Reproductive biology of Pêga Jackasses: characteristics of sperm-rich fraction collected using the split-ejaculate method / Biologia reprodutiva de jumentos da raça Pêga: características da fração rica em espermatozoides coletada de forma fracionada

The aim of this study was to evaluate the physical and morphological characteristics of the sperm-rich fraction of jackass semen. To this end, 130 ejaculates from five Pêga jackasses were collected using an open model artificial vagina. The sperm-rich fraction was collected using the split-ejaculate method and assessed for the number of mounts/ejaculate, for physical and morphological characteristics of the semen and number of doses produced/ejaculate. It was observed that all characteristics evaluated differed among the five jackasses, except for the head defect rates. The mean values obtained for the jackass sperm-rich fraction collected were: number of mounts/ejaculate - 1.27; semen volume - 20.21mL; motility - 84.53%; vigor - 4.46; motility after dilution - 80.10%; sperm concentration/mL - 894.38 x 10 6; total sperm/ejaculate - 16.14 x 10 9; number of insemination doses/ejaculate (400x10 6 motile sptz) - 33.39; number of insemination doses/ejaculate (800 x 10 6 motile sptz) - 16.69; and percentage of normal sperm - 90.46%. Thus, in the present experiment the split-ejaculate method using an open artificial vagina worked well with the jackasses, and the sperm-rich fraction of the ejaculate of Pêga jackasses had high quality and sperm concentration, allowing its use for semen processing without reducing the number of insemination doses produced per ejaculate.(AU)

Foram coletados 130 ejaculados de cinco jumentos da raça Pêga, utilizando-se vagina artificial modelo aberta, com o objetivo de se avaliar as características físicas e morfológicas da fração rica em espermatozoides do ejaculado de asininos. Para tal, a fração rica em espermatozoides, composta pelos três primeiros jatos ejaculados, foi coletada e avaliada quanto ao número de montas/ejaculado, quanto às características físicas e morfológicas do sêmen, bem como quanto ao número de doses inseminantes produzidas/ejaculado. Observou-se que todas as características avaliadas diferiram entre os cinco reprodutores avaliados, com exceção do percentual de defeitos de cabeça. Os valores médios obtidos da coleta da fração rica do ejaculado de jumentos foram: número de montas/ejaculado - 1,27; volume de sêmen - 20,21mL; motilidade - 84,53%; vigor - 4,46; motilidade pós-diluição - 80,10%; espermatozoides/mL - 894,38 x 10 6; espermatozoides/ejaculado - 16,14 x 10 9; número de doses inseminantes/ejaculado (400x10 6 sptz móveis) - 33,39; doses inseminantes/ejaculado (800 x 10 6 sptz/móveis) - 16,69; e percentual de espermatozoides normais - 90,46%. Assim, no presente experimento, observou-se boa aceitação dos reprodutores à coleta fracionada utilizando-se a vagina artificial aberta, sendo a fração rica do ejaculado de jumentos da raça Pêga caracterizada por alta qualidade e concentração espermática, o que viabilizou sua utilização para o processamento do sêmen, sem prejuízos quanto ao número de doses inseminantes produzidas/ejaculado.(AU)

Physical and morphological characteristics of the first three jets of Pêga jackasses sperm-rich fraction / Características físicas e morfológicas dos três primeiros jatos da fração rica do ejaculado de jumentos da raça Pêga

The first three jets of the sperm-rich fraction of Pêga jackasses were collected and assessed separately. Five fertile Pêga jackasses were used as semen donors and underwent fractionated semen collection, using an open model artificial vagina. The first three jets of the semen were collected separately and assessed for volume, sperm motility, vigor, concentration/mL of semen, and sperm morphology. These characteristics were compared between first, second and third jets and between jackasses. It was observed that the jet volume differed (P<0.05) between jackasses, although it was similar (P>0.05) between first, second and third jets. Sperm motility did not differ (P>0.05) between jets and jackasses. Vigor was similar (P>0.05) between jets of the same jackass, and only the first jet differed (P<0.05) between jackasses. The first, second and third jets of the sperm-rich fraction had decreased sperm concentrations (P<0.05) of 955.56, 725.56 and 280.56x 106 sperm/mL of semen, respectively. Sperm morphology differed between the first three jets only for the incidence of mid-piece defect, higher in the third one (4.26%), compared to the first (3.36%) and second (3.38%) ones. When comparing the morphological characteristics of the sperm-rich fraction between five jackasses, regardless of the jet, there were differences in the percentage of normal sperm, proximal cytoplasmic droplet, mid-piece and head defects.(AU)

Objetivou-se, no presente experimento, caracterizar os três primeiros jatos da fração rica do ejaculado de jumentos da raça Pêga. Para tal, cinco jumentos foram submetidos à coleta fracionada do sêmen, utilizando-se vagina artificial modelo aberta. Os três primeiros jatos da fração rica do ejaculado foram coletados separadamente e avaliados quanto ao volume, à motilidade e ao vigor espermáticos, à concentração espermática/mL de sêmen e à morfologia espermática. Comparações foram realizadas entre jatos e entre jumentos. Observou-se que o volume do jato diferiu (P<0,05) entre os jumentos, embora fosse similar (P>0,05) entre os jatos. A motilidade espermática não diferiu (P>0,05) entre jatos nem entre jumentos. O vigor espermático foi similar (P>0,05) entre os jatos de um mesmo jumento, e apenas o vigor do jato 1 diferiu (P<0,05) entre os jumentos. Independentemente do jumento, a fração rica do ejaculado foi composta por três jatos apresentando concentrações espermáticas decrescentes (P<0,05), com 955,56; 725,56 e 280,56 x 106 espermatozoides/mL de sêmen. A morfologia espermática diferiu entre os três jatos avaliados apenas para a incidência de defeitos de peça intermediária, sendo maior no jato três (4,26%), em relação aos jatos um (3,36%) e dois (3,38%). Comparando-se as características morfológicas do sêmen entre os cinco jumentos avaliados, independentemente do jato, observaram-se diferenças entre os reprodutores quanto ao percentual de espermatozoides normais, com gota citoplasmática proximal, com defeitos de peça intermediária e de cabeça.(AU)

The Difference in the Outcomes between Surgically Retrieved and Ejaculated Spermatozoa for Intracytoplasmic Sperm Injection Cycles in Sulaimanyah Province.

Background: Intracytoplasmic sperm injection (ICSI) is introduced as a treatment modality for severe male factors infertility. It is an effective form of infertility treatment. Progress in the micromanipulation, now is bringing a new way in the treatment of severe male factor infertility to achieve acceptable rates of fertilization and pregnancy success. Objective: To evaluate the difference between ejaculated and surgically retrieved sperm on intra cytoplasmic sperm injection (ICSI) outcomes. Design: A Correlative study. Setting: Dwarozh- IVF center. Patients and Methods: One hundred twenty patients have been taken in Dwarozh and International infertility centers in Sulaimani, for male infertility indications with normoovulatory female partners of ages less than 38 years, from 1st of September 2010 to 1st of September 2013. We divided patients into two groups: patients with normal ejaculation;group one (G1), and patients with surgically retrieved spermatozoa (normal sperms); group two (G2), all underwent Intracytoplasmic Sperm Injection (ICSI) cycles. Main Outcome Measures: Fertilization, cleavage, biochemical and clinical pregnancy rates. Results: The fertilization rate has been found 72.6% in G1, while in G2 it is 60.4% and a significant statistical difference was observed in fertilization rates between the two groups with a P value of less than 0.01. Also it has been found that in G1, 90.2% of oocytes were cleaved, while in G2, 89% of oocytes were cleaved, no significant statistical difference was observed in cleavage rates between the two groups, with a P value of more than 0.05. It has been found that in G1, pregnancy was positive (+ve) biochemically in 65% of the cases, while pregnancy was positive (+ve) clinically in 45% of the cases. In G2 pregnancy were (+ve) biochemically in 28.3%, while pregnancy was (+ve) clinically in 15% of the cases, with significant statistical differences in both biochemical and clinical pregnancies between the two groups with a P value of less than 0.01. Conclusions: It has been found, with the use of ICSI cycles in the treatment of male factor infertility, there are significant differences in reproductive outcomes between cycles using ejaculated and surgically retrieved sperm. The data show that ICSI is aacceptable treatment option in oligospermic and azoospermic males. Acceptable rates of fertilization, cleavage and pregnancy success can be attainwith ICSI from patients with Azoospemia, reaching levels comparable with those of patients using ejaculated spermatozoa for ICSI.

Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men / 亚洲男科学杂志(英文版)

This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P < 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P < 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

Efeito do tipo de envase, flatpacks ou palhetas, sobre a criopreservação da fração rica do sêmen asinino / The effect of the type of package, FlatPacks or straws, on cryopreservation of the jackass sperm-rich fraction

Com o objetivo de se estudar o efeito do tipo de envase sobre a criopreservação do sêmen asinino, coletado de forma fracionada, utilizou-se o sêmen de cinco jumentos da raça Pêga, submetido a um protocolo de congelamento envolvendo um diluidor e duas formas de envase, flatpacks ou palhetas de 0,55mL, tendo como ponto final a avaliação dos parâmetros seminais in vitro após o descongelamento do sêmen. Os resultados de motilidade espermática para as amostras envasadas em flatpacks ou em palhetas foram de 24,67% e 35,34%, no momento do descongelamento; de 13,39% e 25,83%, 60 minutos após o descongelamento; e de 7,86% e 13,33%, aos 120 minutos após o descongelamento, respectivamente. Já os valores para o vigor espermático foram de 2,55 e 3,33, no momento do descongelamento; de 1,57 e 2,48, após 60 minutos do descongelamento; e de 1,11 e 1,67, após 120 minutos do descongelamento, na mesma ordem anterior. As características seminais diferiram entre os jumentos, evidenciando uma grande variação individual. O percentual de ejaculados aprovados (motilidade ≥30%, vigor ≥3) foi influenciado pelo reprodutor e pelo tipo de envasamento, estando o melhor resultado (P<0,05), de 86,21%, associado ao sêmen envasado em palhetas, quando comparado ao valor de 56,67%, obtido para o sêmen envasado em flatpacks.

The effect of the type of package on the cryopreservation of jackass semen was studied, using the sperm-rich fraction of five jackasses and a special cryopreservation semen protocol, related to one extender and two different packages, FlatPacks or straws. The characteristics of the in vitro semen were evaluated after thawing. The motility results were 24.67% and 35.34% after thawing; 13.39% and 25.83% 60 minutes after thawing and 7.86% and 13.33%, 120 minutes after thawing. The vigor results were 2.55 and 3.33 after thawing, 1.57 and 2.48 60 minutes after thawing and 1.11 and 1.67 120 minutes after thawing, for FlatPacks or straws packages, respectively. The jackasses used were different regarding seminal characteristics, with great individual variation. The approved post-thaw viability (motility ≥ 30%, vigor ≥ 3) was affected by jackass and type of package, with the best results (P<0.05) for semen packed in straws (86.21%), compared to semen packed in FlatPacks (56.67%).

Efeito da porção do ejaculado e do método de resfriamento sobre as características físicas do sêmen suíno / Effect of ejaculate portion and conservation method on the physical characteristics of boar semen

Duas porções do ejaculado suíno - primeiros 15mL da fração espermática rica (P1) e o restante do ejaculado (P2) - foram coletadas semanalmente de cinco varrões e submetidas a dois protocolos de resfriamento, diluição no diluidor MR-A® e conservação a 17°C (T1) ou no diluidor glicina-gema de ovo e conservação a 5°C (T2). As doses foram avaliadas no que se refere à motilidade, ao vigor e à morfologia espermáticas no sêmen a fresco e em diferentes tempos de estocagem. Todos os tratamentos mantiveram uma motilidade aceitável, superior a 50 por cento, nas primeiras 24 horas de armazenamento. O grupo P2T2 manteve uma motilidade similar (P>0,05) ao longo de todo o período de resfriamento (72 horas), sendo inclusive superior aos demais neste período, enquanto os outros tratamentos apresentaram uma redução da motilidade no decorrer do tempo de armazenamento. Com relação às características morfológicas do sêmen, não se observaram diferenças (P>0,05) quanto às porcentagens de espermatozoides normais entre as duas frações do ejaculado fresco. Ainda, todos os tratamentos mantiveram-se dentro dos limites aceitáveis, independentemente do tempo de armazenamento. A P1 parece ser mais adequada à produção de doses para o transporte em virtude de seu pequeno volume e alta concentração, enquanto o restante do ejaculado (P2) pode ser utilizado com eficiência dentro da própria granja.

Two portions of boar ejaculate - first 15mL of the sperm rich fraction (P1) and the rest of the ejaculate (P2) - were collected weekly from 5 mature boars and submitted to two cooling methods, extended in MR-A® and cooled at 17°C (T1) or extended in glycine-egg yolk and cooled at 5°C (T2). Spermatozoa motility, vigor, and morphological characteristics were evaluated immediately after collection and in different storage times. All treatments kept an acceptable motility, higher than 50 percent, in the first 24h of storage. The P2T2 maintained a similar motility (P>0.05) throughout the cooling storage (72h) and was superior in that period, while the other treatments presented a decrease in motility related to time. There was no difference between the two portions regarding the total number of normal spermatozoa in the fresh semen (P>0.05). All treatments showed morphological abnormalities within the acceptable thresholds, irrespectively of the storage time. Thus, due to low volume and high concentration, P1 seems to be more adequate for sperm dose transportation. Furthermore, this methodology will allow the development of new proposals concerning the transportation of swine semen, while the rest of the ejaculate could be used in farm routines to produce conventional liquid semen doses.

Ejaculate characteristics of rabbits infected with Trypanosoma congolense and changes caused after treatment with Diminazene aceturate (Diminaveto®) / Características eyaculatorias de conejos infectados con Trypanosoma congolense y cambios causados después del tratamiento con Diminazeno aceturato (Diminaveto®)

The effects of experimental Trypanosoma congolense infection on the ejaculate of rabbits and changes caused after treatment with Diminaveto® were investigated using 24 New Zealand White rabbits (bucks). The bucks were housed singly in standard rabbit cages and fed on specialized ration containing 10 percent Protein supplement, grains, legume, salt and fresh water ad libitum during the study. Data on ejaculate characteristics were collected from all the bucks in the first phase (i.e. before infection) and in the second phase (i.e. during infection, with 4.8x10(5) Trypanosoma congolense, intraperitoneally). Similar data were collected from 12 randomly selected bucks treated with 7.0mg/kg Diminaveto® following reconstitution during the third phase. Data collected were analysed using the Paired T- Test and Analysis of Variance. The infection led to significant (P< 0.05) reduction in spermatozoa motility, concentration and mass activity, with a significant (P< 0.05) increase in percentage of sperm cells with morphological abnormalities. Treatment with Diminaveto® led to improvement in all ejaculate parameters investigated. However, it was observed that the ejaculate did not attain the "before-infection" status following treatment with Diminaveto®. The study showed that infection with Trypanosoma congolense in rabbits caused significant reduction in ejaculate characteristics. Treatment with Diminaveto® however led to improvement in the ejaculate though at a rate slower than that at which the infection caused the reduction.

Fue estudiado en 24 conejos Nuevo zelandeses blancos, machos, los efectos de la infección experimental de Trypanosoma congolense sobre la eyaculación y los cambios producidos después del tratamiento con Diminaveto® . Los machos fueron colocados individualmente en jaulas de conejos estándar. Durante el estudio fueron alimentados con ración especializada, con 10 por ciento de suplemento de proteínas, granos, legumbres, sal y agua fresca ad libitum. Los datos sobre las características de la eyaculación se obtuvieron de todos los machos en la primera fase (es decir, antes de la infección) y en la segunda fase (es decir, durante la infección, con 4,8x10(5) Trypanosoma congolense, por vía intraperitoneal). Datos similares se obtuvieron de 12 machos al seleccionados al azar, tratados con Diminaveto® 7,0mg/kg después de la reconstitución durante la tercera fase. Los datos fueron analizados mediante t de student y análisis de varianza. La infección fue significativa (P <0,05) habiendo reducción de la motilidad de los espermatozoides, la concentración y actividad de masas, con un efecto significativo (P <0,05) aumento en el porcentaje de espermatozoides con anomalías morfológicas. El tratamiento con Diminaveto® condujo a una mejoría en todos los parámetros investigados del eyaculado. Sin embargo, se observó que el eyaculado no alcanzó el "antes de la infección" tras el tratamiento con Diminaveto®. Además, el estudio mostró que la infección con Trypanosoma congolense en conejos causó una reducción significativa en las características del eyaculado. Sin embargo, el tratamiento con Diminaveto ® condujo a una mejoría en la eyaculación aunque a un ritmo más lento que en la infección causada por la reducción.