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2. Bovine oocyte activation with bull or human sperm by conventional ICSI and Piezo-ICSI: Its relationship with PLCɀ activity.

Author

Alvarez, Gabriel, Villanueva, Sofia, Breininger, Elizabeth, Geller, Marisa, Ruhlmann, Claudio, Dalvit, Gabriel, Cetica, Pablo, and Masashige Kuwayama

Subjects
SPERMATOZOA, OVUM, INTRACYTOPLASMIC sperm injection, BLASTOCYST, HUMAN embryology, BOS, PHOSPHOLIPASE C, ACTIVATION (Chemistry)
Abstract

Background: The intracytoplasmic sperm injection (ICSI) technique has low efficiency in cattle. This has mainly been attributed to the oocyte activation failure due to oocyte and/or sperm factors. Aim: Our aim was to evaluate the effect of conventional ICSI and Piezo-ICSI with bull or human sperm on bovine oocyte activation and embryo development and to assess its relationship with the phospholipase C zeta (PLCɀ) activity of both species. Methods: In vitro matured bovine oocytes were randomly divided into five groups and were fertilized as follows: conventional ICSI using bovine sperm with chemical activation (control), conventional ICSI using bovine sperm, Piezo-ICSI using bovine sperm, conventional ICSI using human sperm, and Piezo-ICSI using human sperm. PLCɀ activity was determined in bull and human sperm samples. Results: Within the groups using bull sperm, the oocytes fertilized by conventional ICSI had the lowest values of 2 pronuclei (PN) formation and cleavage, Piezo-ICSI increased both percentages and ICSI + chemical activation presented the highest 2 PN, cleavage, and blastocyst rates (p < 0.05). Within the groups using human sperm, the oocytes fertilized by Piezo-ICSI presented higher 2 PN and cleavage rates than those activated by conventional ICSI (p < 0.05). Piezo-ICSI with human sperm increased bovine oocyte activation as much as conventional ICSI + chemical activation with bovine sperm (p < 0.05). Higher values of PLCɀ activity were found in human sperm compared with bovine sperm (p < 0.05). Conclusion: Our results suggest that the higher stability of the bovine sperm in combination with its relatively low content of PLCɀ impairs bovine oocyte activation after ICSI. [ABSTRACT FROM AUTHOR]

Published
2024
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7. Comparing two simple cryopreservation methods for porcine sperm: Effect on sperm quality parameters

Author

Perez, M.C, Petrinelli, A., Rodriguez, Pablo C., Satorre, María Mercedes, and Breininger, Elizabeth

Subjects
criopreservación, calidad seminal, purl.org/becyt/ford/4.4 [https], semen, porcino, purl.org/becyt/ford/4 [https]
Abstract

Cerca del 40% de la carne roja que se consume en el mundo es provista por los cerdos. Los productores porcinos de nuestro país se encuentran ante el enorme desafío de lograr mejoras productivas con un desarrollo económico sustentable y eficiente, así como implementar tecnologías reproductivas, como la inseminación artificial y la criopreservación de semen. Tales adelantos aún presentan enormes problemas en estaespecie. El objetivo de este trabajo fue comparar dos métodos simples de congelamiento para semen porcino, evaluando los parámetros de calidad seminal al momento del descongelado. Las muestras de semen fueron criopreservadas en pajuelas siguiendo el protocolo descripto por Peña y colaboradores en 2003, realizando (grupo tratamiento) o no (grupo control) una modificación en el paso final de congelamiento posterior al envasado en las pajuelas. Las evaluaciones de motilidad y vitalidad se realizaron por microscopía óptica en platina termostatizada y por las técnicas de eosina/nigrosina y la combinación del colorante supravital azul tripán con microscopía de contraste interferencial diferencial, respectivamente. La criocapacitación se evaluó por la técnica fluorescente de clorotetraciclina (CTC). Los resultados de los diferentes experimentos fueron analizados por la prueba t de Student. Las muestras del grupo tratamiento evidenciaron una motilidad significativamente más alta (p

Published
2019

8. Effect of Hydroponic Green Forage Supplementation during Prepartum and Lactation on Sow and Litter Performances.

Author

Velásquez Amores, Soledad, Breininger, Elizabeth, Lorenzo Smirnoff, Alejandra, Caggiano, Nicolás, De Simone, Emilio, Acerbo, Marcelo, and De Luca Sarobe, Verónica

Subjects
*DIETARY supplements, *LACTATION, *PREGNANCY, *SOWS, *NUTRITION
Abstract

The incorporation of fibre in the diet of pregnant sows improves the performance during pregnancy and increases voluntary consumption during lactation. Hydroponic green forage (HGF) constitutes a method of cultivation without soil in controlled environmental conditions. The aim of this study was to evaluate the impact of the supplementation with HGF in lactating sows on productive performance and to investigate the metabolic state through the analysis of different biochemical parameters. Twelve sows of the Porcine Productive Unit of the Faculty of Veterinary Sciences were randomly assigned to 2 feeding groups: control (conventional diet) or HGF (hydroponic green forage diet, conventional diet supplemented with HGF) from 7 days prepartum until day 28 postpartum. Productive parameters in sows (weaning-to-oestrus interval and back fat thickness) and in the litter (number of piglets born and weaned, and piglet weight) were evaluated. The effect of the diet in the metabolism of the sows was evaluated by biochemical parameters (total plasma proteins, albumin, glucose, urea and creatinine). Supplementation with HGF did not significantly affect the litter size nor productive parameters but produced a higher weight of piglets at day 60. Although lactation affected some biochemical parameters, no substantial negative consequences of the HGF supplemented diet were observed. Our results suggest that the use of HGF could be an option to take into account in porcine production with economic and environmental benefits. [ABSTRACT FROM AUTHOR]

Published
2021

10. Study of sperm evaluation parameters to estimate cryopreserved bovine semen fertility

Author

Morado, Sergio Adrián, Pereyra, V., Breininger, Elizabeth, Sara, R., and Cetica, Pablo Daniel

Subjects
Semen fertility, CIENCIAS AGRÍCOLAS, Otras Ciencias Veterinarias, Ciencias Veterinarias, In vitro fertilization, Bull sperm, purl.org/becyt/ford/4.3 [https], Functional parameters, purl.org/becyt/ford/4 [https]
Abstract

In cattle production, reproductive biotechnologies such as artificial insemination with Cryopreserved semen samples show the same performance as insemination with fresh semen. The most representative method to evaluate semen fertilizing capability is through the results obtained by in vivo fertility. However, as it is difficult to apply this method in practice, routine laboratory tests can be used to evaluate semen quality. The aim of this work was to evaluate functional spermatic parameters in cryopreserved bovine semen samples from males presenting high and low field fertility rates. Semen samples were obtained from bulls presenting low or high fertility in a program of artificial insemination. Routine and functional parameters were determined in both groups. As regards routine tests, no significant differences were observed in progressive motility and vigor between samples of animals with high and low fertility in vivo, but differences were detected in vitality and acrosome integrity (p

Published
2015

11. Participation of lactate dehydrogenase in capacitation and acrosome reaction of fresh and cryopreserved, with or without α-tocopherol, boar sperm.

Author

Rodriguez, Pablo, Duarte, Juliana, Duprat, María Paris, Pereyra, Vanina, Satorre, María, Cetica, Pablo, and Breininger, Elizabeth

Subjects
SWINE, ACROSOME reaction, LACTATE dehydrogenase, BOARS, SEMEN, ENZYME inhibitors, SPERM motility, SPERMATOZOA
Abstract

The aim of this work was to study the participation of lactate dehydrogenase (LDH) enzyme in capacitation and acrosome reaction in porcine sperm by determining its enzymatic activity and the effect of its inhibition on these processes. Semen samples, from four crossbred boars (Pietrain x Yorkshire) of proven fertility, 1-1.5 year of age, were analysed. The enzymatic activity of LDH was determined spectrophotometrically in extracts from fresh and cryopreserved, with and without α-tocopherol, porcine sperm. Besides, sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well-known capacitation inducer, follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of sodium oxamate, a specific inhibitor of the enzyme. The activity of LDH in enzymatic extracts of fresh, cryopreserved without or cryopreserved with α-tocopherol porcine sperm were 3.85±0.27, 0.7±0.04 and 1.73±0.23 U/1010 sperm, respectively (p<0.05). The addition of sodium oxamate inhibited capacitation or acrosome reaction in fresh and cryopreserved with or without α-tocopherol porcine sperm, affecting (at different concentrations) motility and sperm viability (P<0.05). Obtained results demonstrate the participation of LDH in bicarbonate-induced capacitation and follicular fluid-induced acrosome reaction in fresh and cryopreserved porcine sperm. [ABSTRACT FROM AUTHOR]

Published
2020

12. Ascorbic acid or pyruvate counteracts peroxidative damage in boar sperm cryopreserved with or without alpha-tocopherol

Author

Breininger, Elizabeth and Beconi, Martha Teresa

Subjects
espermatozoide, CIENCIAS AGRÍCOLAS, Otras Ciencias Veterinarias, Ciencias Veterinarias, porcino, criopreservado
Abstract

Ejaculates of four crossbred boars were frozen with or without 200 μg/mL of alpha-tocopherol. Frozen samples were then thawed in the extender containing either ascorbic acid (2.5, 5 or 10 mM) or pyruvate (1 or 5 mM) to obtain the samples in which sperm quality parameters and lipid peroxidation level were evaluated. Ascorbic acid decreased lipid peroxidation in a dose-dependent manner and increased post-thaw sperm motility. The addition of pyruvate decreased lipid peroxidation level without improving sperm quality parameters. Although the addition of ascorbic acid or pyruvate did not show a synergic effect with alpha-tocopherol, the decrease of lipid peroxidation observed when the antioxidants were added to the thawing extender could be indicating a protective effect on the sperm plasma membrane from peroxidative damage. Since alpha-tocopherol protects from the lipid peroxidation and improves the sperm motility, it would be considered as the recommended antioxidant to use in boar sperm cryopreservation to extend the time of storage for the use in commercial applications. Fil: Breininger, Elizabeth. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Produccion Animal; Argentina Fil: Beconi, Martha Teresa. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina

Published
2014

13. Participation of membrane adenylyl cyclase in heparin-induced capacitation in cryopreserved bovine spermatozoa

Author

Breininger, Elizabeth, Ricart, María Cecilia, Beconi, Martha, and Rodriguez, Pablo Cristian

Subjects
capacitation, spermatozoa, CIENCIAS AGRÍCOLAS, bovine, Ciencias Veterinarias, cryopreserved
Abstract

The aim of this work was to study the participation of membrane adenylyl cyclase in heparin-induced capacitation in cryopreserved bovine spermatozoa. Sperm suspensions were incubated in Tyrode´s albumin lactate pyruvate medium in the presence of heparin (10 IU ml(-1) ) or forskolin (1-75 μm), a well-known membrane adenylyl cyclase activator. The participation of membrane adenylyl cyclase was confirmed using a specific inhibitor, 2´,5´-dideoxyadenosine (6-25 μm). Spermatozoa capacitated with forskolin (25 μm) were incubated with bovine follicular fluid to evaluate their ability to undergo acrosome reaction. Capacitation percentages were determined by the fluorescence technique with chlortetracycline, and true acrosome reaction was determined by trypan blue and differential interferential contrast. The forskolin concentrations employed had no effect on progressive motility or sperm viability. Capacitation values induced by 25-μm forskolin treatment (27.80 ± 2.59%) were significantly higher respect to the control (4.80 ± 1.30%). The inhibitor 2´,5´-dideoxyadenosine prevented forskolin-induced capacitation and significantly diminished capacitation induced by heparin. Follicular fluid induced physiological acrosome reaction in spermatozoa previously capacitated with 25-μm forskolin (P < 0.05). Forskolin acts as a capacitation inducer and involves the participation of membrane adenylyl cyclase as part of the intracellular mechanisms that lead to capacitation in cryopreserved bovine spermatozoa. Fil: Breininger, Elizabeth. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Produccion Animal; Argentina Fil: Ricart, María Cecilia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina Fil: Beconi, Martha. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina Fil: Rodriguez, Pablo Cristian. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina

Published
2014
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14. Ascorbic acid or pyruvate counteracts peroxidative damage in boar sperm cryopreserved with or without α-tocopherol.

Author

Breininger, Elizabeth and Beconi, Martha Teresa

Subjects
*LIPID peroxidation (Biology), *SPERM motility, *BOARS, *VITAMIN C, *PYRUVATES, *FROZEN semen
Abstract

Ejaculates of four crossbred boars were frozen with or without 200 µg/mL of α-tocopherol. Frozen samples were then thawed in the extender containing either ascorbic acid (2.5, 5 or 10 mM) or pyruvate (1 or 5 mM) to obtain the samples in which sperm quality parameters and lipid peroxidation level were evaluated. Ascorbic acid decreased lipid peroxidation in a dose-dependent manner and increased post-thaw sperm motility. The addition of pyruvate decreased lipid peroxidation level without improving sperm quality parameters. Although the addition of ascorbic acid or pyruvate did not show a synergic effect with α-tocopherol, the decrease of lipid peroxidation observed when the antioxidants were added to the thawing extender could be indicating a protective effect on the sperm plasma membrane from peroxidative damage. Since α-tocopherol protects from the lipid peroxidation and improves the sperm motility, it would be considered as the recommended antioxidant to use in boar sperm cryopreservation to extend the time of storage for the use in commercial applications. [ABSTRACT FROM AUTHOR]

Published
2014

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