Novel prospects for evaluation of follicle wall blood flow using color-Dopplerultrasonography

The goal of this study was to develop anobjective method for evaluation of ovarian follicle wallblood flow in cattle. Two subjective methods were used:(I) real-time ultrasound evaluations performed by oneoperator in the barn and (II) video clip evaluationsperformed by four operators in the laboratory. Thefollowing objective methods evaluated in the laboratorywere used for comparison: (I) percentage of follicle wallcircumference under blood flow (WUF) and (II) pixelarea of color-Doppler signals. Cows (n = 21) weresubmitted to a synchronization protocol, follicles ≥7 mmwere measured, and blood flow was evaluated every 12 huntil ovulation using color-Doppler ultrasonography. Nodifference (P > 0.05) was observed among laboratoryoperators from day 2 of training onwards. Therefore,an average score of all operators was used forcomparisons among different methods. Both subjectiveand objective methods of evaluation showed anincrease (P < 0.0001) in follicle blood flow over time.Higher (P < 0.001) correlations were obtained betweenWUF and subjective laboratory evaluation thanbetween WUF and pixel area or WUF and subjectivebarn data. Higher (P < 0.0003) correlation coefficientswere observed for WUF than for the pixel area whencompared with the barn (r = 0.70 vs. r = 0.42) orlaboratory (r = 0.84 vs. r = 0.62) data. Subjectiveevaluations at the laboratory and barn producedstronger correlations with WUF (P < 0.0008) than withpixel area (P < 0.01). In conclusion, WUF is aneffective and reliable method for objective evaluationof follicle wall blood flow in cows.

Novel prospects for evaluation of follicle wall blood flow using color-Dopplerultrasonography

The goal of this study was to develop anobjective method for evaluation of ovarian follicle wallblood flow in cattle. Two subjective methods were used:(I) real-time ultrasound evaluations performed by oneoperator in the barn and (II) video clip evaluationsperformed by four operators in the laboratory. Thefollowing objective methods evaluated in the laboratorywere used for comparison: (I) percentage of follicle wallcircumference under blood flow (WUF) and (II) pixelarea of color-Doppler signals. Cows (n = 21) weresubmitted to a synchronization protocol, follicles ≥7 mmwere measured, and blood flow was evaluated every 12 huntil ovulation using color-Doppler ultrasonography. Nodifference (P > 0.05) was observed among laboratoryoperators from day 2 of training onwards. Therefore,an average score of all operators was used forcomparisons among different methods. Both subjectiveand objective methods of evaluation showed anincrease (P < 0.0001) in follicle blood flow over time.Higher (P < 0.001) correlations were obtained betweenWUF and subjective laboratory evaluation thanbetween WUF and pixel area or WUF and subjectivebarn data. Higher (P < 0.0003) correlation coefficientswere observed for WUF than for the pixel area whencompared with the barn (r = 0.70 vs. r = 0.42) orlaboratory (r = 0.84 vs. r = 0.62) data. Subjectiveevaluations at the laboratory and barn producedstronger correlations with WUF (P < 0.0008) than withpixel area (P < 0.01). In conclusion, WUF is aneffective and reliable method for objective evaluationof follicle wall blood flow in cows.(AU)

Effect of ß-mercaptoetanol and cysteine on post-thawing quality and oxidative activity of ram sperm and on the viability of vitrified sheep embryos / Efeito do ß-mercaptoetanol e da cisteína sobre a qualidade e a atividade oxidativa do sêmen ovino após o descongelamento e sobre a viabilidade de embriões ovinos vitrificados

The effects of ß-mercaptoethanol (BME) and cysteine on the viability and oxidative activity of ram sperm after thawing and on development in vitro and viability of vitrified sheep embryos were evaluated. Ejaculates from four rams were pooled and extended, composing six treatments: no antioxidants; 2mM BME; 5mM BME; 2mM BME and 5mM cysteine; 5mM BME and 5mM cysteine; and 5mM cysteine. Sperm motility, membrane and acrosome integrity, mitochondrial functionality, production of reactive oxygen species and total antioxidant capacity were similar across treatments (P>0.05). A medium with no antioxidant presented cleavage and blastocyst development rates (60.3% and 33.6%, respectively) similar (P>0.05) to those of a medium with 50µM BME and 600µM cysteine (64.3% and 36.6%, respectively). Post-thawing viability of vitrified embryos was similar between media (P>0.05). Cysteine and BME had no influence on the post-thawing viability and oxidative activity of ram sperm and on the viability of vitrified sheep embryos.(AU)

Foram avaliados os efeitos do ß-mercaptoetanol (BME) e da cisteína sobre a viabilidade e a atividade oxidativa após o descongelamento do sêmen ovino e sobre o desenvolvimento in vitro e a viabilidade de embriões ovinos vitrificados. Ejaculados de quatro carneiros foram agrupados e diluídos, compondo seis tratamentos: sem antioxidantes; com BME 2mM; com BME 5mM; com BME 2mM e cisteína 5mM; com BME 5mM e cisteína 5mM; e com cisteína 5mM. Motilidade, integridade da membrana e do acrossoma, função mitocondrial, produção de espécies reativas de oxigênio e capacidade antioxidante total foram semelhantes entre os tratamentos (P>0,05). Em um meio sem antioxidantes, as taxas de clivagem e de desenvolvimento embrionário até blastocisto (60,3%, e 33,6%, respectivamente) foram semelhantes (P>0,05) às obtidas em um meio com BME 50µM e cisteína 600µM (64,3% e 36,6%, respectivamente). A viabilidade pós-descongelamento dos embriões vitrificados não diferiu entre os meios (P>0,05). O BME e a cisteína não influenciaram a viabilidade e a atividade oxidativa do sêmen ovino após o descongelamento e a viabilidade de embriões ovinos vitrificados.(AU)

Effect of ß-mercaptoetanol and cysteine on post-thawing quality and oxidative activity of ram sperm and on the viability of vitrified sheep embryos / Efeito do ß-mercaptoetanol e da cisteína sobre a qualidade e a atividade oxidativa do sêmen ovino após o descongelamento e sobre a viabilidade de embriões ovinos vitrificados

The effects of ß-mercaptoethanol (BME) and cysteine on the viability and oxidative activity of ram sperm after thawing and on development in vitro and viability of vitrified sheep embryos were evaluated. Ejaculates from four rams were pooled and extended, composing six treatments: no antioxidants; 2mM BME; 5mM BME; 2mM BME and 5mM cysteine; 5mM BME and 5mM cysteine; and 5mM cysteine. Sperm motility, membrane and acrosome integrity, mitochondrial functionality, production of reactive oxygen species and total antioxidant capacity were similar across treatments (P>0.05). A medium with no antioxidant presented cleavage and blastocyst development rates (60.3% and 33.6%, respectively) similar (P>0.05) to those of a medium with 50µM BME and 600µM cysteine (64.3% and 36.6%, respectively). Post-thawing viability of vitrified embryos was similar between media (P>0.05). Cysteine and BME had no influence on the post-thawing viability and oxidative activity of ram sperm and on the viability of vitrified sheep embryos.(AU)

Foram avaliados os efeitos do ß-mercaptoetanol (BME) e da cisteína sobre a viabilidade e a atividade oxidativa após o descongelamento do sêmen ovino e sobre o desenvolvimento in vitro e a viabilidade de embriões ovinos vitrificados. Ejaculados de quatro carneiros foram agrupados e diluídos, compondo seis tratamentos: sem antioxidantes; com BME 2mM; com BME 5mM; com BME 2mM e cisteína 5mM; com BME 5mM e cisteína 5mM; e com cisteína 5mM. Motilidade, integridade da membrana e do acrossoma, função mitocondrial, produção de espécies reativas de oxigênio e capacidade antioxidante total foram semelhantes entre os tratamentos (P>0,05). Em um meio sem antioxidantes, as taxas de clivagem e de desenvolvimento embrionário até blastocisto (60,3%, e 33,6%, respectivamente) foram semelhantes (P>0,05) às obtidas em um meio com BME 50µM e cisteína 600µM (64,3% e 36,6%, respectivamente). A viabilidade pós-descongelamento dos embriões vitrificados não diferiu entre os meios (P>0,05). O BME e a cisteína não influenciaram a viabilidade e a atividade oxidativa do sêmen ovino após o descongelamento e a viabilidade de embriões ovinos vitrificados.(AU)

Contribution of boars to reproductive performance and paternity after homospermic and heterospermic artificial insemination.

Heterospermic AI is commonly used in swine despite preventing precise evaluation of individual boar fertility. The present study compared the contribution of four boars (A, B, C and D) for reproductive performance and for paternity using homospermic and heterospermic (AB, AC, AD, BC, BD and CD) AI (n=204 for homospermic AI; n=307 for heterospermic AI). Blood samples from the four boars, from all sows inseminated with heterospermic doses and from the umbilical cords of their piglets, as well as tissue smears from mummified fetuses, were genotyped using single nucleotide polymorphisms (SNPs). Differences among boars were detected for the in vitro oocyte penetration rate and for the number of spermatozoa per oocyte (P<0.05), but not for sperm motility, mitochondrial functionality and integrity of the membrane, acrosome and DNA (P>0.05). Homospermic and heterospermic AI resulted in similar (P>0.05) farrowing rates (90.5% and 89.9%, respectively) and total litter size (12.4±0.4 and 12.7±0.7, respectively). Farrowing rate was lower for Boar B than for Boar C (P<0.05), but no other differences in reproductive performance among boars were observed with homospermic AI. The SNPs determined the paternity of 94.2% of the piglets sired by heterospermic AI. In the AC pool, paternity contribution per boar was similar (P>0.05), but differences between boars occurred in all other pools (P<0.05). Boar D achieved the greatest paternity contribution in all pools and parity categories (nearly 60%), whereas Boar B sired the fewest piglets (at most 40%). Reproductive performance was similar with homospermic and heterospermic AI, but differences in performance among boars undetected with homospermic AI were only evident after genotyping the piglets sired through heterospermic AI.