One of the requirements for a successful insemination in horse breeding is the selection of quality stallions which produced ejaculate in enough quantities and of good quality. For this purpose were discovered methods of dealing with the evaluation of semen quality. In this dissertation was ejaculate evaluated by using an objective computerized method CASA, system SCA. Monitoring the quality of sperm was performed on 11 breeding stallions involved in artificial insemination fresh sperm in ZH Písek. 53 samples of ejaculate were evaluated. Volume, sperm concentration, total and progressive motility and sperm vitality were monitored. Volume of ejaculate was from 20 to 127.5 ml. Average values of sperm concentration for individual stallions were from 104.18 to 384.4 M/ml. Average values of total motility were moving by interval from 51.23 to 89.54 %. Average progressive motility was move from 12.99 to 47.12 %. At sperm vitality were discovered average results from 35.16 to 71.21 %. The average value of individual stallions was identical to the Decree of the Ministry of Agriculture, which sets minimum requirements for quality indicators for short-term preservation of semen. Also the quality of the semen of stallions was compared by age. The first group was made by stallions to 10 years old and the second group were older stallions. It was found that the younger stallions to 10 years had higher semen volume and greater sperm concentration. Stallions older over 10 years had higher levels of total and progressive motility and sperm vitality. Next ejaculate quality was evaluate by the time interval between samples. Regular taken from stallions is doing better ejaculate quality but the time interval between samples is important. The best values were found in samples with interval between the sampling 4 or more days. The worst quality of sperm was in the samples with the interval between 1-2 days.

As biotécnicas da reprodução na espécie equina avançaram na última década e os criadores de equinos começaram a questionar as possibilidades de interferência na determinação do sexo dos potros. A determinação do sexo é importante, pois o sexo do potro possui grande influência no valor comercial do mesmo. O objetivo do presente estudo foi avaliar a influência do momento da cobertura em relação ao momento da ovulação no sexo dos potros e outros fatores como: idade da égua e garanhão; indutor da ovulação; ovário em que ocorreu a ovulação e o diâmetro do folículo pré-ovulatório também foram avaliados. O estudo foi realizado em Haras na região de Bagé e Aceguá (RS) – Brasil (31°24'06.1''S 54°07'47.5''W) e (31°30'16.0''S 54°07'45.5''W) nas estações de monta de 2011 a 2015. Utilizou-se 259 ciclos reprodutivos de 160 éguas e 22 garanhões da raça Puro Sangue de Corrida. Informações como agente indutor de ovulação utilizado (Deslorelina; n = 187 ou hCG; n = 72); data da cobertura (n = 259); momento da ovulação em relação a cobertura (+ 24 horas; n = 69 e - 24 horas; n = 190); idade da égua (G1: até 8 anos; n = 123, G2: 9 até 14 anos n = 110 e G3: >14 anos; n = 26); idade do garanhão (Até 14 anos; n = 11 e >15 anos; n = 11) ; ovário em que a ovulação ocorreu (Direito; n = 122 e Esquerdo; n = 137); foram registrados e avaliados No total de potros nascidos, 136 foram machos (52,51%) e 123 foram fêmeas (47,49%). O tempo decorrido após a cobertura não influenciou o sexo dos produtos, nas éguas que ovularam com – 24h após a indução da ovulação: 104 potros (54,74%) eram machos e 86 (45,26%) eram fêmeas, enquanto que nas éguas que ovularam com + 24h, 32 potros (46,38%) eram machos e 37 (53,62%) eram fêmeas. A porcentagem de fêmeas nascidas em relação a idade da égua foi de 46,34% (n=57), 47,27% (n=52) e 46,15% (n=12) nos grupos G1, G2 e G3 respectivamente. Garanhões com idade até 15 anos tiveram 44,14% (n=49) de fêmeas e com mais de 15 anos 49,66% (n=73) eram fêmeas. Não houve diferença no sexo dos produtos quanto ao agente indutor da ovulação (Deslorelina x hCG) e ovário em que ocorreu a ovulação. O presente estudo concluiu que nenhum dos fatores estudados alterou a proporção macho:fêmea dos potros nascidos. / The biotechnologies of reproduction in equine species have been improved in the last decade and the horse breeders started to inquire about the possibility of intervention regarding to the sex determination in foals. Sex determination is important because the sex of the foal has a great influence on the commercial value of the foal. The aim of the present study is was to evaluate the influence of the time of the breeding in relation to time of ovulation in the sex of the foals, besides analyzing other factors such as: the age of the mare and the stallion; ovulation inductor, ovary and the diameter of the preovulatory follicle. The study was accomplished in studs located Bagé/RS and Aceguá/RS – Brazil (31°24'06.1''S 54°07'47.5''W) and (31°30'16.0''S 54°07'45.5''W) during the breeding seasons from 2011 to 2015, using 259 reproductive cycles of 160 mares and 22 stallions of Thoroughbred breed. Information like the induction of ovulation agent that was used (Deslorelin; n = 187 or hCG; n = 72); date of breeding (n = 259); time of ovulation in relation to the breeding (+24 hours; n = 69 and -24 hours; n = 190); age of the mare (G1: up to the age of 8; n = 123 G2: from the age of 9 to the age of 14; n = 110 and G3: >14 years old; n = 26); age of the stallion (up to the age of 14; n = 11 and >15 years old; n = 11); ovary in which the ovulation occurred (Right; n = 122 and Left; n = 137) were catalogued and evaluated As result, 136 (52,51%) were born colts and 123 (47,49%) were born fillies. The elapsed time from breeding to ovulation did not influence on the sex of the product, on mares that ovulated in less than 24 hours after the ovulation induction: 104 foals (54,74%) were male and 86 (45,26%) were female, whereas in the mares that ovulated in more than 24 hours, 32 foals (46,38%) were male and 37 (53,62%) were female. The percentage of born females regarding to the age of the mare was 46,34% (n = 57), 47,27% (n = 52) and 46,15% (n = 12) in the groups G1, G2 and G3, respectively. From stallions up to the age of 15 years, 44,14% (n = 49) were females and from those which were older than 15 years old, 49,66% (n = 73) were females. There was no difference regarding the sex of the product in relation to the ovulation inducer agent (Deslorelin x hCG) and ovary in which the ovulation occurred. None of the factors studied modified the male:female proportion of the born foals.

Em todas as espécies de mamíferos estudadas, o gene PLCζ é responsável pelo aumento de cálcio no momento da fecundação do oócito. Recentemente, o gene WBP2NL, também esta sendo estudado por sua possível capacidade de iniciar a ativação de oócitos. O gene TNF-α é uma citocina pró-inflamatória que pode ser encontrada no plasma seminal, mas seu efeito na viabilidade espermática ainda não está claro. O objetivo desse trabalho foi investigar a expressão dos genes PLCζ, WPB2NL e TNF-α no espermatozoide e suas relações com a qualidade e fertilidade em equinos. Foram utilizados ejaculados de 40 garanhões da raça Crioula, de criatórios localizados próximos a Porto Alegre. A concentração espermática foi avaliada em câmara de Neubauer. As demais análises microscópicas foram realizadas através do Computer Assisted Sperm Analysis AndroVision. A análise da integridade física da membrana foi realizada utilizando-se sondas fluorescentes A integridade funcional da membrana plasmática foi avaliada por meio do teste hiposmótico. Para análise estatística foi utilizada correlação de Pearson, com nível de significância de P<0,05. Foi encontrada correlação positiva do gene PCLζ com integridade funcional da membrana plasmática (0,449), integridade física da membrana plasmática (0,438), taxa de prenhez (0,454), motilidade total (0,486), motilidade progressiva (0,413); correlação negativa com motilidade lenta (-0,427) e espermatozoides imóveis (-0,405). Não foram encontradas correlações dos genes WBP2NL e TNF-α. Concluí-se que a expressão do gene PCLζ no Espermatozoide equino pode ser utilizada com um marcador para a qualidade seminal e fertilidade do garanhão. / In all species of mammals studied, the PLCζ gene is a factor of calcium increase at the time of oocyte fertilization. Recently, the WBP2NL gene is also being studied for its potential ability to initiate oocyte activation during fertilization. The TNF-α is a pro-inflammatory cytokine found in seminal plasma but it effect on motility and viability sperm still unclear. The aim of this study was to verify the expression of PLCζ, WPB2NL and TNF-α genes in spermatozoa and their relationship with quality and fertility in horses. We used the ejaculates of 40 Crioulo stallions, were used from horse farms located near Porto Alegre. After semen collection, the ejaculate was diluted in a commercially available extender. Microscopic analysis of total and progressive motility was performed using a computer assisted sperm analysis system AndroVision. Membrane physical integrity was assessed by fluorescent probes and membrane functionality was analyzed by the hypo-osmotic test (HOST) Statistical analysis was performed using the Pearson correlation, with a significance level of P<0.05. A moderate positive correlation of the PCLζ gene was found with functional plasma membrane function (0.449), physical organization of the plasma membrane (0.438), conception rate (0.454), total motility (0.486), progressive motility (0.413) and negative correlation with slow motility (-0.427) and spermatozoa properties (-0.405). No correlation on WBP2NL and TNF-α genes were found, showing that those genes expression should not be used as a marker of seminal quality and fertility in stallions. It can be concluded that expression of the PCLζ gene on equine spermatozoa can be used as a marker for seminal quality and fertility in the stallion.

Dois experimentos foram conduzidos para verificar a viabilidade do uso do leite UHT como diluente para sêmen equino refrigerado. O objetivo principal deste estudo foi avaliar o uso de leite UHT nas apresentações desnatado, semidesnatado e integral como diluente de sêmen e posteriormente foi avaliada a viabilidade espermática com o uso de leite como diluente comparado a outros dois diluentes industrializados. No experimento I foram utilizados 31 ejaculados de 3 garanhões, as amostras de cada ejaculado foram diluídas em leite UHT nas apresentações integral, semidesnatado e desnatado, numa proporção de 3:1 (3 ml de diluente e 1ml de sêmen). Foram realizadas análises de viabilidade espermática com testes de motilidade espermática, vigor espermático, integridade de membrana (CFDA/PI) e funcionalidade de membrana espermática (HOST) pós-diluição, os quais foram repetidos nas amostras refrigeradas a 4ºC após 24 e 48h. Neste experimento, não foram observadas diferenças significativas entre os 3 diluentes, quanto a motilidade espermática, vigor, CFDA/PI e HOST. No entanto, houve uma diminuição na qualidade do sêmen armazenado por 48h (P < 0,01), sendo mais acentuada entre 24 e 48h de refrigeração. No experimento II foram utilizados 30 ejaculados de 3 garanhões onde, logo após a coleta do sêmen, as amostras de cada ejaculado foram diluídas com leite UHT desnatado (LD), Botusemen® (BS) e Botusemen Special® (BSP). As amostras foram examinadas a fresco, após a diluição (0h) e em resfriamento de 24h e 48h a 4°C. Foram realizados os mesmos testes de análise espermática do primeiro experimento para avaliação comparativa dos diluentes. As amostras de sêmen refrigeradas com BSP obtiveram maior porcentagem de motilidade total (51,1%) após 24h de refrigeração quando comparado aos outros dois diluentes testados, LD (45,8%) e BS (47,3%). Com base nos resultados obtidos nos dois experimentos, o leite UHT, independente da apresentação, pode ser utilizado como diluente de sêmen para as primeiras 24h de refrigeração, mantendo a viabilidade espermática adequada, sendo também de fácil acesso e manuseio. Se for necessário a manutenção do sêmen por mais de 24h, o BSP se mostrou superior em manter a qualidade do sêmen até 48h. / Two experiments were conducted to verify the feasibility of the use of UHT milk as an extender for chilled equine semen. The main objective of this study was to evaluate the use of UHT milk in skimmed, semi-skimmed and whole milk as extender for semen and subsequently assess sperm viability using milk as a diluent compared to two other industrial extenders. In the first experiment were used 31 ejaculates from 3 stallions of known fertility, samples of each ejaculate were diluted in the whole UHT milk, semi-skimmed milk and skim milk shows a ratio of 3:1 (3ml diluent and 1ml of semen). Sperm viability analyzes were performed, including sperm motility, sperm vigor, membrane integrity (CFDA/PI) functionality and sperm membrane (HOST) post-dilution tests which were repeated in the samples cooled to 4°C after 24 and 48h. In this experiment, no significant differences in sperm motility, vigor, CFDA/IP and HOST were observed among the 3 extenders. However, there was a decrease in the quality of semen stored for 48h (P < 0.01), being more pronounced between 24 and 48h of cooling. In the second experiment, 30 ejaculates from 3 stallions were used soon after collection of semen samples, each ejaculate was diluted with UHT skim milk (LD), Botusemen® (BS) and Botusemen Special® (BSP). The samples were analyzed fresh, immediately after the dilution (0h) and after cooling for 24h and 48h at 4°C. The same tests of sperm analysis used in the first experiment for comparative evaluation of extenders were performed. Semen samples cooled with BSP obtained the highest percentage of total motility (51.1%) after 24h of cooling when compared to the two other extenders tested, LD (45.8%) and BS (47.3%). Based on the results from the two experiments, UHT milk, regardless of the presentation, can be used as a diluent of semen during the first 24h of cooling maintaining adequate sperm viability and is easy to access and handle. If maintenance of semen for more than 24h is required, the BSP was superior in maintaining the quality of semen until 48h.

The thesis evaluates the performance testing system of Czech Warmblood stallions. Data acquired from basic stallion preformance test were analysed. Analysed data covered figures of 302 stallions from 9 test stations, taken in years 2007 to 2010. 12 traits of each stallion were analysed. Only basic statistic characteristics were used. Traits were compared by years, testing stations and especially by preferred sirs. The statistic evaluation of traits pointed the best sirs for breeding. Average marks and variability of offspring were taken into acount. Best sirs were Le Patron (x? = 7,84; variability = 2,5 %, Porter (x? = 7,73; variability = 3,0 %) Catango Z (x? = 7,86; variability = 4,4 %) and Przedswit XVI - 12 (x? = 7,85; variabiliy = 4,7 %). Suggestions for improvment of objectivity in performance testing system of Czech Warmblood were given on basis of evaluation component trait/overal score relations. Frequent usage of foreign stallions in Czech Warmblood breeding is common. It is importatnt, for breeders to be familiar with performance testing of foreign breeds. Therefore short description of stallion performence testing across Europe is included in thesis.

Performance tests are necessary to determine the performance and individual character traits of the horse, which are needed for inclusion in breeding. The aim of the diploma thesis was to assess the performance tests and the method of evaluation. At first, the thesis compares the system and evaluation of tests of cold-blooded horses in the Czech Republic and neighbouring countries. The following section compares the performance requirements for stallions and mares. Then there was compared the performance of different breeds. The following part deals with determining, how the year of performance tests affected the evaluation of individual disciplines. The last part of the thesis deals with observing the influence of the breed on evaluation of individual disciplines.

The aim of this work was to compile a summary of the information provided to the public breeders by the five warm-blood horse breeding organizations in the Czech Republic. Mentioned horses are used for sporting purposes. A similar report was done on the extent and quality of information provided to the public breeders in neighboring European countries. We had prepared a proposal for a system that would provide the necessary information for effective breeding work . In the Czech Republic there is a system for collecting information about the population of horses. However, it is necessary to evaluate and publish information in an understandable form to breeders. As a basis for the preparation of tabular overview of the number of warm-blood horses bred in the Czech Republic, were collected the data from the Central horse register website and data from yearbooks and websites of individual breeders associations. Data were collected for the period of 2006 - 2013. In this thesis the processed data covers these breeds: Czech warmblood, Slovak warmblood, Moravian warmblood , Kinsky horse and Czech Traken. We have suggested the system, configuration and method for disclosure of the breeding information for warm-blood horse breeders and there is also recommended the annual frequency. As a model stallion was chosen Dutch warmblood Great Pleasure and we have created an information portfolio of this stallion. The system of the published information includes identification of the horse, pedigree, exterior evaluation, the results of his performance, comparing features and characteristics of the offspring population and the average breeding value estimation for exterior and performance. An important point is to make the overview of births and evaluated offspring. The proposed system should allow the improvement of the current breeding process and provide objective information to breeders for the right breeding decisions.

Made available in DSpace on 2015-03-26T13:54:49Z (GMT). No. of bitstreams: 1 texto completo.pdf: 996495 bytes, checksum: d0704166d94adb1ce9c77dd01311d0e5 (MD5) Previous issue date: 2009-02-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this study was evaluate the effectiveness of the Mangalarga Marchador stallions semen diluted with glycine-yolk egg based extender or milk-based extender and cooled at 5 °C for 24 hours, assessed by in vitro tests and of fertility in vivo. In Experiment 1, were used 5 ejaculated of 3 stallions. Following semen collection, the experimental group was divided into two treatments: 1 - cooling and dilution of semen with the milk-based extender, 2 - cooling and dilution of semen with extender glycine-egg yolk based extender. Semen was packaged in samples containing 12 mL of diluted semen with 30 x 106 viable sperm / mL concentration and then stored in Equitainer® for 24 hours. The sperm quality was evaluated after its dilution and after its cooling. The following seminal characteristics were evaluated: progressive motility, spermatic vigor, sperm morphology, integrity of the plasma membrane integrity by supravital test and epifluorescent using the dyes carboxyfluorescein diacetate (CFDA) and propidium iodide (PI); plasma membrane functionality by hypoosmotic test and progressive motility and spermatic vigor during the spermatic thermo-resistance test (TTR) for 90 minutes. The vigor of semen diluted with Kenney was higher (p <0.05) than T2 in the assessment of fresh diluted semen. No statistical difference was observed in other seminal characteristics from the fresh diluted semen. In the cooled semen, the Kenney extender had a better motility and spermatic vigor after the 24 hours storage, but Foote extender had higher average values for reactive spermatozoa percentage to the hypoosmotic test and the viabity to the epifluorescent test (p<0,05). In Experiment 2, the objective was to evaluate the fertility of cooled semen with both extenders previously described. Seventeen semen samples of stallion 1 and twenty three samples of stallion 2 were collected. Following semen collection, the semen was diluted with two extenders used in Experiment 1, and adjusted to an insemination dose of 500 million viable sperm in a final volume of 15 mL per dose. Semen was packaged in Equitainer® and cooled down to 5° C and storage for at least 24 hours. The following seminal characteristics were evaluated: progressive motility and vigor of the diluted and cooled semen; the sperm morphology of fresh semen and cooled with both extenders. Thirty one estrous cycles of 26 mares were used in the first period and 43 cycles of 28 mares in the second period. After detection 30 mm follicle size, the mares were randomly assigned into two groups: group A (mares inseminated with semen cooled with Kenney extender) and group B (mares inseminated with semen cooled with dilutive Foote). The stallions semen were used randomly among mares. The inseminations were done after a period of at least 24 hours of storage at 5° C. Semen was deposited in the body uterus, after the detection of a 30-40 mm follicle detection throughout ovulation. The inseminations were performed only on fixed days (Tuesday, Thursday and Saturday). The pregnancy diagnosis was performed by transrectal ultrasonography. As results, Kenney extender had higher progressive motility and spermatic vigor maintance after cooling compared to T2 extender. The pregnancy rate was higher from treatment A than treatment B (p<0,05), 55,26% (21/38) and 30,56% (11/36), respectively. With this data, it is possible to conclude that: the integrity and functionality of sperm cell plasma membrane evaluation tests are considered ancillary assessments in predicting the reproductive potential of stallions, but are not yet conclusive. The glycine-egg yolk based extender was not effective in preserving the seminal characteristics, and its fertility rate was not acceptable. The milk based extender proposed by Kenney et al. (1975) is still an excellent option for use in programs of AI with cooled semen from Mangalarga Marchador breed. / Objetivou-se neste estudo avaliar a eficácia de dois diluidores um à base de licina-gema de ovo e outro à base de leite em pó resfriado a 5 °C por 24 horas na preservação do sêmen de garanhões da raça Mangalarga Marchador, determinada por meio de testes in vitro e avaliação da fertilidade in vivo. No experimento 1, foram utilizados cinco ejaculados de três garanhões da raça Mangalarga Marchador, que foram divididos em dois tratamentos: Kenney - diluição e resfriamento do sêmen com diluidor à base de leite (Kenney et al., 1975); Foote - diluição e resfriamento do sêmen com diluidor de glicina-gema de ovo (Foote, 2002). Amostras de 12 mL de sêmen foram diluídas numa concentração de 30 x 106 espermatozoides viáveis/mL e, depois, armazenadas em Equitainer® por 24 horas. A qualidade seminal foi avaliada após diluição e resfriamento do sêmen. Foram avaliadas as seguintes características seminais: motilidade progressiva; vigor espermático; morfologia espermática; integridade da membrana plasmática pelo teste supravital e da epifluorescência utilizando os corantes diacetato de carboxifluoresceína e iodeto de propídeo; funcionalidade da membrana plasmática pelo teste hiposmótico; e motilidade progressiva e vigor espermáticos durante o teste de termorresistência por 90 minutos. O vigor espermático do sêmen diluído com Kenney foi superior (P<0,05) ao do Foote na avaliação do sêmen fresco diluído. Não houve diferença estatística entre as demais características seminais avaliadas no sêmen fresco diluído. No sêmen resfriado, o Kenney propiciou a manutenção da motilidade e do vigor espermático após as 24 horas de armazenamento, porém o Foote apresentou maiores valores médios (P<0,05) para porcentagem de espermatozoides reativos ao teste hiposmótico e viáveis pelo teste de epifluorescência. Os valores médios da motilidade progressiva e do vigor espermático durante os testes de termorresistência do sêmen resfriado foram inferiores ao preconizado pelo CBRA (1998) aos 30 minutos com o diluidor de Foote e aos 60 minutos com o dilluidor Kenney. No experimento 2, avaliou-se a fertilidade do sêmen resfriado com ambos os diluidores. Foram realizadas 17 coletas do sêmen do garanhão 1 e 23 coletas do garanhão 2. Após as coletas, o sêmen foi diluído com os diluidores utilizados no experimento 1 e ajustado para uma dose inseminante de 500 milhões de espermatozoides viáveis num volume final de 15 mL por dose. O sêmen foi acondicionado no Equitainer®, resfriado até 5 oC e armazenado por no mínimo 24 horas. Foram avaliadas as seguintes características seminais: motilidade progressiva e vigor espermático do sêmen diluído e resfriado; e morfologia espermática do sêmen fresco e resfriado com ambos os diluidores. Foram utilizados 31 ciclos estrais de 26 éguas no primeiro período experimental e 43 ciclos de 28 éguas no segundo período experimental. Após a detecção de folículo de 30 mm, as éguas foram distribuídas ao acaso em dois grupos: grupo A (éguas inseminadas com sêmen resfriado com diluidor Kenney) e grupo B (éguas inseminadas com sêmen resfriado com diluidor Foote). O sêmen dos garanhões foi utilizado aleatoriamente entre as éguas. As inseminações foram realizadas depois de no mínimo 24 horas de armazenamento do sêmen. O sêmen foi depositado no corpo do útero, a partir da detecção de um folículo de 30-40 mm até a observação da ovulação. As inseminações foram feitas somente em dias predeterminados (terça-feira, quinta-feira e sábado) e o diagnóstico de gestação foi realizado por meio de ultrassonografia transretal. O diluidor de Kenney se mostrou superior ao Foote na manutenção da motilidade progressiva e do vigor espermático após o período de resfriamento. A taxa de gestação do grupo A, 55,26% (21/38), foi maior (P<0,05) que a do grupo B, 30,56% (11/36). Os testes de avaliação da integridade e funcionalidade da membrana plasmática da célula espermática auxiliam na predição do potencial fecundante do garanhão, porém ainda não são conclusivos. O diluidor seminal à base de glicina-gema de ovo (Foote, 2002) não é eficaz em preservar as características seminais e não possibilita obter índice de fertilidade aceitável. O diluidor proposto por Kenney et al. (1975) ainda é uma ótima opção para inseminação artificial com sêmen resfriado de garanhões da raça Mangalarga Marchador.

As biotécnicas da reprodução na espécie equina avançaram na última década e os criadores de equinos começaram a questionar as possibilidades de interferência na determinação do sexo dos potros. A determinação do sexo é importante, pois o sexo do potro possui grande influência no valor comercial do mesmo. O objetivo do presente estudo foi avaliar a influência do momento da cobertura em relação ao momento da ovulação no sexo dos potros e outros fatores como: idade da égua e garanhão; indutor da ovulação; ovário em que ocorreu a ovulação e o diâmetro do folículo pré-ovulatório também foram avaliados. O estudo foi realizado em Haras na região de Bagé e Aceguá (RS) – Brasil (31°24'06.1''S 54°07'47.5''W) e (31°30'16.0''S 54°07'45.5''W) nas estações de monta de 2011 a 2015. Utilizou-se 259 ciclos reprodutivos de 160 éguas e 22 garanhões da raça Puro Sangue de Corrida. Informações como agente indutor de ovulação utilizado (Deslorelina; n = 187 ou hCG; n = 72); data da cobertura (n = 259); momento da ovulação em relação a cobertura (+ 24 horas; n = 69 e - 24 horas; n = 190); idade da égua (G1: até 8 anos; n = 123, G2: 9 até 14 anos n = 110 e G3: >14 anos; n = 26); idade do garanhão (Até 14 anos; n = 11 e >15 anos; n = 11) ; ovário em que a ovulação ocorreu (Direito; n = 122 e Esquerdo; n = 137); foram registrados e avaliados No total de potros nascidos, 136 foram machos (52,51%) e 123 foram fêmeas (47,49%). O tempo decorrido após a cobertura não influenciou o sexo dos produtos, nas éguas que ovularam com – 24h após a indução da ovulação: 104 potros (54,74%) eram machos e 86 (45,26%) eram fêmeas, enquanto que nas éguas que ovularam com + 24h, 32 potros (46,38%) eram machos e 37 (53,62%) eram fêmeas. A porcentagem de fêmeas nascidas em relação a idade da égua foi de 46,34% (n=57), 47,27% (n=52) e 46,15% (n=12) nos grupos G1, G2 e G3 respectivamente. Garanhões com idade até 15 anos tiveram 44,14% (n=49) de fêmeas e com mais de 15 anos 49,66% (n=73) eram fêmeas. Não houve diferença no sexo dos produtos quanto ao agente indutor da ovulação (Deslorelina x hCG) e ovário em que ocorreu a ovulação. O presente estudo concluiu que nenhum dos fatores estudados alterou a proporção macho:fêmea dos potros nascidos. / The biotechnologies of reproduction in equine species have been improved in the last decade and the horse breeders started to inquire about the possibility of intervention regarding to the sex determination in foals. Sex determination is important because the sex of the foal has a great influence on the commercial value of the foal. The aim of the present study is was to evaluate the influence of the time of the breeding in relation to time of ovulation in the sex of the foals, besides analyzing other factors such as: the age of the mare and the stallion; ovulation inductor, ovary and the diameter of the preovulatory follicle. The study was accomplished in studs located Bagé/RS and Aceguá/RS – Brazil (31°24'06.1''S 54°07'47.5''W) and (31°30'16.0''S 54°07'45.5''W) during the breeding seasons from 2011 to 2015, using 259 reproductive cycles of 160 mares and 22 stallions of Thoroughbred breed. Information like the induction of ovulation agent that was used (Deslorelin; n = 187 or hCG; n = 72); date of breeding (n = 259); time of ovulation in relation to the breeding (+24 hours; n = 69 and -24 hours; n = 190); age of the mare (G1: up to the age of 8; n = 123 G2: from the age of 9 to the age of 14; n = 110 and G3: >14 years old; n = 26); age of the stallion (up to the age of 14; n = 11 and >15 years old; n = 11); ovary in which the ovulation occurred (Right; n = 122 and Left; n = 137) were catalogued and evaluated As result, 136 (52,51%) were born colts and 123 (47,49%) were born fillies. The elapsed time from breeding to ovulation did not influence on the sex of the product, on mares that ovulated in less than 24 hours after the ovulation induction: 104 foals (54,74%) were male and 86 (45,26%) were female, whereas in the mares that ovulated in more than 24 hours, 32 foals (46,38%) were male and 37 (53,62%) were female. The percentage of born females regarding to the age of the mare was 46,34% (n = 57), 47,27% (n = 52) and 46,15% (n = 12) in the groups G1, G2 and G3, respectively. From stallions up to the age of 15 years, 44,14% (n = 49) were females and from those which were older than 15 years old, 49,66% (n = 73) were females. There was no difference regarding the sex of the product in relation to the ovulation inducer agent (Deslorelin x hCG) and ovary in which the ovulation occurred. None of the factors studied modified the male:female proportion of the born foals.

Em todas as espécies de mamíferos estudadas, o gene PLCζ é responsável pelo aumento de cálcio no momento da fecundação do oócito. Recentemente, o gene WBP2NL, também esta sendo estudado por sua possível capacidade de iniciar a ativação de oócitos. O gene TNF-α é uma citocina pró-inflamatória que pode ser encontrada no plasma seminal, mas seu efeito na viabilidade espermática ainda não está claro. O objetivo desse trabalho foi investigar a expressão dos genes PLCζ, WPB2NL e TNF-α no espermatozoide e suas relações com a qualidade e fertilidade em equinos. Foram utilizados ejaculados de 40 garanhões da raça Crioula, de criatórios localizados próximos a Porto Alegre. A concentração espermática foi avaliada em câmara de Neubauer. As demais análises microscópicas foram realizadas através do Computer Assisted Sperm Analysis AndroVision. A análise da integridade física da membrana foi realizada utilizando-se sondas fluorescentes A integridade funcional da membrana plasmática foi avaliada por meio do teste hiposmótico. Para análise estatística foi utilizada correlação de Pearson, com nível de significância de P<0,05. Foi encontrada correlação positiva do gene PCLζ com integridade funcional da membrana plasmática (0,449), integridade física da membrana plasmática (0,438), taxa de prenhez (0,454), motilidade total (0,486), motilidade progressiva (0,413); correlação negativa com motilidade lenta (-0,427) e espermatozoides imóveis (-0,405). Não foram encontradas correlações dos genes WBP2NL e TNF-α. Concluí-se que a expressão do gene PCLζ no Espermatozoide equino pode ser utilizada com um marcador para a qualidade seminal e fertilidade do garanhão. / In all species of mammals studied, the PLCζ gene is a factor of calcium increase at the time of oocyte fertilization. Recently, the WBP2NL gene is also being studied for its potential ability to initiate oocyte activation during fertilization. The TNF-α is a pro-inflammatory cytokine found in seminal plasma but it effect on motility and viability sperm still unclear. The aim of this study was to verify the expression of PLCζ, WPB2NL and TNF-α genes in spermatozoa and their relationship with quality and fertility in horses. We used the ejaculates of 40 Crioulo stallions, were used from horse farms located near Porto Alegre. After semen collection, the ejaculate was diluted in a commercially available extender. Microscopic analysis of total and progressive motility was performed using a computer assisted sperm analysis system AndroVision. Membrane physical integrity was assessed by fluorescent probes and membrane functionality was analyzed by the hypo-osmotic test (HOST) Statistical analysis was performed using the Pearson correlation, with a significance level of P<0.05. A moderate positive correlation of the PCLζ gene was found with functional plasma membrane function (0.449), physical organization of the plasma membrane (0.438), conception rate (0.454), total motility (0.486), progressive motility (0.413) and negative correlation with slow motility (-0.427) and spermatozoa properties (-0.405). No correlation on WBP2NL and TNF-α genes were found, showing that those genes expression should not be used as a marker of seminal quality and fertility in stallions. It can be concluded that expression of the PCLζ gene on equine spermatozoa can be used as a marker for seminal quality and fertility in the stallion.