Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders / Recuperação de espermatozoides após refrigeração do epidídimo de gatos domésticos utilizando os diluidores ACP-117c e Tris

We aimed to compare fresh sperm and sperm cooled to 4ºC that had been recovered from the epididymides of cats using powdered coconut water (ACP-117c) and Tris extenders. Sixty epididymides were divided into 6 groups: 10 fresh epididymides were recovered using Tris (T0h); 10 were kept at 4°C/2h and recovered using Tris (T2h); 10 were kept at 4°C/4h and recovered using Tris (T4h); 10 fresh were recovered using ACP-117c (A0h); 10 were kept at 4°C/2h and recovered using ACP-117c (A2h), and 10 were kept at 4°C/4h and recovered using ACP-117c (A4h). The testis-epididymis complexes (TEC) control were not cooled. The others were cooled at 4°C for 2 or 4h. The epididymis was separated and the sperm was recovered by the modified flotation method. Sperm kinetic parameters were evaluated by a computer-system analysis, and vigor, viability, concentration, membrane function and morphology of the sperm were assessed under a light microscope. The progressive motility with ACP-117c declined after 2h of cooling, but did not differ between fresh and 4h. The vigor and membrane function were higher in A4h than A0h. The vigor at T2h and T4h were decreased compared to T0h. T0h was higher than A0h for vigor and sperm membrane function. However, after 4h of cooling, ACP-117c maintained a higher percentage of living cells. Feline epididymal sperm quality can be maintained to the degree necessary for artificial breeding programs following cooling and ACP-117c may be successfully used to recover cat sperm that have been cooled for up to 4h.(AU)

Objetivou-se comparar a qualidade de espermatozoides recuperados a fresco e após refrigeração a 4ºC do epidídimo de gatos domésticos utilizando-se os diluidores ACP-117c e Tris. Sessenta epidídimos foram distribuídos em seis grupos: 10 epidídimos a fresco com o Tris (T0h), 10 a 4°C/2h e recuperados com Tris (T2h), 10 a 4°C/4h e recuperados com Tris (T4h), 10 epidídimos a fresco com o ACP-117c (A0h), 10 a 4 °C/2h e recuperados com ACP-117c (A2h), 10 a 4°C/4h e recuperados com ACP-117c (A4h). Os complexos testículo-epidídimo (CTE) do controle não foram refrigerados. Os outros foram refrigerados a 4°C durante duas e quatro horas. Os epidídimos foram separados das demais estruturas, e os espermatozoides recuperados pela técnica de flutuação modificada. Os parâmetros cinéticos foram avaliados em um sistema computadorizado, e o vigor, a viabilidade, a concentração, a funcionalidade de membrana e a morfologia celular foram avaliados em microscopia de luz. A motilidade progressiva com ACP-117c declinou após duas horas de refrigeração, mas não diferiu entre a recuperação a fresco e após refrigeração por quatro horas. Vigor e integridade funcional da membrana celular foram significativamente superiores no grupo A4h em comparação ao A0h. O vigor espermático em T2h e T4h reduziu significativamente em comparação com T0h. T0h foi significativamente superior ao A0h quanto aos parâmetros de vigor e integridade funcional da membrana espermática, entretanto, após quatro horas de refrigeração, o ACP-117c apresentou um maior percentual de células vivas. Os espermatozoides epididimários de felinos domésticos conseguem manter a qualidade necessária para serem utilizados em programas de reprodução artificial após serem refrigerados e recuperados por meio da técnica de flutuação modificada, e o diluidor ACP-117c pode ser utilizado com sucesso para recuperação de células espermáticas refrigeradas de gatos por até quatro horas.(AU)

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.(AU)

Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders / Recuperação de espermatozoides após refrigeração do epidídimo de gatos domésticos utilizando os diluidores ACP-117c e Tris

We aimed to compare fresh sperm and sperm cooled to 4ºC that had been recovered from the epididymides of cats using powdered coconut water (ACP-117c) and Tris extenders. Sixty epididymides were divided into 6 groups: 10 fresh epididymides were recovered using Tris (T0h); 10 were kept at 4°C/2h and recovered using Tris (T2h); 10 were kept at 4°C/4h and recovered using Tris (T4h); 10 fresh were recovered using ACP-117c (A0h); 10 were kept at 4°C/2h and recovered using ACP-117c (A2h), and 10 were kept at 4°C/4h and recovered using ACP-117c (A4h). The testis-epididymis complexes (TEC) control were not cooled. The others were cooled at 4°C for 2 or 4h. The epididymis was separated and the sperm was recovered by the modified flotation method. Sperm kinetic parameters were evaluated by a computer-system analysis, and vigor, viability, concentration, membrane function and morphology of the sperm were assessed under a light microscope. The progressive motility with ACP-117c declined after 2h of cooling, but did not differ between fresh and 4h. The vigor and membrane function were higher in A4h than A0h. The vigor at T2h and T4h were decreased compared to T0h. T0h was higher than A0h for vigor and sperm membrane function. However, after 4h of cooling, ACP-117c maintained a higher percentage of living cells. Feline epididymal sperm quality can be maintained to the degree necessary for artificial breeding programs following cooling and ACP-117c may be successfully used to recover cat sperm that have been cooled for up to 4h.(AU)

Objetivou-se comparar a qualidade de espermatozoides recuperados a fresco e após refrigeração a 4ºC do epidídimo de gatos domésticos utilizando-se os diluidores ACP-117c e Tris. Sessenta epidídimos foram distribuídos em seis grupos: 10 epidídimos a fresco com o Tris (T0h), 10 a 4°C/2h e recuperados com Tris (T2h), 10 a 4°C/4h e recuperados com Tris (T4h), 10 epidídimos a fresco com o ACP-117c (A0h), 10 a 4 °C/2h e recuperados com ACP-117c (A2h), 10 a 4°C/4h e recuperados com ACP-117c (A4h). Os complexos testículo-epidídimo (CTE) do controle não foram refrigerados. Os outros foram refrigerados a 4°C durante duas e quatro horas. Os epidídimos foram separados das demais estruturas, e os espermatozoides recuperados pela técnica de flutuação modificada. Os parâmetros cinéticos foram avaliados em um sistema computadorizado, e o vigor, a viabilidade, a concentração, a funcionalidade de membrana e a morfologia celular foram avaliados em microscopia de luz. A motilidade progressiva com ACP-117c declinou após duas horas de refrigeração, mas não diferiu entre a recuperação a fresco e após refrigeração por quatro horas. Vigor e integridade funcional da membrana celular foram significativamente superiores no grupo A4h em comparação ao A0h. O vigor espermático em T2h e T4h reduziu significativamente em comparação com T0h. T0h foi significativamente superior ao A0h quanto aos parâmetros de vigor e integridade funcional da membrana espermática, entretanto, após quatro horas de refrigeração, o ACP-117c apresentou um maior percentual de células vivas. Os espermatozoides epididimários de felinos domésticos conseguem manter a qualidade necessária para serem utilizados em programas de reprodução artificial após serem refrigerados e recuperados por meio da técnica de flutuação modificada, e o diluidor ACP-117c pode ser utilizado com sucesso para recuperação de células espermáticas refrigeradas de gatos por até quatro horas.(AU)

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.

Efeito da Insulina-N sobre a Integridade das Membranas Plasmáticas em Sêmen Fresco de Ovino, utilizado Gema de Ovo de Codorna / Insulin- N Effects on Plasma Membranes Integrity in Ovine Fresh Semen, using Quail Egg of Yolk

This study aimed to evaluate the effect of insulin on the membrane integrity evaluated by the eosin testand nigrosine in ovine semen. Seven sheep were used, harvest were made with the aid of eletroejaculator. Theyolk-citrate extender was used for dilution of the ejaculate (25x106 sperm / mL) using quail's egg yolk. Eachejaculate was divided into four equal fractions, corresponding to treatments: control group (Gcont) using onlydilutive citrat gem; Group 50 (G50) plus dilutive 50UI/ml insulin type N; Group 100 (G100) plus dilutive 100IU/Ml Insulin type N; Group 200 (G200) plus dilutive 200 IU/Ml Insulin type N. After dilution, the semenremained in water - bath at 45°C to perform the eosin test and nigrosine for evaluation by time (0, 20, 40 and 60minutes) for membrane integrity. Thus, the use of insulin does not preserve the integrity of the membranesduring the incubation period of 45C, with a gradual decrease due to stress caused by time and temperature atwhich the semen was subjected.(AU)

Efeito de diferentes meios e concentrações espermáticas na fecundação in vitro curta sobre a produção de embriões bovinos / Effect of different mediums and sperm concentrations in the short-in vitro fertilization on the bovine embryo production

tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16  106 and 20  106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16  106 and 20  106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20  106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.(AU)

Viabilidade de espermatozoides epidídimários de touros de corte post-mortem refrigerados em diferentes períodos utilizando ACP-111® e TRIS / Epididymal sperm viability cutting bulls post-mortem chilled at different times using ACP- 111® and TRIS

The increase in productivity in the cattle sector can be boosted with the use of biotecnias that can decrease the generation interval and increase the intensity of selection, and facilitate the dissemination of genetic material quality. During the period of November 13 to December 13, 2015, it was used 40 pairs of epididymis coming from bulls without defined breed (SPRD) compared the sperm viability after cooling, using two extenders (ACP and TRIS). Overall, there was no significant difference between the diluent, however, motility use with the ACP is less than TRIS after 12 hours of cooling.

Viabilidade de espermatozoides epidídimários de touros de corte post-mortem refrigerados em diferentes períodos utilizando ACP-111® e TRIS / Epididymal sperm viability cutting bulls post-mortem chilled at different times using ACP- 111® and TRIS

The increase in productivity in the cattle sector can be boosted with the use of biotecnias that can decrease the generation interval and increase the intensity of selection, and facilitate the dissemination of genetic material quality. During the period of November 13 to December 13, 2015, it was used 40 pairs of epididymis coming from bulls without defined breed (SPRD) compared the sperm viability after cooling, using two extenders (ACP and TRIS). Overall, there was no significant difference between the diluent, however, motility use with the ACP is less than TRIS after 12 hours of cooling.(AU)

Análise da integridade de membrana de espermatozoides extraídos de fragmentos frescos de testículos de cães / Sperm membrane integrity analysis from dog fresh testis

To evaluate testicular tissue before cryopreservation is important to quantify and qualify losses ofsperm during the freezing/thawing of testicular tissue. The aim of this study was to analyze the sperm plasmamembrane structural and functional integrity on post-pubertal dogs (n = 7) fresh testis. The sperm fromtesticular tissue were submitted to plasma membrane structural and functional integrity analysis by fluorescentmicroscopy using SYBR 14 and Propidium Iodide and the hiposmotic test, respectively. For statistical analysis,it was used normality test (Shapiro Wilk) and Wilcoxon test (R 3.3.1; P > 0.05). It was observed 94.9% and91.8% spermatozoa presenting structural and functional integrity, respectively. Thus, analysis of the membraneintegrity is an important tool to analyze spermatozoa extracted from testicles. Despite the fluorescence be apractical method, it has a high cost.(AU)

Efeito de diferentes meios e concentrações espermáticas na fecundação in vitro curta sobre a produção de embriões bovinos / Effect of different mediums and sperm concentrations in the short-in vitro fertilization on the bovine embryo production

tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16  106 and 20  106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16  106 and 20  106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20  106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.

Efeito da Insulina-N sobre a Integridade das Membranas Plasmáticas em Sêmen Fresco de Ovino, utilizado Gema de Ovo de Codorna / Insulin- N Effects on Plasma Membranes Integrity in Ovine Fresh Semen, using Quail Egg of Yolk

This study aimed to evaluate the effect of insulin on the membrane integrity evaluated by the eosin testand nigrosine in ovine semen. Seven sheep were used, harvest were made with the aid of eletroejaculator. Theyolk-citrate extender was used for dilution of the ejaculate (25x106 sperm / mL) using quail's egg yolk. Eachejaculate was divided into four equal fractions, corresponding to treatments: control group (Gcont) using onlydilutive citrat gem; Group 50 (G50) plus dilutive 50UI/ml insulin type N; Group 100 (G100) plus dilutive 100IU/Ml Insulin type N; Group 200 (G200) plus dilutive 200 IU/Ml Insulin type N. After dilution, the semenremained in water - bath at 45°C to perform the eosin test and nigrosine for evaluation by time (0, 20, 40 and 60minutes) for membrane integrity. Thus, the use of insulin does not preserve the integrity of the membranesduring the incubation period of 45C, with a gradual decrease due to stress caused by time and temperature atwhich the semen was subjected.

Análise da integridade de membrana de espermatozoides extraídos de fragmentos frescos de testículos de cães / Sperm membrane integrity analysis from dog fresh testis

To evaluate testicular tissue before cryopreservation is important to quantify and qualify losses ofsperm during the freezing/thawing of testicular tissue. The aim of this study was to analyze the sperm plasmamembrane structural and functional integrity on post-pubertal dogs (n = 7) fresh testis. The sperm fromtesticular tissue were submitted to plasma membrane structural and functional integrity analysis by fluorescentmicroscopy using SYBR 14 and Propidium Iodide and the hiposmotic test, respectively. For statistical analysis,it was used normality test (Shapiro Wilk) and Wilcoxon test (R 3.3.1; P > 0.05). It was observed 94.9% and91.8% spermatozoa presenting structural and functional integrity, respectively. Thus, analysis of the membraneintegrity is an important tool to analyze spermatozoa extracted from testicles. Despite the fluorescence be apractical method, it has a high cost.

Avaliação automática da morfometria de espermatozoides ovinos com uso do programa ImageJ / Automatic morphometric evaluation of ram sperm using the ImageJ software

The objective of this study was to evaluate the ImageJ software in automatic analysis of morphometricparameters of ram sperm head. Semen smears from three Santa Inês rams were stained with fast panoptic anddigital images of sperms from these smears were analyzed in ImageJ software for morphometric parameters.The mean dimensions of sperm heads analyzed were 8,4±0,6 µm; 4,4±0,3 µm; 21,8±1,6 µm and 29,0±3,9 µm2for length, width, perimeter and area, respectively. Individual variations for these parameters were observed.The ImageJ software was suitable for automatic sperm morphometric analysis with results consistent with thosefound in the literature.(AU)

Avaliação automática da morfometria de espermatozoides ovinos com uso do programa ImageJ / Automatic morphometric evaluation of ram sperm using the ImageJ software

The objective of this study was to evaluate the ImageJ software in automatic analysis of morphometricparameters of ram sperm head. Semen smears from three Santa Inês rams were stained with fast panoptic anddigital images of sperms from these smears were analyzed in ImageJ software for morphometric parameters.The mean dimensions of sperm heads analyzed were 8,4±0,6 µm; 4,4±0,3 µm; 21,8±1,6 µm and 29,0±3,9 µm2for length, width, perimeter and area, respectively. Individual variations for these parameters were observed.The ImageJ software was suitable for automatic sperm morphometric analysis with results consistent with thosefound in the literature.

Morfologia espermática de touros: interpretação e impacto na fertilidade / Bull sperm morphology: interpretation and impact on fertility

Embora nas últimas décadas tenham sido desenvolvidas diversas técnicas laboratoriais que aumentaram a possibilidade de uma análise mais criteriosa da integridade estrutural dos espermatozoides, o estudo e aplicação das alterações da forma espermática (patologia espermática) continuam sendo componentes essenciais para o exame do sêmen, pois fornecem uma estimativa do percentual de espermatozoides normais ou estruturalmente íntegros, assim como a distribuição dos diferentes defeitos morfológicos. Neste trabalho são apresentados, entre outros, os aspectos de técnicas utilizadas para a avaliação e classificação da morfologia espermática. Ainda, quanto à morfologia espermática, discutiu-se a origem, interpretação e impacto na fertilidade. Por fim, foram apresentados os padrões normalmente utilizados para a avaliação do potencial de fertilidade do sêmen bovino congelado de forma convencional e sexado.(AU)

Although in recent decades it has been developed various laboratory techniques that increased the possibility of a more detailed analysis of the structural integrity of sperm, the study and application of changes in the sperm morphology (sperm pathology) continue to be essential components for the semen examination, it gives an estimative of the percentage of normal sperm or structural integrity, as well as the distribution of different morphological defects. This work presents, among others, aspects of techniques used for the assessment and classification of sperm morphology. Moreover, regarding the sperm morphology, the origin, interpretation and impact on fertility were discussed. Finally, some patterns used for assessing the potential fertility of cryopreserved bovine semen (conventional and sexed) are presented.(AU)

Morfologia espermática de touros: interpretação e impacto na fertilidade / Bull sperm morphology: interpretation and impact on fertility

Embora nas últimas décadas tenham sido desenvolvidas diversas técnicas laboratoriais que aumentaram a possibilidade de uma análise mais criteriosa da integridade estrutural dos espermatozoides, o estudo e aplicação das alterações da forma espermática (patologia espermática) continuam sendo componentes essenciais para o exame do sêmen, pois fornecem uma estimativa do percentual de espermatozoides normais ou estruturalmente íntegros, assim como a distribuição dos diferentes defeitos morfológicos. Neste trabalho são apresentados, entre outros, os aspectos de técnicas utilizadas para a avaliação e classificação da morfologia espermática. Ainda, quanto à morfologia espermática, discutiu-se a origem, interpretação e impacto na fertilidade. Por fim, foram apresentados os padrões normalmente utilizados para a avaliação do potencial de fertilidade do sêmen bovino congelado de forma convencional e sexado.

Although in recent decades it has been developed various laboratory techniques that increased the possibility of a more detailed analysis of the structural integrity of sperm, the study and application of changes in the sperm morphology (sperm pathology) continue to be essential components for the semen examination, it gives an estimative of the percentage of normal sperm or structural integrity, as well as the distribution of different morphological defects. This work presents, among others, aspects of techniques used for the assessment and classification of sperm morphology. Moreover, regarding the sperm morphology, the origin, interpretation and impact on fertility were discussed. Finally, some patterns used for assessing the potential fertility of cryopreserved bovine semen (conventional and sexed) are presented.

Sperm biotechnologies in domestic species: state of the art

Modern livestock breeding is basically dependent on the proper use of semen for artificial insemination of femeles and of other reproductive biotechnologies such as the production of embryos in vitro for embryo transfer (IVP). Both these techniques have made possible not only the wide dissemmation of genetic material onto breeding populations but also anhanced the selection of best sires, owing to the development of better diagnostic techniques for sperm function and of preservation of seminal material over time. Although use ofliquid semen cooled to room temperature, to intermediate temperatures (+16-20ºC) or chilled (+5ºC) dominates in different species cryopreservation is preferred in bovine A1 and it is advancing in other species by the design of new containers, freezing methods and the use of better insemination strategies. Techniques to separate the aliquot of most robust spermatozoa from an ejaculate have shown a renascent particularly for sires with low sperm quality, and technological advances in separating spermatozoa for c hromosomal sex make the technique suitable for commercial use, following applicat ion of novel findings in sperm and seminal plasma (SP) diagnostics and function. Alongside, knowledge of the epigenome and signalling c apabilities of the semen (sperm and SP) call s for further studies regarding transgene production via ICSI for IVP or AI.

Sperm biotechnologies in domestic species: state of the art

Modern livestock breeding is basically dependent on the proper use of semen for artificial insemination (AU) of femeles and of other reproductive biotechnologies such as the production of embryos in vitro for embryo transfer (IVP). Both these techniques have made possible not only the wide dissemmation of genetic material onto breeding populations but also anhanced the selection of best sires, owing to the development of better diagnostic techniques for sperm function and of preservation of seminal material over time. Although use ofliquid semen cooled to room temperature, to intermediate temperatures (+16-20ºC) or chilled (+5ºC) dominates in different species cryopreservation is preferred in bovine A1 and it is advancing in other species by the design of new containers, freezing methods and the use of better insemination strategies. Techniques to separate the aliquot of most robust spermatozoa from an ejaculate have shown a renascent particularly for sires with low sperm quality, and technological advances in separating spermatozoa for c hromosomal sex make the technique suitable for commercial use, following applicat ion of novel findings in sperm and seminal plasma (SP) diagnostics and function. Alongside, knowledge of the epigenome and signalling c apabilities of the semen (sperm and SP) call s for further studies regarding transgene production via ICSI for IVP or AI.(AU)

Insemination of four cows per dose of frozen semen with a fixed-time artificial insemination protocol

The aim of the study was to evaluate the practicality of a 1: 4 dilution of doses of semen in fixed - time artificial insemination ( FTAI ) programs. Nelore cows (n = 803 ) were synchronized by a conventional FTAI protocol. For AI in the dilut ed group (n = 392) , 0.5 ml semen straws were thawed and 0.5 ml of extender conta ining amino acids and methylxanthine derivatives was added. The straws were fractionated into four straws of 0.25 ml each (dilution 1:4) with approximately 2.5 x 10 6 spermatozoa each and the insemination was performed in the ipsilateral horn to the ovary c ontaining the dominant follicle . In the control group (n = 411), AI was performed in the uterine body with one straw of 0.5 ml (10 x 10 6 spermatozoa) containing seme n from the same bull and batch . The conception rate was 49.2% (193/392) for the diluted gro up and 50.1% (206/411) for the control group, with 1.97 and 0.50 pregnancies per dose of semen , respectively . T he dilution of semen provide d a pregnancy index similar to the control group ; however, the technique increased the number of pregnancies per dos e , allowing for the best use of semen of high genetic value and reducing the cost per pregnancy.

Semen evaluation techniques and their relationship with fertility

This review summarizes those methods - established and emerging of semen assessment whose outcom e intents revealing its potential fertility and, as a carry over concept, that of the sire whose semen we examined. The review does not, however, focus on the wide display of current techn iques designed to explore specific or multiple sets of sperm attributes essential for fertilization but on two basic con cerns present: the heterogeneity of the sperm suspension and the multitude of attributes required for each spermatozoon to be fertile; concepts that shadow our diagnostic capabilities. The review points out advancements in the exploration of the genome, the transcriptome, and the proteome of both spermatozoa and the seminal plasma which unveil how spermatozoa modulate their own survival an d signal to the environment when displaying degenerative changes. Specific seminal plasma components, both among individuals and portions of the ejaculate, not only relate to survival but also signal differential immune tolerance by the female with a previ ously unattended linkage to fertility. Lastly it foresees how Cytomics, combining novel designed motility analyzers, flow cytometers and enhanced digital imaging shall dominate the landscape of andrological laboratories and enable quick determinations on huge sperm numbers for markers highly relevant to sperm function and hence, for fertility.