Effects of Egg Yolk Source on the Cryopreservation of Stallion Spermatozoa

Three experiments were conducted to determine whether replacement of chicken egg yolk, as a component of freezing extenders, with egg yolk from other avian species would improve the post-thaw motility and percentage of intact acrosomes of stallion spermatozoa. In the first experiment, substitution of chicken egg yolk with chukar egg yolk, as a component of the lactose-ethylenediaminetetraacetic acid extender, improved (P ≤ .05) the post-thaw motility of stallion spermatozoa. These results were not replicated in (IMV Technologies, Maple Grove, MN, USA) a more expansive study comparing 2%, 4%, 6%, or 8% egg yolk combined with INRA 96 when a “slow freeze” method was used, or the same substitution at levels ranging from 13% to 22% when egg yolk was combined with lactose-ethylenediaminetetraacetic acid for diluents used for a “fast freeze” method of cryopreservation. In the third study, egg yolks from regular and high omega-3 chicken eggs as well as from turkey, chukar, and mallard duck eggs were analyzed for lipid content and fatty acid profile. The yolk from the turkey eggs was higher (1,300 mg/100 g) and that from mallard ducks was lower (560 mg/100 g) in cholesterol as compared with the two types of chicken eggs and chukar egg yolk (range, 1,046-1,094 mg/100 g). In addition, the high omega-3 eggs did test higher for fatty acids (4.51 g/100 g) than other types of eggs (range, 0.28-0.73 g/100 g). Substitution of chicken egg yolk with turkey, but not duck, egg yolk resulted in higher post-thaw total motility (P ≤ .05) for spermatozoa obtained from two of the three stallions used in the third experiment.     

In Vitro Evaluation of Liquid‐stored Buffalo Semen at 5°C Diluted in Soya Lecithin Based Extender (Bioxcell®), Tris‐Citric Egg Yolk,Skim Milk and Egg Yolk‐Citrate Extenders

This study was designed to compare the quality of liquid‐stored buffalo bull spermatozoa in soya lecithin based extender Bioxcell^® (BIOX), milk (MILK), tris‐citric egg yolk (TEY) and egg yolk‐citrate (EYC) extender at 5°C. Semen was collected from five Nili‐Ravi buffalo (Bubalus bubalis) bulls of 6–7 years of age with artificial vagina over a period of 3 weeks (two consecutive ejaculates once in a week). Semen ejaculates having more than 60% motility were pooled, split into four aliquots, diluted (37°C; 10 × 10⁶ motile spermatozoa/ml), cooled from 37 to 5°C in 2 h (0.275°C/min) and stored for 5 days. Sperm motility, viability, plasma membrane integrity (PMI) and normal acrosomal ridge were studied at first, third and fifth day of storage. Higher values of progressive sperm motility (%), sperm viability (%), sperm PMI (%) and normal apical ridge (%) were observed in BIOX, MILK and TEY extenders at first, third and fifth day of storage than EYC extender. Progressive sperm motility, sperm viability and sperm PMI in BIOX^® extender were not different from MILK and TEY extenders at 1st and third day storage period. However, at fifth day of storage, the values for these parameters remained significantly higher (p < 0.05) in BIOX^® compared with MILK, TEY and EYC extenders. At fifth day of storage, the semen quality parameters for Bioxcell^® were comparable to those with MILK and TEY extenders at third day of storage. In conclusion, motility, viability and PMI of buffalo bull spermatozoa remained similar in Bioxcell^®, milk and TEY extender at first and third days of storage at 5°C. Yet, the values for the aforementioned parameters in Bioxcell^® were higher compared with milk, TEY and EYC extender at fifth day of storage at 5°C.     

Quantification,Morphology and Ultrastructure of Preantral Follicles of Buffalo (Bubalus bubalis) Foetuses

The objective of this study was to determine the number, morphology and ultrastructure of preantral ovarian follicles of buffalo (Bubalus bubalis) foetuses at different ages. Quantification revealed number of primordial, primary and secondary follicles of 48 857 ± 17 506, 26 000 ± 20 452, 18 428 ± 10 875 and 18 375 ± 19 690, 225 ± 349, 326 ± 288 at 12–34 cm and 35–60 cm crown rump length (CRL), respectively. Follicular diameter values were 28.9 (±3.4), 34.7 (±5.9) and 59.4 (±12.6) μm; oocyte diameters were 21.7 (±2.8), 24.3 (±3.4) and 33.0 (±7.7) μm, and the numbers of follicular cells in the follicle equatorial section were 7.1 (±1.4), 12.0 (±2.4) and 13.8 (±2.4) for primordial, primary and secondary follicles, respectively. The primordial follicle consisted of an oocyte surrounded by a layer of flattened follicular cells with a normally eccentric oocyte nucleus. Dispersed Golgi complex, smooth endoplasmic reticulum, rounded mitochondria and several lipid vesicles were observed in the cytoplasm and cell junctions between the follicle cell membranes and the oocyte. This work describes the number, morphometry and ultrastructure of preantral follicles of buffalo foetuses, concluding that folliculogenesis is established between 8 and 34 cm CRL and that follicle number varies individually and according to age and that further studies are needed in this species.     

Evaluation of gastric pressures as an indirect method for measurement of intraabdominal pressures in the horse

Objective – To develop an indirect method for measurement of intraabdominal pressures in the standing horse using measurement of gastric pressures as a less invasive technique, and to compare this method with direct intraabdominal pressures obtained from the peritoneal cavity. Design – Prospective, experimental study. Setting – University‐based equine research facility. Animals – Ten healthy adult horses, 7 geldings and 3 mares. Interventions – Gastric pressures were measured using a nasogastric tube with a U‐tube manometry technique, while intraperitoneal pressures were measured with a peritoneal cannula. Measurements of intraabdominal pressure were obtained by both methods, simultaneously, and were evaluated using 5 increasing volumes of fluid infused into the stomach (0, 400, 1,000, 2,000, and 3,000 mL). Bias and agreement between the 2 methods were determined using Bland‐Altman analysis and Lin's concordance correlation coefficients. Measurements and Main Results – Mean gastric pressure was 14.44±4.69 cm H₂O and ranged from 0 to 25.8 cm H₂O. Intraperitoneal pressure measurements were generally subatmospheric, and ranged from ?6.6 to 3.1 cm H₂O (mean±SD, ?1.59±2.09 cm H₂O). Measurements of intraperitoneal pressures were repeatable; however, intra‐ and interindividual variance was significantly larger for measurements of gastric pressures. The mean and relative bias for comparison between the 2 techniques was 15.9±5.3 cm H₂O and 244.3±199.2%, respectively. The Lin's concordance correlation coefficient between gastric and intraperitoneal pressures was ?0.003 but this was not statistically significant (P=0.75). Conclusions – There was no statistical concordance between measurements of intraabdominal pressure using gastric and intraperitoneal pressure measurement, indicating that gastric pressures cannot be substituted for intraperitoneal pressure measurement. Direct measurement of intraperitoneal pressures may be a more consistent method for comparison of intraabdominal pressures between horses, due to less variability within and between individuals.     

Acute aldicarb toxicity in dogs: 15 cases (2001–2009)

Objective – To describe the common clinical signs, laboratory abnormalities, treatment, and prognosis associated with acute aldicarb toxicosis in dogs. Design – Retrospective observational study from 2001 to 2009. Setting – Urban referral hospital. Animals – Fifteen client‐owned dogs. Interventions – None. Measurements and Main Results – The most common clinical signs associated with acute aldicarb toxicosis were vomiting, ptyalism, diarrhea, and tremors. Of the 15 dogs, 11 were admitted to the hospital for treatment, 2 were euthanized at presentation and 2 were discharged against medical advice following minimal treatment and lost to follow‐up. Laboratory abnormalities included lactic acidosis and hyperglycemia in 12 and 9 patients, respectively. Treatment of hospitalized dogs included induction of emesis with apomorphine (4 dogs), activated charcoal (5), IV fluids (11), atropine (7), methocarbamol (3), diazepam (1), pralidoxime (1) and diphenhydramine (1). Ten of 11 hospitalized dogs survived to discharge; 1 was euthanized following a respiratory arrest after 36 hours of hospitalization. One patient received mechanical ventilation and treatment for pneumonia before discharge from the hospital. The median duration of hospitalization was 22 hours (range 12–168 h). Conclusions – Acute aldicarb toxicosis carries a good prognosis for survival and hospital discharge with treatment. Supportive care should be considered for at least 18–24 hours to monitor for response to therapy and development of respiratory failure.