Orientador: Frederico Ozanam Papa / Banca: João Carlos Pinheiro Ferreira / Banca: Alício Martins Junior / Resumo: A despeito das inúmeras variáveis que influenciam direta e indiretamente a fertilidade das fêmeas bovinas, a qualidade das amostras seminais exerce um papel importante na determinação das taxas de concepção dos programas de inseminação artificial. Os objetivos dessa pesquisa foram comparar a efetividade de dois diluidores de criopreservação de sêmen bovino no processamento de amostras seminais apresentando diferentes concentrações espermáticas em relação aos índices de congelabilidade determinados laboratorialmente (Experimento I) e as taxas de concepção proporcionadas por cada metodologia quando utilizada em programas de inseminação artificial em tempo fixo (IATF) em bovinos (Experimento II). No Experimento I foram utilizados 14 ejaculados de diferentes touros da raça Nelore. Cada ejaculado foi fracionado em oito alíquotas iguais, submetidas a criopreservação com os diluidores Tris-gema de ovo-frutose (meio TRIS) e MKA nas concentrações de 12, 25, 50 e 100 milhões de espermatozóides totais por mililitro de meio, formando oito grupos experimentais em função das variáveis diluidor e concentração. As amostras foram descongeladas a 46 ºC por 20 segundos, avaliando-se os padrões de motilidade através do método computadorizado (CASA), integridade de membrana plasmática (IMP), resistência ao teste de termorresistência rápido (TTR) e taxa de recuperação e IMP após seleção espermática pela técnica de swim-up. Para o Experimento II foram selecionados sete touros utilizados no Experimento I, obtendo-se um ejaculado de cada animal por eletroejaculação...(Resumo completo, clicar acesso eletrõnico abaixo) / Abstract: Although there are many variables which directly or indirectly influence female bovine fertility, the quality of sperm samples plays a important role in the determination of conception rates in artificial insemination programs. The aim of the present study was to compare the efficiency of two bovine semen extenders for sperm freezing with different spermatic concentrations in the freezability determined by lab tests (Experiment I), and conception rates after fixed time artificial insemination (FTAI; Experiment II). In Experiment I 14 ejaculates of different Nelore bulls were used. Each ejaculate was splitsampled in to eight equal parts and then submitted to cryopreservation with Tris-egg yolk fructose (TRIS) and MKA extenders, at concentrations of 12, 25, 50 and 100 millions spermatozoa per milliliter forming eight experimental groups. The samples were thawed at 46 ºC for 20 seconds, and the following parameters were evaluated: sperm motility and movement (by computer-assisted semen analysis - CASA), sperm membrane integrity (SMI), resistance to the fast thermoresistance test (TT), recovery rate and sperm membrane integrity after sperm selection through swim-up technique. Seven of 14 bulls used in Experiment I were selected for Experiment II, and semen was collected from each of the animals by electroejaculation. The seven ejaculates obtained were mixed (semen pool) and cryopreserved, thus forming eight experimental groups according to the freezing extenders and sperm concentrations/straws: TRIS 12, 25, 50 and 100, and MKA 12, 25, 50 and 100...(Complete abstract, click electronic address below) / Mestre

Call number: LD2668 .T4 1985 M52 / Master of Science

Thesis (M. Tech.) -- Central University of Technology, Free State, 2007 / Four different South African indigenous (Naked Neck (NN), Ovambo (OVB), Venda (VD) and Potchefstroom Koekoek (PK) chicken breeds were used in this study. From each of the four breeds of chicken, 40 hens and 8 cocks were selected randomly. Two groups each of sixteen cocks were subsequently formed: high performing (HP) and low performing (LP) groups to determine the relationships between cock semen viability and the fertility of artificially inseminated South African indigenous layer breeds. Semen was collected following five minutes of sexual massage (5SM) and evaluated for semen volume (ml), sperm motility (%), live sperm (%) and total sperm (x109/ml). Semen from each cock was then used to inseminate five hens per breed, in each treatment. Each hen was inseminated twice a week throughout the duration of the trial. During the experimental period, each hen was inseminated with 0.05 ml diluted semen. The artificially inseminated hens were examined for average egg weight (g), fertility (%), hatchability of set eggs (%), live chicks (%), normal chicks (%) and chick weight (g). A total of 1600 eggs, i.e. 400 eggs from each breed were collected in three batches following artificial insemination from individually caged hens and were hatched to compare hatching parameters among breeds. The hatchability traits of hens of the four breeds (NN, OVB, PK and VD) were compared. Hatching egg weight had significant (P < 0.05) difference among the four breeds. The results of this study indicate that semen viability exemplified by ejaculate volume, sperm motility; live sperm and total sperm per ejaculate were significantly (P < 0.01) superior in the HP cocks compared to the LP cocks. Hens inseminated with semen from the HP cocks in each experimental group resulted in higher egg weight (g), fertility (%), hatchability of set eggs (%), live chicks (%), normal chicks (%) and chick weight (g). Significant positive relationships existed between semen volume and sperm motility (P < 0.05), semen volume and live sperm cells (P < 0.01), semen volume and total sperm (P < 0.01) in NN, OVB and VD, with negative correlations in PK. Some positive correlations were found between sperm motility and live spermatozoa (P < 0.01), sperm motility and total sperm (P < 0.01), live sperm and total sperm (P< 0.01) in NN, OVB, PK and VND. Fertility was the highest in the HP group. Fertility was also the highest in PK, intermediate and similar in OVB and NN and lowest in VD (P<0.05). Breed had a significant effect on hatchability of fertile eggs (P<0.05). Hatchability of total eggs set was highest in PK and NN, intermediate in OVB and lowest in VD (P<0.05). Breed had a significant effect on live, normal chicks and chick weight (P<0.05). Live chick was the highest in NN, whereas at day-old, normal chick and chick weight at hatching were the highest (23.50 ± 0.11) (P<0.05) in PK (98.14 ± 0.67 vs. 37.90 ± 0.28 g), intermediate and similar in NN (87.90 ± 0.63 vs. 23.50 ± 0.11) and OVB (87.75 ± 0.45 vs. 32.81 ± 0.49 g) and the lowest but with an acceptable value in VD (76.85 ± 0.46 vs. 26.90 ± 0.36 g). There were some correlations among different hatchability traits depending on breed. The correlations were more profound among PK. It was clear that chick weight as percent of egg weight was not just a function of egg weight, and that genotype also played an important role favouring the heavier breeds. The results obtained in this study on the relationships between cock semen viability and the fertility of artificially inseminated South African indigenous layer breeds elucidate that the use of high performing (HP) cocks following five minutes of sexual massage, prior to semen collection and artificial insemination of layers is a practical method for optimising sperm viability and subsequent fertility of hens. The results of this study suggest that the Potchefstroom Koekoek (PK) cocks and hens are superior to the Naked Necks (NN), Ovambo (OVB) and Venda (VD). The Ovambo and Naked Neck cocks ranked second in donating quality semen as well as in improving the fertility and hatchability traits of the indigenous chicken breeds. Thus selection of high performing cocks through five minutes sexual massage prior to semen collection and use is recommended for poultry AI breeding programmes.

Thesis (MScMedSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Male factor infertility is a general term that describes couples in which an inability to conceive is associated with a problem identified in the male partner. Intrauterine insemination (IUI) together with ovulation induction has been shown to be an effective treatment method for male factor infertility. Oocyte production by the ovaries is stimulated by the use of fertility drugs. A prepared sperm sample is then injected into the uterus through the vagina using an IUI catheter which brings the oocytes and spermatozoa into close proximity. Semen preparation is an integral part of an IUI cycle. In a developing country, a simple inexpensive semen preparation method for IUI procedures, not necessitating a lot of equipment, is essential. An example of such a method, the Sep-D Kit (Surelife Sep-D Kit, Surelife Media Technologies Pty Ltd, Singapore) has been proposed as a possible preparation method. In a pilot study performed by the principal investigator (Roxanne Gentis), comparing the Sep-D Kit and standard swim-up preparation methods, it was found that the Sep-D Kit compared very well with the swim-up method regarding most pre- and post-preparation semen parameters. The Sep-D Kit method, however, still needed further testing to see whether or not pregnancy rates resulting from the method are comparable with that resulting from the standard swim-up method, as this ultimately is the required result of an IUI. The primary aim of this study was to compare the Sep-D Kit method to the standard swim-up method with regards to biochemical pregnancy outcome, post-preparation sperm count, motility, total motile count (TMC), morphology, DNA compaction and fragmentation (CMA3 and TUNEL). The secondary aim was to evaluate which variables, male and female, affect biochemical pregnancy outcome. The study took place at Drs Aevitas Fertility Clinic, Vincent Pallotti Hospital, Pinelands. The study was a prospective analytical study and was conducted from December 2010 until October 2012. A total of 473 IUI cycles were evaluated. Results showed that the Sep-D Kit semen preparation method was non-inferior to the standard swim-up method with regards to biochemical pregnancy rates, post-preparation count and TMC. The swim-up method produced samples with a significantly higher post-preparation motility compared to the Sep-D Kit method, however both methods still managed to produce similar biochemical pregnancy rates (10.39% for the swim-up group versus 11.57% for Sep-D Kit group). For the total cohort of cycles analysed the only female parameter which significantly predicted biochemical pregnancy outcome in this study was age. Sperm motility (post-preparation) was the only male parameter that significantly affected biochemical pregnancy outcome. The Sep-D Kit method is more cost effective and also time saving compared to the swim-up method. There is also no need for expensive laboratory equipment or a trained embryologist using the Sep-D Kit preparation method. The Sep-D Kit may therefore be used with confidence as a standard semen preparation method, and may be implemented in developing countries for use in routine IUI procedures. / AFRIKAANSE OPSOMMING: Manlike faktor infertiliteit is 'n algemene term wat gebruik word om paartjies te beskryf wat 'n onvermoë toon om swanger te raak as gevolg van 'n probleem wat geassosieer word met die man. Die kombinasie van intra-uteriene inseminasie (IUI) en ovulasie induksie kan doeltreffend gebruik word om manlike faktor infertiliteit te behandel. Vrugbaarheidsmiddels word gebruik om oösietproduksie in die die eierstokke te stimuleer en 'n voorbereide spermmonster word dan transvaginaal in die baarmoeder ingespuit om sodoende die spermatozoa en oösiete na-aan mekaar te bring. Semenvoorbereiding is 'n integrale deel van 'n IUI siklus en in 'n ontwikkelende land is 'n eenvoudige, goedkoop semenvoorbereidingsmetode – wat die gebruik van duur toerusting uitsluit – noodsaaklik. Die Sep-D Kit metode (Surelife Sep-D Kit, Surelife Media Technologies Pty Ltd, Singapore) is 'n voorbeeld van so 'n voorbereidingsmetode. 'n Loodsstudie, uitgevoer deur die hoofnavorser, (Roxanne Gentis), het gewys dat die Sep-D Kit en standaard opswem voorbereidingmetodes goed vergelyk ten opsigte van meeste semenparameters voor- en na voorbereiding. Dit is egter ook noodsaaklikheid vir verdere navorsing om vas te stel of swangerskapuitkoms na die gebruik van die twee semenvoorbereidingsmetodess vergelykbaar is, aangesien dit die uiteindelike, verlangde uitkoms van 'n IUI is. Die primêre doel van hierdie studie was om die Sep-D Kit metode te vergelyk met die standaard opswemmetode met betrekking tot biochemiese swangerskapuitkoms asook spermtelling, motiliteit, totale motiele spermtelling (TMS), morfologie, DNA kompaksie en fragmentering (CMA3 en TUNEL) na spermvoorbereiding. Die sekondêre doel was om te evalueer watter veranderlikes, manlik en vroulik, die bichemiese swangerskapuitkoms beïnvloed. Die studie is uitgevoer by die Drs Aevitas Fertiliteitskliniek, Vincent Pallotti Hospitaal, Pinelands. Die studie was prospektief analities en het gestrek vanaf Desember 2010 tot en met Oktober 2012. 'n Totaal van 473 IUI siklusse is evalueer en ontleed. Die resultate van die studie het getoon dat die Sep-D Kit semenvoorbereidingsmetode nie ondergeskik aan die opswemmetode was ten opsigte van biochemiese swangerskap, spermtelling en TMS na semenvoorbereiding nie, Spermmotiliteit was betekenisvol hoër vir die opswemmetode vergelykend met die Sep-D Kit, maar ten spite van die verskil was die biochemiese swangerskapsyfers in die twee groepe nie verskillend nie (10.39% in die opswem groep en 11.57% in Sep-D Kit groep). In die totale kohort siklusse wat ontleed is was dit net die ouderdom van die vrou wat 'n betekenisvolle effek op biochemiese swangerskapuitkoms gehad het. Die enigste manlike faktor wat 'n betekenisvolle effek op biochemiese swangerskapuitkoms gehad het was die motiliteit na semenvoorbereiding. Die Sep-D Kit metode is meer koste-effektief en tydbesparend as die standard opswemmetode. Die uitvoer van die Sep-D Kit metode vereis ook ook geen duur apparaat of 'n opgeleide embrioloog nie. Die Sep-D Kit metode kan dus met vertroue gebruik word as 'n standaard semenvoorbereidingsmetode en kan in ontwikkelende lande vir gebruik tydens roetine IUI prosedures geïmplementeer word.

Název diplomové práce v anglickém jazyce: Legal aspects of abortion, sterilization and assisted reproduction Summary: The thesis deals with the three topics from the field of medical law, particularly abortion, sterilization and assisted reproduction. The issue of giving birth is the key here. The new Special Medical Services Act regulating, inter alia, some aspects of sterilization and assisted reproduction came into effect in April 2012. The previous regulation of these matters was outdated and some legal issues were not tackled at all. The first part of the thesis focuses on legal regulation of abortion, which is contained in the act from 1986 and in the implementing legislation. Especially the conditions of intervention itself, payments and intervention on juveniles (under 18) and foreigners in the Czech Republic are tackled. The regulation of other countries is mentioned and compared as well. The second part of the thesis deals with the legal regulation of sterilization, conditions of sterilization and contraindication as well. The specific problem of illegal sterilizations of gypsy women in the second half of the twentieth century is pointed out. These disputes were present even before the European Court for Human Rights in Strasbourg. The methods of assisted reproduction, as a modern way of resolving...

Master of Science / Department of Animal Sciences and Industry / Jeffrey S. Stevenson / We hypothesized that pregnancy outcomes may be improved by inducing luteal regression, ovulation, or both before a control CO-Synch + CIDR program (100 mcg GnRH i.m. [GnRH-1] and insertion of a progesterone-impregnated intravaginal controlled internal drug release [CIDR] insert on d -10, 25 mg PGF2alpha (PG) i.m. and CIDR insert removal on d -3, and 100 mcg GnRH i.m. [GnRH-2] and timed AI [TAI] on d 0) in suckled beef cows. This hypothesis was tested in 2 experiments, in which cows were treated with either PG or PG + GnRH before initiating a control CO-Synch + CIDR program to increase the proportion of cows starting the program in a low (< 1 ng/mL; Exp. 1) or high (≥ 1 ng/mL; Exp. 2) progesterone status, respectively. Blood was collected before each injection for later progesterone analyses. In Exp. 1, cows at 9 locations (n = 1,537) were assigned to either: (1) control or (2) PrePG (same as control with a PG injection on d -13). The PrePG cows had larger (P < 0.05) follicles on d -10 and more (P < 0.05) ovulated after GnRH-1 than controls (60.6 vs. 36.5%). Incidence of estrus between d -3 and 0 was greater (P < 0.05) for treated multiparous cows than multiparous controls and treated and control primiparous cows (74.1 vs. 64.3, 58.6, and 59.1%, respectively). In Exp. 2, cows at 4 locations (n = 803) were assigned to: (1) control (same as Exp. 1) or (2) PrePGG (same as control with PG injection on d -20 and GnRH injection on d -17. Cows with BCS > 5.0 or ≥ 70 d postpartum at TAI were more (P < 0.05) likely to become pregnant than thinner cows or those with fewer days postpartum. Treated cows in both experiments were more (P < 0.05) likely than controls to have luteolysis after initial PG injections and reduced (P < 0.05) serum progesterone. In both experiments, pregnancy rates at d 35 did not differ between treatment and control; however, cows classified as anestrous before d -10, but with elevated progesterone on d -10, had increased (P < 0.05) pregnancy outcomes than remaining anestrous cows with low progesterone concentrations. In summary, luteal regression and ovulation were enhanced by treatments before the 7 d CO-Synch + CIDR program; however, pregnancy per TAI was not improved.

Master of Science / Department of Animal Sciences and Industry / Jeffrey S. Stevenson / Our objectives were to determine: the effectiveness of upfront PGF2α injection to regress the corpus luteum; ovulation response to GnRH; and pregnancy outcomes. Dairy heifers (n = 545) from three locations (Florida, Kansas, and Mississippi) were assigned randomly to each of two treatments: 1) 25 mg of PGF2α injection and insertion of previously used autoclaved CIDR on d 7 followed by 100 µg of GnRH administered on d 5, and a 25 mg PGF2α injection at CIDR removal (7D) on d 0; 2) 100 µg of GnRH and insertion of previously used autoclaved CIDR on d 5 and 25 mg of PGF2α injection at CIDR removal (5D) on d 0. Artificial insemination (AI) occurred after detected estrus from d 0 to 3. Those heifers not detected in estrus were inseminated on d 3 and given a second 100 µg of GnRH. Blood collected on d 7 and 5 was assayed to determine concentrations of progesterone, presence of a CL (progesterone ≥1 ng/mL) on d 7, and whether luteolysis occurred in 7D heifers. Blood progesterone concentration from d 0 and 3 determined if luteolysis occurred in all heifers. Ovarian structure maps on d 5 and 0 were used to determine ovulation in response to GnRH on d 5. Pregnancy was determined on d 32 and 60 and intervening pregnancy loss was calculated. Of those heifers in the 7D treatment having progesterone ≥1 ng/mL on d 7, the proportion having progesterone <1 ng/mL 2 d later (luteolysis) was greater (P < 0.05) than that in the 5D treatment (43.0 vs. 22.9%, respectively). Total proportion of follicles that ovulated per heifer was numerically greater in the 7D treatment but only differed (P < 0.05) between locations. A treatment x location interaction was detected for pregnancy rates per AI. The Kansas location had no detectable treatment differences. In contrast, the 7D treatment produced greater (P < 0.05) pregnancy rates in the first replicate of the Florida location and at the Mississippi location. We concluded that the 5D protocol was not effective in producing acceptable luteolysis, pregnancy, and ovulation rates in comparison with the modified 7D protocol.

Orientador : Flávio Abranches Pinheiro / Banca: Izabel Cristina Takitane / Banca: Ciro Moraes Barros / Banca: Geraldo Sant'Ana C. de Barros / Banca: Evaristo Marzabal Neves / Resumo: A bovinocultura de corte é de reconhecida importância no agronegócio brasileiro (mercado interno e externo), sendo que o país tem o segundo rebanho bovino mundial, com cerca de 80% destinados a produção de carne (maioria de zebuínos). Um dos limitantes desta atividade é a reprodução dos animais, que permite que o rebanho cresça em número e qualidade. Atuando-se sobre a reprodução, pode-se melhorar a conversão alimentar, diminuindo o custo. A inseminação artificial (IA) tem grande importância na obtenção de ganhos quantitativos e qualitativos (e.g. ganho genético), além de permitir um melhor controle do rebanho. Novas tecnologias de inseminação estão sendo aperfeiçoadas, inclusive tratamentos hormonais que possibilitam a IA com tempo fixo (IATF). Considerando a questão energética, é importante o desenvolvimento de tecnologias que otimizem os gastos energéticos na bovinocultura. Este trabalho tem por objetivo avaliar os custos econômicos (valores em reais, julho de 2003) nos manejos reprodutivos: IA convencional (IAC), IATF (GnRH-PGF2 -GnRH, GnRH-PGF2 -BE), GP (controle, GnRH-PGF2 e IA) e Monta; em vacas Nelore (zebuínos), além de fazer considerações sobre o gasto energético biológico destes manejos. A partir de dados obtidos em uma fazenda comercial da região de Botucatu, SP, foram realizadas análises de custos de reprodução de bezerros, nas tecnologias de IAC, IATF (grupos GPG-9, GPG-12, GPE-15 e GPE-18) e GP (GP-21 e GP-24) associados à Monta; e Monta-Simulado. O melhor Custo Total Unitário de reprodução por bezerro (CTU) foi observado na IAC (R$ 127,00) com taxa de prenhez (TxPr) de 84%. Na IATF, o melhor resultado foi do grupo GPE-15 (R$ 130,00, TxPr 74%)...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Beef cattle is a very important sector in Brazilian agribusiness, both in domestic and international market. Brazil has the second largest bovine herd in the world, with 80% of beef cattle (mostly Bos indicus). A limiting factor of this activity is the animal reproduction, which allows herd improvement and growth. Acting on reproduction may allow improvements on feed conversion, diminishing costs. Artificial insemination (AI) is a way of obtaining quantitative and qualitative gains (genetic improvement), in addition to a better herd management. New technologies on AI are been developed, including hormonal treatments which allow timed fixed AI (TAI). Besides, on the energetic approach, the development of new technologies optimizing energy expenses is also very important. The aim of this study was to evaluate the economic costs (in Reais, July 2003) on several reproductive handlings: Conventional AI (IAC), Timed AI (TAI; GnRH-PGF2 -GnRH; GnRH-PGF2 -BE), Synchronized estrus GP (control, GnRH-PGF2 and AI) and Natural Service; in Nelore cows (Bos indicus), in addition to making short considerations regarding energetic expenses on those handlings. Reproduction costs analysis were performed on data obtained on a commercial farm in Botucatu, SP, on IAC and TAI (groups GPG-9, GPG-12, GPE-15 and GPE-18) and GP (GP-21 e GP-24) associated to Natural Service; and simulated Natural Service group. The best total reproduction cost per calf (CTU, R$ 127,00) was observed in IAC with the pregnancy rate (TxPr) of 84%; otherwise, in TAI, the best result was from GPE-15 (R$ 130,00, TxPr 74%). The simulated Natural Service group had a CTU of R$ 116,00, with TxPr of 87%. The other groups obtained CTUs of R$163,00 (GPG-12, TxPr 69%), R$174,00 (GPE-18, TxPr 68%), R$181,00 (GP-21, TxPr 59%), R$ 189,00 (GPG-9, TxPr 67%) e R$ 338,00 (GP-24 TxPr 31%)...(Complete abstract click electronic access below) / Doutor

O objetivo do presente estudo foi avaliar a viabilidade da refrigeração do sêmen bovino comparada com a criopreservação. Este estudo foi realizado em dois experimentos. No primeiro experimento foram comparados os efeitos in vitro do sêmen refrigerado em três meios diluidores comerciais a 5&deg; C por até 48 horas, e criopreservado em dois meios. Para este experimento foram utilizados ejaculados de 10 touros da raça Nelore. Cada ejaculado foi dividido em três alíquotas, sendo diluídas nos meios Botubov&reg;, Steridyl&reg; e Bovidyl&reg;. Após a diluição o sêmen foi envasado em palhetas e refrigerado nos três diluidores e criopreservado utilizando somente os meios Botubov&reg; e Steridyl&reg;. A refrigeração do sêmen foi realizada a 5&deg; C por até 48 horas em sistema passivo de refrigeração BotuFlex&reg; e a criopreservação realizada em sistema automatizado TK 4.000&reg;. O sêmen foi avaliado nos tempos 0, 24, 36 e 48 horas após a refrigeração e após a descongelação, para cada diluidor. Foram realizadas análises dos padrões de cinética espermática pelo sistema computadorizado de análise espermática (CASA, programa SCA - Sperm Class Analyser), integridade das membranas plasmática e acrossomal, potencial de membrana mitocondrial e estresse oxidativo por sondas fluorescentes, sob microscopia de epifluorescência e morfologia espermática por microspcopia de contraste de interferência diferencial (DIC). Notou-se efeito de tempo de refrigeração para os três diluidores para de 0 para 24h, se mantendo semelhante até 48 h. Os diluiores Botubov&reg; e Steridyl&reg; preservaram as características espermáticas de forma semelhante até 48 horas de refrigeração diferindo apenas na variável de velocidade curvilianear; no entanto, ambos foram superiores ao diluidor Bovidyl&reg;, para as variáveis, motilidade total, motilidade progressiva, células rápidas, velocidade curvilinear, velocidade progressiva, velocidade do trajeto, retilinearidade, integridade da membrana plasmática, alto potencial de mitocondrial e espermatozoides com membranas plasmática e acrossomal íntegras e alto potencial mitocondrial. O segundo experimento foi realizado, baseado nos resultados do primeiro experimento, para avaliar os efeitos da refrigeração e da criopreservação do sêmen sobre a fertilidade in vivo. Foram utilizados ejaculados de três touros das raças Brangus, Braford e Angus, com idade entre 5 e 7 anos. O sêmen foi colhido por meio de vagina artificial, o ejaculado foi dividido em três alíquotas, sendo duas alíquotas para refrigeração e uma para criopreservação. Para a refrigeração o sêmen foi diluído nas concentrações de 15x106 (R15) e 30x106 (R30) espermatozoides/palheta e para a criopreservação diluído na concentração de 30x106 espermatozoides/palheta (CRIO). Para todas as diluições foi utilizado o meio Botubov&reg; e o sêmen armazenado em palhetas de 0,5 mL. A refrigeração foi realizada a temperatura de 5&deg; C por 48 horas em sistema passivo de refrigeração BotuFlex&reg; e a criopreservação em sistema automatizado TK&reg;. Foram sincronizadas 552 vacas da raça Brangus em programas de inseminação artificial em tempo fixo. Os resultados da taxa de prenhez para os grupos de vacas inseminadas foram de 49,4% para R15, 43,38% para R30 e 47,59% para o sêmen criopreservado. Pode-se concluir que a refrigeração do sêmen a 5&deg; C por 48 horas resulta em taxa de prenhez semelhante às obtidas com o sêmen criopreservado, sendo que esses resultados indicam que a refrigeração por até 48 horas pode ser uma opção de uso para IATF. / The objective of the present study was to evaluate the viability of cooling bovine semen compared to cryopreservation. This study was carried out in two experiments. In the first experiment, the in vitro effects of cooled semen were compared in three commercial extenders at 5&deg; C for up to 48 hours, and cryopreserved in two extender. For this experiment were used ejaculates of 10 Nellore bulls. Each ejaculate was divided into three aliquots, being diluted in the Botubov&reg;, Steridyl&reg; and Bovidyl&reg; extenders. After dilution, the semen was cooled into three extenders and cryopreserved using the Botubov&reg; and Steridyl&reg;. Semen refrigeration was performed at 5&deg; C for up to 48 hours in BotuFlex&reg; passive refrigeration system and cryopreservation performed in TK 3.000&reg; automated system. Semen was evaluated at 0, 24, 36 and 48 hours after refrigeration and after thawing, for each diluent. The sperm kinetics of the spermatic kinetics (CASA, SCA - Sperm Class Analyzer), plasma and acrosomal membrane integrity, mitochondrial membrane potential and oxidative stress by fluorescent probes were analyzed under epifluorescence microscopy and sperm morphology By Differential Interference Contrast Microscopy (DIC). The Botubov&reg; and Steridyl&reg; diluents were very similar after 48 hours of cooling differing significantly only in the Curvilianear Velocity (VCL) 106.04 m/s and 124.56 m/s. The Bovidyl&reg; diluent yielded results significantly lower than the 48 hours of refrigeration for the variables: total motility (MT), progressive motility (MPRO), fast cells (RAP), curvilinear velocity (VCL), progressive velocity (AP), plasma membrane integrity (PI), high mitochondrial potential (AP), and spermatozoa with intact plasma and acrosomal membranes and high mitochondrial potential (PIAIA). The second experiment was carried out, based on the results of the first experiment I, to evaluate the effects of cooled and cryopreservation of semen on in vivo fertility. We used ejaculates of three bulls of the Brangus, Braford and Angus breeds of a IA center, aged between 5 and 7 years. The semen was collected by artificial vagina, the ejaculate was divided into three aliquots, two aliquots for refrigeration and one for cryopreservation. For cooling, the semen was diluted in the concentrations of 15x106 (R15) and 30x106 (R30) spermatozoa/straw, for cryopreservation diluted in the concentration of 30x106 spermatozoa / vane (CRIO). For all dilutions, the Botubov&reg; medium and the semen stored in 0.5 mL vial were used. Refrigeration was carried out at a temperature of 5&deg; C for 48 hours in BotuFlex&reg; passive refrigeration system and cryopreservation in an automated TK&reg; system. 552 Brangus cows were synchronized in fixed-time artificial insemination programs. The results of the pregnancy rate for the groups of inseminated cows were 49.4% for R15, 43.38% for R30 and 47.59% for cryopreserved semen. It can be concluded that the cooling of the semen at 5&deg; C for 48 hours results in pregnancy rate similar to those obtained with cryopreserved semen, and these results indicate that refrigeration for up to 48 hours may be an option of use for FTAI.

In recent years, in dairy cattle, while it was observed a gradual increase in productivity, a decrease occurred in the reproductive efficiency. Several factors, such as increased incidence of diseases, higher susceptibility to heat stress and increase of dry matter intake, have been awarded as possible causes for the decrease in fertility. Increased dry matter intake is associated with increased liver blood flow, which is associated with an increase in liver metabolism of steroid hormones. Given the high metabolism of steroid hormones in high producing dairy cows, six studies were carried out, which in this thesis are divided in three chapters, involving hormone supplementation in lactating dairy cows. The first study aimed to increase the synchronization rate of dairy cows submitted to a fixed time artificial insemination (FTAI) estradiol (E2)/progesterone (P4)-based protocol. For this purpose, two experiments were performed, the first (n = 44 cows) compared a 2.0 vs 3.0 mg of estradiol benzoate (EB) associated to a P4 implant at the beginning of the protocol. The second experiment (n = 82 cows) performed presynchronization with GnRH prior to the onset of a FTAI protocol to produced different follicular development stages at the time of E2/P4: emergence vs. dominance. Daily ultrasound and hormone evaluations were performed. Other four experiments are described in the second (n = 1070 cows) and third (n = 1498 cows) chapter, which have been developed to evaluate the effect of P4 supplementation after ovulation in lactating dairy cows. In general, these studies evaluated the effect of supplementation on the corpus luteum (CL) development and function, mRNA abundance for interferon stimulated genes (ISG), on fertility of cows subjected to AI after estrus detection or FTAI protocol, or to embryo transfer. Increasing the EB dose from 2.0 for 3.0 mg did not improve emergence wave synchronization. In fact, it induced luteolysis in a larger number of cows. Altering the stage of the estrous cycle of the cows at the beginning of the E2/P4-based FTAI protocol did not improve synchronization of wave emergence. Post ovulation P4 supplementation did not affect CL development and function, and did not increase the mRNA abundance for ISG. Cows subjected to AI after estrus detection or after an E2/P4-based FTAI protocol did not have increased fertility. However when P4-supplemented cows were subjected to a GnRH-based FTAI protocol there was an improvement in the fertility of about 8%. Thus, we can concluded that regardless of the EB dose or stage of the estrous cycle at beginning of the E2/P4-based FTAI protocol, still there are cows that fail to have a synchronized emergence of a new wave and/or to ovulate at the end protocol. Additionally, depending on the protocol used, P4 supplementation may increase the fertility of dairy cows, but compromises the fertility when embryos are transferred. / Nos últimos anos, em rebanhos leiteiros, foi observado um aumento acentuado da produtividade acompanhado de uma diminuição da eficiência reprodutiva das vacas lactantes. Diversos fatores, como o aumento da incidência de doenças, da suscetibilidade ao estresse térmico e da ingestão de matéria seca, têm sido atribuídos como possíveis causas para esse decréscimo na fertilidade. Aumento da ingestão de matéria seca está associado maior fluxo sanguíneo hepático, que por sua vez está relacionado com aumento da metabolização hepática de diversas moléculas, entre elas os hormônios esteroides. Tendo em vista o alto metabolismo destes hormônios nas vacas leiteiras, foram realizados seis estudos, que na presente tese estão divididos em três capítulos, envolvendo suplementação hormonal em vacas leiteiras lactantes. O primeiro capítulo teve como objetivo aumentar a taxa de sincronização de vacas leiteiras ao protocolo de inseminação artificial em tempo fixo (IATF) à base de estradiol (E2)/progesterona (P4). Para isso foram realizados dois experimentos, sendo o primeiro (n = 44 vacas) utilizando dose de 2.0 ou 3.0 mg de benzoato de estradiol (BE) associado a um dispositivo de P4, no início do protocolo. No segundo (n = 82 vacas), foi realizada uma pré-sincronização com GnRH para iniciar o protocolo de IATF em diferentes estágios foliculares: recrutamento vs. dominância. Avaliações ultrassonográficas e dosagens hormonais foram feitas. Outros 4 experimentos, descritos no segundo (n = 1070 vacas) e no terceiro (n = 1498 vacas) capítulo, foram desenvolvidos para avaliar o efeito da suplementação de P4 após a ovulação em vacas leiteiras. Nesses estudos, foram avaliados os efeitos desta suplementação na formação e função do corpo lúteo (CL), na expressão de genes estimulados por interferon (ISG), na fertilidade de vacas submetidas a IA, através da observação de estro ou de protocolo de IATF, e em receptoras de embrião. A dose de 3.0 mg de BE, além de não aumentar a taxa de sincronização da emergência de uma nova onda folicular, induziu luteólise em um maior número de vacas que a dose de 2.0 mg. Independente da fase do ciclo estral, no início do protocolo a base de E2/P4, houve falhas na indução na sincronização da emergência e de ovulação. A suplementação com P4 após a ovulação não alterou a formação e função do CL, mas também não aumentou a expressão de ISG. Vacas submetidas a IA após detecção de estro ou submetidas à IATF em protocolos a base de E2/P4 não apresentaram aumento na fertilidade, no entanto quando submetidas ao protocolo de IATF à base de GnRH foi observado em torno de 8% de incremento de fertilidade. Contudo, receptoras de embrião suplementadas com P4 tiveram menor fertilidade. Assim, concluiu-se que independente da dose de EB ou do momento do ciclo estral em que se inicia o protocolo de IATF à base de E2/P4 há falhas de emergência de uma nova onda e/ou de ovulação ao final do protocolo. Além disso, dependendo do protocolo utilizado, a suplementação com P4 pósovulação pode aumentar a fertilidade de vacas submetidas à IATF, contudo compromete a fertilidade de receptoras de embrião.