Sperm Mass Motility Research Articles

Use of grayscale histogram to assess testicular changes in ram caused by heat stress

Because it detects discrete changes in tissue density, ultrasound allows an early diagnosis of physiologic changes, especially when dealing with subclinical changes. aimed to evaluate the use of quantitative analysis of the gray scale of ultrasound images of ovine testicles as a method for early diagnosis of testicular changes caused by heat stress. Testicles from ten ovine were insulated and then evaluated regarding echogenicity, echotexture, anatomical measurements, and seminal characteristics. Echogenicity was the first variable to show changes during the insulation period, as well as the first one to show evidence of regeneration of those changes. There was a correlation (p < 0.05) between echogenicity and sperm vigor, motility, and mass sperm motility, as well as a correlation (p < 0.05) between echotexture and motility. Echogenicity, motility, vigor, and mass sperm motility decreased (p < 0.05) within four days of testicular insulation, and there was no change (p > 0.05) in echotexture. Testicular measurements were only altered (p < 0.05) after insulation; however, the structural integrity of the seminiferous tubules was not altered. Echogenicity is an efficient ultrasound parameter for the early diagnosis of a testicular degenerative process as well as for the early diagnosis of its regeneration.

Incorporation of fresh leaves of wormwood (Artemisia herba alba) and/or rosemary (Rosmarinus officinalis) in the diet of rams: Effect on testicular function, sexual behavior, and blood parameters.

This study aimed to investigate the effect of wormwood and rosemary supplementation on some reproductive traits of Barbarine rams. The experiment lasted 2 months. Twenty-four adult rams were divided into four groups (n=6) balanced for the weight (53.3 ± 1.2 kg body weight [BW] ± SD). All rams received 1200 g of straw and 600 g of barley. Control rams (C) without aromatic medicinal plant (AMP), while experimental rams received 20 g of fresh rosemary leaves (R), 20 g of fresh wormwood leaves (A), and 10 g of fresh rosemary leaves plus 10 g of fresh wormwood leaves (RA). The results revealed that the live weight of all rams increased (p < .05) in the RA group compared to the C, A, and R groups. Scrotal circumference increased in the R rams when compared to the controls rams (p < .05). For sperm parameters we showed that the A rams had higher sperm concentrations (p < .05). But, the sperm volume decreased in the R rams (p > .05). However, when the rams received rosemary plus wormwood, their sperm volume increased (p > .05). The sperm mass motility was higher for the A, R and AR rams in comparison to the C rams (p=.05). On the other hand, biochemical analysis of the seminal fluid showed no effect of diets on calcium and total proteins concentration. But the measurement of glucose and seminal insulin showed a decrease (p < .05) in these two biochemical markers in group A rams and a decrease (p < .05) in insulin without modification of the glucose concentration in R rams. Blood glucose and insulin decreased in the animals on AMP diet compared to the other groups (p < .05) while aspartate aminotransferase (AST) increased (p < .05). Rosemary leaves (R and RA groups) increased (p < .05) plasma cortisol compared to the other groups. It can be concluded that the addition of Rosmarinus officinalis and/or Artemisia herba alba in ram diet can have a positive effect on the reproductive function by increasing the concentration and motility of sperm, plasma testosterone, and sexual behavior.

Effect of Dietary Ginger Rhizome Powder Addition on Reproductive Performance of Rabbit Bucks

This study was conducted to evaluate the effect of the addition of ginger rhizome powder (GRP) to the rabbit bucks ration on semen quality and semen plasma traits. A total of 28 male New Zealand rabbits (7- month- old), divided into four groups; the first treatment served as a control (Con; 0% ginger); the second, third, and fourth treatments were supplemented with 0.5%, 1.0%, and 1.5% ginger powder in the diet, respectively. The results revealed no change in ejaculate volume, a significant increase in the mass sperm motility, sperm concentration, total sperm output, total motile sperm, live sperm, and Total functional sperm fraction. Dietary ginger reduced the reaction time, semen pH, and the percentage of abnormal sperm. Bucks that received 1.0% or 1.5% dietary ginger had a higher seminal plasma total proteins, albumin, globulin concentrations, alkaline phosphatase and acid phosphatase activities. Conversely, seminal plasma aspartate aminotransferase and alanine aminotransferase activities were significantly decreased compared to the control group. Practically, it can be concluded that ginger is a suitable supplement for improving semen quality and semen plasma in adult male rabbits.

Mass Sperm Motility Is Correlated to Sperm Motility as Measured by Computer-Aided Sperm Analysis (CASA) Technology in Farmed Ostriches.

Simple SummarySemen quality in ostriches is still being studied to optimize the reproductive performance of these birds in a commercial farming environment. Various methods such as mass sperm motility scoring and computer-aided sperm analysis (CASA) technology have been employed successfully to evaluate semen quality characteristics such as sperm motility. The relationship between these methods in assessing sperm motility in ostriches is unknown. This study evaluated the relationship between the CASA and the mass sperm motility scoring method when analyzing ostrich sperm motility. Positive significant correlations between these methods for sperm motility traits were recorded, suggesting that farmers could utilize the affordable mass sperm motility scoring to screen ostrich males for sperm quality.Semen analyses have gained momentum in various livestock industries. However, in farmed ostriches, semen analysis is still in its experimental stage, and males are not screened for sperm quality before breeding. This study investigated the correlations between computer-aided sperm analysis (CASA) technology and the traditional, yet affordable, mass sperm motility score. Semen was collected from nine South African Black ostrich males (mean age ± SD: 5.25 ± 1.21 years), using the dummy female method for 5 consecutive days monthly, for 8 months. Mass sperm motility scores were recorded on a scale of 1–5 (1: little to no sperm movement; 5: rapid sperm movement). The CASA traits recorded were: total motility (MOT), progressive motility (PMOT), curve–linear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), amplitude of lateral head displacement (ALH), linearity (LIN), straightness (STR), wobble (WOB), and beat-cross frequency (BCF). The results revealed positive correlations between mass sperm motility and PMOT, MOT, VCL, and VAP ranging from 0.34 to 0.59 (p < 0.0001). In contrast, negative correlations were recorded between mass sperm motility and LIN, STR, and BCF, with correlations ranging from −0.20 to −0.39 (p < 0.0001). VSL, ALH, and WOB were not correlated to mass sperm motility (p > 0.05). Ostrich farmers may thus be able to evaluate sperm motility reliably and potentially select breeding males by using the affordable mass sperm motility scoring method. Determining the correlation between these methods and fertility after artificial insemination or natural mating is however needed.

Using transrectal ultrasound-guided massage of the accessory sex glands for buck semen collection yields semen with greater cryoresistance than electroejaculation alone during the breeding season

The cryoresistance of bucks’ semen collected by transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) and electroejaculation (EE) was compared during the breeding season (Study 1) or the late non-breeding season (Study 2). Semen was collected from 10 Gabon bucks with both methods in each season and then frozen according to a standardized protocol. The time required for ejaculation, the number of electrical pulses applied, the sum of the electrical pulses applied∗voltage and the total number of vocalizations emitted during the collection by each animal were recorded. In Study 1, TUMASG required a longer time (P = 0.0006) but fewer electrical pulses and a lower sum of pulses∗voltage than EE (P < 0.0001 for both comparisons). Bucks vocalized fewer times during TUMASG than during EE (P < 0001). Semen collected with TUMASG had greater sperm concentration, sperm mass motility, total number of motile sperm and of sperm with progressive motility, and tended to have greater total number of sperm with functional membrane than semen collected with EE (P = 0.02; P = 0.003; P = 0.02; P = 0.02; P = 0.06, respectively). After the freezing-thawing process, sperm collected with TUMASG had a greater quality of the motility, a total number of motile sperm and sperm with progressive motility, a greater percentage of motile sperm and sperm with functional membrane, and tended to have a greater total number of sperm with normal morphology than semen collected with EE (P = 0.04; P = 0.04; P = 0.03; P = 0.02; P = 0.04; P = 0.06, respectively). In Study 1 sperm collected with TUMASG had greater cryoresistance for almost all the variables considered than when it was collected with EE. In Study 2, the number of electrical pulses and the sum of pulses∗voltage were greater with EE than with TUMASG (P < 0001; P = 0.0002, respectively). There were no differences in any sperm variable in fresh or thawed samples collected with TUMASG or EE. However, the other seminal characteristics evaluated did not differ according to the method, and there were no differences in the cryoresistance of any variable. Overall, TUMASG affected animal welfare less than EE and was suitable for collecting semen of good quality with high cryoresistance during the breeding season in buck.

Proteomic Changes Associated With Sperm Fertilizing Ability in Meat-Type Roosters.

The molecular basis of male fertility remains unclear, especially in chickens, where decades of genetic selection increased male fertility variability as a side effect. As transcription and translation are highly limited in sperm, proteins are key molecules defining their functionality, making proteomic approaches one of the most adequate methods to investigate sperm capacity. In this context, it is interesting to combine complementary proteomic approaches to maximize the identification of proteins related to sperm-fertilizing ability. In the present study, we aimed at identifying proteins related to fertility in meat-type roosters, showing fertility variability. Fertile roosters (fertility rates higher than 70% after artificial insemination) differed from subfertile roosters (fertility rates lower than 40%) in their sperm mass motility. Fertile and subfertile sperm protein contents were compared using two complementary label-free quantitative proteomic methods: Intact Cell MALDI-TOF-Mass Spectrometry and GeLC-MS/MS. Combining the two strategies, 57 proteins were identified as differentially abundant. Most of them were described for the first time as differentially abundant according to fertility in this species. These proteins were involved in various molecular pathways including flagellum integrity and movement, mitochondrial functions, sperm maturation, and storage in female tract as well as oocyte–sperm interaction. Collectively, our data improved our understanding of chicken sperm biology by revealing new actors involved in the complexity of male fertility that depends on multiple cell functions to reach optimal rates. This explains the inability of reductionist in vitro fertility testing in predicting male fertility and suggests that the use of a combination of markers is a promising approach.

Seasonality of the stress response in goat bucks when there is use of electroejaculation for semen collection

Electroejaculation (EE) is frequently used to collect semen, but this procedure is both stressful and painful. In seasonal-breeding species the sensitivity to stressors might vary with season of the year when semen collection occurs, therefore, the present study was conducted to compare the responses when imposing the EE procedure, stress response to EE and fresh semen characteristics in goat bucks throughout the year. Semen was collected using EE from goat bucks once in autumn (mid-breeding season), winter (transition to the non-breeding season), spring (non-breeding season), and summer (beginning of the breeding season). The number of pulses applied during ejaculation was larger in the autumn and spring than winter and summer months. The number of vocalizations/pulse while bucks were ejaculating was greater in the summer than winter and spring months. The increase in heart rate was greater in spring than summer months. Cortisol concentration was least in the autumn, and concentration of creatine kinase (CK) was greatest in summer. Sperm mass motility was greater in autumn than spring months. Spermatozoa concentration, total number of spermatozoa ejaculated, motile spermatozoa ejaculated, and progressively motile spermatozoa were greater in the autumn and spring than in the winter and summer months. The bucks were less stressed by imposing EE procedures in the autumn, and buck responses were greater in the spring-summer period. Bucks had a greater incremental increase in serum CK and number of vocalizations in the summer months, indicating there was a greater muscular damage and quite likely pain associated with lesser semen quality.

Effect of season and genotype on values for bull semen variables under tropical conditions

To assess the effects of season and genotype on fresh semen quality for freezing and subsequent use for AI, 32 healthy, mature (>4 to <6 years old) and habituated bulls (Bos indicus n = 14, Bos taurus n = 5, Crossbred taurus x indicus n = 6, and tropically adapted Bos taurus composites n = 7) were evaluated at a Venezuelan AI center for 12 months in which four distinct seasons (Hot-dry, Transition, Hot-humid, Cool-rainy) occur. Ejaculates were collected weekly from Bos taurus (n = 260), Bos indicus (n = 669), tropically adapted Bos taurus composites (n = 389), and crossbred Bos taurus x Bos indicus (n = 340) bulls. Routine AI Center assessments were conducted i.e., ejaculate volume (EV), sperm mass-motility (MM), total sperm number/ejaculate (TSE), sperm concentration/mL (SC), pre-freezing (PREF), and post-freezing minimum criteria rate for AI use (POSTF). Genotype affected EV (P < 0.0001), TSE (P < 0.0001), and SC (P < 0.0001) but not MM (P>0.05). Season affected EV (P < 0.001), TSE (P < 0.0001), SC (P < 0.01), and MM (P < 0.05). There were genotype x season interactions for EV, MM, TSE, and SC. The PREF averaged 74.0% during the study, although was less (P < 0.0001) during the hot-humid season than the other seasons. Even though, percent ejaculates considered unsuitable for freezing differed (P < 0.03) among the Hot-dry (20.2%), transition (30.9%), Hot-humid (32.4%), and Cool-rainy (24%) seasons. For POSTF, there were no seasonal differences (P>0.05). It is concluded that in tropical regions, season and genotype can affect bull semen variables, particularly those which affect the success of semen freezing and AI.

A Comparative Study of Semen Characteristics and Testosterone Concentration in Ram Breeds of Saudi Arabia

The goal of this study was to evaluate and compare the semen characteristics, and testosterone concentration for three breeds raised under hot, desert conditions in Saudi Arabia. A total of 150 rams (Najdi, Naemi, and Harri) were used, aged 12, 18, 24, 30 and 36 months (ten rams from each breed in each age group). Body weight and testicular parameters were measured. In addition, testosterone concentration (TC), ejaculate volume (EV), sperm concentration (SC), total sperm output per ejaculate (TSO), live sperm percentage (LSP), sperm progressive motility (SPM) and sperm mass motility (SMM). The results show that TC and EV differed significantly (P less than 0.01) between breeds. At 36 months, semen characteristics (TSO, SMM and SPM) differed significantly among breeds. Naemi rams were superior to Najdi and Harri in terms of the mean live sperm percentage (LSP) for all age groups. These results may assist in ram selection for breeding programs based on high reproductive performance.

Age-Dependent Changes in Fresh Semen Quality of Swamp Buffalo (Bubalus bubalis)

The purpose of this study was to evaluate the fresh semen quality of swamp buffalo raised in Lembang Artificial Insemination Center with respect to different age. The semen was collected from one swamp buffalo bull at three different stages (5, 6, and 7 years old). The evaluation of semen quality was conducted directly upon collection. Semen color, consistency, and mass sperm motility were analyzed descriptively, while semen volume, pH, sperm concentration and individual sperm motility were analyzed using analysis of variance followed by least significant difference test. The results showed that the dominant physical characteristics of swamp buffalo’s semen were milky white color with moderate consistency and fast mass sperm motility (++). There was a highly significant effect (P<0.01) of age on semen volume, sperm concentration, and individual sperm motility, whereas semen pH was not affected (P>0.05) by age. Swamp buffalo had higher (P<0.01) semen volume and sperm concentration at 7 years old, but with lower (P<0.01) individual sperm motility as compared to 5 and 6 years old. In brief, this study reveals that semen production of swamp buffalo is dependent of age with 7 years old become the most superior age.

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance.

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1hr with 0, 20 and 100µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p<.05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8±1.5 and 69.1±1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p<.05), while the percentage of spermatozoa with high ΔΨm increased (p<.05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4±5.8 and 83.7.1±1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p<.05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p<.05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0±11.4). In conclusion, the treatment with 100µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.

Can mineral supplementation modify the characteristics of young buffalo ejaculate?

This study aimed to determine the influence of mineral supplementation on the concentration of macroelements and microelements in the seminal plasma and the semen characteristics of young buffaloes. To this end, 60 animals were kept under rotational grazing and were randomly divided into two groups: G-Bov (conventional mineral supplement recommended for bovine cattle; n = 30) and G-Buf (mineral supplement recommended for buffaloes; n = 30). Mineral supplement consumption was calculated from the difference between the amount of supplement offered and the leftover bunk, calculated every 28 days. Eight ejaculates from each animal were collected. The means of the response variables were analyzed using the Kruskal-Wallis test, with a significance level of 5%, and Pearson’s correlation was analyzed between the concentration of minerals in seminal plasma and fresh semen characteristics. Animals in the G-Buf group had lower mineral supplement consumption (G-Bov = 83.18 vs. G-Buf = 77.14 g AU-1 day-1, P &lt; 0.05) and had higher ejaculate volume, sperm motility, sperm vigor, and sperm mass motility than did the animals in the G-Bov group. This research presented the concentration of macroelements and microelements in the seminal plasma of buffaloes receiving different mineral supplements. In addition, the study concluded that the physical and morphological characteristics of the semen of young buffaloes are influenced by the formulation of the mineral supplement, which resulted in better sperm quality in the animals receiving a species-specific supplement. Among the minerals present in buffalo seminal plasma, phosphorus is the element that shows the highest positive correlation with semen characteristics.

Serum and seminal plasma IGF-1 associations with semen variables and effect of IGF-1 supplementation on semen freezing capacity in buffalo bulls

The objective of the study was to establish correlation of seminal and serum IGF-1 with seminal attributes, estimate antioxidant potential of IGF-1 by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays and to study the effect of IGF-1 supplementation on semen cryopreservation. For this study, buffalo bulls were divided into sub-fertile (n = 2) and normal (n = 5) on the basis of sperm mass motility and individual motility. The serum IGF-1 concentration of normal bulls was greater than in sub-fertile bulls, but there was no difference in the seminal IGF-1 concentration among the groups. The values from correlation analyses indicated that serum IGF-1 concentration is positively correlated with semen mass motility and sperm concentration. In the second experiment, IGF-1 did not have antioxidant activities when assessed with DPPH and FRAP assays. In the third experiment, the ejaculates of normal and sub-fertile bulls were cryopreserved using semen extender in which there was IGF-1 supplementation at 0 (control), 50, 100, 150, 200, 250, 350 and 450 ng/mL of extender. Supplementation of IGF-1 at 250 ng/ml resulted in improved sperm motility, longevity and membrane intactness as compared to control after cryopreservation of semen from normal buffalo bulls, but not sub-fertile bulls. In summary, serum IGF-1 concentration was correlated with sperm mass motility and concentration in buffalo bulls and supplementation of IGF-1 protected sperm during the cryopreservation process but effects were not due to direct antioxidant activity.

Response of Seminal Traits of Horro Rams to Different Levels of Supplementation under Grazing of Natural Pasture

The fertility of the male animals is often reported to have a greater influence on stock performance than does the fertility of females, which further greatly affected by nutritional factors. An experiment was conducted to evaluate the response of seminal traits of Horo rams to nutritional supplementation of Noug seed cake and Wheat bran mixture under grazing of natural pasture in western Ethiopian, where 18 uniform intact yearling Horro rams with average initial body weight of 18.55 ± 0.99kg (mean ± SD) were involved in the experiment. The treatments consisted grazing natural pasture alone (T1;Control), or T1 supplemented with concentrate mixture at 0.9% of body weight (T2) or T1 supplemented with concentrate mixture at 1.5% of Body weight (T3). The experimental design used was complete randomize where six rams per treatment were randomly allocated to three dietary treatments. Supplementation significantly (P&lt;0.05- P&lt;0.00) improved most seminal traits. With the exception of semen volume (SV), sperm concentration per ml (SCO), sperm mass motility(SMM), and sperm progressive individual motility(SPIM), there was no significant (P&gt;0.05) difference between T2 and T3, with respect to parameters considered. In general, since supplementation at 1.5% of body weight (T3) significantly improved SV, SCO, SMM and SIM, compared to T2, T3 could be recommended as appropriate feeding strategy to improve the reproductive performance and exploit the genetic potential of the Horro rams, especially for Artificial Insemination program.

Daily administration of a GnRH analogue enhances sperm quality in bucks during the non-breeding season

The aim of this study was to determine if daily administration of a GnRH analogue (buserelin acetate) to bucks during the non-breeding season increases testosterone concentration and improves sperm quality. Five bucks received a daily dose of buserelin for 10 days, starting on Day 0 (first administration), and another five bucks remained as controls. Testosterone concentrations were greater in treated than in control bucks during the first hours after buserelin administration (P = 0.05), but greater in controls 10 h later (P < 0.01). Sperm mass motility and percentage of motile sperm were greater in treated (3.9 ± 0.6 and 70.1 ± 7.9%, respectively) than in control bucks (1.0 ± 0.6, P < 0.01; 45.0 ± 7.9%, P < 0.05 respectively) on Day 4. Percentage of sperm with normal morphology tended to be greater in treated than in control bucks (81.8 ± 6.2% compared with 63.5 ± 6.4% respectively, P = 0.08). The treatment decreased the percentage of sperm with mid piece defect and with bent tail (7.0 ± 1.5% compared with 12.0 ± 1.5%; 8.0 ± 1.7% compared with 13.5 ± 1.7%, treated and control bucks, respectively, P = 0.05 for both). The square root percentage of sperm with loose but heads with normal structures tended to be less in treated than control bucks (1.3 ± 0.3% compared with 0.4 ± 0.3% respectively, P = 0.06). It was concluded that daily administration of buserelin during the non-breeding season led to a rapid increase in testosterone concentration and improved sperm quality.

Relationship of organic mineral supplementation and spermatozoa/white blood cells mRNA in goats

The antioxidant properties and the protective role of organic zinc (Zn) and copper (Cu) in white blood cells (WBCs) and spermatozoa were analyzed through quantification of superoxide dismutase 1 (SOD1), catalase (CAT), glutathione peroxidase 4 (GPx4) and nuclear factor erythroid 2-like 2 (NFE2L2) and correlations were determined with sperm functional characteristics in Osmanabadi bucks. Bucks (aged 5 months; n = 40) were divided into ten groups, and the dietary treatments comprised of a control and nine treatment groups as follows: organic Zn as Zn 20, Zn 40 and Zn 60, organic Cu as Cu 12.5, Cu 25, Cu 37.5 and combined organic Zn and Cu as Zn 20+Cu 12.5, Zn 40+Cu 25, Zn 60+Cu 37.5, respectively per kg dry matter for a period of 8 months. The blood (120 and 240 days) and semen (240 days: 40 × 4 = 160) samples were collected from 40 bucks. In WBCs: the relative abundance of mRNA for SOD1, CAT, GPx4, NFE2L2 was greater (P < 0.05) in (120 and 240 days) in majority of the mineral supplemented animals. In spermatozoa: the relative abundance of SOD1, NFE2L2, GPx4 and CAT mRNA was greater (P < 0.05) in selected treatment groups. The abundance of SOD1 mRNA in WBCs was positively correlated (P < 0.05) with sperm mass motility (r = 0.692, P = 0.027). The abundance of GPx4 mRNA was negatively correlated (P < 0.05) with type A sperm (straightness; STR) > 85% and amplitude of lateral head displacement (ALH) > 2.5 μm/ s) (r = −0.711, P = 0.021) and (P < 0.05) positively correlated with sperm viability (r = 0.669, P = 0.035). Organic Zn and Cu supplementation was associated with an increase in the expression of antioxidant defense enzyme genes in bucks.

Effect of different proteolytic enzymes on liquefaction of semen of dromedary camel

In most of domestic animals, semen biology is well studied, but it is not so well studied in camelids particularly in dromedary camels. In camelids, the ejaculated semen is highly viscous, so before its evaluation and processing for preservation, it is necessary to be liquefied. Our study was designed to evaluate the effect of three different enzymes (0.1% Collagenase, 0.5 X Accutase, 0.1% Trypsin-EDTA - Ethylene-diamine-tetra-acetic acid) on seminal viscosity at different time intervals along with its effects on seminal parameters. Semen was collected from the six adult healthy male camels using female camel as dummy. Bovine artificial vagina was used for the semen collection and ejaculate was kept at 37ºC for further processing after its collection. All three enzymes were diluted in 1x phosphate buffered saline (PBS) and the semen samples were kept at 37ºC. The samples were evaluated at different time intervals 0, 5, 10, 15, 30, 60, 120 and 240 min (T0–T240) for viscosity and it was evaluated by pipette method. Immediately after collection, the viscosity was very high, and no sperm mass motility was seen. However, after enzyme treatment viscosity was showed declining trend in all treatment groups compared to control. In trypsin treated group maximum motility was reached in 15 minutes. Live-dead percent was observed to be similar among treatment groups. Observations on liquefaction of semen revealed that use of enzymes decreasing the viscosity but with different rates.

Sedation or anaesthesia decrease the stress response to electroejaculation and improve the quality of the collected semen in goat bucks

Electroejaculation (EE) is stressful and probably painful; thus the administration of anaesthesia is recommended to decrease those negative effects. However, anaesthesia has a direct risk of provoking death, but sedation is less risky than anaesthesia. At the same time, α2-adrenergic agonists may improve semen quality. Therefore, the aim of the present study was to compare the physiological and behavioural responses indicative of stress and possibly pain, and the semen quality in electroejaculated untreated, anaesthetised or sedated goat bucks. Semen was collected from eight bucks using three different procedures in all them (EE in untreated bucks, EE under sedation or EE under general anaesthesia). The number of vocalizations during EE and the behavioural pattern before and after procedures were recorded. Pain visual analogue scale (VAS) score was also determined during EE. Rectal temperature, heart rate, serum cortisol concentration, biochemical and haematological parameters were measured before and after each procedure, and sperm characteristics were determined. Bucks vocalised more often when untreated than sedated or anaesthetised (P<0.02). The pain VAS score was greater when bucks were untreated than sedated or anaesthetised (P<0.002). The rectal temperature, heart rate, total protein, albumin and haemoglobin concentrations were greater when bucks were untreated than anaesthetised or sedated (P<0.02). Serum cortisol increased after EE (P=0.0006), without differences between procedures. The frequency and duration of lying down after EE were greater when bucks were anaesthetised than sedated or untreated (P<0.05), and were also greater when bucks were sedated than untreated (P<0.05). The number of times that the animal tried to stand up after EE was greater when bucks were anaesthetised than sedated or untreated (P<0.02). The sperm mass motility was greater when bucks were anaesthetised or sedated than when they were untreated (P=0.048). When animals were sedated, the ejaculate contained more spermatozoa with functional plasma membrane (P=0.03) and morphologically normal (P=0.05) than when they were untreated. In conclusion, general anaesthesia and sedation decreased the stress and probably the pain response provoked by EE and especially sedation improved the quality of the semen collected.

Sperm collection by transrectal ultrasound-guided massage of the accessory sex glands is less stressful than electroejaculation without altering sperm characteristics in conscious goat bucks

In anesthetized non-domestic ruminants transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) is an alternative method to collect semen slightly less stressful than electroejaculation (EE). However, some sperm characteristics are better when semen is collected with EE than with TUMASG. As anesthesia reduces the response to stressors, the advantages of TUMASG may be reduced in anesthetized animals, and thus, TUMASG may be even more advantageous in conscious animals. Therefore, the aim of the present study was to compare the stress response and the characteristics of the sperm collected with TUMASG and EE in conscious goat bucks. Semen was collected in 10 bucks with both procedures. During each procedure, the time required for ejaculation, the number of electric pulses applied and the number of vocalizations were recorded. Rectal temperature, heart rate, serum cortisol concentration, biochemical and hematological parameters were measured before and after each procedure. Sperm characteristics [ejaculated volume, sperm concentration, sperm mass motility (scale 0–5), sperm vigor (scale 0–5), the percentages of motile and progressive motile sperm, of sperm vitality, of sperm with plasma membrane integrity, and with acrosome damage and morphological abnormalities] were also determined. Electroejaculation required more electric pulses than TUMASG (P < 0.0001), but TUMASG took more time than EE (P < 0.0001). The EE provoked more vocalizations (P = 0.02) and a greater increase of cortisol concentrations than TUMASG (P = 0.04). Heart rate also tended to be greater with EE than with TUMASG (P = 0.07). The sperm characteristics did not differ between TUMASG and EE. In conclusion, TUMASG was less stressful and probably less painful than EE without affecting the semen quality. Thus, although it required more time, TUMASG is an alternative procedure to decrease the welfare concerns raised by EE in conscious goat bucks.

Continuous contact with females in estrus throughout the year enhances testicular activity and improves seminal traits of male goats

The “female effect” consists in the stimulation of males' reproductive activity by different signals emitted by females. This stimulation leads to endocrine and behavioral changes that may modify the seasonal pattern of male ruminants. The aims of this experiment were (1) to describe the local reproductive seasonal pattern of Gabon bucks and (2) to determine if continuous chemical, auditory, and visual contact with does in estrus enhances bucks' testicular activity and improves seminal traits throughout the year and modify their seasonal pattern. We used 16 adult Gabon bucks assigned to two experimental groups: nine bucks remained continuously isolated from females (isolated bucks, group IB) and seven bucks were in continuous chemical, auditory, and visual contact through a fence line with does in estrus (stimulated bucks, group SB). During 13 months, scrotal circumference and testosterone concentration were measured weekly and testicular echogenicity was measured every 2 weeks. Also, sperm motility mass and percentage of abnormal spermatozoa were determined, and sperm concentration and total number of motile spermatozoa were calculated every 2 weeks. Testicular echogenicity was greater in IB than that in SB bucks (P < 0.0001), but there were no differences in scrotal circumference. Overall, testosterone concentration was greater in IB than that in SB bucks (P = 0.04), but from late winter to mid-summer, when testosterone concentration presented basal concentrations, SB bucks had greater values than IB bucks (P = 0.004). Sperm concentration (P = 0.05) and sperm mass motility (P = 0.01) were greater in SB than that in IB bucks, and the total number of progressive motile spermatozoa tended to be greater in SB than in that IB bucks (P = 0.1). The percentage of abnormal spermatozoa was lower in SB than in IB bucks in several time points (P < 0.0001). Testicular and seminal traits were better from the end of the spring until mid-autumn. We concluded that does in estrus stimulated bucks' testicular activity, including better seminal quality and a greater increase of testicular fluid content than bucks isolated from females. However, the general seasonal pattern was not modified by stimulation with does in estrus.