Avaliação inicial da gestação equina por ultrassonografia 3D Relato de caso / Initial evaluation of equine pregnancy by 3D ultrasound Case report

O presente trabalho objetivou o acompanhamento da gestação inicial na égua entre os dias 12 a35 pós-ovulação por meio da ultrassonografia tridimensional (3D). Após o diagnóstico de gestação no dia12 pós-ovulação, a reprodutora foi examinada diariamente por palpação retal e ultrassonografia paraadquirir imagens no modo B e em 3D. Nas avaliações diárias e com as imagens construídas pela técnicade ultrassonografia tridimensional foi possível acompanhar o crescimento do concepto, a identificação doembrião propriamente dito e a diferenciação de estruturas e membranas (saco vitelino e saco alantoide)permitindo visualizações em 360 graus. Pode-se concluir que na técnica de processamento da imagemtridimensional as imagens geradas fornecem excelente material educacional e acadêmico, precisando demais estudos para permitir uma efetiva aplicação clínica na perda embrionária.(AU)

The present study aimed at monitoring the initial pregnancy in the mare between days 12 to 35after ovulation using three-dimensional (3D) ultrasonography. After the diagnosis of gestation on day 12after ovulation, the reproductive tract was examined daily by rectal palpation and ultrasonography toacquire images in B mode and in 3D. In the daily evaluations and with the images constructed by thetechnique of three-dimensional ultrasonography, it was possible to follow the growth of the concept, theidentification of the embryo proper and the differentiation of structures and membranes (yolk andallantoic sac) allowing visualizations in 360 degrees. It can be concluded that the 3D generated imagesprovide excellent educational and academic material however, more studies are needed to allow aneffective application in embryonic loss.(AU)

Utilização de leite com diferentes concentrações de gordura como diluentes para sêmen equino refrigerado / Use of milk with different fat concentrations as extenders for equine cooled semen

Dois experimentos foram realizados para verificar a viabilidade de diluentes para sêmen equino refrigerado. O objetivo do primeiro experimento foi verificar a eficiência do leite UHT integral e semidesnatado, comparado ao leite desnatado. Amostras foram analisadas a fresco e posteriormente diluídas em leite UHT integral, semidesnatado e desnatado. Foram realizados exames de motilidade espermática e vigor, CFDA/PI e HOST pós-diluição (0h), e no sêmen refrigerado a 5 ºC após 24 e 48 horas. Não houve diferença entre os três diluentes, quanto à motilidade espermática (p=0,9880), vigor (p=0,7249), CFDA/PI (p=0,3382) e HOST (p > 0,01). Houve uma diminuição na qualidade do sêmen, no decorrer do tempo (p < 0,01), independente do diluente utilizado. Pode-se afirmar que, na falta de leite UHT desnatado, o semidesnatado e o integral podem ser usados, uma vez que eles não provocam prejuízo na qualidade do sêmen refrigerado armazenado por até 48 horas. No segundo experimento, o objetivo foi verificar a eficácia do leite desnatado em comparação com diluentes comerciais (Botusemen® (BS) e Botusemen Special® (BSS)). Logo após a coleta do sêmen, as amostras foram diluídas com leite UHT desnatado (LD), Botusemen® (BS) e Botusemen Special®(BSS). A análise de sêmen seguiu o mesmo protocolo do primeiro experimento. As amostras de sêmen refrigeradas com BSS apresentaram HOST e CFDA/PI (p< 0,001) maiores nas 24h e 48 h o que indica que o diluente BSS promove aumento da longevidade dos espermatozoides em comparação com os outros analisados.

Two trials were performed to verify the viability of extenders equine cooled semen. The objective of the first trial was to verify the efficacy of whole and semi-skimmed UHT milk compared to skim milk. Fresh semen samples were analyzed and later diluted in UHT whole milk, semi skimmed and skimmed milk. Sperm motility and vigor, CFDA / PI and HOST tests were performed post-dilution (0h), and in semen cooled at 5oC after 24 and 48h. There was no difference between the 3 extenders, for sperm motility (p = 0.9880), vigor (p = 0.7249), CFDA / PI (p = 0.3882) and HOST (p> 0.01). There was a decrease in semen quality over time (p <0.01), regardless of the extender used. It can be stated that in the absence of skimmed UHT milk, semi-skimmed and whole milk can be used since they do not impair the quality of refrigerated semen stored for up to 48 hours. In the second trial the objective was to verify the efficacy of the skim milk compared to commercial extenders (Botusemen® (BS) and Botusemen Special® (BSS)). Soon after semen collection, the samples were diluted with UHT skim milk (LD), Botusemen® (BS) and Botusemen Special® (BSS). Semen analysis followed the same protocol as the first experiment. BSS-cooled semen showed higher HOST and CFDA / PI (p <0.001) in 24h and 48h indicating that BSS diluent promotes increased sperm longevity compared to the others analyzed.

Utilização de leite com diferentes concentrações de gordura como diluentes para sêmen equino refrigerado / Use of milk with different fat concentrations as extenders for equine cooled semen

Dois experimentos foram realizados para verificar a viabilidade de diluentes para sêmen equino refrigerado. O objetivo do primeiro experimento foi verificar a eficiência do leite UHT integral e semidesnatado, comparado ao leite desnatado. Amostras foram analisadas a fresco e posteriormente diluídas em leite UHT integral, semidesnatado e desnatado. Foram realizados exames de motilidade espermática e vigor, CFDA/PI e HOST pós-diluição (0h), e no sêmen refrigerado a 5 ºC após 24 e 48 horas. Não houve diferença entre os três diluentes, quanto à motilidade espermática (p=0,9880), vigor (p=0,7249), CFDA/PI (p=0,3382) e HOST (p > 0,01). Houve uma diminuição na qualidade do sêmen, no decorrer do tempo (p < 0,01), independente do diluente utilizado. Pode-se afirmar que, na falta de leite UHT desnatado, o semidesnatado e o integral podem ser usados, uma vez que eles não provocam prejuízo na qualidade do sêmen refrigerado armazenado por até 48 horas. No segundo experimento, o objetivo foi verificar a eficácia do leite desnatado em comparação com diluentes comerciais (Botusemen® (BS) e Botusemen Special® (BSS)). Logo após a coleta do sêmen, as amostras foram diluídas com leite UHT desnatado (LD), Botusemen® (BS) e Botusemen Special®(BSS). A análise de sêmen seguiu o mesmo protocolo do primeiro experimento. As amostras de sêmen refrigeradas com BSS apresentaram HOST e CFDA/PI (p< 0,001) maiores nas 24h e 48 h o que indica que o diluente BSS promove aumento da longevidade dos espermatozoides em comparação com os outros analisados.(AU)

Two trials were performed to verify the viability of extenders equine cooled semen. The objective of the first trial was to verify the efficacy of whole and semi-skimmed UHT milk compared to skim milk. Fresh semen samples were analyzed and later diluted in UHT whole milk, semi skimmed and skimmed milk. Sperm motility and vigor, CFDA / PI and HOST tests were performed post-dilution (0h), and in semen cooled at 5oC after 24 and 48h. There was no difference between the 3 extenders, for sperm motility (p = 0.9880), vigor (p = 0.7249), CFDA / PI (p = 0.3882) and HOST (p> 0.01). There was a decrease in semen quality over time (p <0.01), regardless of the extender used. It can be stated that in the absence of skimmed UHT milk, semi-skimmed and whole milk can be used since they do not impair the quality of refrigerated semen stored for up to 48 hours. In the second trial the objective was to verify the efficacy of the skim milk compared to commercial extenders (Botusemen® (BS) and Botusemen Special® (BSS)). Soon after semen collection, the samples were diluted with UHT skim milk (LD), Botusemen® (BS) and Botusemen Special® (BSS). Semen analysis followed the same protocol as the first experiment. BSS-cooled semen showed higher HOST and CFDA / PI (p <0.001) in 24h and 48h indicating that BSS diluent promotes increased sperm longevity compared to the others analyzed.(AU)

Avaliação inicial da gestação equina por ultrassonografia 3D – Relato de caso / Initial evaluation of equine pregnancy by 3D ultrasound – Case report

O presente trabalho objetivou o acompanhamento da gestação inicial na égua entre os dias 12 a35 pós-ovulação por meio da ultrassonografia tridimensional (3D). Após o diagnóstico de gestação no dia12 pós-ovulação, a reprodutora foi examinada diariamente por palpação retal e ultrassonografia paraadquirir imagens no modo B e em 3D. Nas avaliações diárias e com as imagens construídas pela técnicade ultrassonografia tridimensional foi possível acompanhar o crescimento do concepto, a identificação doembrião propriamente dito e a diferenciação de estruturas e membranas (saco vitelino e saco alantoide)permitindo visualizações em 360 graus. Pode-se concluir que na técnica de processamento da imagemtridimensional as imagens geradas fornecem excelente material educacional e acadêmico, precisando demais estudos para permitir uma efetiva aplicação clínica na perda embrionária.

The present study aimed at monitoring the initial pregnancy in the mare between days 12 to 35after ovulation using three-dimensional (3D) ultrasonography. After the diagnosis of gestation on day 12after ovulation, the reproductive tract was examined daily by rectal palpation and ultrasonography toacquire images in B mode and in 3D. In the daily evaluations and with the images constructed by thetechnique of three-dimensional ultrasonography, it was possible to follow the growth of the concept, theidentification of the embryo proper and the differentiation of structures and membranes (yolk andallantoic sac) allowing visualizations in 360 degrees. It can be concluded that the 3D generated imagesprovide excellent educational and academic material however, more studies are needed to allow aneffective application in embryonic loss.

Histomorphometric and vascular changes in equine endometrium after the infusion of conceptus fragments

This experiment aimed to verify if the proteins present in a 13th day conceptus induce changes in the equine endometrial ultra-structure, histology, and vascularization, two days after its infusion. Ten healthy cyclic mares were used. Once estrus was confirmed, mares were examined daily to detect ovulation (day 0). After ovulation, mares were examined daily until day seven by transrectal palpation and B-mode and Doppler ultrasonography. In this first cycle, intrauterine biopsies were collected at day seven after ovulation, constituting the Cyclic group (n = 10). In the second cycle, the same mares daily were examined until ovulation was detected. After ovulation, mares were examined daily by transrectal palpation and B-mode and Doppler ultrasonography until day 7. On day 5, after ovulation, fragments from previously collected 13-day-old concepti were infused into the uterus of each mare. Intrauterine biopsies were collected at day 7 in all mares (n = 10), constituting the Fragment group. The percentage of ciliated and flattened cells decreased in the Fragment group. Protruded cells, superficial and intraglandular secretion, glandular lumen and diameter, blood vessel diameter, endometrial vascularization, and immune cells were higher in the Fragment group than in the Cyclic group. In summary, proteins of 13th day equine conceptus fragments infused at day five after ovulation signaled histological and vascular changes in the endometrium at the 7th day after ovulation.

Histomorphometric and vascular changes in equine endometrium after the infusion of conceptus fragments

This experiment aimed to verify if the proteins present in a 13th day conceptus induce changes in the equine endometrial ultra-structure, histology, and vascularization, two days after its infusion. Ten healthy cyclic mares were used. Once estrus was confirmed, mares were examined daily to detect ovulation (day 0). After ovulation, mares were examined daily until day seven by transrectal palpation and B-mode and Doppler ultrasonography. In this first cycle, intrauterine biopsies were collected at day seven after ovulation, constituting the Cyclic group (n = 10). In the second cycle, the same mares daily were examined until ovulation was detected. After ovulation, mares were examined daily by transrectal palpation and B-mode and Doppler ultrasonography until day 7. On day 5, after ovulation, fragments from previously collected 13-day-old concepti were infused into the uterus of each mare. Intrauterine biopsies were collected at day 7 in all mares (n = 10), constituting the Fragment group. The percentage of ciliated and flattened cells decreased in the Fragment group. Protruded cells, superficial and intraglandular secretion, glandular lumen and diameter, blood vessel diameter, endometrial vascularization, and immune cells were higher in the Fragment group than in the Cyclic group. In summary, proteins of 13th day equine conceptus fragments infused at day five after ovulation signaled histological and vascular changes in the endometrium at the 7th day after ovulation.(AU)

Uterine infusion of conceptus fragments changes the protein profile from cyclic mares

This experiment aimed to compare at day seven after ovulation, the protein profile of uterine fluid in cyclic mares with mares infused two days before with Day 13 conceptus fragments. Experimental animals were ten healthy cyclic mares, examined daily to detect ovulation (Day 0) as soon as estrus was confirmed. On day seven, after ovulation, uterine fluid was collected, constituting the Cyclic group (n = 10). The same mares were examined in the second cycle until ovulation was detected. On day five, after ovulation, fragments from a previously collected concepti were infused into each mare's uterus. Two days after infusion, uterine fluid was collected, constituting the Fragment group (n = 10). Two-dimensional electrophoresis technique processed uterine fluid samples. A total of 373 spots were detected. MALDI-TOF/TOF and NanoUHPLC-QTOF mass spectrometry identified twenty spots with differences in abundance between the Cyclic and Fragment group. Thirteen proteins were identified, with different abundance between groups. Identified proteins may be related to embryo-maternal communication, which involves adhesion, nutrition, endothelial cell proliferation, transport, and immunological tolerance. In conclusion, conceptus fragments signalized changes in the protein profile of uterine fluid seven days after ovulation in comparison to the observed at Day 7 in the same cyclic mares.

Uterine infusion of conceptus fragments changes the protein profile from cyclic mares

This experiment aimed to compare at day seven after ovulation, the protein profile of uterine fluid in cyclic mares with mares infused two days before with Day 13 conceptus fragments. Experimental animals were ten healthy cyclic mares, examined daily to detect ovulation (Day 0) as soon as estrus was confirmed. On day seven, after ovulation, uterine fluid was collected, constituting the Cyclic group (n = 10). The same mares were examined in the second cycle until ovulation was detected. On day five, after ovulation, fragments from a previously collected concepti were infused into each mare's uterus. Two days after infusion, uterine fluid was collected, constituting the Fragment group (n = 10). Two-dimensional electrophoresis technique processed uterine fluid samples. A total of 373 spots were detected. MALDI-TOF/TOF and NanoUHPLC-QTOF mass spectrometry identified twenty spots with differences in abundance between the Cyclic and Fragment group. Thirteen proteins were identified, with different abundance between groups. Identified proteins may be related to embryo-maternal communication, which involves adhesion, nutrition, endothelial cell proliferation, transport, and immunological tolerance. In conclusion, conceptus fragments signalized changes in the protein profile of uterine fluid seven days after ovulation in comparison to the observed at Day 7 in the same cyclic mares.(AU)

Desempenho reprodutivo de novilhas acasaladas aos 14 e 27 meses de idade submetidas a diferentes protocolos para a inseminação artificial / Reproductive performance of beef cattle heifers mated to 14 or 27 months of age subject to different protocols for artificial insemination

Avaliaram-se os efeitos de diferentes protocolos de inseminação sobre o desempenho reprodutivo em 249 novilhas Britânicas e cruzas, acasaladas aos 14 e 27 meses de idade. Sessenta e duas novilhas de 27 meses foram submetidas à IATF após o tratamento OvSynch; 61 novilhas de 27 meses e 32 novilhas de 14 meses foram submetidas à IATF após o tratamento Crestar; 62 novilhas de 27 meses e 32 novilhas de 14 meses foram submetidas à inseminação artificial (IA) após detecção de estros por 7 dias, no 7º dia foi aplicado PGF2a nos animais não observados em estro postergando-se IA por mais 5 dias (grupo PGF2a). Foram avaliados o peso vivo (PIA), o escore de condição corporal (ECC), o escore de trato reprodutivo (ETR) ao início da estação reprodutiva, a taxa de concepção e a taxa de prenhez à IATF, a taxa de prenhez ao término da estação. Os tratamentos OvSynch e Crestar tiveram 100% dos animais inseminados (IATF), enquanto os grupos PGF2a-14 meses e PGF2a-27 meses tiveram, respectivamente apenas 21,9% e 43,5% dos animais inseminados, devido à ausência ou falhas na identificação de cio. Não houve diferença nas taxas de concepção entre tratamentos hormonais e idades (P>0,05), com valores respectivos de 46,5%, 56,3%, 64,7% para os tratamentos Crestar, OvSynch, PGF2a e de 44,4% e 57,1%, para as idades 14 e 27 meses. A taxa de prenhez à IA (TF) foi de 46,5%, 56,3%, 23,4% para os tratamentos Crestar, OvSynch, PGF2a, com superioridade significativa (P<0,05) a favor dos grupos Crestar e OvSynch. A taxa de prenhez à IA (TF) foi de 18,6% e 41,3% (P<0,01) para as idades 14 e 27 meses. A taxa de prenhez final, após o repasse com touros foi respectivamente de 93,0%, 96,9%, 90,4% para os tratamentos Crestar, OvSynch, PGF2a e de 97,7% e 90,5%, para as idades 14 e 27 meses, sem diferença significativa. O ETR influenciou a taxa de prenhez após a inseminação, respectivamente de 0%, 25,6% e 48,8% para os ETR 1, 2 e 3. Os tratamentos hormonais para a IATF resultaram em maior taxa de prenhez...(AU)

The effects of three insemination protocols were evaluated on the reproductive performance in 249 British and crosses beef heifers, mated to the 14 or 27 months of age. Sixty two heifers with 27 months were submitted to FTAI after the treatment OvSynch; 61 heifers with 27 months and 32 heifers with 14 months were submitted to FTAI after the treatment to Crestar; 62 heifers with 27 months and 32 heifers with 14 months were submitted to the artificial insemination (AI) after oestrus detection for 7 days, in the 7th day PGF2a was applied in the animals without oestrus observed being postponed AI during more 5 days (PGF2a group). The live weight and body condition score (BCS), the reproductive tract score (RTS) to the beginning of mating season, the conception and the pregnancy to FTAI and the pregnancy to the end of the mating season were evaluated. The treatments OvSynch and Crestar had 100% of the inseminated animals (FTAI), while the groups PGF2a-14 months and PGF2a-27 months had respectively, only 21.9% and 43.5% of the inseminated animals, due to the absence or fails in the oestrus identification. There was no difference in the conception rates between hormonal treatments and ages (P>0.05), with respective values of 46.5%, 56.3%, 64.7% for the treatments Crestar, OvSynch, PGF2a and of 44.4% and 57.1%, for the ages 14 and 27 months. The pregnancy rate to FTAI/AI it was of 46.5%, 56.3%, 23.4% for the treatments to Crestar, OvSynch, PGF2a, with significant superiority (P<0.05) in favor of the groups Crestar and OvSynch. The pregnancy rate to AI (FTAI) it was of 18.6% and 41.3% (P <0,01) for the ages 14 and 27 months. The final pregnancy rate, after clean-up bulls was respectively of 93.0%, 96.9%, 90.4% for the treatments to Crestar, OvSynch, PGF2a and of 97.7% and 90.5%, for the ages 14 and 27 months, without significant difference. RTS influenced the pregnancy rate after the insemination, respectively of 0%, 25,6% and 48,8% for RTS 1, 2 and 3. The hormonal...(AU)

Desempenho reprodutivo de novilhas acasaladas aos 14 e 27 meses de idade submetidas a diferentes protocolos para a inseminação artificial / Reproductive performance of beef cattle heifers mated to 14 or 27 months of age subject to different protocols for artificial insemination

Avaliaram-se os efeitos de diferentes protocolos de inseminação sobre o desempenho reprodutivo em 249 novilhas Britânicas e cruzas, acasaladas aos 14 e 27 meses de idade. Sessenta e duas novilhas de 27 meses foram submetidas à IATF após o tratamento OvSynch; 61 novilhas de 27 meses e 32 novilhas de 14 meses foram submetidas à IATF após o tratamento Crestar; 62 novilhas de 27 meses e 32 novilhas de 14 meses foram submetidas à inseminação artificial (IA) após detecção de estros por 7 dias, no 7º dia foi aplicado PGF2a nos animais não observados em estro postergando-se IA por mais 5 dias (grupo PGF2a). Foram avaliados o peso vivo (PIA), o escore de condição corporal (ECC), o escore de trato reprodutivo (ETR) ao início da estação reprodutiva, a taxa de concepção e a taxa de prenhez à IATF, a taxa de prenhez ao término da estação. Os tratamentos OvSynch e Crestar tiveram 100% dos animais inseminados (IATF), enquanto os grupos PGF2a-14 meses e PGF2a-27 meses tiveram, respectivamente apenas 21,9% e 43,5% dos animais inseminados, devido à ausência ou falhas na identificação de cio. Não houve diferença nas taxas de concepção entre tratamentos hormonais e idades (P>0,05), com valores respectivos de 46,5%, 56,3%, 64,7% para os tratamentos Crestar, OvSynch, PGF2a e de 44,4% e 57,1%, para as idades 14 e 27 meses. A taxa de prenhez à IA (TF) foi de 46,5%, 56,3%, 23,4% para os tratamentos Crestar, OvSynch, PGF2a, com superioridade significativa (P<0,05) a favor dos grupos Crestar e OvSynch. A taxa de prenhez à IA (TF) foi de 18,6% e 41,3% (P<0,01) para as idades 14 e 27 meses. A taxa de prenhez final, após o repasse com touros foi respectivamente de 93,0%, 96,9%, 90,4% para os tratamentos Crestar, OvSynch, PGF2a e de 97,7% e 90,5%, para as idades 14 e 27 meses, sem diferença significativa. O ETR influenciou a taxa de prenhez após a inseminação, respectivamente de 0%, 25,6% e 48,8% para os ETR 1, 2 e 3. Os tratamentos hormonais para a IATF resultaram em maior taxa de prenhez...

The effects of three insemination protocols were evaluated on the reproductive performance in 249 British and crosses beef heifers, mated to the 14 or 27 months of age. Sixty two heifers with 27 months were submitted to FTAI after the treatment OvSynch; 61 heifers with 27 months and 32 heifers with 14 months were submitted to FTAI after the treatment to Crestar; 62 heifers with 27 months and 32 heifers with 14 months were submitted to the artificial insemination (AI) after oestrus detection for 7 days, in the 7th day PGF2a was applied in the animals without oestrus observed being postponed AI during more 5 days (PGF2a group). The live weight and body condition score (BCS), the reproductive tract score (RTS) to the beginning of mating season, the conception and the pregnancy to FTAI and the pregnancy to the end of the mating season were evaluated. The treatments OvSynch and Crestar had 100% of the inseminated animals (FTAI), while the groups PGF2a-14 months and PGF2a-27 months had respectively, only 21.9% and 43.5% of the inseminated animals, due to the absence or fails in the oestrus identification. There was no difference in the conception rates between hormonal treatments and ages (P>0.05), with respective values of 46.5%, 56.3%, 64.7% for the treatments Crestar, OvSynch, PGF2a and of 44.4% and 57.1%, for the ages 14 and 27 months. The pregnancy rate to FTAI/AI it was of 46.5%, 56.3%, 23.4% for the treatments to Crestar, OvSynch, PGF2a, with significant superiority (P<0.05) in favor of the groups Crestar and OvSynch. The pregnancy rate to AI (FTAI) it was of 18.6% and 41.3% (P <0,01) for the ages 14 and 27 months. The final pregnancy rate, after clean-up bulls was respectively of 93.0%, 96.9%, 90.4% for the treatments to Crestar, OvSynch, PGF2a and of 97.7% and 90.5%, for the ages 14 and 27 months, without significant difference. RTS influenced the pregnancy rate after the insemination, respectively of 0%, 25,6% and 48,8% for RTS 1, 2 and 3. The hormonal...

Viability of equine semen stored in a polyethylene system for transport for eight hours / Viabilidade do sêmen equino armazenado em sistema de polietileno para transporte por oito horas

Background: Equine semen storage and shipment, being it colled or frozen, allows the veterinarian to direct matings, providing the use of genetically superior stallions, which are mostly located in breeding stations or training centers. Achieving good pregnancy rates depends, beyond the moment of artificial insemination (AI), on factors related to the semen cooling, such as: system used for transport, cooling rate, final storage temperature, storage time and individual variation among stallions, such as age and resistance to cooling. Based on these aspects, this experiment was conducted in order to test a polyethylene system to ship equine semen. Materials, Methods & Results: A total of 87 ejaculates from five stallions with known fertility were used. The stallions aged between 6 to 14 years old, being three Thoroughbred and two Miniature Pony horse. The ejaculates were collected twice a week using a Hannover artificial vagina. After each collection, the semen sample was macroscopically evaluated for appearance, color and smell. A semen sample was used to evaluate the parameters of total motility, vigor and concentration, being these last three parameters assessed by counting 100 sperm cells for analysis. These analysis were performed using an optical microscope, being the concentration taken with a Neubauer chamber after dilution of 1:20 (semen: citrate formol). Subsequently to this, the semen was diluted 1+1 (diluent+semen) with skim UHT milk, and divided into four aliquots of equal volume, yielding a total of four groups. In the control group (GC) the semen was analyzed immediately after dilution (zero h - 0-h). Samples from other groups were stored for eight h (8-h) in three different devices: Equitainer® (GE), BotuFLEX® (GB) or Polyethylene System - Cooled (SP) and non-Cooled (SPN). [...]

Viability of equine semen stored in a polyethylene system for transport for eight hours / Viabilidade do sêmen equino armazenado em sistema de polietileno para transporte por oito horas

Background: Equine semen storage and shipment, being it colled or frozen, allows the veterinarian to direct matings, providing the use of genetically superior stallions, which are mostly located in breeding stations or training centers. Achieving good pregnancy rates depends, beyond the moment of artificial insemination (AI), on factors related to the semen cooling, such as: system used for transport, cooling rate, final storage temperature, storage time and individual variation among stallions, such as age and resistance to cooling. Based on these aspects, this experiment was conducted in order to test a polyethylene system to ship equine semen. Materials, Methods & Results: A total of 87 ejaculates from five stallions with known fertility were used. The stallions aged between 6 to 14 years old, being three Thoroughbred and two Miniature Pony horse. The ejaculates were collected twice a week using a Hannover artificial vagina. After each collection, the semen sample was macroscopically evaluated for appearance, color and smell. A semen sample was used to evaluate the parameters of total motility, vigor and concentration, being these last three parameters assessed by counting 100 sperm cells for analysis. These analysis were performed using an optical microscope, being the concentration taken with a Neubauer chamber after dilution of 1:20 (semen: citrate formol). Subsequently to this, the semen was diluted 1+1 (diluent+semen) with skim UHT milk, and divided into four aliquots of equal volume, yielding a total of four groups. In the control group (GC) the semen was analyzed immediately after dilution (zero h - 0-h). Samples from other groups were stored for eight h (8-h) in three different devices: Equitainer® (GE), BotuFLEX® (GB) or Polyethylene System - Cooled (SP) and non-Cooled (SPN). [...](AU)

Efeito da endometrose no potro / The influence of endometrosis on Foal Health

O presente artigo teve por objetivo revisar o papel da endometrose no ambiente uterino e seus efeitos sobre o potencial atlético dos potros gerado neste ambiente. Possibilidades de tratamento para a endometrose, como a terapia celular e a utilização da transferência de embriões são discutidos.(AU)

This paper aimed to review the role of endometrosis in the uterine environmentand its effects on the athletic potential of foals generated in this environment. Possibilities for treatment, such as cell therapy and the use of embryo transfer are discussed.(AU)

Efeito da endometrose no potro / The influence of endometrosis on Foal Health

O presente artigo teve por objetivo revisar o papel da endometrose no ambiente uterino e seus efeitos sobre o potencial atlético dos potros gerado neste ambiente. Possibilidades de tratamento para a endometrose, como a terapia celular e a utilização da transferência de embriões são discutidos.

This paper aimed to review the role of endometrosis in the uterine environmentand its effects on the athletic potential of foals generated in this environment. Possibilities for treatment, such as cell therapy and the use of embryo transfer are discussed.

O endométrio no transporte espermático e durante a gestação precoce / The endometrium in the sperm transport and during early gestation

Este trabalho objetivou revisar as ações do endométrio durante o transporte espermático após a cobertura ou inseminação artificial e suas alterações após a entrada do embrião no útero até a formação da placenta.(AU)

This paper aimed to review the endometrial actions during the sperm transport after breeding or artificial insemination and the changes occurred after the embryo enter in the uterus until placenta formation.(AU)

O endométrio no transporte espermático e durante a gestação precoce / The endometrium in the sperm transport and during early gestation

Este trabalho objetivou revisar as ações do endométrio durante o transporte espermático após a cobertura ou inseminação artificial e suas alterações após a entrada do embrião no útero até a formação da placenta.

This paper aimed to review the endometrial actions during the sperm transport after breeding or artificial insemination and the changes occurred after the embryo enter in the uterus until placenta formation.

Effect of intramuscular injection of butafosfan and cobalamin on the quality of fresh and cooled stallion semen / Efeito da administração intramuscular de butafosfan e cobalamina na qualidade do sêmen fresco e resfriado de garanhões

The use of butafosfan in combination with cobalamin modulates many cellular metabolic functions in several species. Its use enhances productive and reproductive performance and reduces stress responses in animals. Despite all these attributes, so far there have been no controlled studies to evaluate the effects of butafosfan and cobalamin on the quality of stallion semen. The purpose of this study was to evaluate the action of butafosfan in combination with cobalamin on the quality of fresh and cooled stallion semen. Four healthy stallions were kept in the same place and under the same management conditions during the entire experiment. Stallions were randomly assigned to two treatment groups in a 2x2 crossover design. Group A stallions were treated with an intramuscular injection of butafosfan twice a week for 80 days, while group B did not receive any treatment. After that, both groups were not treated for another 80 days allowing a washout period for the treated group. Then, the groups were reversed, and group B was treated with butafosfan and group A acted as the control for another 80 days. Semen was collected twice a week, diluted in skim milk and evaluated for total sperm count, total and progressive sperm motility, membrane integrity (CFDA/PI staining) and membrane functionality (HOS test) at 0 and 24 hours after preservation at 5ºC. Data were analyzed by comparing the...(AU)

A utilização de Butafosfan em combinação com cobalamina modula uma série de funções metabólicas celulares em várias espécies. Seu uso melhora o desempenho produtivo e reprodutivo e reduz as respostas ao estresse em animais. Apesar de todos esses atributos, até agora não houve estudos controlados para avaliar os efeitos da substância Butafosfan e cobalamina sobre a qualidade do sêmen de garanhões. O objetivo deste estudo foi avaliar a ação do Butafosfan em combinação com cobalamina sobre a qualidade do sêmen fresco e resfriado de garanhões. Quatro garanhões saudáveis foram mantidos no mesmo lugar e sob as mesmas condições de manejo durante todo o experimento. Garanhões foram divididos aleatoriamente em dois grupos de tratamento em um desenho cruzado 2x2. Garanhões do grupo A foram tratados com uma injeção intramuscular de Butafosfan duas vezes por semana durante 80 dias, enquanto o grupo B não recebeu qualquer tratamento. Depois disso, ambos os grupos não foram tratados durante mais 80 dias, permitindo um período de washout para o grupo tratado. Em seguida, os grupos foram invertidos, o grupo B foi tratado com Butafosfan enquanto o grupo A agiu como controle por mais 80 dias. O sêmen foi coletado duas vezes por semana, diluído em leite desnatado e avaliado para a contagem total de espermatozoides, motilidade progressiva e total, integridade da membrana (coloração CFDA/PI) e...(AU)

Resposta reprodutiva de novilhas de corte associada a marcadores moleculares relacionados à fertilidade / Reproductive Performance of Beef Cattle Heifers Associated with Molecular Markers Related to the Fertility

Background: The marker assisted selection (MAS) is indicated as an auxiliary strategy to increase the productive performance in cattle. The principle consists in the identifi cation and selection of molecular markers associated with genes involved directly or indirectly with the expression of a desired trait, usually multifactorial and polygenic. The pregnancy rate is a form of expression fertility, with great economic importance in cattle production systems. The pregnancy rate is a multifactorial trait, which is infl uence by genetics, environment and their interactions. In the current study was search for an association between the reproductive response, expressed by pregnancy rate at insemination and pregnancy rate at the end of the mating season, and molecular markers of genes related to IGF-1 receptor, LHß, Leptin, receptors of FSH and LH. Materials, Methods & Results: Data were evaluated from 249 beef heifers of British breeds, Angus, Devon and crosses. The animals at fourteen or twenty seven months were subjected to different protocols for artificial insemination (AI) and fixed-time artificial insemination (TAI), followed by clean up bulls. The protocols AI and TAI, called AI/TAI, were: Group 1 -Ovsynch protocol (27th months). At day zero the animals received an injection of a GnRH at a dose of 10 µg buserelin, at seventh day received 500 µg of cloprostenol sodium, at ninth day the animals received another dose of 10 µg of buserelin. TAI was performed 10 h after the application of buserelin; Group 2 - Protocol Crestar® with ½ dose of estradiol valerate injectable (27th months). At eighth day the implant was removed, followed by 375 µg of cloprostenol sodium. At ninth day, was administered 1 mg of estradiol benzoate. TAI was performed in tenth day, 52 h after implant removal; Group 3 - Control group (27th months), with AI 12 h after estrus during the fi rst seven days. Seventh day, was applied 375 µg of cloprostenol sodium on all heifers not inseminated, followed by AI for fi ve days more; Group 4 (14th months) - Same protocol of the Group 2; Group 5 (14th months) - Same protocol of the Group 3. During the group formation, blood samples for extraction of DNA were obtained individually from all experimental animals. Microsatellite markers (short tandem repeats, STR) and/or single nucleotide polymorphisms (SNPs) were investigated by means of polymerase chain reaction (PCR). The STR investigated were the AFZ-1, HEL5, ILST002, IDVGA51, FSHR and LHR. We considered the effects of the treatment groups, age, body condition score (BCS), reproductive tract score (RTS), weight at the beginning of the breeding season, daily weight gain on pregnancy rate to AI/TAI and pregnancy rate to final. Molecular markers investigated did not show association with pregnancy rate. The mean pregnancy rate at insemination was 41.0% and the pregnancy rate final was 91.6%. There were no differences in pregnancy between the age of the animals. The average weight at the beginning of the mating was 313 kg without differentiate between animal that become pregnant or empty during the reproductive season. The RTS influenced the pregnancy rate after insemination in respectively 0%, 32.1% and 49.5% (P < 0.05) for RTS 1, 2 and 3. Discussion: Molecular markers AFZ-1, HEL5, ILST002, IDVGA51, FSHR and LHR could not be associated with pregnancy rate at insemination and pregnancy in beef heifers, perhaps because the high degree of reproductive selection that these animals are subjected. The nutritional status of the herd, expressed by weight and body condition score at mating of animals contributed to the achievement of significant pregnancy rates. The reproductive tract score can be considered as a predictor of fertility in herds of beef heifers.

Avaliação do desempenho reprodutivo de vacas de corte lactantes submetidas à IATF a partir da aplicação do GnRH, da manifestação estral, da reutilização de dispositivos intravaginais e da condição corporal / Evaluation of reproductive performance of lactating beef cows subjected to FTAI from the application of Gnrh, estrus manifestation, re-use of intravaginal devices and body condition

Background: The success of fixed-time artificial insemination (FTAI) is linked to several factors that affect pregnancy rates. The use of different hormones, estrus expression and body condition score (BCS) are variables that can influence the results of the technique. The increased production of calves provides greater amount of steers to slaughter. The productive intensification of the beef cows breeding depends on factors related to reproductive technique used and the physiological conditioning of animals. The objective of this study was to evaluate the effect of the application of GnRH, estrus manifestation, re-use of progesterone intravaginal devices and BCS on pregnancy rates of cows subjected to FTAI. Materials, Methods & Results: Were analyzed the effects of GnRH injection and re-use of intravaginal device, estrus manifestation and body condition score (BCS) on reproductive performance of 222 Montana beef cows subjected to fixedtime artificial insemination (FTAI). The FTAI protocol was based on the placement of intravaginal progesterone device (1st, 2nd or 3rd use) and application of estradiol benzoate (E.B.) (2 mg) on day 0. On day 8 was applied sodium cloprostenol (500 µg), removed the intravaginal device and performed the separation of calves. On day 9 was applied E.B. (1 mg) and started the observation of estrus in the morning, afternoon and morning of day 10. On the afternoon of day 10 was realized the insemination, part of the animals showed estrus received buserelin acetate (GnRH) (100µg); the same was proceeded with the animals without estrus. After FTAI the calves returned to cows. Thus, it was formed four groups: Without-Estrus/Without-GnRH; Without-Estrus/With-GnRH; With-Estrus/Without-GnRH; With-Estrus/With-GnRH. The evaluation alone of application of GnRH (with and without) and estrus manifestation (with and without) had no effect on pregnancy rates to FTAI and pregnancy final (P > 0.05). The evaluation of the interaction between GnRH and estrus had no effect on the FTAI pregnancy (P > 0.05): cows Without-Estrus/Without-GnRH did not differ those with GnRH, 33.3% and 48.4% respectively; cows With-Estrus/Without-GnRH did not differ those with GnRH, 53.2% and 53.8% respectively. Intravaginal device 1st, 2nd and 3rd use did not influence pregnancy rates to FTAI and final pregnancy, 50.0%, 57.7%, 41.7% and 91.7%, 88.5%, 93 8% respectively (P > 0.05). The BCS influenced the pregnancy rate to FTAI and final pregnancy, animals with BCS ≥ 3.0 had a higher pregnancy rate to FTAI and animals with BCS ≥ 2.5 had a higher pregnancy to final pregnancy (P < 0.05). The BCS also influenced the estrus manifestation, animals with BCS ≥ 2.5 had higher rates of estrus (P < 0.05). The intravaginal device 1st use promoted greater estrus manifestation in relation to the 2nd and 3rd use (P < 0.05). Discussion: The pregnancy rates did not have the effect of application of GnRH. Some studies report that effect of GnRH is related to estrus phase of the cow. Although 11 percentage points above to cows with estrus, the manifestation estrus did not influence the pregnancy and also did not interact with GnRH. However, it is reported that estrus before at insemination may be conducive to conception. The intravaginal devices used did not influence the pregnancy rates, showing that it is possible re-use them. The results showed at importance of BCS to pregnancy, studies report that cows with low BCS have difficulty to pregnant at FTAI and clean-up bulls. The estrus manifestation was influenced by the BCS and the intravaginal device used.

Suplementação de touros com sabões cálcicos de ácidos graxos poli-insaturados e qualidade seminal pré e pós-congelação / Bulls supplemented with calcium soaps of polyunsaturated fatty acids and semen quality before and after freezing

O objetivo deste experimento foi avaliar a qualidade seminal de touros suplementados com sabões cálcicos de ácidos graxos poli-insaturados (PUFA). Vinte touros das raças Angus, Brangus, Hereford e Braford foram distribuídos aleatoriamente em dois grupos, cada um com dez animais. Cada grupo recebeu uma dieta composta de volumoso, ração comercial, sal mineral e um de dois suplementos: funcional com a incorporação de sabões cálcicos de PUFA ou energético, com a incorporação de raspa de mandioca (Manihot esculenta, Crantz). Durante o período experimental, a qualidade seminal foi avaliada utilizando testes rotineiros (volume, concentração, motilidade em massa, motilidade individual e avaliação da morfologia espermática) e testes complementares (teste hiposmótico, coloração dupla giemsa/tripan-azul e teste de termorresistência). O sêmen in natura do grupo de touros suplementados com sabões cálcicos de PUFA comercial (Megalac-E®) apresentou aumento na motilidade espermática (83,3% vs. 75,3%), na percentagem de espermatozoides com acrossoma íntegro (94,8% vs. 91,8%) e na percentagem de espermatozoides viáveis (98,0% vs. 96,6%) quando comparado com o sêmen do grupo de touros suplementados com raspa de mandioca (P<0,05). O tipo de suplemento não afetou o volume seminal, a concentração espermática e concentração de testosterona em sangue (P>0,05). Foram encontradas interações entre tratamento e número de coleta para percentagem de espermatozoides morfologicamente normais e percentagem de espermatozoides positivos ao teste hipo-osmótico (P<0,05). Após a descongelação, o tipo de suplemento não afetou a motilidade dos espermatozoides, o teste hipo-osmótico e o teste de termoresistência rápido (P>0,05). A suplementação com sabões cálcicos de PUFA aumentou as percentagens de espermatozoides com acrossoma íntegro (48,0% vs. 39,2%) e de espermatozoides com membrana plasmática integra (51,5% v.s 42,2%) pós-descongelação.(AU)

The objective of this experiment was to evaluate semen quality of bulls supplemented with calcium soaps of fatty acids polyunsaturated. Twenty Angus, Brangus, Hereford and Braford bulls were randomly allocated into two groups, each with ten animals. Each group received a diet consisting of forage, commercial ration, mineral supplements and supplement. Two supplements types were evaluated: functional supplement, with fatty acids polyunsaturated (PUFA) incorporation or energy supplement, with cassava meal (Manihot esculenta, Crantz) incorporation. During trial period the semen quality was evaluated using routine test (volume, concentration, mass motility, individual motility, sperm morphology) and complementary test: (hypo-osmotic test, giemsa/tripan-azul coloration and thermotolerance test). The semen of group bulls supplemented with commercial calcium soaps of PUFA (Megalac-E®) showed greater sperm motility (83.3% vs. 75.3%), percentage of sperm with intact acrosome (94.8% vs. 91.8%) and percentage of sperm with intact plasma membrane (98.0% vs. 96.6%) when compared with semen of group bulls supplemented with cassava (P<0.05). The supplement type did not affect semen volume, sperm concentration and testosterone concentration in blood (P>0.05). There were interactions between treatment and collection period for percentage of normal sperm and percentage of spermatozoa with hypo-osmotic positive test (P<0.05). The type of supplement did not affect sperm motility, the hypo-osmotic test and fast thermotolerance test after thawing (P>0.05). PUFA calcium soaps supplementation increased percentage of spermatozoa with plasma membrane integrity (51.5 vs. 42.2%) and spermatozoa intact acrosome (48.0% vs. 39.2%) after thawing.(AU)