Purpose - The purpose of the present study is to compare intrauterine msemination (IUI) pregnancy rates (PR) as a function of diagnosis and ovulation protocol utilizing an extended semen transport time. This allowed clients to conveniently collect IUI specimens in the comfort and privacy of their home. A single IUI per treatment cycle was performed. Basic Procedures - Three-hundred-ten consecutive infertilty couples having unexplained, male factor, ovulatory dysfunction, endometriosis, tubal factor or combined diagnostic factors receiving a total of 584 cycles of IUI were included. Ovulation protocols included LH surge, clomiphene citrate (CC)-hCG, CC-gonadotropins(Gn)-hCG, Gn-hCG or leuprolide acetate (L)-Gn-hCG followed 36-42 hours by a single IUI. Pregnancy rates per cycle (fecundity) and per couple (fertility) as a function of diagnosis, ovulation protocol and cycle number were evaluated. In each cycle the couples processed the specimen by adding sperm washing medium at room temperature to the specimen 30 min following collection and allowed it to incubate for two hours prior to IUI during transport. Main Findings - Overall, fecundity was 11.8% (69/584) and fertility was 22.3% (69/310); respectively by diagnosis was: unexplained 22.6%,38.8%; male factor 18.8%,42.9%; ovulatory dysfunction 12.4,22.6%; endometriosis 5.3%,11.1%; tubal factor 7.6%,13.3%; and combined factors 9.7%, 20.0%. Unexplained vs endometriosis (P < 0.0001, P < 0.005), tubal factor (fecundity P < 0.008) and ovulatory dysfunction (fecundity P < 0.027) was statistically different. Male factor vs endometriosis (P < 0.011, P < 0.036) was significantly different. Ovulatory dysfunction vs endometriosis was significantly different (fecundity P < 0.027). Pregnancies by ovulation protocol: LH surge 4.5%,10.5%; CC-hCG 9.4%,14.9%; CC-Gn-hCG 13.7%,23.7%; Gn-hCG 17.5%,45.3%; L-Gn-hCG 3.5%,6.7%. For Gn-hCG vs L-Gn-hCG (P < 0.009, P < 0.030) and LH surge (fecundity P < 0.033). CC-Gn-hCG vs CC-hCG (fertility P < 0.050) and L-Gn-hCG (P < 0.033, P < 0.034). Gn-hCG vs CC-hCG (fecundity P < 0.043). Conclusions - We conclude that IUI is effective when utilizing an extended transport time allowing most couples to collect the specimen at home and is most effective when utilizing Gn-hCG therapy. Based on our analysis, endometriosis, tubal factor and combined diagnostic categories should proceed earlier to higher level assisted reproductive technologies.

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.

Background: This thesis is based on a randomised controlled trial comparing the effectiveness of intrauterine insemination (IUI) plus Controlled Ovarian Hyperstimulation (COH) versus in vitro fertilisation (IVF) as the first line treatment option for couples with unexplained subfertility. Subfertility of a couple is classed as unexplained when they fail to conceive after one year of regular unprotected intercourse and when all the standard investigations for ovulation, tubal patency and semen analysis have been found to be normal. It affects 30-40% of couples. The age-old methods of treating these couples have included the empirical use of clomiphene or gonadotrophins to correct any possible subtle defects in ovulation with or without IUI (to overcome any existing cervical barrier to natural conception) or IVF. However, the best treatment options for these couples have yet to be determined. The matter has been made even more controversial by the issue of NICE (National Institute for Health and Care Excellence) guidelines in the UK that suggest IUI be abandoned completely for these women in favour of IVF after 2 years of expectant management. A systematic review of the available literature comparing IUI + COH versus IVF for unexplained subfertility revealed limited numbers of available studies and high clinical and statistical heterogeneity among them. An online survey was also conducted among fertility specialists to establish the general consensus regarding management of such couples. The results revealed a lack of agreement among fertility specialists with regards to the first line treatment of couples with unexplained subfertility. The mixed 8 response to this survey demonstrated the ongoing dilemma among practitioners, much of which was due to the lack of robust evidence. A randomised controlled trial was then designed to examine the effectiveness of COH with gonadotrophins + IUI versus IVF as the first line approach to the treatment of unexplained subfertility (Figure 1). This was the first UK-based randomised controlled trial comparing these two first-line management options for unexplained subfertility.

Orientador: Maria Denise Lopes / Resumo: O objetivo deste estudo foi o de avaliar os efeitos da criopreservação sobre a qualidade e fertilidade do sêmen de cão. Para isso foram realizados dois experimentos. No primeiro experimento foram colhidos 4 ejaculados de 5 cães (n=20), os quais foram avaliados no sêmen fresco, resfriado e descongelado. A motilidade total (MT), progressiva (MP) e porcentagem de rápidos (RAP) por sistema computadorizado e por citometria de fluxo, a fluidez da membrana plasmática [Yo-Pro 1 (YP) e merocianina 540 (M540)], a translocação da fosfatilidil serina (FS) (anexina V e FITC-PSA), integridade da membrana plasmática e acrossomal (iodeto de propídeo (IP) combinado ao FITC-PSA), potencial da membrana mitocondrial (JC1), lipoperoxidação dos lipídeos de membrana (LPO, C11-BODIPY) e apoptose (CellEvent®). O sêmen do cão que apresentou melhores resultados numéricos em todas as análises foi escolhido para ser utilizado separadamente para as inseminações (cão 1) e o sêmen dos demais 4 cães foram utilizados em pool. Foram inseminadas 20 cadelas (2 IAs/cadela), por via transcervical (TCIA) sendo 10 delas com o cão 1 e 10 com o pool. Foram utilizadas 2 concentrações espermáticas (160 e 450 x 106 espermatozoides/TCIA). Houve diferença para os índices de motilidade (p < 0,01) entre o sêmen fresco e resfriado quando comparados ao descongelado para todos os índices avaliados, (MT = 87,00 ± 1,24; 88,15 ± 1,38; 72,55 ± 6,26); (MP = 69,95 ± 1,28 ; 71,75 ± 1,91; 56,30 ± 6,00) e (RAP = 83,15 ± 1,94; 81,55 ±... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

The objective of the thesis was to analyse the reproductive traits of sows on a particular farm with regard to the method used, as observed during the period of one year. The highest rate of live-born piglets per the first litter (12.4) was reached in nucleus breeding. Regarding the mating method, the highest number of piglets born in the first litter (12.7) was reached through natural mating, followed by intrauterine insemination (12.3) and cervical insemination (11.6). During the one-year period, the rate of successful conception at first service was 83.3 per cent of sows in case of intrauterine insemination, 80.6 per cent of sows in case of natural mating and 79.2 per cent of sows in case of cervical insemination. When calculating the number of all born piglets per 100 inseminations, it was found out that 1 028 piglets were born in case of intrauterine insemination, 1 026 piglets in case of natural mating and only 921 piglets were born in case of cervical insemination, that is 107 (or 105) less. The highest number of born piglets (11.8) was reached in sows serviced when they were 256270 days old, followed by sows serviced when 241255 days old (11.1 piglets). With the gestation length under 114 days, the sows farrowed 1.2 piglets more than when the gestation length was over 115 days. With the weaning-to-conception interval under 5 days, the sows farrowed more (12.4) piglets than with the weaning-to-conception interval lasting over 6 days, which resulted in only 11.9 piglets being born.

Made available in DSpace on 2015-03-26T13:47:29Z (GMT). No. of bitstreams: 1 texto completo.pdf: 142651 bytes, checksum: 3f0b67aa44d733223926c3e7c4aa8f20 (MD5) Previous issue date: 2007-04-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This research was carried out into a commercial sows farm, located on Zona da Mata of state Minas Gerais, in order to evaluate the decreasing effect of the volume and the spermatic content between the traditional (ICAI) and the intrauterine insemination (IUAI) procedure, assess the introduction of the intrauterine insemination catheter, estimate the seminal backflow and the bleeding occurrence in the inseminated females by the both techniques. In a completely design, 300 females randomly distributed into 60 females in each one of the 5 treatments: T1 (control) intracervical with 3x109 spermatozoa/100 mL; T2 intrauterine with 1x109 spermatozoa/100 mL; T3 intrauterine with 1x109 spermatozoa/50 mL; T4 intrauterine with 5x108 spermatozoa/100 mL and T5 intrauterine with 5x108 spermatozoa/50 mL. The females inseminated by the IUAI technique showed the pregnancy general media rate of 95,0%, 90,8% farrowing rate and 9,2% return to the estrus. This results revealed none significant difference in comparison to the ICAI technique (91,7%, 90,0% and 10,0% respectively). The total litter size did not differ (p>0,05) between the IUAI and ICAI, and the average rates were 11,5, 11,7, 11,4, 11,9 and 11, 4 piglets in the treatments 1, 2, 3, 4 and 5 respectively. Although 4,6% of the achieved IUAI presented a subtle difficulty in the passage by cervix, the catheter was successfully inserted in 100% of females. There was no difference in the sperm backflow occurrence between the both methods. However, the ICAI demonstrated semen backflow rate of 85,8% of the injected volume, containing about 26% of the inseminated sperm content while in the IUAI the backflow diverge from 83,0% to 90,6% of the volume presenting 11,1% - 16,4% of the spermatozoa content. The percentual of the backflow volume did not show difference (p>0,05) between the treatments, on the other hand the spermatozoa content in the ICAI backflow collected was higher than in the IUAI. There was no difference (p>0,05) about the presence of blood in the both insemination procedure. Thus, these results indicate that the two treatments, applying a reduced volume and spermatozoa content, by the IUAI technique would be adopted in the commercial farm daily routine without compromising the female swine reproductive performance. / Este trabalho foi realizado em uma granja comercial localizada na Zona da Mata do Estado de Minas Gerais, com os objetivos de avaliar o efeito da diminuição do volume e concentração espermática inseminante na comparação do desempenho reprodutivo de matrizes suínas inseminadas pela técnica intra-uterina (IAIU) versus inseminação intracervical (IAIC); avaliar a possibilidade de passagem do cateter de inseminação intra-uterina; e, por fim, analisar o refluxo vulvar e a ocorrência de sangramento nos animais inseminados por essas técnicas. Foram utilizadas 300 fêmeas, 60 por tratamento, distribuídas aleatoriamente em cinco tratamentos: T1 (controle), em que os animais foram inseminados com três bilhões de espermatozóides por dose, em um volume de diluente q.s.p. para 100 mL; T2, inseminados com um bilhão de espermatozóides por dose, em um volume de diluente q.s.p. para 100 mL; T3, inseminados com um bilhão de espermatozóides por dose, em um volume de diluente q.s.p. para 50 mL; T4, inseminados com 500 milhões de espermatozóides por dose, em um volume de diluente q.s.p. para 100 mL; e T5, em que as matrizes foram inseminadas com 500 milhões de espermatozóides por dose, em um volume q.s.p. para 50 mL. As matrizes do tratamento 1 foram inseminadas pela técnica de IAIC, enquanto as dos tratamentos 2, 3, 4 e 5, pela técnica de IAIU. As fêmeas inseminadas pela técnica intra- uterina apresentaram taxa média geral de concepção de 95,0%, de parto de 90,8% e de repetição de estro de 9,2%, não apresentando diferença significativa em comparação com a IAIC, que obteve valores de 91,7%, 90,0% e 10,0%, nas taxas de concepção, parição e repetição de estro, respectivamente. Com relação ao resultado do total de leitões nascidos por parto, foram obtidas médias de 11,5; 11,7; 11,4; 11,9; e 11,4 leitões, nos tratamentos 1, 2, 3, 4 e 5, respectivamente. O total de leitões nascidos não foi diferente (p>0,05) entre os animais inseminados pela técnica IAIU, em comparação com a IAIC. Neste trabalho, a inseminação intra-uterina foi possível de ser realizada em 100% dos animais testados. Porém, 4,6% das inseminações realizadas por essa técnica apresentaram pequeno grau de dificuldade na passagem do cateter interno pela cérvix do animal. A avaliação da ocorrência de refluxo de sêmen nos animais inseminados pelas duas técnicas constituiu outro objetivo deste experimento e indicou não haver diferença significativa entre a inseminação intra-uterina e a tradicional. Com o emprego da inseminação artificial tradicional, as fêmeas inseminadas obtiveram um porcentual de refluxo de 85,8% do volume da dose infundida, contendo em torno de 26% do número de espermatozóides inseminados. Nos animais inseminados pela técnica intra-uterina, os resultados variaram de 83% a 90,6% do volume e de 11,1% a 16,4% dos espermatozóides infundidos. O porcentual de volume coletado no refluxo não apresentou diferença (p>0,05) entre os tratamentos estudados, no entanto o total de espermatozóides refluídos foi maior nas fêmeas submetidas à IAIC do que à IAIU. Por fim, na análise de ocorrência de sangramento não houve diferença significativa entre ambas as técnicas. Pelos resultados deste trabalho, é possível concluir que qualquer um dos tratamentos testados com a inseminação intra-uterina pode substituir a inseminação intracervical na rotina de uma granja comercial, sem comprometimento do desempenho reprodutivo dos animais.

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Agradecemos a compreensão. on 2016-09-28T16:27:18Z (GMT) / Submitted by CARMEN CECÍLIA SICHERLE null (carmensicherle@terra.com.br) on 2016-09-28T16:38:54Z No. of bitstreams: 1 Carmen C Sicherle Tese completa.pdf: 1312570 bytes, checksum: 9e105448cb368db57c02753ea8a7cf32 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-09-28T17:14:42Z (GMT) No. of bitstreams: 1 sicherle_cc_dr_bot.pdf: 1312570 bytes, checksum: 9e105448cb368db57c02753ea8a7cf32 (MD5) / Made available in DSpace on 2016-09-28T17:14:42Z (GMT). No. of bitstreams: 1 sicherle_cc_dr_bot.pdf: 1312570 bytes, checksum: 9e105448cb368db57c02753ea8a7cf32 (MD5) Previous issue date: 2016-08-03 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo deste estudo foi o de avaliar os efeitos da criopreservação sobre a qualidade e fertilidade do sêmen de cão. Para isso foram realizados dois experimentos. No primeiro experimento foram colhidos 4 ejaculados de 5 cães (n=20), os quais foram avaliados no sêmen fresco, resfriado e descongelado. A motilidade total (MT), progressiva (MP) e porcentagem de rápidos (RAP) por sistema computadorizado e por citometria de fluxo, a fluidez da membrana plasmática [Yo-Pro 1 (YP) e merocianina 540 (M540)], a translocação da fosfatilidil serina (FS) (anexina V e FITC-PSA), integridade da membrana plasmática e acrossomal (iodeto de propídeo (IP) combinado ao FITC-PSA), potencial da membrana mitocondrial (JC1), lipoperoxidação dos lipídeos de membrana (LPO, C11-BODIPY) e apoptose (CellEvent®). O sêmen do cão que apresentou melhores resultados numéricos em todas as análises foi escolhido para ser utilizado separadamente para as inseminações (cão 1) e o sêmen dos demais 4 cães foram utilizados em pool. Foram inseminadas 20 cadelas (2 IAs/cadela), por via transcervical (TCIA) sendo 10 delas com o cão 1 e 10 com o pool. Foram utilizadas 2 concentrações espermáticas (160 e 450 x 106 espermatozoides/TCIA). Houve diferença para os índices de motilidade (p < 0,01) entre o sêmen fresco e resfriado quando comparados ao descongelado para todos os índices avaliados, (MT = 87,00 ± 1,24; 88,15 ± 1,38; 72,55 ± 6,26); (MP = 69,95 ± 1,28 ; 71,75 ± 1,91; 56,30 ± 6,00) e (RAP = 83,15 ± 1,94; 81,55 ± 1,53; 64,30 ± 7,68). Em relação a fluidez da membrana espermática houve diferença na porcentagem da população de células íntegras (CI) e de células com aumento de fluidez de membrana (CIMF) (p < 0,01) entre o sêmen fresco e resfriado quando comparados ao descongelado, (CI = 78,29 ± 6,22; 75,76 ± 6,60; 23,02 ± 9,12); (CIMF = 18,33 ± 5,54; 22,55 ± 6,49; 71,78 ± 9,85). Sobre a identificação da translocação da FS houve diferença (p < 0,01) na porcentagem de células íntegras nos 3 momentos avaliados (CI = 79,90 ± 7,95; 63,50 ± 4,66; 27,19 ± 9,22). A porcentagem de células apresentado a membrana plasmática e acrossomal íntegras foi diferente (p < 0,01) também entre o sêmen fresco e resfriado quando comparados ao descongelado (MPAI = 85,71 ± 6,22; 70,37 ± 6,43; 30,87 ± 9,90). Quanto ao potencial da membrana mitocondrial houve diferença (p < 0,01) na porcentagem de células com alto potencial mitocondrial nos 3 momentos avaliados (APM = 85,15 ± 2,76; 60,52 ± 3,31; 38,06 ± 6,40). A LPO da membrana plasmática foi diferente (p < 0,05) no sêmen congelado quando comprado ao resfriado mas não teve diferença entre o sêmen fresco e resfriado e do fresco ao congelado. Não foi observada qualquer diferença, nos momentos estudados, na atividade da caspase. O cão 1 foi significamente melhor para os índices de integridade de membrana plasmática e acrossomal e potencial mitocondrial. Pelo calculo da resposta média do animal e limites mínimos e máximos, apesar de não haver diferença, observamos que o animal 1 apresentou melhores valores pós descongelação para todas os testes realizados. O aumento na concentração espermática foi positivo e observamos 50% de prenhez no grupo inseminado com o pool de sêmen utilizando a concentração de 450 x 106. Nenhuma gestação foi observada nas TCIA com o sêmen do cão 1. Em conclusão a criopreservação leva a alterações estruturais e funcionais da célula espermática, o que explica a sua sobrevivência curta após a descongelação. As mudanças semelhantes a crio-capacitação podem estar relacionadas a outros fatores, como as proteínas do plasma seminal que ainda não são totalmente conhecidas nesta espécie. O processo de apoptose pode ser iniciado com a abertura dos poros mitocondriais durante o processo de congelação/descongelação, o que libera fatores pro-apoptóticos no citoplasma celular. O potencial mitocondrial não está relacionado com a motilidade dos espermatozoides, mas sim com a sobrevivência do espermatozoide. Melhores resultados de gestação puderam ser obtidos quando mais de 100 x 106 de espermatozoides móveis e morfologicamente íntegros for utilizados por TCIA. Não houve relação entre a qualidade seminal, avaliada pelas técnicas aqui descritas, e a fertilidade. / The aims of the study were to assess individual characteristics and sperm concentration of cryopreserved semen characteristics using different structural and functional analysis and dog fertility. Twenty ejaculates were collected from 5 dogs and sperm were cryopreserved by one-step protocol. To better understand dog semen quality after thawing five healthy mature dogs were used to the one-step sperm cryopreservation protocol. Sperm analysis was performed at three time points: after collection, refrigeration, and after thawing, by computer sperm motility analysis (total [TM] and progressive motility [PM], and rapid sperm), membrane damage (membrane fluidity and viability - Yo-Pro 1 and merocianina 540, phosphatidyl serine translocation – annexin – V – propidium iodide - PI, acrosomal and membrane integrity FITC-PSA – PI), mitochondrial potential (JC-1), membrane lipid peroxidation, LPO (BODIPY 581/591 C11) and apoptosis (kit CellEventTM Caspase-3/7- FITC Green Flow Cytometry), evaluated by flow cytometry. Twenty bitches were inseminated by transcervical intrauterine (TCAI), twice using 2 sperm concentration (160 and 450 x 106 spermatozoa/TCAI). A decrease on frozen/thawed semen comparing to fresh and cooled semen occurred for the parameters of total and progressive motility (TM 87.00 ± 1.24, 88.15 ± 1.38, 72.55 ± 6.26; PM 69.95 ± 1.28, 71.75 ± 1.91, 56.30 ± 6.00, fresh cooled and frozen/thawed respectively P < 0.01), percentage of rapids (83.15 ± 1.94, 81.55 ± 1.53, 64.30 ± 7.68 P < 0.01), membrane fluidity and cell viability (78.29 ± 6.22, 75.76 ± 6.60, 23.02 ± 9.12 P < 0.01) and acrosomal and membrane integrity (85.71 ± 6.22, 70.37 ± 6.43, 30.87 ± 9.90 P < 0.01). Sperm cells were affect for cooling and freezing process when analyzed for the translocation of phosphatidyl serine (79.90 ± 7.95, 63.50 ± 4.66, 27.19 ± 9.22 P < 0.01) and high mitochondrial potential (85.15 ± 2.76, 60.52 ± 3.31, 38.06 ± 6.40 P < 0.01). For LPO significant difference was observed in semen after thawing only comparing to cooled semen (16.57 ± 5.44, 16.22 ± 2.43, 23.00 ± 4.13 P < 0.05). No difference on caspase activity was observed. Dog 1 was significantly better for the parameters of plasma membrane and acrosome integrity and mitochondrial potential. By calculating the average of the animal response and minimum and maximum limits, although there is no difference, we observed that the semen from Dog 1 displayed better post thaw values for all analyzes. The pregnancy rate was 0% for dog 1 and 50% for pool at concentration 450 x 106 spermatozoa. In conclusion, cryopreservation leads to structural and functional changes in sperm cell, which explains their short survival after thawing. The cryo-capacitation like changes can be related to others factors, as the seminal plasma proteins that are not know yet on this species. Apoptosis process can be initiated with the opening of mitochondrial pores during the freezing/thaw process, which releases pro-apoptotic factor on the cytoplasm. The mitochondrial potential is not related to sperm motility but sperm survival. Although the dog 1 displayed better sperm quality, no pregnancy was observed, which makes us to rethink the prediction of fertility of semen samples by commonly used parameters. The concentration dose used for AI procedures must consider motility and sperm viability to satisfactory pregnancy rates. / FAPESP: 2013/02050-5

Philosophiae Doctor - PhD / Infertility is currently a serious problem that is escalating, not only in South Africa, but also worldwide. In Cape Town, a culturally diverse, urban community of approximately 1000 couples are referred to the Groote Schuur Hospital Infertility Clinic annually. Although infertility is primarily regarded as a medical condition, the treatments have emotional effects on infertile couples due to the recurring highs and lows that often accompany treatments. This study aimed to qualitatively explore and understand the emotional and psychological experiences of infertility and its treatments (specifically In Vitro Fertilisation and Embryo Transfer). Social constructionism is based on the premise that realities are not constructed in a vacuum but rather undergo a process whereby the subjective and inter-subjective experiences over time and through cultural processes come to be regarded as truths. These truths become internalised and function as lenses through which we see ourselves, compose and invent ourselves accordingly, making sense of what would otherwise have been chaotic and meaningless experiences. Additional aims were to examine women's experiences of infertility care whilst undergoing treatment and describe their experiences of coping with infertility and In Vitro Fertilisation and Embryo Transfer (IVF-ET). Semi-structured, in-depth individual interviews were conducted with 21 women presenting with primary infertility at a fertility clinic. This study utilised an ethnographic case study design. The results of the study suggested that women perceived themselves as not conforming to a dominant belief system that promotes motherhood as the most important role for women. The women described their 'failure' to fulfill socio-cultural expectations as emotionally turbulent. Some of the psychological responses to infertility included feelings of disappointment, shock, denial, devastation, anger, frustration, sadness, inadequacy, poor self-image and self-esteem. The women's personal accounts of their experiences of In Vitro Fertilisation and Embryo Transfer (IVF-ET) revealed that they found the treatment to be highly stressful, with emotional bouts of anxiety, nervousness, excitement and optimism. A psychological synopsis of infertility and IVF-ET is infertility and IVF-ET presents. The fmdings in this study suggest the need for the incorporation of presented. This diagrammatic representation shows the intensity of the emotional rollercoaster that psychosocial intervention into infertility management. Greater attention to the psychological and emotional repercussions of infertility treatment could lead to a more personalised approach which, in turn, would optimise patient satisfaction and also prepare couples for the demands of the program by informing them about better ways of coping.

L’infertilité touche environ 11 à 15 % des couples au Canada. Les modalités de la procréation médicalement assistée (PMA) incluent la stimulation ovarienne (SO), l’insémination intra-utérine (IIU) et les techniques de la reproduction assistée (TRA). Les grossesses multiples sont parmi les effets indésirables les plus communs de l’utilisation des TRA. Toutefois, il n’y a pas de consensus sur l’ampleur du risque de grossesses multiples associé à l’utilisation des modalités de la PMA. De plus, il n’est pas clair si les modalités de la PMA sont associées à un risque élevé de malformations congénitales majeures (MCM) d’un ou plusieurs systèmes et organes du corps humain. Un projet de recherche en trois volets a été développé afin de répondre à ces questions. Dans un premier volet, une étude cas-témoin a été conduite afin de quantifier le risque de naissances multiples associé à l’utilisation des modalités de la PMA. Comparativement à une conception spontanée (CS), le recours à la SO seule, l’IIU, et les TRA étaient associés à une augmentation significative de plus de quatre, neuf et 31 fois du risque de naissances multiples, respectivement. Les femmes ayant fait appel à une IIU+SO ou aux TRA avaient un risque accru de naissances multiples (odds ratio (OR)ajusté, 1,98; IC95%, 1,12-3,49; ORajusté, 6,81; IC95%, 3,72-12,49, respectivement) comparativement à l’utilisation de la SO seule. Dans un deuxième volet, une analyse cas-témoin a été conduite afin de quantifier le risque de MCM associés à l’utilisation des modalités de la PMA. Comparée à une CS, l'utilisation des TRA était associée à une augmentation du risque des malformations du système urogénitale (ORajusté, 3,11; IC95%, 1,33–7,27) et l'utilisation de l'IIU était associée à un risque accru des malformations du système musculosquelettique (ORajusté, 2,02; IC95%, 1,10–3,71). L'utilisation des TRA était associée à une augmentation du risque des malformations du système urogénitale (ORajusté, 7,18; IC95%, 1,59-32,53) par rapport à l’utilisation de la SO seule. Dans un troisième volet, les résultats de la revue systématique et méta-analyse ont démontré que l'utilisation de la SO seule et l’IIU±SO étaient associée à une augmentation significative de presque 9 fois du risque de grossesses multiples par rapport à une CS. Des augmentations similaires ont été observées suite à l’utilisation du citrate de clomifène seul. Par rapport à une CS, l'utilisation de la SO seule était associée à une augmentation significative de 48% du risque des malformations du système musculosquelettique, de 73% le risque des malformations du système nerveux, de 76% le risque de malformations congénitales du système digestif, de 68% le risque de malformations des yeux, des oreilles, du visage et du cou, et plus de deux fois le risque de malformations congénitales du système respiratoire. L'utilisation de l'IIU était associée à une augmentation significative de 52% du risque de malformations du système urogénitales et de 54% du risque de malformations musculosquelettiques par rapport à une CS. Bien que le risque de grossesses multiples associées aux TRA puisse être contrôlé par le transfert d'un embryon unique, un suivi particulier devrait être accordée au risque de grossesses multiples associé à l’utilisation des traitements n’impliquant pas un transfert d’embryons. L’effet tératogène potentiel associé à l’utilisation des modalités de la PMA doit être considéré dans la prise des décisions thérapeutiques. La mise en place d’un registre de la PMA pour la surveillance des effets périnataux indésirables devient nécessaire / Infertility affects 11 to 15% of couples in Canada. The modalities of medically assisted reproduction (MAR) include ovarian stimulation (OS), intrauterine insemination (IUI) and assisted reproduction techniques (ART). Multiple pregnancies are among the most common side effects of using ART. However, there is no consensus on the magnitude of the risk of multiple pregnancies associated with the use of MAR modalities. In addition, it is unclear whether the modalities of MAR are associated with a high risk of major congenital malformations (MCM) affecting one or more specific organ system. A three-part research project was developed to answer these questions. In the first part, a case-control study was conducted to quantify the risk of multiple births associated with the use of the three MAR modalities. Compared to spontaneous conception (SC), the use of OS alone, IUI, and ART was associated with a significant increase of more than four, nine, and 31 times the risk of multiple births, respectively. Women who used IUI with OS or ART had an increased risk of multiple births (adjusted odds ratio (OR), 1.98; 95%CI, 1.12-3.49; adjusted OR, 6.81; 95%CI, 3.72-12.49, respectively) compared to the use of OS alone. In a second part, a case-control analysis was conducted in order to quantify the risk of MCM associated with the use of MAR modalities. Compared to SC, the use of ART was associated with an increased risk of urogenital malformations (adjusted OR, 3.11; 95%CI, 1.33–7.27) and the use of IUI was associated with increased risk of musculoskeletal malformations (adjusted OR, 2.02; 95% CI, 1.10–3.71). The use of ART was associated with an increased risk of urogenital malformations (adjusted OR, 7.18; 95% CI 1.59-32.53) compared to the use of OS alone. In a third part, results of the systematic review and meta-analysis demonstrated that the use of OS alone and IUI ± OS were associated with a significant increase of nine times the risk of multiple pregnancies compared to a SC. Similar increases have been found following the use of clomiphene citrate alone. Compared to SC, the use of OS alone was associated with a significant 48% increased risk of musculoskeletal malformations, 73% increased risk of malformations of the nervous system, 76% increased risk of digestive system malformations, 68% increased risk of malformations of the eye, ear, face and neck, and more than twice the risk of congenital respiratory system malformations. The use of IUI was associated with a significant 52% increased risk of urogenital malformations and 54% increased risk of musculoskeletal malformations compared to SC. Although the risk of multiple pregnancies associated with ART can be controlled using a single embryo transfer in IVF cycles, monitoring the risk of multiple pregnancies associated with the use of treatments that do not involve embryo transfer is essential. The potential teratogenic effect associated with the use of MAR modalities should be considered when making therapeutic decisions. The establishment of a registry for the surveillance of MAR adverse perinatal outcomes becomes necessary.