Hyperactivated Sperm Motility: Are Equine Sperm Different?

A recent study has suggested a central role for hyperactivated sperm motility in successful equine in vitro fertilization, as the highest apparent fertilization rates reported yet were obtained using exogenous stimulation of hyperactivated motility in sperm that had been incubated for capacitation. Hyperactivated motility has been studied intensively in other species, but little data are available in this area in the horse. Hyperactivated motility is associated with an influx of calcium from the extracellular space, and in other species this occurs because of an increase in intracellular pH during capacitation. Influx of calcium seems to trigger changes in flagellar function through calcium–calmodulin–calmodulin kinase-related effects on dynein function, but the exact mechanisms resulting in hyperactivated motility are still unclear. We have been exploring the physiology of hyperactivated motility in stallion spermatozoa and have found many of the mechanisms present in other species to be in place in the stallion. Further research is needed to determine why stallion spermatozoa seem to fail to undergo hyperactivation in response to capacitating environments that support this activity in other species.     

Addition of Seminal Plasma to Post‐thawing Equine Semen: What is the Effect on Sperm Cell Viability?

Effect of seminal plasma addition after thawing on viability or cryocapacitation is not definitively established. This experiment was performed to verify the effect of adding seminal plasma, autologous or homologous (from an animal with good semen freezability). Five ejaculates from each of four stallions with proven fertility were collected and cryopreserved. The semen was subsequently thawed and divided into the following three treatment groups: no seminal plasma addition after semen thawing (NOSP); the addition of homologous seminal plasma after semen thawing (HSP) and the addition of autologous seminal plasma after semen thawing (ASP). The addition of 20% of seminal plasma led to an increase in the cell population that simultaneously show plasma and acrosomal membrane integrity (p < 0.05). The addition of seminal plasma did not alter the total motility, the amount of cells with mitochondrial membrane potential or the sperm velocities (average path velocity, straight-line velocity and curvilinear velocity). However, the beat/cross-frequency, straightness and linearity were reduced in ASP and HSP groups (p < 0.05). Unexpectedly, the addition of homologous seminal plasma reduced the proportion of cells with progressive motility (p < 0.05) and the addition of autologous seminal plasma reduced the amplitude of the lateral head displacement (p < 0.05). Based on the increase in the cell populations that had the plasma and acrosomal membrane integrity simultaneously identified in this study, we proposed that the addition of seminal plasma (autologous or homologous) into post-thawed semen before insemination could increase semen fertility.     

Effect of Freezing Rates and Supplementation of α-Tocopherol in the Freezing Extender in Equine Sperm Cryosurvival

This study was conducted to determine the impact of antioxidant (α-tocopherol) supplementation in the cryopreservation extender and three different freezing rates (FRs) on quality of post-thaw semen to elaborate a new protocol for stallion semen cryopreservation. Six ejaculates from each of four stallions were subjected to cryopreservation with a commercial extender (Gent, Minitub Iberia, Spain), without any supplementation (control) or supplemented with 2 mM α-tocopherol. The semen was exposed to three different freezing rates between 5°C and −15°C: slow (5°C/min), moderate (10°C/min), and fast (20°C/min). After thawing, the sperm viability (Sybr-14 and propidium iodide [PI]), mitochondrial membrane potential (MMP) (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine), lipid membrane peroxidation (LPO; C₁₁-BODIPY^581/591), and apoptosis status (fluorescein isothiocyanate–conjugated annexin V and PI) of the plasmatic membrane of each sample were determined by flow cytometry. For both extenders, the percentage of viable cells was higher for spermatozoa cooled at 5°C/min than at 10°C/min and 20°C/min (P ≤ .05). The FR of 20°C/min demonstrated a lower value of MMP than the FR of 5°C/min and 10°C/min (P ≤ .05). The α-tocopherol extender improved (P ≤ .05) post-thaw membrane LPO; however, it did not improved viability and the apoptosis status of the sperm plasmatic membrane after thawing. In conclusion, results clearly indicate that the cryosurvival of stallion spermatozoa is improved when FR of 5°C/min is used from 5°C to −15°C.     

Effect of Fatigue on Equine Metacarpophalangeal Joint Kinematics—A Single Horse Pilot Study

The objective was to validate a scientific method for characterizing equine metacarpophalangeal joint (MCPJ) motion in the nonfatigued and fatigued states using a single horse at trot, slow canter, and fast canter. One healthy Thoroughbred gelding exercised on a treadmill to exhaustion (fatigued state) (heart rate >190 BPM and blood lactate >10 mmol/L) while bilateral MCPJ angular data were acquired using electrogoniometry. Blood lactate and heart rate reflected transition from nonfatigued to fatigued states with increasing exercise duration and treadmill speed. Electrogoniometry consistently demonstrated: increase in mean MCPJ maximum extension angle with onset of fatigue; altered extension and flexion angular velocities with onset of fatigue; and increasing stride duration and decreasing stride frequency with onset of fatigue. The method allowed a preliminary but comprehensive characterization of the dynamic relationship between MCPJ kinematics and fatigue, prompting the need for multisubject studies that may enhance our ability to moderate exercise-related distal limb injury in equine athletes.     

The Effect of Coenzyme Q10 and α-Tocopherol in Skim Milk–Based Extender for Preservation of Caspian Stallion Semen in Cool Condition

The purpose of this study was to determine the effects of different concentrations of coenzyme Q₁₀ (CoQ₁₀) and α-tocopherol (T) along with their interaction effects on the quality of preserved stallion semen at 5°C for a period of 48 hours. Semen was collected and diluted with skim milk–based extender that was supplemented with different antioxidants: no antioxidant (negative control [NC]), 0.9% (vol/vol) dimethyl sulfoxide (positive control [PC]), α-tocopherol (5 [T5] or 10 [T10] mM), CoQ₁₀ (1 [C1] or 2 [C2] μM), 1 μM CoQ₁₀ + 5 mM α-tocopherol (C1T5), 1 μM CoQ₁₀ + 10 mM α-tocopherol (C1T10), 2 μM CoQ₁₀ + 5 mM α-tocopherol (C2T5), and 2 μM CoQ₁₀ + 10 mM α-tocopherol (C2T10), then kept at 5°C. The results showed that C1 extender resulted in higher total motility (62.44 ± 3.82) and plasma membrane integrity (65.16 ± 3.63%) compared with NC after 48 hours of storage (P < .05). Different concentrations of α-tocopherol had no significant effects on sperm quality, with the exception of plasma membrane integrity, compared with NC and PC extenders (P > .05). Also, C1T5 extender improved total and progressive motility, plasma membrane integrity and functionality, and decreased lipid peroxidation compared with NC and C2T10 extenders over 48 hours of storage at 5°C (P < .05). The C1T5 extender was similar to C1 and T5 extenders in all semen parameters evaluated during storage time. In conclusion, between previously mentioned extenders, C1T5 could improve stallion sperm quality during 48 hours of storage. In the present study, none of extenders had effect on sperm quality until 24-hour storage.     

Does the Microbial Flora in the Ejaculate Affect the Freezeability of Stallion Sperm?

In an attempt to evaluate the possible relationship between the microbial flora in the stallion ejaculate and its ability to freeze, three ejaculates from five stallions were frozen using a standard protocol. Before freezing, an aliquot was removed for bacteriological analysis. Bacterial growth was observed in all the ejaculates studied. The isolated microorganisms were: Staphylococcus spp. and Micrococcus spp. (in all the stallions), β-haemolytic Streptococcus (in stallions 3 and 4), Corynebacterium spp. (in stallions 1, 3–5), Rhodococcus spp. (in stallion number 2), Pseudomonas spp. (in stallion number 1) and Klebsiella spp. (in stallions 1, 3 and 5). The presence and richness of Klebsiella and β-haemolytic Streptococcus in the ejaculate were related to two sperm variables post-thaw, namely the proportion of dead spermatozoa (ethidium+ cells; r = 0.55, p < 0.05) and the amplitude of lateral displacement of the sperm head (ALH, μm; r = −0.56, p < 0.05), respectively. The degree of growth of Corynebacterium spp. in the ejaculate was positively correlated with the percentage of spermatozoa showing high caspase activity post-thaw (r = 0.62, p < 0.05). The presence and number of colonies of β-haemolytic Streptococcus were negatively correlated (r = −0.55, p < 0.05) with low sperm caspase activity. It is concluded that the microbial flora of the equine ejaculate may be responsible for some of the sublethal damage experimented by the spermatozoa during cryopreservation.     

Effect of β-carotene Supplementation on Italian Trotter Mare Peripartum

When the mare’s estrous cycle resumes in winter, the β-carotene content of hay is depleted. Sixty Italian trotter mares were randomly assigned to a Control or a Treated Group. Treated Group received 1g/d synthetic β-carotene for 15 days from parturition. Blood samples collected at parturition and on days 5, 10 and 15 after partum were analysed for β-carotene, vitamins A, progesterone, 17 β-estradiol, the energy parameters (glucose, cholesterol, NEFA), the protein profile (total protein, albumin, urea) and LDH. Some changes in these measures were attributable to treatment, which significantly affected β-carotene and 17 β-estradiol concentrations. A significant effect was also found on the resumption of estrous activity (χ² test=P<0.052).     

Effect of Biodegradable Gelatin β-Tri Calcium Phosphate Sponges Containing Mesenchymal Stem Cells and Bone Morphogenetic Protein-2 on Equine Bone Defect

This study evaluated the osteoanagenetic effects of administering biodegradable gelatin β-tri calcium phosphate sponges containing mesenchymal stem cells (MSCs) and bone morphogenetic protein-2 (BMP-2) on equine bone defect. Six healthy Thoroughbred horses were used in this study. Horses were anesthetized, and skin incisions were made on all the limbs. Splint bones were exposed and a 1-cm bone defect was created in each exposed bone. Gelatin β-tri calcium phosphate sponges containing MSC and BMP-2 (MSC + BMP-2 sponge), MSC only (MSC sponge), BMP-2 only (BMP-2 sponge), or saline (saline sponge) were implanted into each bone defect at random. Defects were monitored for 16 weeks by radiography followed by computed tomography (CT) and histologic analyses. At 16 weeks, radiographic scores of MSC + BMP-2 sponge-treated defects were significantly higher than those of saline-treated defects (P = .027). Moreover, the CT value of the MSC + BMP-2 sponge group was significantly higher than that of the other groups (P = .027; P = .046; and P = .027, respectively), and the histologic score of the MSC + BMP-2 sponge group was significantly greater than that of the saline sponge group (P = .041). We conclude that MSC + BMP-2 sponge administration to bone defects accelerates bone regeneration in equines.     

Does meatiness of pigs depend on the level of gastro-intestinal parasites infection?

The aim of the present paper was to determine an influence of the presence and a level of intestine parasites infection on the quality of pork carcass expressed by the content of meat in carcass (meatiness) in pigs. The experimental part of the study was conducted on pigs farm produced in a closed cycle. The population in the study included 120 fattening pigs maintained in two keeping systems: group I--60 individuals kept on slatted floor, and group II--60 individuals kept on deep litter. All the experimental animals were treated in the same manner. The analysed fatteners were slaughtered in Meat Processing Plant when their body mass reached 110 kg, and the post-slaughter assessment was conducted according to the EUROP classification of pigs carcass using the Ultra-Fom 300 device. The study concerning the internal parasites were conducted basing on coproscopic quantitative McMaster method. As a results, the eggs of three nematode taxa were isolated and identified: Oesophagostomum spp., Ascaris suum and Strongyloides ransomi. Overall prevalence of infection of fatteners kept on litter was lower (25%±11.2) as compared to those kept on slatted floor (38.3%±12.6), however the differences were not statistically significant (χ(2)=2.465; df=1; P=0.116). The mean value of meatiness for pigs free from parasites was 53.68, while in the case of infected pigs the meatiness was statistically lower and was 52.12 (t=2.35; P=0.02). The analysed pigs were classified into three categories and conducted analysis of an influence of parasites on meatiness demonstrate the relationship that is statistically significant. The analysis of correlation between meatiness and an average number of helminth eggs also demonstrated the negative, statistically significant, relationship (F=5.52; P=0.020), i.e. in fatteners with higher EPG value the meatiness was lower.     

Does Age Matter? The Influence of Age on Response Rates in a Mixed-Mode Survey

The appeal of cost savings and faster results has fish and wildlife management agencies considering the use of Internet surveys instead of traditional mail surveys to collect information from their constituents. Internet surveys, however, may suffer from differential age-related response rates, potentially producing biased results if certain age groups respond to Internet surveys differently than they do to mail surveys. We examined this concern using data from a mixed-mode angler survey conducted in South Dakota following the 2011 fishing season. Results indicated that young anglers (16–18) had the lowest return rates and senior anglers (65+) had the highest, regardless of survey mode. Despite this consistency in response rates, we note two concerns: (a) lower Internet response rates and (b) different age groups represented by the Internet and mail survey samples differed dramatically. Findings indicate that constituent groups may be represented differently with the use of various survey modes.     

Effect on fertility of storing turkey semen for 24 hours at 10 °C in fluids of different pH

1. A study was undertaken into the storage of turkey semen in vitro for 24 h.

2. Turkey semen storage for 24 h at 10 °C was not noticeably improved by using a buffered diluent compared with an unbuffered (pH 7.3) diluent. This is different from the case of the fowl and probably reflects differences in the metabolism of spermatozoa between the species.

3. Adverse effects on the fertilising ability of turkey spermatozoa were observed with diluents buffered around pH 7.4 and 5.8 when compared with the use of diluent either buffered at pH 7.1 or unbuffered.     

A Systematic Review and Meta-analysis on Sodium Bicarbonate Administration and Equine Running Performance: Is it Time to Stop Horsing Around With Baking Soda?

Sodium bicarbonate administration in the hours prior to exercise has been used as a performance-enhancing substance in horses since the late 1980s. Although sodium bicarbonate administration to racehorses 24 hours before racing is a banned practice in most racing industries, whether or not it improves running performance in racehorses is currently unclear. The aim of this systematic review and meta-analysis was to establish whether or not acute sodium bicarbonate administration improves running performance in trained Standardbred and Thoroughbred horses. Seven randomized controlled trials, including eight experimental (exercise) trials and 74 horses, were included after a comprehensive search for relevant studies that met the inclusion criteria. Results indicated that sodium bicarbonate administration at 2.5–5 hours prior to a standardized treadmill test to exhaustion or simulated race (time-trial) does not influence running performance (number of horses, the overall effect [95% CI]: 32, –0.13 [–0.64 to 0.37] and 42, 0.01 [–0.42 to 0.44], respectively, both P > .05). The included studies demonstrated minimal heterogeneity (I² = 0%–2%), low risks of bias according to the Cochrane Risk of Bias Tool and a lack of publication bias. On the basis of these findings, there is high-quality evidence to suggest that sodium bicarbonate administration does not improve running performance in trained Standardbred or Thoroughbred horses.