Individual Progressive Motility Research Articles

Uso da técnica de swim-up para a remoção do Vírus da Artrite Encefalite Caprina do sêmen de reprodutores infectados

Neste estudo, 67 ejaculados foram avaliados, antes e depois da técnica de swim-up,em relação à qualidade seminal e à presença do CAEV. Das 67 amostras testadas por PCRn, antes do swim-up,47 (70,15%) foram positivas para o DNA pró-viral. No entanto, quatro amostras adicionais foram positivas ao RT-nested PCR após o swim-up, o que permite dizer que, pelo menos, 76,12% (51/67) delas estavam infectadas antes da lavagem. Todavia, em 23,88% (16/67) das amostras não foi detectada a presença do CAEV. Após a aplicação da técnica de swim-up, constatou-se, pela PCRn e RT-nested PCR, que houve uma redução significativa (χ²= 9,078; p<0,001) da presença do CAEV nas amostras seminais, pois 28 de 51 amostras positivas resultaram livres do vírus (54,90%), tanto para DNA pró-viral quanto para o vírus livre. Em relação à motilidade individual progressiva (MIP) e vigor espermático obtidos antes e depois da técnica de swim-up,observou-se uma diminuição significativa em suas médias, sendo o MIP de 86,42% para 71,49%, já o vigor espermático de 4,16 para 3,93. Conclui-se que a eliminação do CAEV no sêmen é de caráter intermitente, e que a associação da PCRn e RT-nested PCR é uma opção segura para a certificação sanitária individual das amostras seminais quanto à presença ou ausência do CAEV. Finalmente, a técnica de swim-uppromove uma redução na infectividade de amostras de sêmen contaminadas, e, além disso, é possível promover a recuperação de espermatozoides viáveis.

Egg yolk powder an alternative to fresh-egg yolk for buffalo semen cryopreservation

The present work was carried out to evaluate the effect of commercial egg yolk (EY) powder as an alternative to fresh EY on buffalo semen freezability. Ejaculates (12) from 3 bulls (4 from each bull) were collected as per standard procedure. After initial selection of ejaculates on the basis of mass activity (+3 and above) and individual progressive motility (70% and above), ejaculates were divided into 3 parts and diluted with tris dilutor, G1, containing 5% EY powder; G2, containing 10% EY powder, and G3, containing fresh EY (20%). Ejaculates were processed and cryopreserved using automated biological cell freezer followed by examination of frozen samples after 24 h. Semen quality variables (individual progressive motility, viability using propidium iodide, acrosome integrity using FITC-PSA and membrane integrity by HOST) were examined at fresh and post-thaw stage. At fresh stage semen quality variables did not vary among 3 groups.

Seminal PDC-109 protein vis-à-vis cholesterol content and freezability of buffalo Spermatozoa

Advancements in reproductive technologies have shown seminal plasma (SP) as a nutritive-protective medium for spermatozoa metabolism, function and transport. At the same time quality variables and thus freezability of spermatozoa are influenced by SP proteins originating from male reproductive tract. One such protein, viz. PDC-109 is reported to influence freezability of spermatozoa in cattle. Thus the present investigation was designed to evaluate effect of seminal PDC-109 protein concentration on post-thaw cholesterol content and semen quality variables (SQP) as an indicator of membrane integrity and freezability, respectively of buffalo spermatozoa. Ejaculates (n=42) selected on the basis of mass activity and individual motility were divided into three parts, first part for SP proteins isolation, second for cholesterol estimation and third part was cryo-preserved to evaluate freezability based on post-thaw SQP, viz. individual progressive motility, viability and acrosome integrity of spermatozoa. A total of 28 (66.7%) and 14 (33.3%) ejaculates from four bulls were found as freezable or non-freezable, respectively. Though total seminal plasma protein (TSPP) concentration was found similar in freezable and non-freezable ejaculates, the heparin binding proteins (HBP) content in non-freezable semen was greater (P<0.01) than freezable ejaculates. There was a similar trend for the PDC-109 protein content in respective ejaculates. Cholesterol content of spermatozoa and SQP were greater (P<0.05 and 0.01, respectively) in freezable as compared to non-freezable ejaculates of each bull at post-thaw stage. This study showed that concentrations of HBP and PDC-109 in non-freezable semen might be responsible for greater cryo-damage reflecting in poor freezability of buffalo spermatozoa.

VIABILIDAD ESPERMÁTICA E INTEGRIDAD DEL ACROSOMA EN SEMEN CONGELADO DE TOROS NACIONALES.

The deterioration of the sperm membrane and acrosome integrity as a method for predicting fertility in bulls was evaluated. Four bulls (2 Holstein and 2 Brown Swiss) from the National Bank of Semen, Lima-Peru were used. Acrosome integrity, sperm membrane integrity, motility, live sperm cells, volume, and sperm concentration during cooling, freezing and thawing was evaluated in 10 ejaculates per sire. In fresh semen, volume was 4.33 ml; sperm concentration was 922.5 x 106/ml and 78.5% of live cells. The individual progressive motility in diluted semen was 82.7 to 86.0% with significant difference between bulls (p&lt;0.05). Acrosome integrity in chilled semen varied from 59.3 to 69.2% and without statistical difference between bulls. Percentage of sperm with intact acrosome decreased from 65.2 in chilled semen to 48.6% in frozen/thawed semen, without statistical difference between bulls. None significant correlation was found between motility and sperm membrane integrity or acrosome integrity.

Hypo-osmotic test in cat spermatozoa

The hypo-osmotic (HOS) test has been used in other species as an indicator of the fertilising capacity of spermatozoa. The aims of this study were to assess the response of domestic cat spermatozoa to the hypo-osmotic test, to determine the type of solution, concentration and time of incubation needed to obtain a maximum percentage of swelling, to correlate the selected combination with the percentages of progressive motility and to evaluate whether dilution of the ejaculate alters the results. Incubation for 30 and 45 min in solutions of fructose and of citrate of 50 and 100 mOsmol kg⁻¹ was evaluated. The highest percentage of swelling was obtained using the 50 mOsmol kg⁻¹ solution, and no significant differences were observed between the times of exposure to the solutions. A positive correlation was observed between the percentage of individual progressive motility and the percentage of sperm swelling in a 50 mOsmol kg⁻¹ fructose solution, with no significant differences being observed between raw and diluted semen samples. The results of this study suggest that the HOS test could be useful for evaluating membrane function in domestic cat spermatozoa, both in raw semen and in samples diluted in the EZ Mixin® commercial extender, and thus could be incorporated into routine semen evaluation protocols.

Impact of selection for residual feed intake on breeding soundness and reproductive performance of bulls on pasture-based multisire mating1

There is concern in the beef industry that selecting bulls for feed efficiency based on residual feed intake (RFI) may have a negative impact on bull reproductive performance and fertility. Here we investigated the impact of selection of bulls for low RFI on breeding soundness evaluation (BSE), reproductive performance, and fertility of bulls under natural service in multisire mating groups on pasture. Of the 412 RFI-tested bulls available, 98 (23.8%) were culled for performance, type, temperament, or other reasons, and 88 (21.4%) were culled for failing BSE, for an overall cull rate of 45.1%. From among the 314 bulls subjected to BSE, 32 (10.2%), 20 (6.4%), and 36 (11.4%) were culled for poor feet and legs, scrotal circumference, and semen quality, respectively. The BSE traits were not different (P > 0.10) between bulls categorized as either inefficient (+RFI) or efficient (-RFI), but the proportion of bulls that failed to meet the 60% minimum sperm motility requirement tended (P = 0.07) to be greater in the -RFI group than in the +RFI group (10.2% vs. 4.4%, respectively). In a subpopulation of 115 bulls, individual progressive sperm motility was greater (P < 0.05) in +RFI (85%) than -RFI (80%) bulls. A multisire natural mating experiment was conducted during 2 consecutive breeding seasons (2006 to 2007 and 2007 to 2008) using 18 +RFI and 18 -RFI bulls. The overall calving rate (calves born/cows exposed) was 72.9%. Mean number of progeny per sire was significantly greater (P < 0.01) in -RFI bulls (18.3) than in +RFI bulls (11.8). Selection for feed efficiency based on RFI appears to have no detrimental impact on reproductive performance and fertility in beef bulls bred in multisire groups on pasture. However, the decreased sperm motility and the greater number of progeny per sire associated with -RFI status need further investigation.

EFECTO DE DOS DILUTORES SOBRE LA MOTILIDAD E INTEGRIDAD DE LA MEMBRANA ESPERMÁTICA EN SEMEN CONGELADO DE OVINOS.

The objective of the study was to evaluate the effects of two semen extenders: Tris- Fructose-egg yolk (Tris) and Citrate-Glucose-egg yolk (citrate) on motility and sperm membrane integrity (HOST) in ovine frozen semen in pellets. The study was carried out at the Semen Bank of the Agrarian University La Molina, in Lima, Peru, using 4 rams (2 Assaf and 2 Blackbelly) of 3.5 to 4 years old. A covariance analysis was used to evaluate the effect of the treatment and breed on Individual Progressive Motility (IPM), and randomized block design to evaluate the effect of extenders on sperm membrane integrity. Semen was frozen of dry ice and thawing was done in test tubes at 38 °C. In the Assaf breed, IPM of thawed semen was 63.77 and 61.11% when using Tris and citrate respectively, showing statistical difference (p<0.05). In the Blackbelly breed the IPM was 62.33 and 61.33%, when used Tris and citrate respectively, and without significant difference. In the Assaf the HOST values of thawed semen were 43.56 and 40.38%, while in Blackbelly were 40.19 and 38.16% respectively, both without significant differences. It is concluded that Tris showed better effect on individual motility but both of them had similar effect on sperm membrane integrity.

Viabilidade in vitro de células espermáticas ovinas submetidas a diferentes diluentes e a refrigeração

Avaliou-se a viabilidade in vitro de células espermáticas após a adição de três diluentes comerciais, que foram comparados com o diluente tradicional Tris-gema, utilizados no processo de refrigeração do sêmen ovino, em até 48h de armazenamento. Foram utilizados nove ejaculados diários, obtidos de três reprodutores Dorper, com vagina artificial, em três repetições com intervalo de três dias. O sêmen foi mantido a 30°C, e foram avaliadas suas características macro e microscópicas. Após formação do pool, repetiram-se as avaliações acrescidas da concentração espermática e da integridade do DNA e do acrossoma. Dividiu-se o pool em cinco tratamento, cada um constituído de uma parte de sêmen para três partes dos respectivos diluentes: Equimix (DI), Laiciphos Green Ovine (DII), FR 4 (DIII), Equimix-Gema- Equimix com gema de ovo (DIV) e Tris-Gema (DV). O material obtido em cada tratamento foi subdividido em quadruplicata, refrigerado e mantido a 4°C até as avaliações da motilidade individual progressiva (MIP), do vigor e da integridade do DNA e do acrossoma, correspondendo a zero, 12, 24, 36 e 48h. Nas avaliações do sêmen, com o DI ocorreu a maior queda de MIP às 12h em relação aos demais grupos (P&lt;0,05). Às 24h, os tratamentos DII, DIV e DV apresentaram a melhor MIP (P&lt;0,05), que não divergiram (P&gt;0,05) entre si; às 48h, o DII e o DV foram melhores (P&lt;0,05) que os demais. Com relação ao vigor, os tratamentos DII e DV apresentaram valores mais elevados (P&lt;0,05) em relação ao DI e DIII, a partir das 12h, e ao DIV, a partir das 24h (P&lt;0,05). Concluiu-se que o diluente Laiciphos Green Ovine, da mesma forma que o Tris-gema, pode ser utilizado na conservação do sêmen a 4°C por 48h, enquanto o Equimix, acrescido de 20% de gema de ovo, pode ser seja utilizado no armazenamento do sêmen (4°C) por até 24h.

Effect of Supplementation with Agro‐industrial By‐products and Khat (Catha edulis) Leftovers on Testicular Growth and Sperm Production in Ogaden Bucks

The aim of this study was to characterize growth and sperm production parameters in Ogaden bucks fed a basal diet of hay and supplemented with agro-industrial by-products and Khat leftovers in Ethiopia. Thirty-five bucks with a mean (+/-SD) initial live body weight (BW) of 15.5 +/- 1.5 kg were randomly assigned to one of four dietary treatments for a period of 13 weeks. Treatments consisted of native hay fed ad libitum (control; C), native hay supplemented with a 1% of BW agro-industrial by-products (treatment 1; T1), native hay supplemented with Khat (Catha edulis) leftovers at a rate of 1% of BW (treatment 2; T2) and Khat leftovers fed ad libitum (treatment 3; T3). Bucks fed on T1-T3 had higher BW, body condition score, scrotal circumference (SC), testicular width and testicular length, compared to controls (P < 0.05). Also, bucks in T1-T3 had higher sperm progressive motility, sperm concentration per ml and total number of spermatozoa per ejaculate compared to controls (P < 0.05). Between treatments, bucks in T3 recorded the highest BW (17.2 +/- 0.16) and testicular size (21.1 +/- 0.17 cm). Both testicular and epididymal weight and dimensions were significantly affected (P < 0.05) by supplementation compared to controls. Testicular size was positively correlated to live BW (r = 0.53, P < 0.001). SC was positively correlated with ejaculate volume (r = 0.37, P < 0.001), sperm mass activity (r = 0.65, P < 0.001) and individual sperm progressive motility (r = 0.40; P < 0.001). Supplementation with Khat leftovers induced the highest improvement in live BW, testicular size, semen production and sperm motility in Ogaden bucks and can possibly be considered as a feed supplement to enhance goat production under smallholder livestock farming system in Ethiopia.

The onset of puberty in North American bison ( Bison bison) bulls

Prepubertal testicular development was evaluated in young North American bison ( Bison bison) bulls over a 12 months period, by following various parameters of semen quality intended to measure sexual maturity level. The study goals were to characterize bison male reproductive development and to define the age and onset of puberty. Semen was collected by electro-ejaculation from 12 bulls once a month for a period of 12 months, from 13 through 24 months of age. Volume, density, gross motility, individual motility, morphology, live/dead ratio, and concentration were used to judge the quality of the ejaculates. Abnormal sperm were classified into categories according to descriptions used for bovine evaluations and guidelines created by the Western Canadian Association of Bovine Practitioners were used for determining satisfactory semen quality. Minimums of 60% progressive motility and 70% normal sperm cells identified samples as satisfactory. Puberty was defined as the point in testicular development where an ejaculate contains a minimum of 50 × 10 6 sperm cells, showing at least 10% progressive motility. Fecal samples collected from bulls prior to electro-ejaculation for testosterone determination by a commercial testosterone specific antibody radioimmunoassay showed no significant changes in mean fecal testosterone concentrations over time. Changes in mean body weights over the study period were calculated, the average body weight at 13 and 24 months of age were 263 ± 18.6 and 475 ± 52.2 kg, respectively. Under these experimental conditions, age was determined to be the most significant factor in determining the onset of puberty. On average, bulls reached puberty at 16.5 ± 2.5 months. There was a significant increase in the number of normal sperm with increasing bull age as well as significant decreases in proximal cytoplasmic droplets and head defects, but no significant changes in midpiece and principal piece defects. There were significant increases with age in individual progressive motility, gross motility, concentration, density, and volume but not in the percent of live sperm. Bison bulls used for breeding should be capable of passing a breeding soundness evaluation by 24 months of age. The commercial bison industry semen tests, and possibly rejects, many developmentally incomplete bulls between 19 and 21 months of age. We suggest that these may actually be developing normally and should not be hastily discarded; it may be worth testing again at 24 months.

Comparison of four antibiotics for inactivating leptospires in bull semen diluted in egg yolk extender and experimentally inoculated with Leptospira santarosai serovar guaricura

Inactivation of leptospires in pools of semen from three Holstein Friesian bulls, collected in an artificial vagina, was investigated. Spermatic concentration was adjusted in egg yolk citrate extender, submitted to the following treatments: A (control; without antibiotics); B (penicillin, 1,000 UI/mL - streptomycin, 1,000 µg/mL); C (amoxicillin, 1,000 µg/mL); D (ceptiofur sodium, 1,000 µg/mL); E (amoxicillin 1,000 µg/mL - ceptiofur sodium 1,000 µg/mL). Leptospires (2.0 x 106 leptospires/mL) were added into the diluted semen. Recovery of leptospires was obtained in modified EMJH semi-solid medium with and without antibiotics. The antibiotics in the concentrations used did not affect means of percentage of progressive motility and individual progressive motility of spermatozoids. Penicillin-streptomycin presented the best results in leptospire inactivation (97.1%). Amoxicillin, ceptiofur sodium and their combination at the concentrations studied presented poor results: 59.29%; 32.5% and 60.36% of inactivation, being less effective in leptospire inactivation than penicillin-streptomycin.

Observations on semen quality of dogs in the tropics.

Semen was obtained from eight mongrels and 17 purebred dogs kept in a tropical climate. The mean spermatozoal output per ejaculate was 303.6 +/- 138.2 and 602.9 +/- 466.2 million, respectively. Values for individual progressive motility, live and abnormal spermatozoa were comparable with those obtained in temperate regions. The type and frequency of spermatozoal abnormalities are illustrated.