Percentage Of Normal Spermatozoa Research Articles

Comparative efficacy of two extenders on post-thaw sperm characteristics of cryopreserved Black Bengal buck semen

The research work was conducted at the Artificial Insemination (AI) Center under the Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh to compare the efficacy between Egg Yolk Citrate (EYC) and Tris extenders for cryopreserved semen production in Black Bengal buck. The parameters of semen characteristics included percentage of individual motility (progressive), normal and live sperm and sperm concentration (billion/ml). Statistical analysis of fresh semen of individual buck effect was found to be significant (p<0.01) on normal sperm percentage but insignificant on individual motility, sperm concentration and live sperm percentage. After dilution with EYC extender, individual bucks had significant effect (p<0.05) on sperm motility and but insignificant on motility after cooling with and without glycerol. In the same way with Tris extender, it was insignificant (p>0.05) on diluted semen motility and motility after cooling with and without glycerol. Motility and morphology of the sperm after equilibration and thawing showed insignificant difference among the bucks using EYC extender. On the other hand, variation in the motility after equilibration and thawing was found significant (p<0.05) using Tris extender and insignificant on normal and live sperm percentages.
 Res. Agric., Livest. Fish.6(1): 119-125, April 2019

Effect of Age and Frequency of Collection in Quality of Jersey Bulls Semen at National Livestock Breeding Center, Nepal

The present study was undertaken to determine the effect of age and frequency of collection on quality parameters of Jersey bull semen at National Livestock Breeding Center (NLBC), Pokhara, Kaski district. Nine Jersey bulls were selected randomly from NLBC. To obtain the effect of age, six bulls of three different age interval (3- 4, 5-7 and 8-9 years) were selected randomly. Remaining three bulls were used to determine the effect of frequency of collection on semen quality. Studies were conducted for three months (Nov/Dec to Dec/Jan 2017). Semen quality was analyzed using CASA system and SMILE software. ANOVA and paired t- tests were applied to find out level of significance. All the statistical analyses were done using SPSS 17.0. Sperm concentration (0.551 ±0.004 to 1.084 ± 0.014x109), post- thaw motility (42.44±0.11 to 51.72±0.03 %), live sperm (45.62 ± 0.13 to 51.23±0.15 %), and abnormal sperm (11.57±0.12 to 19.59±0.24%) were significantly affected by the age of the bulls. Semen quality was found superior in the bulls belonging to 5-7 years of age intervals. On the other hand, volume (3.98±0.09 to 6.72±0.06 ml), initial motility (72.68±0.02 to 78.64±0.15 %) and pre-filling motility (71.34±0.17 to 77.62±0.02 %) did not differ significantly among different age intervals. Significantly, higher value for semen volume (3.98±0.12 to 6.02±0.14 ml), sperm concentration (0.572±0.119 to 1.095±0.015) and post- thaw motility (40.34±0.21 to 48.54±0.06% was obtained on second collection. However, initial motility (70.58±0.06 to 75.64±0.13 %), prefilling motility (68.11±0.12 to 71.62±0.02 %), live sperm (53.27±0.54 to 56.17±0.02 %) and abnormal sperm (12.31±0.12 to 14.3±0.47 %) did not differ significantly between the frequencies of collection. Different quality parameters of semen like volume, concentration, initial motility, pre-filling motility, post thaw motility, live and normal spermatozoa percentage were higher in 5-7 years old bulls and in second collection of semen.

Relationship between pre-pubertal biometrical measures and sperm parameters for the selection of high genetic merit pure and crossbred boars

The determination of specific reproductive parameters, which allow the early selection of high genetic merit boars from different lines, is critical as it may avoid the high maintenance costs of keeping unfertile males in the production system. The aim of this work was to evaluate body and testicular measurements during the pre-pubertal phase, and associate them with reproductive characteristics, such as precocity and libido, and seminal characteristics during puberty in two different lines. Two pure breeds used in the crossing of female lines (FL) and two crossbred terminal lines (TL) were evaluated through body and testicular biometrical measures and seminal characteristics. Terminal line boars presented greater body weights and testicular measures (P < 0.01) compared to FL animals throughout the pre-pubertal period. TL males had their first collection at 24.0 ± 0.3 weeks, while their FL counterparts started about one week later (25.0 ± 0.3 weeks, P < 0.05) and age of selection presented a two-week delay in FL males (29.0 ± 0.7 versus 27.0 ± 0.6 weeks). Overall, 57% of FL boars were selected up to 31 weeks of age, while 90% of TL males were selected during the same period (P < 0.05). It was observed that genotype did not affect the seminal characteristics evaluated: volume, concentration, number of total spermatozoa, number of viable spermatozoa or sperm motility and kinetics. However, TL crossbred males showed higher percentage of normal spermatozoa and lower percentage of tail and head defects (P < 0.05). Sperm concentration in the 28th week was positively correlated with body weight in the 1st (r = 0.58, P < 0.001) and 15th (r = 0.39, P < 0.05) weeks of age. Moreover, sperm concentration in the 28th week was also correlated with right testicle length (RTL) in the 3rd week (r = 0.40, P < 0.05), and right testicle width (RTW) at week 15 (r = 0.36, P < 0.05). There was a negative correlation between RTW at week 15 and age of selection (r = −0.32, P < 0.05). Therefore, body and testicular measurements in the pre-pubertal period may predict semen concentration and can be used as early classification criteria for the selection of boars from different lines, without the risk of culling high value boars.

EFFECT OF FEEDING WITH DRIED DISTILLERS GRAINS WITH SOLUBLES AND RICE POLISHINGS ON SCROTAL CIRCUMFERENCE AND SEMEN PARAMETERS OF RAM LAMBS

&lt;p&gt;The effects of level of dried distillers grains with solubles (DDGS) and rice polishings (RP) on the scrotal circumference and semen parameters were examined in 22 crossbred (Pelibuey x Dorper) ram lambs (average live weight 24.84 ± 2.07 kg, approx. 5 months of age). The lambs were randomly allotted to four treatments in a 2 x 2 factorial design. The two factors were dietary levels of RP (0 and 15%) and of DDGS (0 and 30%). The experiment lasted 52 days. The animals were weighed on days 0, 34 and 52, and semen variables were measured on days 34 and 52. Semen evaluation included sperm morphology, mass and progressive motility, and sperm concentration. On day 34, sperm concentration showed an interaction of both effects (P=0.01) and was greater in rams in group 4 (both RP and DDGS) than in those in groups T2 (RP alone) or T3 (DDGS alone). Sperm morphology (% normal cells) was higher (main treatment effect) in rams fed with 30% DDGS than in those not fed with DDGS (P&amp;lt;0.05). The other variables showed no treatment effect. On day 52, none of the ejaculate variables varied with treatments. In conclusion, the diets did not affect body weight of ram lambs, but did affect the percentage of normal spermatozoa at 34 d of the trial, although this effect was not observed after 52 d of the trial; thus, in the long run, these diets do not alter scrotal circumference or semen parameters of young Pelibuey x Dorper ram lambs.&lt;/p&gt;

Effect of season on sexual behavior, testicular measurements and seminal characteristics in "Beni arouss" North Moroccan bucks

Changes in anatomical, behavioural and seminal characteristics were measured throughout the year in seven Beni Arouss bucks maintained at a latitude of 35 °N. Testicular size varied significantly. There were the greatest values during the summer and autumn for scrotal circumference and least in the winter for testicular length. Values for most variables related to sexual behavior were not affected by season except the number of mounts before first ejaculation which was least in the summer. No differences were detected between seasons in terms of semen volume. The sperm concentration, viability and percentage of normal sperm, however, were less during the winter period. The size of sperm heads was larger in the autumn and motility of spermatozoa was greater in the summer and autumn. Results from the correlation analysis indicated that the majority of conventional seminal characteristics were correlated with testis measurements (P < 0.05; r < 0.5). Values for conventional and CASA motility variables indicated there were positive correlations between viability and progressive motility (r = 0.4; P < 0.05) and between the percentage of normal sperm and straight line and average path velocity (r = 0.4; P < 0.05). A global reproduction performance score was established for each buck, which allowed for assessment of magnitude of seasonal changes for each individual. These global score values indicated there was a greater reproductive performance for all bucks during summer and autumn. This study described for the first time seasonal variations of reproductive characteristics of Beni Arouss bucks and results indicate that even though there is a maximal capacity for reproductive performance during the summer and autumn, breeding should be possible throughout the year.

PENGARUH PUASA TERHADAP JUMLAH DAN MORFOLOGI SPERMATOZOA MENCIT (Mus musculus)

One of the benefits of fasting is being able to control testosterone levels in the blood so as not to exceed normal. Normal testosterone levels will produce good quality (morphology, viability, motility) and good quantity (number) of spermatozoa. Therefore, this study was conducted to understand the effect of fasting on the number and morphology of spermatozoa in mice. This study is an experimental study with a completely randomized design trial (CRD) using5 treatments and 3 replications. The type of treatment given is not fasting (control); middle fasting month of Hijriah for 3 days (13th, 14th and 15th); Monday and Thursday fasting; fasting Daud (a day of fasting a day is not); and fasting every day for 36 days. Male mice used in this study were Balb C strains with an initial weight of 23-26 grams and in the age range of 8-10 weeks. Calculation of the number of epididymal spermatozoa carried out according to WHO procedures in Syamrizal (1995) and morphological observations of spermatozoa carried out included normal and abnormal. Data analysis used is one-way ANOVA with a significance level of 5%. The statistical program used for the analysis was the SPSS 20 program. The results showed that fasting could increase the number and percentage of normal spermatozoa in Balb C strain mice eventhough statistically the increase was not significantly different (sig.> 0.05). Based on the type of fasting that was done, fasting on Thursday Thursday had the highest effect on the increase in the number and percentage of normal spermatozoa. Then followed by fasting every day and fasting of Daud. While the mid-fasting month of Hijriah actually decreases the number and percentage of normal spermatozoa compared to controls.

The role of flavonoid antioxidant, morin in improving procarbazine-induced oxidative stress on testicular function in rat.

Background:Procarbazine (PCZ) is an effective chemotherapeutic drug used in the treatment of lymphoma; however, oxidative stress–mediated testicular toxicity is a major side effect. Recently, therapeutic intervention using flavonoids against oxidative stress–related pathologies is gaining more attention. Morin (MOR) is a natural flavonoid with proven antioxidant activity. This study was designed therefore to evaluate the potential role of MOR in ameliorating PCZ-induced testicular oxidative stress and altered sperm quality in rat model.Methods:A total of 24 male Wistar rats (170–180 g) were randomly assigned into 4 treatment groups: I, control; II, PCZ (2 mg/kg b.w.); III, PCZ (2 mg/kg b.w.) + MOR (100 mg/kg b.w.) simultaneously administered and IV, MOR (100 mg/kg b.w.), and all treatments lasted 14 days.Results:PCZ treatment displayed significant reduction in sperm number, sperm motility, percentage normal sperm cells, and daily sperm production rate. Meanwhile the activities of testicular enzymes: gamma-glutamyl transferase, acid phosphatase, and lactate dehydrogenase were significantly altered in the PCZ group compared to control. Furthermore, PCZ caused a significant reduction in levels of glutathione and ascorbic acid as well as activities superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase in the testes of PCZ-treated rats. A significant increase in testicular malondialdehyde level was also observed in the PCZ group. MOR treatment, however, significantly restored the altered sperm parameters and biochemical markers in the testis.Conclusions:Our data suggest that MOR administration protected against PCZ-induced testicular and spermatotoxicity in rat, by improving testicular antioxidant system.

Sperm abnormalities in matrinxã Brycon amazonicus after hormonal treatment with carp pituitary extract and Ovaprim™.

SummaryThe aim of this study was to evaluate if hormonal treatment causes changes in the morphology of matrinxã Brycon amazonicus spermatozoa. Twelve males were randomized into three treatment groups: 1, Ovaprim™ (0.5 ml/kg); 2, carp pituitary extract (CPE; 2.0 mg/kg); and 3, NaCl solution 0.9% - control group (0.5 ml/kg); with four replicates. Morphological sperm analysis was performed using an optical and scanning electron microscope. The percentage of normal spermatozoa (49.6±4.6% to 60.8±2.8%), with primary (26.1±6.4% to 45.3±4.5%) and secondary abnormalities (4.4±2.9% to 13.9±3.5%) did not differ significantly between treatment groups. There were no significant differences between treatments in relation to the primary abnormalities found in the head (10.5±3.8% to 25.5±6.3%), the midpiece (0.1±0.1% to 0.2±0.2%) and in the flagellum (18.9±3.8% to 15.5±3.1%), as well as in the secondary abnormalities, located in the head (0.8±0.4% to 7.8±4.6%) and in the flagellum (2.2±1.1% to 6.1±1.5%). When each abnormality was evaluated individually, only the percentage of degenerated head was higher in the CPE-induced group (24.9±5.9%) than the control group (7.2±3.1%). We concluded that the use of pituitary extract of carp or Ovaprim™ under the conditions of this experiment does not influence the percentage of sperm abnormalities in B. amazonicus.

Sperm cryopreservation in the Far-Eastern wildcat (Prionailurus bengalensis euptilurus).

The Far-Eastern wildcat (Prionailurus bengalensis euptilurus) is a rare and poorly investigated nondomestic felid species. An attempt of freezing and cryopreserving Far-Eastern wildcat spermatozoa in CaniPlus Freeze (CPF) medium is reported. Sperm was collected by electroejaculation from five adult Far-Eastern wildcat captive-born males. Epididymal spermatozoa from five adult randomly bred domestic cat males were used as a reference. The viability of frozen-thawed spermatozoa evaluated by double staining with SYBR Green I and PI followed by the subsequent confocal laser scanning microscopy (CLSM) was 38.2%±3.0% for the domestic cat and 38.0%±10.2% for the Far-Eastern wildcat. The motility of frozen-thawed spermatozoa was 30.8%±9.8% for the domestic cat and 33.7%±15.1% for the Far-Eastern wildcat. Sperm morphology was assessed by light microscopy. The total percentage of normal spermatozoa after freezing and thawing was 51.9±5.9 for the domestic cat and 55.0%±6.4% for the Far-Eastern wildcat. Defects of flagella were the most frequently observed abnormalities in both species (32.2%±4.8% and 30.8%±4.4% of all reported anomalies for the domestic cat and Far-Eastern wildcat, respectively). Domestic cat epididymal and Far-Eastern ejaculatory spermatozoa fertilized in vitro-matured oocytes of the domestic cat (30.0%±5.5% and 35.5%±15.0%, respectively). Taken together, these results suggest that the freezing of Far-Eastern wildcat spermatozoa with CPF medium is a suitable method for Felidae cryopreservation.

SEASONAL VARIATION IN SEMINAL QUALITY IN BRAZILIAN BOCACHICO (TELEOSTEI, CHARACIFORMES)

ABSTRACT The Brazilian bocachico, Prochilodus brevis, is a rheophilic fish. Although there is evidence that this species shows reproductive seasonality in the wild, in captivity hormonal induction techniques allow semen sampling in different seasons. This study aimed to compare the kinetics, morphology and biochemical composition of the semen of Brazilian bocachico in captivity when hormonally induced to breed in the reproductive and non-reproductive seasons. During sampling spermiation was hormonally induced in breeders. The concentrations of total protein, glucose, fructose, triglyceride, calcium and chloride were evaluated with biochemical kits. The pH data (6.5 to 8.5) suggest semen requires alkaline conditions, as expected for freshwater fish. Seminal plasma contained more protein (1.51 ± 0.06 dL g-1), glucose (79.44 ± 1.88 mg dL-1) and triglycerides (61.59 ± 8.10 mg dL-1) in the non-reproductive than the reproductive season, but calcium ions (15.98 ± 1.02 mg dL-1) showed the opposite pattern. There was a significant seasonal difference in sperm morphology, with a higher percentage of normal sperm in the reproductive season. From these data it can be concluded that the physical, kinetic, morphological and biochemical characteristics of semen of captive Prochilodus brevis are influenced by reproductive season.

An illustration of human sperm morphology and their functional ability among different group of subfertile males.

Condensed sperm chromatin is a prerequisite for natural fertilization. Some reports suggested the prevalence of chromatin condensation defects in teratozoospermia cases with head anomalies; conversely, earlier studies exemplified its occurrence in morphologically normal spermatozoa too. The aim of this study was to compare the condensation defects in correlation with head anomalies among different groups of subfertile males and its impact on the rate of fertilization in assisted reproduction procedures. Ultrastructure analysis of spermatozoa through scanning electron microscopy and atomic force microscopy could facilitate an in-depth evaluation of sperm morphology. Nuclear condensation defects (%) in spermatozoa were analyzed in 666 subjects, and its effect on the rate of fertilization was analyzed in 116 IVF and 90 intracytoplasmic sperm injection cases. There was no correlation of condensation defects with head anomalies (%). Student's t-test showed no significant changes in mean values of condensation defects in abnormal semen samples in comparison with the normal group. Condensation defects were observed in normal spermatozoa too, which was negatively associated with the rate of fertilization in IVF (p<0.01), but intracytoplasmic sperm injection outcome remained unaffected. Ultrastructure study revealed sperm morphological features in height, amplitude, and three-dimensional views in atomic force microscopy images presenting surface topography, roughness property of head, and compact arrangement of mitochondria over axoneme with height profile at nanoscale. In pathological forms, surface roughness and nuclear thickness were marked higher than the normal spermatozoa. Thus, percentage of normal spermatozoa with condensation defects could be a predictive factor for the rate of fertilization in IVF. From diverse shapes of nucleus in AFM imaging, it could be predicted that defective nuclear shaping might be impeding the activity of some proteins/ biological motors, those regulate the proper Golgi spreading over peri-nuclear theca.

Technical aspects of mesenchymal stem cells transplantation into the stallion testes

Changes in anatomical, behavioural and seminal characteristics were measured throughout the year in seven Beni Arouss bucks maintained at a latitude of 35 °N. Testicular size varied significantly. There were the greatest values during the summer and autumn for scrotal circumference and least in the winter for testicular length. Values for most variables related to sexual behavior were not affected by season except the number of mounts before first ejaculation which was least in the summer. No differences were detected between seasons in terms of semen volume. The sperm concentration, viability and percentage of normal sperm, however, were less during the winter period. The size of sperm heads was larger in the autumn and motility of spermatozoa was greater in the summer and autumn. Results from the correlation analysis indicated that the majority of conventional seminal characteristics were correlated with testis measurements (P < 0.05; r < 0.5). Values for conventional and CASA motility variables indicated there were positive correlations between viability and progressive motility (r = 0.4; P < 0.05) and between the percentage of normal sperm and straight line and average path velocity (r = 0.4; P < 0.05). A global reproduction performance score was established for each buck, which allowed for assessment of magnitude of seasonal changes for each individual. These global score values indicated there was a greater reproductive performance for all bucks during summer and autumn. This study described for the first time seasonal variations of reproductive characteristics of Beni Arouss bucks and results indicate that even though there is a maximal capacity for reproductive performance during the summer and autumn, breeding should be possible throughout the year.

Semen analysis parameters from a captive population of the endangered Chacoan peccary (Catagonus wagneri) in Paraguay

Baseline information about the reproductive physiology of an endangered species is vital to captive breeding programs. This study analyzed reproductive parameters from eleven captive Chacoan peccaries (Catagonus wagneri) (mean age: 10.5 ± 1.2 years old) in Paraguay. After immobilization, testis length, width and firmness (hard to flaccid, 1–3) were assessed, followed by electroejaculation and analysis of semen. Samples were examined for percentage and progressive motility, total cell count, percentage live spermatozoa and morphology. Mean (±SEM) testis volume and firmness were 24.7 ± 1.8 cm3 and 2.1 ± 0.1, respectively. Mean ejaculate volume was 2.9 ± 0.7 ml with a pH of 7.7 ± 0.3. Few male peccaries had motile spermatozoa (n = 5/9) with a mean percentage and progressive motility of 18.3 ± 8.5% and 0.6 ± 0.3, respectively. The mean percentage of live spermatozoa was 25.1 ± 5.6%. Male peccaries had a low percentage of normal spermatozoa (12.4 ± 2.5%). The mean total count of spermatozoa per ejaculate was also quite low at 1.58 ± 1.01 million total spermatozoa per ejaculate. Spermatozoa defects were predominantly primary (77.7%) with the most common spermatozoa defects being tapered head (19.0 ± 7.4%), diadem/crater (17.7 ± 2.8%), and excess residual cytoplasm (9.6 ± 2.5%). Male age was not correlated with semen parameters (percent live: r=-0.19; motility percentage: r = 0.01; percent normal spermatozoa: r = 0.38; total count: r = 0.29; p>0.05). Evaluation of additional males from this population as well as other captive populations at various time points during the year is warranted.

Physiological variations in reproductive and metabolic features of white-tailed deer (Odocoileus virginianus) bucks throughout the rutting season

White-tailed deer farming is a growing industry in the United States, yet there is still a need to improve breeding practices in captive cervids. The objective of this study was to compare reproductive and somatic characteristics of white-tailed bucks early in the breeding season (September), at peak rut (December), and late breeding season (March). We hypothesized that reproductive parameters would improve at the peak of the breeding season. Young, mature bucks (n = 7–11, ages 2–3 yr) were anesthetized via projector with tiletamine-zolazepam and xylazine. Semen was collected by electroejaculation and evaluated for sperm output, motility, and morphology. The bulbourethral, vesicular, and prostate glands were measured by transrectal ultrasound. Body condition score, thoracic circumference, rump fat thickness, scrotal circumference, and testicular measurements were also recorded. Serum testosterone, estradiol, and IGF-I concentrations were measured. Body condition score and rump fat thickness were highest in September, intermediate in December, and lowest in March (p < 0.01), whereas thoracic circumference was lowest in September (p = 0.02). In September, serum IGF-I was highest (p < 0.01) and estradiol was lowest (p ≤ 0.02). Serum testosterone levels were lowest in March and highest in December (p < 0.01). Testicular volume and scrotal circumference were highest in September, intermediate in December, and lowest in March (p < 0.01). Bulbourethral gland length was greatest in March (p ≤ 0.05), whereas prostate length was greatest in September (p ≤ 0.02). Vesicular gland length and width were highest in December, intermediate in September, and lowest in March (p ≤ 0.02). Both overall and progressive sperm motilities were lowest in March (p < 0.01), but did not differ between September and December (p ≥ 0.12). In December, bucks had the highest percentage of normal sperm and lowest percentage of primary sperm defects (p ≤ 0.01). Collectively, our results demonstrated variation in reproductive and metabolic characteristics of white-tailed deer bucks throughout the breeding season at 40°N. The physiological variations described here will be useful for veterinarians and researchers performing reproductive evaluations in white-tailed deer bucks.

Effects of supplemental betaine to semen extenders on semen quality in boars.

Two experiments were conducted to evaluate the use of supplemental betaine in commercially available semen extenders. In experiment 1 (Exp1), semen was collected from six mature boars once weekly for 6 wk (3 wk in summer and 3 wk in winter) and diluted into a commercial extender with the following betaine concentrations: 0, 51, 102, and 205 mM. Semen samples were analyzed on the day of collection (D0) and after 72 h of storage (D3). In experiment 2 (Exp2), semen was collected from four mature boars for 3 wk and was diluted into three commercially available semen extenders (short term, ST; long term with bovine serum albumin, BSA; and long term without BSA, LT), with and without supplemental betaine (0 and 70 mM), and analyzed on D0 and D3. Semen was analyzed using computer-assisted sperm assessment (Ceros II, IMV) and morphology using phase contrast microscopy. In Exp1, total motility on D0 was less for 0 mM than that for 102 mM (P = 0.038) and was substantially reduced for 205 mM compared with 102 mM (P < 0.001). Supplementation with 205 mM betaine resulted in a significant reduction in the percentage of morphologically normal sperm (P < 0.001). In Exp2, 70 mM betaine reduced the total motility compared with 0 mM (P = 0.010) but did not impact percentage of normal sperm (P = 0.942). The use of supplemental betaine may partially alleviate the dilution effect on sperm, though boar genetics may impact its efficacy. Further research is needed to make a definitive conclusion.

Cut-off values for normal sperm morphology and toxicology for automated analysis of rat sperm morphology and morphometry

We used automated sperm morphology analysis to investigate rat sperm morphometry and morphology in Sprague-Dawley and Wistar rats in three research centers to develop normal baseline values for sperm morphometry and to quantify the percentage of morphologically normal sperm in healthy rats. The participating centers were IRSN in Paris, France (Sprague-Dawley rats), University of the Western Cape, South Africa (Wistar rats) and Stellenbosch University (Wistar rats), South Africa. All three centers used identical sperm isolation techniques from the cauda epididymis, the same staining protocols, identical computer-aided sperm morphometry analysis (CASMA) software and microscopes with similar optics. With CASMA, fully automated analysis of the different parts of stained sperm, e.g., head, acrosome, mid-piece, can be performed, many sperm morphometric features can be measured accurately and eventually normal sperm morphology can be defined. We found that it is possible to distinguish sperm morphometric characteristics of Sprague-Dawley and Wistar rats. We also developed cut-off values for evaluating the percentage normal sperm in these two rat strains using the automatic analysis mode. Normal sperm morphology varied between 67 and 74% by contrast with previous findings of > 90%.

Characteristics and fertility of domestic cat epididymal spermatozoa cryopreserved with two different freezing media

The study represents a comparison of cryopreservation of domestic cat epididymal spermatozoa with two commercially available freezing media: CaniPlus Freeze (CPF) and SpermFreeze (SF). The viability of nonfrozen spermatozoa evaluated by the VitalScreen test was 68.7 ± 3.0%. These figures were lower for the frozen-thawed spermatozoa: 51.2 ± 6.3% for CPF group and 54.4 ± 3.1% for SF group. The motility of nonfrozen spermatozoa was 57.2 ± 4.5%. These figures were reduced in both frozen-thawed groups; however, there was no significant difference in these parameters between CPF (30.8 ± 7.1%) and SF (27.4 ± 8.1%) groups. The percentage of nonprogressively moving motile spermatozoa after freezing-thawing was decreased in both frozen-thawed groups (23.5 ± 5.9 and 12.0 ± 2.4 for CPF and SF frozen correspondingly) as compared with nonfrozen controls (42.1 ± 4.1%). Morphology of spermatozoa was assessed by light microscopy. The mean percentages of normal spermatozoa were 28.5 ± 4.1% for nonfrozen group, 26.0 ± 2.3% for CPF frozen group, and 23.9 ± 1.9% for SF frozen group. The most frequent anomalies in all the three groups were flagella and combined defects. In vitro fertilization (IVF) of domestic cat oocytes with nonfrozen and frozen-thawed spermatozoa produced developing embryos. The percentage of in-vitro-derived embryos was 43.6% after using nonfrozen spermatozoa. Frozen-thawed spermatozoa developed at a similar rate (44.0%) after using SF. However, the rate of embryo development was lower (20.1%) when CPF was used. The in-vitro-derived embryos in the nonfrozen group consisted of 46.9 ± 2.5 cells after 5-day culturing. After cryopreservation with SF and CPF the cell numbers per embryo were 39.9 ± 2.7 and 31.8 ± 3.4 correspondingly. In CPF group these numbers were lower than in nonfrozen controls. Cryopreservation of spermatozoa with either of two freezing media led to a decrease in post-thaw viability and motility of spermatozoa but did not affect the rate or spectrum of their morphological anomalies. The use of CPF, but not SF led to a decrease of sperm fertilizing abilities.

46 Effect of Bioxcell® and Triladyl® Extenders on Washed Semen of South African Indigenous Bucks

Semen extenders and seminal plasma are vital for cryopreservation of buck semen. The objectives of the study were to evaluate the effect of 2 extenders: Triladyl® (Minitube, Tiefenbach, Bavaria) and Bioxcell® (IMV, L’Aigle, France) and the removal of seminal plasma on buck semen. Six indigenous bucks were used in this study and 6 ejaculates were collected from individual bucks. The semen was pooled and then randomly allocated into 6 groups: (1) raw-washed, (2) raw-non-washed, (3) Triladyl®-washed, 4) Triladyl®-non-washed, (5) Bioxcell®-washed, and (6) Bioxcell®-non-washed. Spermatozoa viability was assessed using Eosin-Nigrosin and morphology using Spermac® (Vitrolife, Göteborg, Sweden) stains. The washed semen samples were all diluted into (1:4 v/v) with PBS and centrifuged at 1500 × g for 10 min. Semen samples were then extended with Triladyl® or Bioxcell® per treatment groups and equilibrated for 2 h at 5°C. The semen samples were loaded into straws per treatment groups and placed 5 cm above a liquid nitrogen vapour for 10 min and then stored at –196°C until use. After 1 month of storage, frozen semen straws per treatment group were thawed at 37°C for 30 s, and spermatozoa parameters were analysed post-thaw. Significant differences among the mean values of semen parameters were determined by Tukey’s test using ANOVA, GLM procedure of SAS version 12.1 of 2010 (SAS Institute Inc., Cary, NC, USA). There was a higher (P &lt; 0.05) live and normal spermatozoa percentage in non-washed semen extended with Bioxcell® (45.7 ± 21.2) than the semen extended with Triladyl® (24.5 ± 22.2%). Live and normal spermatozoa percentages were drastically reduced in the Bioxcell® (5.2 ± 4.9) and Triladyl® (6.9 ± 8.6%) washed semen groups. There was a higher (P &lt; 0.05) percentage of spermatozoa with head abnormalities in non-washed semen extended with Triladyl® (20.4 ± 10.2), compared with the semen extended with Bioxcell® (18.3 ± 12.4%) following freeze-thawing. There was a higher (P &lt; 0.05) percentage of spermatozoa with head abnormalities in washed semen samples extended with Triladyl® (34.0 ± 16.0) compared with the semen extended with Bioxcell® (10.1 ± 7.0%). There were higher (P &lt; 0.05) percentages of spermatozoa with coiled tail abnormalities in washed semen extended with Bioxcell® (65.4 ± 25.0) compared with Triladyl® (35.9 ± 21.6%). In conclusion, the liveability of spermatozoa was negatively affected by washing of semen extended with Bioxcell® and Triladyl® extender. Bioxcell® significantly increased tail abnormalities and Triladyl® gave less protection against head abnormalities following cryopreservation of South African unimproved indigenous bucks’ semen.

94 Semen Parameters from a Captive Population of the Endangered Chacoan Peccary (Catagonus wagneri) in Paraguay

Priority conservation actions for the endangered Chacoan peccary (Catagonus wagneri) include improvement of captive breeding initiatives. Eleven captive male peccaries housed in mixed-sex groups of 17 to 35 animals at the Centro Chaqueño para la Conservación e Investigación in Paraguay were immobilised with intramuscular tiletamine/zolazepam (30 mg; Zoletil® 50, Virbac, Forth Worth, TX, USA), azaperone (10 mg; 50 mg mL−1; ZooPharm, Windsor, CO, USA), and medetomidine (1 mg;10 mg mL−1; ZooPharm). Animals were administered supplemental intranasal oxygen (1-2 L min−1) and reversed with atipamazole (5 mg; 25 mg mL−1; ZooPharm). Testis length, width, and firmness (1-3 scale; 1 = hard, 2 = normal, 3 = flaccid) were recorded. The preputial sheath was clipped, cleaned, and flushed with sterile saline using a Foley catheter. A lubricated 13-mm-diameter electroejaculator probe (Beltron Instruments, Longmont, CO, USA) was inserted into the rectum to a depth of 5 cm. The electroejaculation protocol consisted of 4 cycles of 10 stimuli each (1-4 V/cycle). Collection tubes were changed frequently during electroejaculation to avoid urine or gel fraction contamination of the sperm-rich fraction. Collected semen was evaluated for volume, color/appearance, and pH. Percent motility was determined as the average motility of spermatozoa from 10 fields (40×). Progressive motility of spermatozoa was scored on a scale of 0-5 (0 = dead sperm; 5 = rapid forward progression). Semen samples were assessed for total spermatozoa. Percent of live spermatozoa and spermatozoa morphology were evaluated with Eosin-Negrosin smears. The mean ± standard error of the mean of each parameter is presented. A Pearson correlation coefficient between age and semen parameters was calculated. Males assessed in this study were 10.5 ± 1.2 years old. The mean testis volume was 1005.7 ± 50.9 cm3 and mean testis firmness was 2.1 ± 0.1. The mean ejaculate volume was 2.9 ± 0.7 mL and pH was 7.7 ± 0.3. Few males had motile spermatozoa (n = 5/9), which yielded a mean percent motility of 18.3 ± 8.5 (range: 0-50%) and a mean progressive motility of 0.6 ± 0.3 (range: 0-2). The mean percentage of live spermatozoa was 25.1 ± 5.6 (range: 4-44%). Male Chacoan peccaries in this population had a low percentage of normal spermatozoa (12.4 ± 2.5%; range: 2.8-24.0%). The mean total count of spermatozoa per ejaculate was 1.58 ± 1.01 million (range: 0.09-8.28 million). Spermatozoa defects were predominantly primary (67.1%), with the most common defects being tapered head (15.4 ± 6.2%), diadem/crater (14.8 ± 2.6%), and excess residual cytoplasm (8.5 ± 2.7%). There was no correlation between age and semen parameters (percent live: r = –0.19; motility percentage: r = 0.01; percent normal spermatozoa: r = 0.38; total count: r = 0.29; P &gt; 0.05). This is the first report of semen parameters in the Chacoan peccary. It is unknown whether the poor semen quality is inherent to the species or this population. Reproductive assessment of males from other captive populations and of wild male Chacoan peccaries is warranted.

Influenza outbreak causes reduction in semen quality of boars

An influenza outbreak occurred at Purdue University’s swine barn, resulting in infection of 28 boars with influenza A virus (H3N2) and causing the death of two boars. The 28 boars, approximately 35 weeks of age, were enrolled in a study at the time of the outbreak and the case report herein describes the effects of the unintended influenza outbreak on sperm production. Semen was collected from the boars once a week and evaluated for total sperm production and concentration, semen volume, and relative motility. Compared to previous collections, total sperm production was substantially decreased (26% reduction) approximately 4 weeks after the first observed clinical signs and remained low for 6 subsequent weeks. Semen production then returned to pre-outbreak levels and was maintained for the duration of the observation period. Sperm motility and percent normal sperm production were also slightly reduced 2 weeks after infection