Freezing equine semen: the effect of combinations of semen extenders and glycerol on post-thaw motility

Objective   We evaluated combinations of two commercial semen extenders and three concentrations of glycerol to determine the combination that yielded the highest post-thaw sperm motility.
Design   A randomised 2 × 3 block design was used.
Procedure   Semen was collected from four stallions (6 collections per stallion). The sample was diluted with either a dried skim-milk glucose extender (EZ Mixin Original Formula) or a chemically defined, milk-free diluent (INRA 96), and each was used in combination with 2%, 3% or 4% glycerol in standard commercial freezing medium. Sperm motility was assessed by microscopy in fresh and post-thaw semen.
Results   There was a significant difference between the two extenders in the motility of spermatozoa after cryopreservation (48.9% for INRA 96; 38.6% for EZ Mixin OF; P < 0.0001). Glycerol at 4% in freezing medium yielded the highest post-thaw motility, significantly better than 2% ( P < 0.05). Three of four stallions had significantly higher post-thaw motility using INRA 96 relative to EZ Mixin OF ( P < 0.01), and two of four stallions had significantly higher post-thaw motility using 4% glycerol ( P < 0.05). The combination of INRA 96 and 4% glycerol in freezing medium gave the highest average post-thaw motility of 51.5%.
Conclusion   In this study, INRA 96 combined with 4% glycerol yielded an average recovery of progressively motile sperm consistently above the 35% target.     

Nitrogen uptake and allocation by field‐grown ‘Arapaho’ Thornless blackberry

Spring surface applications of ¹⁵nitrogen (N)‐labeled urea were made in March 1995 and 1996 on one and two‐year‐old, field‐grown ‘Arapaho’ blackberry (Rubus subgenus Rubus) plants. Individual whole‐plant samples were collected at pre‐harvest (late May), post‐harvest (mid‐July), and early dormancy (late October). Plant parts separated for analysis were roots, primocanes, floricanes, primocane leaves, floricane leaves; and fruits. Soil samples were also taken from within the plant drip line at each sample date. Plant tissues were washed, dry weights measured, ground for acid digestion, total N determined, and ¹⁵N analyzed. Plants collected in October had more total dry matter, with roots, primocanes, and primocane leaves contributing most to this total. Total N content decreased in all vegetative tissues (leaves and canes) from May to October. Compared to other plant tissues, floricane leaves and primocanes recovered significantly more fertilizer N in May while roots and primocane leaves contained more in October of each year. Floricanes and fruits did not increase in fertilizer N levels during the sampling period. Overall, the lowest percent fertilizer N recovery for whole plants was 12.8% for May 1995 and the highest was 32.4% for October 1996. Recovery of fertilizer N in the topsoil ranged from a low of 12.9% in October 1995 to a high of 38.6% in May 1996. There were no statistical differences in percent recovery of fertilizer N from topsoil among sample dates. Recovery of fertilizer N from subsoil in the October sample was much lower than that in May, probably due most to plant uptake, but also possibly to a downward movement out of the sample area by leaching or from other loss mechanisms. Averaging all sample dates, recovery was 59.5% in 1995 and 75.8% in 1996 for the plant and soil combined, with the remainder probably lost via volatilization, leaching, and/or denitrification.     

Agreement among three different equine venipuncture sites with regard to measurement of packed cell volume and total solids

The primary objective of this study was to determine the agreement between the packed cell volume (PCV) and total solids (TS) values in blood collected from the jugular vein (JV) in comparison with the cephalic vein (CV) and the transverse facial venous sinus (TFVS) in healthy adult horses. A total of 72 samples were taken from 24 horses. We found high agreement that reflects no clinically relevant differences between the PCV and TS values obtained from the CV or the TFVS in comparison with the JV in standing healthy adult horses.     

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.     

Advancements in Large Animal Embryo Transfer and Related Biotechnologies

Embryo transfer has been an inherent part of cattle breeding for more than 35 years and has also gained remarkable interest from the equine industry after several breeds allowed registration of more than one foal per year. In both large animal species, non‐surgical embryo recovery and transfer are well‐established techniques. However, success rates after superovulation and cryopreservation of embryos in horses are still lagging behind those of cattle, and more research is needed to address these areas. To address the problem of freezing large equine embryos, we offer a preliminary demonstration of a new cryopreservation method which involves reduction of the blastocoelic volume and microinjection of cryopreservative. Successful cryopreservation will improve the ability of practitioners to preserve and implant embryos in recipient mares. Recent advances in the use of equine FSH to induce superovulation in mares brings to the forefront the issue of how to best preserve the large number of embryos that are produced. Finally, the use of sexed semen after superovulation will provide the bovine and equine breeding industry the offspring of the desired sex.     

Assessment of Orchidometry and Scrotal Circumference in Coatis (Nasua nasua)

This study describes the morphometry of the testicles of captive coatis (Nasua nasua) and verifies the existence of correlations between the testicles and the semen producing characteristics. Twelve ejaculates were obtained from five adult specimens that were restraint by anaesthesia. Visual and morphometric evaluation of the external genitalia was performed by using a caliper. The animals presented a scrotal circumference of 9.4 ± 0.7 cm. The right testicle presented a width of 1.7 ± 0.2 cm, length of 2.6 ± 0.2 cm and volume of 3.9 ± 1.1 cm³. The left testicle presented a width of 1.7 ± 0.1 cm, length of 2.5 ± 0.3 cm and volume of 3.6 ± 0.7 cm³. No significant correlations were obtained between the scrotal circumference or total testicular volume and the semen producing characteristics (volume and sperm concentration). For the first time in this field, this study presents an important contribution to the knowledge of the reproductive morphophysiology of coatis.     

Influence of environmental temperature on experimental infection of redfin perch (Perca fluviatilis) and rainbow trout (Oncorhynchus mykiss) with epizootic haematopoietic necrosis virus, an Australian iridovirus

SUMMARY Experimental transmission of epizootic haematopoietic necrosis virus (EHNV) to adult redfin perch Perca fluviatilis and juvenile rainbow trout Oncorhynchus mykiss was undertaken at different water temperatures using intraperitoneal (IP) and bath inoculation. Redfin perch were highly susceptible to EHNV by both routes of infection. Bath inoculation with as few as 0.08 TCID₅₀. _mL^-1 was lethal. The incubation period in redfin perch was about 11 days at a water temperature of 19–21°C but was longer at colder temperatures and disease did not occur at temperatures below 12°C. The longest incubation period recorded in redfin perch was 28 days. Rainbow trout were not susceptible to infection by bath inoculation but the disease was reproduced after IP inoculation with 10^5.6 TCID₅₀ at water temperatures ranging from 8–21°C. The incubation period was 3–10 days at 19–21°C, but was up to 32 days at 8–10°C. Persistent infection with EHNV was detected by virus isolation in a clinically unaffected rainbow trout after 63 days. The implications of these findings in the understanding of the epidemiology of EHNV infection are discussed.     

Effects of alternate-furrow irrigation: Water conservation on the yields of two soybean cultivars

A 4-year (1980–1983) field study was conducted at the South Central Research Station, Chickasha, OK on a McLain silty clay loam (Fine, Mixed, Thermic, Pachic Argiustoll) to evaluate agronomic characteristics and yields of Essex and Sohoma soybeans (Glycine max (L.) Merr.) when irrigated by alternate- and every-furrow irrigation methods.When averaged over 4 years, Essex yielded 1983 and 2338 kg ha^?1 compared to 2128 and 2595 kg ha^?1 for Sohoma when both soybean cultivars were alternate-furrow and every-furrow-irrigated, respectively. The lower yields for the alternate-furrow irrigation treatment can be attributed to a smaller number of pods, less seeds, and smaller seed weight for both cultivars. Although, yields of soybeans grown on a fine-textured soil and alternate-furrow-irrigated were statistically significantly lower, acceptable yields were still produced with 40–50% less supplemental irrigation water. The magnitude in water savings with alternate-furrow irrigation and the resulting lower yields seems to be acceptable trade-offs and should have application for much of the subhumid and/or semiarid regions of the world.