holstein cows can be used with similar results to the previous works
Competencia del ovocito bovino obtenido por ovum pick-up valorado mediante el azul brillante de cresilo
(2018) Ayala Guanga, Luis Eduardo; Samaniego Campoverde, Jorge Xavier; Nieto Escandón, Pedro Emilio; Rodas Carpio, Ermes Ramiro; Dután Sanango, Jorge Bolívar; Calle Ortiz, Guido Rigoberto; Murillo Apolo, Yury Agapito; Vázquez Mosquera, Juan Mesías; Argudo Garzón, Daniel Ernesto; Perea Ganchou, Fernando Pedro
The objective of the present study was to evaluate the bright blue cresyl (BCB) test as an indirect method to select competent oocytes for the in vitro production of embryos (IVP). The cumulus-oocyte complexes (COC) were obtained from two Creole heifers subjected to two treatments: T1 = COC recovered by OPU (ovum pick-up) previous stimulation with FSH-LH; T2 = COC recovered from non-stimulated animals (control).The two heifers were alternated in the two treatments and five repetitions were done. Recovered COCs were classified into types A, B, C and D. Then the BCB test was applied to each of the COC types to determine if they were BCB+ or BCB-. T1 allowed to recover 5.2 more COC than T2 (p<0.05). When applying the BCB test, it was determined that all type A oocytes of T1 and T2 were BCB+; that is, they finished their growth and were ready to start the process of in vitro maturation; however, about 50% of the type B, C and D COCs of T1 and T2 were BCB+. It is concluded that the selection of COC based on morphological characteristics is a reliable method only for type A and has a 50% error for COC type B, C and D and, therefore, the application of the BCB test allows to improve this selection non-invasively
El empleo de la ozonoterapia en ganadería de leche como alternativa de tratamiento para la mastitis clínica
(2012) Arichábala Cabrera, Natalie; Argudo Garzón, Daniel Ernesto; Soria Parra, Carlos Alonso
Evaluación de dos diluyentes en la congelación de espermatozoides epididimarios de perros (alcohol polivinílico y yema de huevo)
(2022) López Zhindón, Noelia Natali; Duma Pauta, José Mauricio; Samaniego Campoverde, Jorge Xavier; Galarza Lucero, Diego Andrés; Argudo Garzón, Daniel Ernesto
Egg yolk (EY) is a common component of semen extenders due to its effectiveness in protecting the sperm membrane from cold shock during cryopreservation and it is also rich in proteins, vitamins, antioxidants, and phospholipids. However, the EY is an undefined component and at the same time dangerous due to the risk of transmission of pathogenic agents. For this reason, it is necessary to evaluate the use of other components that reduce this problem. An understudied alternative is polyvinyl alcohol (PVA). For this reason, this investigation evaluated the effect of PVA and EY supplemented with TCG diluent (tris, citric acid, glucose) on the cryosurvival of dog epididymal spermatozoa (ES). For this, ES were recovered using the retrograde flow technique in 15 orchiectomized adult dogs to form five pools (4-6 epididymal samples/pool). Each pooled sample was divided into two aliquots, which were subsequently diluted and frozen using two additives supplemented to the TCG diluent: 1% (w/v) PVA [TCG–PVA] and 20% (v/v) egg yolk [TCG–EY]. After thawing, morphological abnormalities, vitality (eosin/nigrosin), plasma membrane integrity (propidion iodide), and kinematic variables (CASA system, SCA-2018 ) were analyzed. The results demonstrated that the vitality and integrity of the plasma membrane were superior (P<0.05) in samples frozen with TCG–EY, compared to those samples frozen with TCG–PVA. Likewise, the samples frozen with TCG–EY obtained higher post-thaw values (P<0.05) of kinematic variables such as total and progressive motility, average and rectilinear velocities, straightness index and flagellum beat frequency, compared to with frozen samples with TCG–PVA. In conclusion, enhancement of EY to TCG medium was effective in freezing dog ES, as it improved vitality, plasma membrane body, and sperm kinetics; however, the addition of 1% PVA did not improve the response to cryopreservation.
The time interval between ovarian stimulation with FSH/LH and ovum-pick up affects the quantity, quality and developmental competence of oocytes retrieved from ecuadorian creole heifers
(2020) Ayala Guanga, Luis Eduardo; Samaniego Campoverde, Jorge Xavier; Argudo Garzón, Daniel Ernesto; Perea Brugal, Mariana Sofia; Perea Ganchou, Fernando Pedro; Rodas Carpio, Ermes Ramiro; Nieto Escandón, Pedro Emilio
The aim of this study was to evaluate the effects of gonadotropin treatment and the interval between ovarian stimulation with FSH/LH and the follicular aspiration session, on the quantity and quality of the cumulus oocyte complexes (COCs) obtained by ovum pick up (OPU)) in Ecuadorian Creole heifers. Four heifers were randomly assigned to four groups (FHS/LH 40h; FSH/LH 48h; control 40h; control 48h). In two groups a dose (0.011 mg, IM) of GnRH was applied on day 0, followed by 500 IU, IM, of FSH/LH on day 2, and the OPU session at 40 (FSH/LH 40 h) or 48 hours (FSH/LH 48h) after the application of gonadotropin. In the other two groups, FSH/LH was replaced by a physiological solution. There was no statistical difference in the recovery rate of COCs between FSH/LH 40h and FSH/LH 48h, but approximately twice as many COCs (p<0.05) were obtained than in those who did not receive ovarian stimulation. In addition, 2.4 times more COCs of quality A were achieved in the treatment of FSH/LH 48h than in that of FSH/LH 40h. All (100%) of these structures reacted positively to Brilliant Cresyl Blue (BCB+), which means that they finished the growth and were competent to initiate in vitro maturation. It is concluded that the treatment with FSH/LH 48h allowed to obtain a greater number of COCs competent for in vitro maturation.
Uso del azul brillante de cresilo en la selección de ovocitos competentes para la producción in vitro de embriones
(2019) Duma Pauta, José Mauricio; Argudo Garzón, Daniel Ernesto; Ochoa Calderón, René Rafael; Alvarado Ulloa, Jhonatan Marcelo; Ayala Guanga, Luis Eduardo
The objective was to evaluate the Brilliant Cresyl Blue (BCB) in the selection of oocytes for the production of embryos in vitro (PIV). Three treatments were evaluated. G1 = Witness without BCB exposure; G2 = staining positive oocytes (BCB +) and G3 = staining negative oocytes (BCB-). Morphometry, maturation (IVM), meiotic configuration, cleavage and embryos were determined. 2,340 cumulus oocyte complexes (COCs) were obtained, classified as suitable and not suitable for IVP. The 2/3 parts were exposed for 90 min. To BCB and the rest incubated for 90 min in H-SOF. 25% of COCs from each group were taken to assess the morphometry. The remaining 75% was placed in maturing medium for 24h. After IVM, 25% of COCs were used for morphometry and meiotic progression. The remaining 50% was in vitro fertilized (IVF) and cultivated. Cleavage was evaluated 48h after IVF and embryos on day 8. 62.3% of G2 oocytes were BCB + and had a larger diameter before (135, 9 μm) and after (134, 8 μm) of IVM. (P <0.05) vs group 3 BCB- (128, 1 and 127, 6 μm) and the Witness (129, 0 and 128, 2 μm). Maturation and meiotic configuration were not different between groups (P> 0.05). Cleavage was higher (P <0.05) in the Control (43.4%) than the BCB + group (33.5%). Finally, the global percentage of embryos was higher in BCB + (52.6%) than the Controlling (32.6%) and BCB- (33.5%). It is concluded that the BCB allowed to select oocytes with a larger diameter, although it did not allow to increase the percentage of maturation and cleavage, it contributed to improving the overall percentage of embryos obtained in vitro.