Testicular Volume and Daily Sperm Output in Ragusano Donkeys

A careful evaluation of daily sperm output (DSO) and seminal parameters is indicative of the potential fertility of the animal and at the same time is useful for maximization of the reproductive activity. The aim of this article was to evaluate the seminal parameters, testicular volume (TV) calculated by ultrasound measurements, DSO, and any eventual relationship between these in Ragusano donkey jackasses. Eight Ragusano donkey stallions underwent a morphometric evaluation of the testes by ultrasound to calculate the TV, and daily semen collection for 10 consecutive days. TV ranged from 250 to 500 cm³ and no significant differences could be observed between the left and right testes. Only ejaculates from 4 to 10 days were analyzed, given that there was evidence that the sperm extragonadal reserve interfered with the total number of sperm in ejaculates during the early days. DSO in the Ragusano donkey ranged from 7 to 23 × 10⁹ sperms. The seminal parameters showed high values of motility, viability, and low abnormality percentages. Age, TVs, and sperm parameters were not significantly correlated. The correlation between TV and DSO was poor (P = .1) in this study and this may suggest that TV and DSO might simply not be highly correlated in the donkey.     

A Comparison of Actual and Suggested Digestible Energy Intakes of Miniature Horses Derived from Different Feeding Recommendations

Despite being largely studied as models of equine nutrition, few published data exist on actual nutritional requirements of Miniature horses. Small equines are particularly prone to obesity, insulin resistance, and the metabolic disorders for which these are risk factors. Improper feed rationing, overfeeding in particular, puts horses at risk for these metabolic problems. Feed manufacturers, however, generally supply feeding recommendations for horses weighing ≥360 kg, whereas the average Miniature horse weighs around 100 kg. Lack of nutritional guidelines for small equines makes it difficult for owners to properly estimate the amount of hay and grain to feed their Miniature horse. The purpose of this study was to determine whether feeding recommendations provided by feed manufacturers for average-sized horses could be linearly extrapolated to Miniature horses. This study surveyed nutritional management of 12 mature Miniature horses that represented the ideal body condition of the breed. Each horse’s hay, grain, and pasture intake was recorded and the average daily digestible energy (DE) intake was calculated from those data. The daily DE intake of the surveyed horses was compared with the extrapolated recommendations published by several different sources. When linearly extrapolated to fit the body weight of a Miniature horse, recommendations significantly (P < .002) misestimated the amount of daily DE that the surveyed horses were consuming. The majority of recommendations overestimated the DE consumed by the Miniature horses per day. However, one source that was designed specifically for Miniature horses, severely (P < .001) underestimated their required DE.     

Genetic-diet interactions in the Hyperkalemic Periodic Paralysis syndrome in quarter horses fed varying amounts of potassium: IV. Pre-cecal and post-ileal absorption of potassium and sodium

During an experiment to determine the genetic-diet interactions in genetically predisposed Quarter Horses, biphasic symptoms of Hyperkalemic Periodic Paralysis (HYPP) were observed when the HYPP H/N broodmares were fed the higher potassium rations. ¹ The cause of those symptoms was assumed to be: 1) the difference in absorption times of the highly absorbable potassium citrate in the concentrates and the potassium contained in the cells of the grains and hays in the rest of the rations, 2) partial absorption of potassium in the small intestine followed by no absorption from the cecum and further absorption in the large colon or 3) a combination of both of the above explanations. Since previously cannulated ponies were available in this laboratory, an experiment was designed to determine which of those possibilities was the most likely.

Three ponies, weighing an average of 132 kg, with posterior-ileal cannulas were used in a 3×3 Latin square experiment to determine the amounts of potassium and sodium that would be absorbed pre-cecally and post-ilealy. The ponies were fed the same rations as the mares in the previously described experiment (see part I), which provided approximately 1.1, 1.9 and 2.9% potassium by weight as fed. Samples of water, feces, ileal contents and blood were collected. Water, feed, ileal and fecal samples were analyzed for sodium and potassium concentration.

Apparent absorption of potassium prior to the cecum was 97% when the ponies were fed the low potassium diet and 98% when they were fed the higher potassium diets. Three percent and 2% of the potassium was absorbed from the hindgut when the ponies were fed the low potassium diet and the higher potassium diets, respectively. Nine percent and 16% of sodium was absorbed pre-cecally when the ponies were fed the low potassium diet and the higher potassium diets, respectively. Ninety-one and 84% of the sodium was absorbed from the hindgut, when the ponies were fed the low potassium diet and the higher potassium diets, respectively for 99–100% total absorption of both sodium and potassium from all diets.

These results indicate that the potassium from the three diets was absorbed pre-cecally, and that the process took up to nine hours, resulting in bi-phasic HYPP symptoms in the HYPP H/N mares in the companion experiment. This information can be used to better understand the relationship between dietary potassium, plasma K⁺ concentration and HYPP symptoms.     

Oestrous Behaviour and Timing of Ovulation in Relation to Onset of Oestrus and LH Peak in Mithun (Bos frontalis) Cows

The objectives of the study were to evaluate the oestrus behaviour and to determine the timing of ovulation in relation to onset of oestrus and the pre-ovulatory LH surge in mithun (Bos frontalis). For this purpose, the blood samples collected at 15-min intervals for 9 h period following onset of oestrus and thereafter, at an interval of 2 h till 4 h post-ovulation for three consecutive cycles from 12 mithun cows were assayed for plasma LH and progesterone. Ovulation was confirmed by palpation of ovaries per rectum at hourly intervals. Various signs of behavioural oestrus were also recorded. The common signs of oestrus and their frequency of occurrence in mithuns were following and mounting by male mithuns (100%), standing to be mounted (100%), frequent urination (62.33%), raising of tail (65.23%), swelling of vulva (54.26%) and congestion of vulvar mucous membrane (69.87%). The pre-ovulatory LH surges consisted of several pulses (2.92 +/- 0.26 pulses/animal; range, 1-4). The mean (+/-SEM) peak level of LH for individual mithun varied from 6.99 +/- 0.44 to 12.69 +/- 2.10 ng/ml and the mean pooled LH peak concentration was 9.10 +/- 0.60 ng/ml. The highest peak (highest amplitude of LH during LH surge) was 10.83 +/- 0.76 ng/ml (range, 8.07-16.49 ng/ml). The duration of LH surge was 6.98 +/- 0.22 h (6-8 h). Onset of LH surge was at 1.23 +/- 0.17 h post-oestrus onset (range, 0.25-2.25 h). Mean plasma progesterone stayed low (<0.24 ng/ml) during the entire duration of sampling. Ovulation occurred at 26.92 +/- 0.31 (range, 26-29 h) after the onset of oestrus and 18.63 +/- 0.35 h (range, 17-20.75 h) after the end of LH surge. The occurrence of the highest LH peaks within a narrow time frame of 2- to 5-h post-oestrus onset in mithuns could have contributed to the animals ovulating within a narrow time interval. These results are very promising from a practical standpoint of potential success when AI program in this species is implemented in a big way. Furthermore, the results of the occurrence of LH pulses during pre-ovulatory LH surges, which are required for ovulation in this species of animals, is unique and species specific.     

Determination of cardiac output in neonatal foals by ultrasound velocity dilution and its comparison to the lithium dilution method

Objective – To compare cardiac output (CO) measured by use of lithium dilution (LiDCO) and ultrasound velocity dilution (UDCO) in conditions of high, intermediate, and low CO in anesthetized foals.
Design – Original prospective study.
Setting – University teaching hospital.
Animals – Six foals 1–3 days of age (38–45 kg).
Interventions – Neonatal foals were anesthetized and instrumented to measure direct blood pressure, heart rate, arterial blood gases, and CO. The CO was measured by use of LiDCO and UDCO techniques. Measurements were obtained from each foal at baseline and during low, intermediate, and high CO states. Measurements were converted to cardiac index (cardiac index=CO/body weight) values for statistical analysis. Agreement between the 2 methods was determined using Bland and Altman analysis and concordance correlation coefficients.
Measurements and Main Results – LiDCO determinations of CO ranged between 4.0 and 14.0 L/min resulting in cardiac index ranging between 75.5 and 310 mL/kg/min. There was no significant effect of blood pressure variation on bias or relative bias ( P =0.62 and 0.93, respectively). The mean bias and relative bias of UDCO (±SD) compared with LiDCO were −20.1±39.2 mL/kg/min and −7.7±23.4%, respectively. Concordance correlation coefficient between LiDCO and UDCO was 0.833.
Conclusions – When compared with LiDCO, the UDCO technique has acceptable clinical utility for measuring CO in healthy anesthetized newborn foals.     

Strategies for Using eFSH for Superovulating Mares

The standard treatment for superovulation of mares is to administer equine follicle-stimulating hormone (eFSH) for 4 to 5 days to stimulate multiple follicles and human chorionic gonadotropin (hCG) to induce synchronous ovulations. Objectives of this study were: (1) to determine whether a short-term (3-day) eFSH treatment protocol would result in similar ovulation and embryo recovery rates compared with the standard eFSH protocol; (2) to determine the efficacy of a decreasing dose of eFSH (step-down protocol) on ovulation rate and embryo recovery; (3) to compare the efficacy of hCG and recombinant equine luteinizing hormone (reLH) for inducing ovulation in FSH-treated mares; and (4) to compare embryo recovery rates and embryo size when mares are flushed at 6.5 or 7.0 days after ovulation. Forty light-horse mares were used in 2005 (experiment 1) and 20 different mares were used in 2006 (experiment 2). In experiment 1, mares were randomly assigned to one of three treatment groups: (1) untreated controls, (2) standard eFSH treatment (12.5 mg intramuscularly twice daily), and (3) 3-day eFSH treatment. In experiment 2, mares were randomly assigned to one of four treatments: (1) untreated controls, (2) standard eFSH protocol, (3) 3-day eFSH treatment, and (4) step-down eFSH treatment (12.5 mg twice daily day 1, 8.0 mg twice daily day 2, 4.0 mg twice daily day 3). Within each treatment, mares were given either hCG (2,500 IU) or equine LH (750 mg, EquiPure LH; reLH) to induce synchronized ovulations. Embryo recovery was performed either 6.5 or 7.0 days after ovulation. In experiment 1, numbers of preovulatory follicles and ovulations were less for mares in the 3-day treatment group than the standard group, but were greater than for controls. Embryo recovery per flush was higher in the standard group (2.6) than the 3-day eFSH treatment (0.8) or control groups (0.8). In experiment 2, the number of preovulatory follicles and number of ovulations were greater in the standard and 3-day treatment groups than in control and step-down groups. The percent embryo recovery per ovulation and mean embryo grade were similar for all groups; however, the embryo recovery per flush was higher for mares in the standard treatment than controls (1.3 vs 0.6) but was similar to the 3-day (1.1) and step-down (0.8) treatments. Embryo recovery was similar for flushes performed on days 6.5 and 7.0 post-ovulation. The percentage of control mares ovulating within 48 hours in response to hCG or reLH was similar. In contrast, a higher percentage of eFSH-treated mares ovulated within 48 hours in response to reLH than hCG (92% vs 71%). In both years, the 3-day eFSH treatment protocol resulted in a greater number of preovulatory follicles and a greater number of ovulations than untreated controls. Unfortunately, the increased ovulation rate for mares administered eFSH for 3 days did not result in a greater number of embryos recovered per flush in either year. Use of a step-down eFSH treatment protocol resulted in fewer preovulatory follicles, fewer ovulations, and fewer embryos as compared with the standard eFSH treatment. In conclusion, the standard eFSH treatment resulted in a greater embryo recovery rate per cycle than either the 3-day or step-down treatment protocols. Recombinant equine LH was more effective than hCG in causing ovulation in eFSH-treated mares.     

Comparison of Various Extenders for Storage of Cooled Stallion Spermatozoa for 72 Hours

Multiple extenders have been developed to preserve cooled stallion semen. Comparisons of some extenders have been made but there is need for further research in this area. Extenders tested included EZ Mixin (Animal Reproduction Systems, Chino, CA), Kenney's, Universal (NASCO, Fort Atkinson, WI), EquiPro, EquiPro CellGuard (Minitube of America, Verona, WI), and INRA 96 (IMV, Maple Grove, MN). Semen was collected and each ejaculate was divided and extended in each of the aforementioned extenders and stored at 4°C. Motility measures were determined using computer-assisted sperm analysis at 0, 24, 48, and 72 hours after collection. Samples were evaluated for total motility, progressive motility (PM), straight-line velocity, curvilinear velocity, straight-line distance, and curvilinear distance. Total motility and PM decreased over time in storage (P < .05). Sperm stored in INRA 96, EquiPro, and EquiPro Cell Guard retained the most total motility and PM over the 72 hour period (P < .05). Universal, EquiPro, and EquiPro Cell Guard had the highest measurements for curvilinear velocity, straight-line velocity, and curvilinear distance (P < .05). There were no significant differences among the extenders for straight-line distance.     

Field comparison of the interferon-gamma assay and the intradermal tuberculin test for the diagnosis of bovine tuberculosis

An extensive field comparison of the gamma interferon (IFN-gamma) assay and the single intradermal tuberculin test for the diagnosis of bovine tuberculosis was conducted in Australia. The specificity of the IFN-gamma assay was determined by testing more than 6000 cattle from tuberculosis-free herds and varied from 96.2% to 98.1%, depending on the cut-off point chosen to define a positive reactor. For the sensitivity trial, cattle from herds being de-populated because of bovine tuberculosis were examined with both assays. The sensitivity of the IFN-gamma assay was shown to be significantly higher than the single intradermal tuberculin test and varied from 76.8% to 93.6% depending on the method of interpretation. A maximum overall sensitivity of 95.2% was obtained by testing with the IFN-gamma and the tuberculin test in parallel. The superior sensitivity of the IFN-gamma assay and the ability to adjust the sensitivity of the system depending on the task involved, will provide the Australian Tuberculosis Eradication Campaign with a valuable additional test to enable it to accomplish its goals.