Sperm Motility Values Research Articles

Associations of dietary inflammatory indices (DII and E-DII) with sperm parameters.

This study aimed to explore the ambiguous link between dietary inflammatory indices and sperm parameters. Specifically, it investigated the associations between the dietary inflammatory index (DII) and the energy-adjusted dietary inflammatory index (E-DII) with sperm motility, morphology, and count in men undergoing routine semen analysis. A cross-sectional study was conducted with 144 men enrolled, where semen samples were collected and evaluated according to the 2010 World Health Organization guidelines. Dietary data were gathered using a 147-item semi-quantitative food frequency questionnaire developed by the researchers. Pearson correlation analysis was employed to assess the relationships of the DII and E-DII with sperm parameters. The mean DII and E-DII scores were 1.23±1.1 and 0.49±0.43, respectively. The mean values for sperm motility, morphology, and count were 43.08%±19.30%, 78.03%±26.99%, and 48.12±44.41 million, respectively. Both motility (r=-0.353) and count (r=-0.348) were found to be inversely and significantly correlated with DII. Similarly, Pearson correlation tests revealed strong and significant inverse correlations of motility (r=-0.389) and count (r=-0.372) with E-DII. The findings suggest that a diet with a higher anti-inflammatory potential may be associated with increased sperm count and motility, but not with changes in morphology. Further research is necessary to confirm these findings, elucidate the underlying mechanisms, and identify dietary modifications that could improve male fertility.

Effect of age, season and sire on semen quality traits in Frieswal breeding bulls

The present study was undertaken to assess the effect of season of semen collection, age and sire on semen quality of Frieswal breeding bulls. The data on 8113 semen ejaculates of 155 bulls born to 34 Frieswal sires were utilized for the study. The effect of age of bulls on semen volume, sperm concentration (million) per ml and sperm motility (%) in fresh semen (directly after collection) was highly significant. Additionally, it was discovered that the sire effect of bulls on different semen quality parameters and the influence of season on sperm concentration and motility both were statistically significant. Heritability estimates of different semen quality traits studied were moderately high and exceptionally significant. For semen volume, the estimate was found to be 0.336±0.078 while for concentration and pre freezing sperm motility values were 0.296±0.068 and 0.407±0.089, respectively. From the results, it may be concluded that the age and sire of bulls significantly affected the semen volume, sperm concentration and sperm motility in Frieswal bulls. The season of semen collection caused significant variations in sperm concentration and motility but not the semen volume. The semen volume and concentration were superior in ejaculates collected during winter season while the summer season was conducive for increased sperm initial motility. The Frieswal bulls up to 4-5 years are able to produce semen of better quality. The moderately high heritability estimates of semen quality traits provide indication for genetic improvement of the traits through selection.

High-coverage targeted lipidomics revealed dramatic lipid compositional changes in asthenozoospermic spermatozoa and inverse correlation of ganglioside GM3 with sperm motility

BackgroundIt has been previously demonstrated that cholesterol content and cholesterol/phospholipid ratio were significantly higher in asthenozoospermia and oligoasthenoteratozoospermia. The majority of published studies have investigated the fatty acid composition of phospholipids rather than lipids themselves. This study evaluated the lipid composition of asthenozoospermic and normozoospermic spermatozoa, and identified the exact lipid species that correlated with sperm motility.MethodsA total of 12 infertile asthenozoospermia patients and 12 normozoospermia subjects with normal sperm motility values were tested for semen volume, sperm concentration, count, motility, vitality and morphology. High-coverage targeted lipidomics with 25 individual lipid classes was performed to analyze the sperm lipid components and establish the exact lipid species that correlated with sperm motility.ResultsA total of 25 individual lipid classes and 479 lipid molecular species were identified and quantified. Asthenozoospermic spermatozoa showed an increase in the level of four lipid classes, including Cho, PE, LPI and GM3. A total of 48 lipid molecular species were significantly altered between normozoospermic and asthenozoospermic spermatozoa. Furthermore, the levels of total GM3 and six GM3 molecular species, which were altered in normozoospermic spermatozoa versus asthenozoospermic spermatozoa, were inversely correlated with sperm progressive and total motility.ConclusionsSeveral unique lipid classes and lipid molecular species were significantly altered between asthenozoospermic and normozoospermic spermatozoa, revealing new possibilities for further mechanistic pursuits and highlighting the development needs of culture medium formulations to improve sperm motility.

Semen characteristics of the endangered cyprinid lake minnow, Eupallasella percnurus (Pall.), from different Polish populations

Abstract The aim of the study was to compare the quality and quantity parameters of semen in male lake minnow, Eupallasella percnurus (Pallas, 1814), from six Polish populations. Substantial differences among the populations were found in milt volume (36–112 µl) and sperm concentration (4.7–8.0 × 109 ml−1). Mean values for sperm motility were similar; however, there were population-related differences in sperm velocity measured with VSL (96.3–127 µm s−1). Despite this, the quality of semen in all cases was high and appropriate for both artificial reproduction and cryopreservation.

Identification of circular RNAs in porcine sperm and evaluation of their relation to sperm motility

Circular RNAs (circRNAs) are emerging as a novel class of noncoding RNAs which potential role as gene regulators is quickly gaining interest. circRNAs have been studied in different tissues and cell types across several animal species. However, a thorough characterization of the circRNAome in ejaculated sperm remains unexplored. In this study, we profiled the sperm circRNA catalogue using 40 porcine ejaculates. A complex population of 1,598 circRNAs was shared in at least 30 of the 40 samples. Generally speaking, the predicted circRNAs presented low abundances and were tissue-specific. Around 80% of the circRNAs identified in the boar sperm were reported as novel. Results from abundance correlation between circRNAs and miRNAs together with the prediction of microRNA (miRNA) target sites in circRNAs suggested that circRNAs may act as miRNA sponges. Moreover, we found significant correlations between the abundance of 148 exonic circRNAs and sperm motility parameters. Two of these correlations, involving ssc_circ_1458 and ssc_circ_1321, were confirmed by RT-qPCR using 36 additional samples with extreme and opposite sperm motility values. Our study provides a thorough characterization of circRNAs in sperm and suggests that circRNAs hold potential as noninvasive biomarkers for sperm quality and male fertility.

Aryl Hydrocarbon Receptor as a Target for Lycopene Preventing DEHP-Induced Spermatogenic Disorders.

Di(2-ethylhexyl)phthalate (DEHP) is widely used as a plasticizer to improve product flexibility and workability. Lycopene (LYC) is a natural compound and has promising preventive potentials, especially antireproductive toxicity, but the specific underlying mechanism is yet to be fully defined. Our study investigated the effect of LYC on DEHP-induced spermatogenesis disorders. Male ICR mice were treated with DEHP (500 or 1000 mg/kg BW/day) and/or LYC (5 mg/kg BW/day) for 28 days. Our results indicated that LYC could relieve the DEHP-induced injury of seminiferous tubules and spermatogenic cells, swelling of endoplasmic reticulum (ER), and an increase of mitochondria. LYC prevented increased levels of nuclear damage to DNA and the deformity rate and decreased values of sperm motility, number, and density. Moreover, LYC treatment decreased DEHP-induced nuclear accumulation of aryl hydrocarbon receptor (AHR) and AHR nuclear translocator (ARNT), and the expressions of their downstream target genes such as cytochrome P450-dependent monooxygenases (CYP) 1A1, 1A2, and 1B1 were markedly reduced to normal in the LYC treatment group. Our study showed that LYC can prevent DEHP-induced spermatogenic disorders via an AHR/ARNT signaling system. This study provided new evidence of AHR as a target for LYC, which can prevent DEHP-induced toxicity.

Effects of glucose, methanol concentration, and time of equilibration on post-thaw sperm motility of rainbow trout semen

The objective of this work was to develop a complex standardisation of cryopreservation procedure for rainbow trout semen. The specific objectives were to determine the effects of the final glucose concentration (0.11–0.23 M), final methanol concentration in the extender (0–20%), equilibration time (5–180 min), and post-thaw storage time of semen (0–180 min) on sperm motility parameters. The final glucose and methanol concentrations significantly affected sperm motility after thawing. The highest sperm motility (63–75%) was found for glucose concentrations ranging from 0.13 to 0.17 M and for 5.0 to 10.0% methanol, with the highest motility (70%) at 7.5% methanol. Concerning post-thaw storage equilibration times, sperm motility of the equilibrated semen was not affected by 120 min, while a significant decrease was recorded after 180 min of equilibration. The highest values of sperm motility immediately after thawing were observed in samples equilibrated for 5 to 120 min. The sperm motility after thawing was not significantly affected up to 60 min for samples equilibrated for 5, 15, and 30 min. However, for samples equilibrated for 60, 120, and 180 min, significant decreases were observed at 60 min after thawing. Our results indicate the importance of the standardisation of concentrations of cryoprotectants as well as cryopreservation equilibration time. The recommended cryopreservation procedure includes final sperm concentration (1.0 × 109 spermatozoa ml−1), as we determined previously, 0.15 M glucose, 7.5% methanol, 15 min equilibration time, and post-thaw storage up to 60 min. The developed standardised procedure has potential for the cryopreservation of rainbow trout semen for practical purposes.

Relationship of sperm plasma membrane and acrosomal integrities with sperm morphometry in Bos taurus.

To date, sperm morphometric studies have assessed whole sperm populations without considering sperm function. The aim of this study was to evaluate the relationship of sperm membrane and acrosomal integrity with sperm morphometry in liquid semen samples collected from bulls. To this end, sperm morphometry was performed on cryopreserved semen samples from 16 bulls by a combination of fluorescent dyes, including Hoechst 33343, carboxyfluorescein diacetate, and propidium iodide. This allowed discrimination of different subpopulations on the basis of sperm membrane and acrosomal integrity and analysis of the morphometrics of the sperm head, nucleus, and acrosome using a specific plug-in module created on ImageJ. Acrosomal integrity was related to sperm morphometry as the heads of spermatozoa with a damaged acrosome were significantly smaller than those with a normal acrosome (P < 0.001). In the case of spermatozoa with an intact acrosome, those with a damaged plasma membrane had a larger sperm head and acrosome than spermatozoa with an intact plasma membrane (P < 0.001). No significant differences in the sperm head size were observed between sperm subpopulations without an acrosome or in the nuclear sperm morphometry of the different subpopulations. There was a positive correlation between the sperm motility values of the samples and the morphometric parameters for intact spermatozoa. These correlations were particularly strong for the morphometric parameters of the sperm acrosome. We conclude that there are clear differences in the sperm morphometry depending on the status of the sperm membrane and acrosome and this should be considered when performing this kind of analysis.

Technical Note: The use of iSperm technology for on-farm measurement of equine sperm motility and concentration

The iSperm is a newly released semen analysis tool from Aidmics Biotechnology Co. LTD, which allows an iPad Mini to be transformed into a handheld microscope with objective semen analysis software for equine available through the Apple Store (version 4.5.2). The aim of this study was to compare iSperm values for sperm motility and sperm concentration to current acceptable methods for semen analysis and to determine the agreement with these methods using statistical methods. Two ejaculates from each of five Standardbred stallions were used to compare sperm motility (computer-assisted semen analysis [CASA] vs. iSperm) and concentration (NucleoCounter SP-100 [NC] vs. hemocytometer vs. iSperm). Data were analyzed by first testing for the differences between the means of each method using a linear mixed-effects model. The agreement between the two continuous measurements for each method was then investigated by computing Lin’s concordance correlation coefficient (CCC), with a value of 1 indicating perfect agreement between methods. Results are reported as the CCC with the associated 95% confidence interval in parentheses. Means for both total motility (TM) and progressive motility (PM) were equal between CASA and iSperm values (P = 0.0741 and P = 0.725, respectively). However, means for all velocity measurements were significantly different between CASA and iSperm readings (P < 0.001). For concentration, means were equal between NC and iSperm values (P = 0.748) and for hemocytometer and iSperm values (P = 0.953). The CCC for TM was 0.871 (0.788, 0.923) and for PM was 0.916 (0.847, 0.955) indicating good agreement between methods. Low levels of agreement were observed for all velocity measurements. Finally, the CCC for concentration compared by iSperm and NC was 0.970 (0.949, 0.982) and for iSperm and hemocytometer it was 0.962 (0.934, 0.978), both close to the line of perfect concordance. Although more work is needed to improve the iSperm software for velocity measurements to be acceptable by research standards, in its present form the iSperm will introduce a low-cost and affordable method for on-farm semen analysis (TM, PM, concentration) for breeders and veterinarians. As a result, more farms will have access to accurate sperm analysis tools which will help to standardize semen processing procedures leading to better overall quality of semen used for artificial insemination.

Changes in sperm parameters of sex-reversed female mandarin fish Siniperca chuatsi during cryopreservation process

This works studies the physiological characteristics (sperm motility characteristics) and biochemical characteristics (DNA damage, enzymes activities and fertilization ability) of fresh and freeze-thawed semen of sex-reversed female mandarin fish Siniperca chuatsi (n = 30) obtained with the application of 17α-methyl testosterone for sex reversal. The fresh sperm motility and fertilization rate of sex-reversed females were about 83% and 70% respectively which had no significant difference with normal males (p > 0.05). Except for the value of DNA damage, other values of sperm motility, related enzymes activities and fertilization rate of sex-reversed female sperm declined after a process of cryopreservation (p < 0.05). But the frozen sperm can still get nearly 60% of the fertilization rate. This study identified the physiological and biochemical characteristics of the fresh and cryopreserved sperm from sex-reversed female mandarin fish, and the sex-reversed female spermatozoa can be used for actual production.

Inbreeding causes decreased seminal quality affecting pregnancy and litter size in the endangered black‐footed ferret

AbstractEx situ conservation breeding for species recovery often must address small founder number. Here we investigated the long‐term effects of small founder size in the endangered black‐footed ferret Mustela nigripes using 20 years of reproductive data. Specifically, we studied seminal traits, pregnancy success and litter size in the context of the population's low genetic diversity. Detailed evaluation of 539 electroejaculates from 262 pedigreed donors with varied inbreeding coefficients (F = 0.000–0.250) revealed a wide range in sperm motility (0.0–100.0%), forward progression (type of forward movement on a scale 0–5; 5 = best; 0.0–4.5), cell concentration (7.2–6348.0 × 106 mL−1 of ejaculate), structural normality (0.0–89.0%) and normal apical ridges (critical for fertilization; 4.8–100.0%). Over this two decade interval, there was an increase (P &lt; 0.05) in F and a decrease (P &lt; 0.05) in sperm forward progression and structurally normal cell morphology. From analysis of artificial insemination (AI) data, higher values for sperm motility, forward progression and structural normality or spermatozoa that retained a normal apical ridge had a positive influence (P &lt; 0.05) on litter size. Males producing higher values for these four traits also sired more kits. Results demonstrate that sperm motility and morphology are decreasing in black‐footed ferrets managed intensively ex situ. These two metrics appear associated with an increasing F and likely are at least partially responsible for lowered reproductive success in this closed, endangered population. These results illustrate the potential physiological and offspring production impact of having few founders available in an ex situ conservation breeding program.

Influence of bull age, ejaculate number, and season of collection on semen production and sperm motility parameters in Holstein Friesian bulls in a commercial artificial insemination centre.

In the current era of genomic selection, there is an increased demand to collect semen from genomically selected sires at a young age. The objective of this study was to assess the effect of bull age, ejaculate number, and season of collection on semen production (ejaculate volume, sperm concentration, and total sperm number; TSN) and sperm motility (prefreeze and post-thaw total and gross motility) parameters in Holstein Friesian bulls in a commercial artificial insemination (AI) center. The study involved the interrogation of a large dataset collected over a 4-yr period, (n = 8,983 ejaculates; n = 176 Holstein Friesian bulls aged between 9 mo and 8 yr). Bulls aged less than 1 yr had the poorest semen production and sperm motility values for all parameters assessed compared with bulls older than 1 yr (P < 0.01). First ejaculates had greater semen production and greater prefreeze motility values than second consecutive ejaculates (P < 0.01), but despite this, there was no difference in post-thaw motility. When subsequent ejaculates were collected from bulls aged less than 1 yr, semen production and sperm motility did not differ compared with mature bulls. Semen collected in winter was poorest in terms of sperm concentration and TSN, but best in terms of post-thaw motility (P < 0.01). In conclusion, second ejaculates can be collected, particularly from bulls aged less than 1 yr, without a significant decrease in post-thaw sperm motility, thus may be a useful strategy to increase semen availability from young genomically selected AI bulls in high demand.

Effect of cooling rate on sperm quality of cryopreserved Andalusian donkey spermatozoa

The aim of this study was to evaluate the effect of different cooling rates on post-thaw quality of cryopreserved donkey spermatozoa. Eighteen ejaculates from six adult Andalusian donkeys (three ejaculates per donkey) were collected using an artificial vagina. Pooled semen samples (two ejaculates per pool) were divided into three aliquots, and frozen in Gent freezing extender using three different cryopreservation protocols (P): P1 (conventional slow freezing, as control): semen pre-cooled in an Equitainer for 2 h and frozen in liquid nitrogen (LN2) vapour; P2 (controlled pre-freeze cooling rate): semen pre-cooled at a controlled rate for 73 min and frozen in LN2 vapour; and P3 (rapid freezing) semen frozen immediately in LN2 vapour. After thawing at 37 °C for 30 s, semen samples were assessed for motility, morphology, acrosome and plasma membrane integrity; spermatozoa were also tested for DNA integrity. Significant (P < 0.01) differences were found between the cryopreservation protocols for all sperm parameters evaluated, except for DNA integrity. Semen samples frozen using P2 showed significantly (P < 0.01) higher values for sperm motility, morphology, sperm membrane integrity, and acrosome integrity. On the contrary, P3 reduced sperm motility (P < 0.01) and increased the percentage of spermatozoa with damaged plasma membrane (P < 0.001). In our study, we demonstrated that the sperm of Andalusian donkey is particularly sensitive to the cooling rate used before freezing. Furthermore, Andalusian donkey semen can be successfully cryopreserved using controlled cooling rates combined with freezing in LN2 vapour.

Dietary effect of Cissus populnea and Securidaca longepedunculata aqueous leave extracts on reproductive, haematological and biochemical parameters of African catfish Clarias gariepinus (Burchell, 1822) broodstocks

The objectives of the present study were to evaluated gametes quality, haematological and biochemical parameters of the Clarias gariepinus brood stocks fed with varying concentrations of Cissus populnea (CP) and Securidaca longepedunculata (SL). Fish were fed with Diet 1 (control, 0 ml plant extract), Diet 2(0.5 ml/kg CP), Diet 3 (1.0 ml/kg CP), Diet 4 (0.5 ml/kg SL), Diet 5 (1.0 ml/kg SL) and Diet 6 ­ (0.5 ml/kg CP + 0.5 ml/kg SL) for 90 days. The results of haematocrit and red blood cell of treated groups were relatively similar to the control. The white blood cell and haemoglobin values were lower than the control group while the mean corpuscular volume, mean corpuscular haemoglobin were significantly difference across treatments however; an elevated glucose level was recorded in the treated groups. Also, the mean platelet volume and platelet distribution width were relatively similar across treatments. The serum testosterone, progesterone and estrogen were found to be higher in fish fed diet 2 (0.5ml/kg CP). C. populnea at 0.5 ml/kg significantly (p<0.05) improved eggs weight, fecundity, gonadosomatic index, % fertilization and hatchability. Male brood stocks fed diet 2 (0.5 ml/kg CP) recorded the highest values for sperm motility (56.67 ± 0.33%), milt count and volume (820.33 ± 0.33×10 6 /ml and 1.80 ± 0.05 ml respectively) across all groups. Similarly, fish fed diet 2 (0.5ml/kg CP) had the highest values for eggs weight (283.7±102.4g), fecundity (168,286 ± 57157), gonadosomatic index (32.59 ± 2.72), fertilization (62 ± 20.4%) and hatchability (62.92 ± 19.75%).The dietary supplementation of 0.5 ml/kg C. populnea extract highly enhanced the reproductive profiles of male and female C gariepinus brood stocks.

Selection of red deer spermatozoa with different cryoresistance using density gradients.

The objective of sperm selection media is selecting the best spermatozoa and to remove seminal plasma and diluent for using them in assisted reproductive techniques. It is known that individuals show different cryoresistance in response to the same freezing procedure. Our hypothesis was that the efficacy of selection media could be dissimilar for samples with different sperm quality after thawing. Epididymal sperm samples from mature Iberian red deer were collected and frozen. Males were classified as with high post-thaw sperm quality when sperm motility (SM)≥70%, or as with low post-thaw sperm quality when SM≤69%. Samples were centrifuged using the following density gradients (DG): Percoll® , Puresperm® and Bovipure™ , and several functional sperm parameters were assessed after sperm selecting and washing. Males classified with high sperm quality had higher post-thawing values (p>.05) for all parameters evaluated, except for linearity index, than those categorized as low sperm quality. After selection, some sperm characteristics improved (viability, apoptosis and mitochondrial activity) for both groups, showing the males with high sperm quality higher values in all sperm parameters except for kinematic traits and DNA fragmentation index (%DFI), regardless of DG. Bovipure™ yield lower values of sperm motility, viability, apoptosis and mitochondrial activity in relation to Percoll® and Puresperm® considering both quality groups. There was an interaction between the type of DG and sperm quality group for sperm viability (p=.040) and apoptosis (p=.003). Thus, Percoll® selected less live and more apoptotic spermatozoa than Puresperm® and Bovipure™ for males with low sperm quality. In conclusion, the DG are more efficient selecting spermatozoa from samples with high sperm quality, acting differently depending on initial sperm quality.

Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants.

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P<0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P<0.001), whereas DMSO affected sperm motility and membrane integrity (P<0.001). DMFA 2.5% yielded higher (P<0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

Interactions Among Different Devices and Electrical Stimulus on the Electroejaculation of Captive Agoutis (Dasyprocta leporina).

The interactions among different electroejaculation devices associated with serial or continuous stimuli were investigated to improve the efficiency of the electroejaculation for semen collection in agoutis. Ten sexually matured male Dasyprocta leporina were restrained by the intramuscular administration of xylazine-ketamine association. Each individual was randomly subjected to four electroejaculation protocols, by combining two devices (one presenting longitudinal electrodes emitting square waves and other presenting ring electrodes emitting sine waves) and two electrical stimuli protocols (serial or continuous). A total of 40 attempts for electroejaculation were conducted in agoutis, being 10 per treatment. The most efficient treatment in providing ejaculates containing sperm (p<0.05) was that using and electroejaculator connected to a probe with ring electrodes and associated with serial stimuli (4/7; 57%). In spite of semen parameters obtained by sine waves were adequate for using the samples for assisted reproduction, higher values for sperm motility and functional membrane integrity were obtained in the use of the square wave, independently of the electric stimulation protocol used (p<0.05). In conclusion, we verified that the use of a device presenting a probe with ring electrodes and emitting sine waves, associated with a serial stimuli protocol, improves the efficiency for semen obtaining by electroejaculation in adults D.leporina.

Testicular, sperm and haematological toxicity of ridomil and chlorpyriphos in single and combined exposure

This study was carried out to investigate the effect of Ridomil and Chlorpyriphos, singly and in combination, on testes histology, sperm parameters, and haematological indices in male albino rat model. In a completely randomized design, thirty five experimental subjects were constituted into seven equal treatment groups: A (control); CH1 (25 mg of Chlorpyriphos/kg bw); CH2 (50 mg of Chlorpyriphos/kg bw); R1 (25 mg of Ridomil/kg bw); R2 (50 mg of Ridomil/kg bw); COM1 (12.5 mg of Chlorpyriphos/kg bw+12.5 mg of Ridomil/kg bw); and COM2 (25 mg of Chlorpyriphos/kg bw+25 mg of Ridomil/kg bw); which orally received the stated pesticide treatments, thrice a week, at forty eight hour intervals for eight weeks; after which the animals were sacrificed and tissues obtained. Analysis of variance revealed significantly (P<0.05) lower RBC and PCV values in groups CH1 and CH2, as well as in groups COM1 and COM2. Sperm count in group CH2 was significantly lower than in all the other experimental groups. CH1 and CH2 also had the lowest sperm motility values, although not significant. Testes micrographs revealed necrosis, atrophic widely spaced seminiferous tubules with eroded basement membrane in group CH2. Chlorpyriphos alone, as well as the combination of both pesticides (to a lesser extent); demonstrated testicular, sperm, and haemo-toxicity. For the affected parameters, the toxic effects noticed in the pesticides combination groups were not more than that observed due to Chlorpyriphos alone.

Standardization of CASA sperm motility assessment in the ram

This study was designed to evaluate the effect of different viewing chambers or slides, drop sample volume, and number of sperm per field on CASA sperm motility parameters in the ram. In the first trial, the effect of two different drop sample volumes, 5 and 10μl, on the parameters of sperm motility were compared using slide-coverslip and Makler chamber. No differences between the two drop sample volumes in the two devices were found in the different sperm motility parameters. In the second experiment, significant differences (P<0.001) were observed for all the sperm motility parameters studied among three devices evaluated: slide-coverslip, Makler and ISAS chambers. A higher proportion of motile spermatozoa was observed in Makler chamber than in other devices, while the ISAS chamber caused an important decrease in the percentage of motile and progressive sperm. Sperm velocity variables were different in the three systems with slide-coverslip>Makler>ISAS. Similar results were recorded in Makler and ISAS chambers, but lower than in the slide-coverslip in the parameters related to the quality of movement. Makler chamber showed to be less variable, whereas the greatest variability was recorded in slide-coverslip, where significant differences between central and peripheral fields were found. In the third trial, fresh semen samples were mixed with different volumes of forced dead semen to determine proportions of non-motile spermatozoa in the samples, which were evaluated for CASA sperm motility and membrane integrity. Slide-coverslip was more accurate, as the percentages of motile sperm were more correlated to the sperm viability results (r=0.95, 0.90, 0.92 for slide-coverslip, Makler and ISAS chamber, respectively, P<0.001) and to the theoretical values of sperm motility after adding known percentages of dead sperm (R2=0.84, 0.79. 0.79, respectively, P<0.001). It was concluded that the type of viewing chamber used for CASA sperm motility analysis in the ram had a considerable influence on results, precision and accuracy, whereas the drop volume had little effect. The final goal of sperm motility assessment is the evaluation of male fertility, the rigorous standardization and validation of the CASA protocol being a pre-requisite to obtain reliable results. Results of the present study contribute to this standardization and validation process in the ram.

Effect of Different Extenders onIn VitroCharacteristics of Feline Epididymal Sperm During Cryopreservation

To evaluate and compare the efficacy of various extenders for the cryopreservation of epididymal cat spermatozoa, two experiments were planned. Bovine and equine commercial extenders in the experiment 1 and TRIS-egg yolk-based extenders in experiment 2 were separately studied since the number of sperm collected per cat is reduced. Epididymal sperm samples were packaged into 0.25-ml straws and frozen. Vigour, motility, morphology, acrosome status, sperm viability and functional membrane integrity were assessed at collection, after cooling and after thawing, while DNA integrity was evaluated at 0- and 6-h post-thaw. Experiment 1 compared the effect of three non-feline commercial extenders - based on TRIS-egg yolk (Triladyl), egg-yolk-free medium (AndroMed) and skimmed milk-egg yolk (Gent) - on the quality of frozen-thawed epididymal cat sperm. Values for sperm motility and functional membrane integrity in cooled sperm diluted in Triladyl were higher (p<0.001) than those recorded for Andromed and Gent. Except sperm morphology, the other assessed characteristics showed significant higher values in frozen-thawed sperm diluted in Triladyl than in Andromed and Gent extenders. Experiment 2 analysed the effects of three TRIS-egg yolk-based extenders, one non-feline commercial (Triladyl) and the other two prepared using different monosaccharides (glucose and fructose), on freezing-thawed sperm. Results showed that specifically prepared extenders for cryopreservation of feline spermatozoa performed better than the commercial extender Triladyl, although sperm quality during the freezing-thawing process did not significantly differ associated with the type of monosaccharide (glucose vs fructose) added to the mentioned extenders. Although TRIS-egg yolk-based extenders prepared in experiment 2 improved sperm cryoprotection, Triladyl remains a good option for practitioners who, for ease of use and availability, prefer to work with commercial extenders.