Assessment of Progesterone Concentration Using Enzymeimmunoassay, for Early Pregnancy Diagnosis in Sheep and Goats

The objective of this study was to determine a value of serum progesterone (P₄) concentration, assessed using an enzymeimmunoassay (EIA), for the early distinction between pregnant and non‐pregnant ewes and goats. Adult, non‐lactating ewes of Chios (n=53), Berrichon (n=30) and Sfakia (n=45) breeds were synchronized during the breeding season with progestagens and gonadotrophins and mated to fertile rams (Experiment I). Adult, lactating goats of Swiss breeds (Alpine and Saanen, n=104) and indigenous Greek breed (n=45) were synchronized during the transitional season with progestagens, PGF₂α and gonadotrophins. Cervical artificial insemination (AI) with fresh semen was applied once, 42–44 h after sponge removal (Experiment II). Jugular blood samples were collected on day 19 after sponge removal (ewes) or on day 21 after AI (goats) and serum P₄ concentration was determined by EIA. Progesterone concentrations ≥1.0, ≥1.5, ≥2.5 and ≥4.0 ng/ml were tested as indicative of pregnancy. Pregnancy diagnosis was verified on birth. In the case of sheep, using a discriminatory level of 2.5 ng/ml, overall accuracy of pregnancy diagnosis was 91.4% and predictive value of negative and positive diagnoses were 98.3 and 85.3%, respectively. In the case of goats, predictive value of negative diagnosis was 95.8 and 94.0% and predictive value of positive diagnosis 71.3 and 71.7%, for 1.5 and 2.5 ng/ml, respectively; overall accuracy was 79.2% using either level. The other discriminatory levels tested did not improve these results. A significant positive correlation was observed between P₄ concentration and the number of lambs or kids born, and further analysis indicated that this relationship is not a simple linear function. Based on the results of this study, P₄ concentrations of 2.5 ng/ml in the case of ewes and 1.5–2.5 ng/ml in the case of goats, determined with EIA, are proposed as discriminatory levels between pregnant and non‐pregnant animals, at an interval of one oestrous cycle after service.     

A survey of neurological diseases in horses

SUMMARY: Case records of 450 horses with signs of neurological disease are reviewed. One hundred and nineteen horses with neurological disease due to trauma were examined, of which 60 were due to spinal cord trauma, 47 to brain or cranial nerve trauma and 12 to peripheral nerve trauma. Cervical vertebral fractures/trauma were the most common injury. Basisphenold/basloccipital bone fractures were the most common form of cranial trauma and facial nerve paralysis the most common cranial nerve injury. Eighty-nine horses with neurological disease due to malformation were examined. Cervical vertebral malformation occurred in 83 horses and congenital defects in 6 foals. Neurological disease due to inflammation or infection occurred in 30 horses. The most common disease of this type was meningitis, which occurred in 11 horses and foals. Neoplasms in the CNS caused neurological disease in 8 horses. The final category was miscellaneous neurological disease, which was diagnosed in 204 horses. Diseases in this category included neonatal (28 cases), toxic/metabolic (27 cases), idiopathic (133 cases), degenerative (3 cases) and other neurological diseases (13 cases). The most common condition was idiopathic laryngeal hemiplegia (116 cases).
Where possible, diagnosis relied on a thorough neurological examination with use of ancillary tests in selected cases including rhinolaryngoscopy, radiography, myelography, ophthalmoscopy and cerebrospinal fluid analysis when indicated. In many cases necropsy and histopathological confirmation or diagnosis was necessary.     

Production responses and cost-benefit of long-acting pre-lambing anthelmintic treatment of yearling ewes in two commercial flocks in New Zealand

AIMS: To investigate the production responses and cost-benefit of administering a controlled-release anthelmintic capsule (CRC) to pregnant yearling ewes prior to lambing.

METHODS: Yearling ewes from two commercial sheep flocks (A, n=489; B, n=248) in the North Island of New Zealand were enrolled in the study. Prior to lambing, CRC containing albendazole and abamectin were administered to half the ewes while the other half remained untreated. Ewe liveweights and body condition scores were measured prior to lambing, at weaning and, for Flock B, prior to subsequent mating. Lambs were matched to dams shortly after birth and the weight and number of lamb weaned per ewe were determined. A cost-benefit analysis was undertaken for Flock B considering the increased weight of lamb weaned per ewe, and the weight of ewes at the next mating and the benefit in terms of lambs born.

RESULTS: The mean weight at weaning of treated ewes was greater for treated than untreated ewes by 2.76 (95% CI 0.64–4.88)?kg in Flock A (p<0.001) and 2.35 (95% CI ?0.41–5.12)?kg in Flock B (p=0.003); the weight of lamb weaned per ewe was greater for treated than untreated ewes by 1.43 (95% CI ?0.71 to –3.49)?kg in Flock A (p=0.041) and 3.97 (95% CI 1.59–6.37)?kg in Flock B (p<0.001), and ewe liveweight prior to subsequent mating was greater for treated than untreated ewes in Flock B by 4.60 (95% CI 3.6–5.6)?kg (p<0.001). There was no difference in the percentage of lambs reared to weaning between treated and untreated ewes in either flock (p>0.8). The overall cost-benefit of treatment for Flock B was NZ$9.44 per treated ewe.

CONCLUSIONS AND CLINICAL RELEVANCE: Pre-lambing CRC administration to yearling ewes resulted in increased ewe weaning weights and weight of lamb weaned in both the flocks studied. There was an economic benefit in the one flock where this was assessed.     

Effect of astaxanthin on the quality of boar sperm stored at 17°C,incubated at 37°C or under in vitro conditions

The aim of the study was to investigate the effect of the antioxidant astaxanthin on boar semen. Twenty ejaculates from 10 boars (two ejaculates/boar) were extended and split in three groups: semen control (SC), solvent control (C; semen with dimethyl sulfoxide, the diluent of astaxanthin) and semen with astaxanthin (A) in concentration 0.5 μmol/L. Sperm quality parameters (motility and kinetics, morphology, viability, functional integrity of sperm plasma membrane by Hypo‐Osmotic Swelling Test [HOST] and DNA integrity) were assessed at 0, 24 and 48 hr of storage at 17°C (experiment I), before (0 hr) and after (1 hr) of sperm thermal resistance assay at 37°C (experiment II) and finally before (0 hr) and after (1 hr) sperm in vitro incubation (38.5°C, 5% CO₂, maximum humidity [experiment III]). In experiment I, group A performed overall better than group SC and as a tendency better than group C regarding viability. Total motility, rapid spermatozoa and HOST remained constant across time in group A, whereas they decreased in the remaining groups. In experiment II, regarding motility and viability, group A displayed better results across time than the other two groups. In experiment III, viability and total motility decreased in groups SC and C, while in group A, these parameters were not significantly different between the examination time points. In conclusion, astaxanthin has a beneficial and protective effect on boar semen quality under the investigated conditions.     

Effect of reducing inspired oxygen concentration on oxygenation parameters during general anaesthesia in horses in lateral or dorsal recumbency

Objective

To compare the effects of two concentrations of oxygen delivered to the anaesthetic breathing circuit on oxygenation in mechanically ventilated horses anaesthetised with isoflurane and positioned in dorsal or lateral recumbency.

Methods

Selected respiratory parameters and blood lactate were measured and oxygenation indices calculated, before and during general anaesthesia, in 24 laterally or dorsally recumbent horses. Horses were randomly assigned to receive 100% or 60% oxygen during anaesthesia. All horses were anaesthetised using the same protocol and intermittent positive pressure ventilation (IPPV) was commenced immediately following anaesthetic induction and endotracheal intubation. Arterial blood gas analysis was performed and oxygenation indices calculated before premedication, immediately after induction, at 10 and 45 min after the commencement of mechanical ventilation, and in recovery.

Results

During anaesthesia, the arterial partial pressure of oxygen was adequate in all horses, regardless of position of recumbency or the concentration of oxygen provided. At 10 and 45 min after commencing IPPV, the arterial partial pressure of oxygen was lower in horses in dorsal recumbency compared with those in lateral recumbency, irrespective of the concentration of oxygen supplied. Based on oxygenation indices, pulmonary function during general anaesthesia in horses placed in dorsal recumbency was more compromised than in horses in lateral recumbency, irrespective of the concentration of oxygen provided.

Conclusion

During general anaesthesia, using oxygen at a concentration of 60% instead of 100% maintains adequate arterial oxygenation in horses in dorsal or lateral recumbency. However, it will not reduce pulmonary function abnormalities induced by anaesthesia and recumbency.     

Flow Cytometric Chromosomal Sex Sorting of Stallion Spermatozoa Induces Oxidative Stress on Mitochondria and Genomic DNA

To date, the only repeatable method to select spermatozoa for chromosomal sex is the Beltsville sorting technology using flow cytometry. Improvement of this technology in the equine species requires increasing awareness of the modifications that the sorting procedure induces on sperm intactness. Oxidative stress is regarded as the major damaging phenomenon, and increasing evidence regards handling of spermatozoa – including sex sorting – as basic ground for oxidative damage. The aim of this study was to disclose whether the flow cytometric sorting procedure increases the production of reactive oxygen species (ROS), and to identify if ROS production relates to DNA damage in sorted spermatozoa using specific flow cytometry‐based assays. After sorting, oxidative stress increased from 26% to 33% in pre‐ and post‐incubation controls, to 46% after sex sorting (p < 0.05). Proportions of DNA fragmentation index post‐sorting were approximately 10% higher (31.3%); an effect apparently conduced via oxidative DNA damage as revealed by the oxyDNA assay. The probable origin of this increased oxidative stress owes the removal of enough seminal plasma due to the unphysiological sperm extension, alongside a deleterious effect of high pressure on mitochondria during the sorting procedure.     

Effects of local anaesthesia of 4 to 8 hours duration on the acute cortisol response to scoop dehorning in calves

Objective To determine the effect of a long-acting local anaesthetic on the plasma cortisol response in calves dehorned using a scoop.
Design A physiological study with controls.
Procedure Seventy female calves, 3 to 4 months old, were divided into seven groups and dehorned by scoop with or without a long acting local anaesthetic (bupivacaine) which provides local anaesthesia for 3 to 4 h. In some treatment groups the local anaesthetic was given a second time, after 4 h, to give 8 h of anaesthesia. Blood samples were taken before and for 9 h after treatment and plasma cortisol concentrations were measured.
Results Scoop dehorning caused a marked rise in plasma cortisol concentration, which returned to pre-dehorning values after 7 h. The cortisol concentrations of calves given bupiva-caine were similar to those of control animals for 4 h after dehorning, but then a significant and protracted rise occurred, with concentrations returning to pretreatment values at about 9 h after dehorning. In calves given a second treatment of bupivacaine 4 h after dehorning, the plasma cortisol concentration remained similar to control values for 8 h after dehorning but then increased sharply.
Conclusions During its period of activity long-acting local anaesthetic prevented the rise usually seen after scoop dehorning in plasma cortisol concentrations; by inference it also alleviated the pain-induced distress caused by this procedure. However, when its effects had worn off the plasma cortisol values rose sharply indicating that animals still experienced pain-induced distress despite local anaesthetic being administered.     

Factors affecting the relationship between arterial and end-tidal carbon dioxide pressures in the anaesthetised horse

Objective To assess the effects of the duration of anaesthesia, position of recumbency, mode of ventilation, anaesthetic drug protocol, patient age and type of surgical procedure on the usefulness of capnometry as a measure of the partial pressure of arterial carbon dioxide (P_aco ₂) during general anaesthesia in horses. Design A prospective study compared the P_aco ₂ values with those of partial pressure of end-tidal carbon dioxide (ETco ₂) in horses anaesthetised for elective or emergency surgical procedures. The difference between P_aco ₂ and ETco ₂ (P_aco ₂− ETco ₂) and the physiological dead space to tidal volume ratio (V_D/V_T) were calculated. The effects of the study parameters on these variables was determined. Results The agreement between P_aco ₂ and ETco ₂ was poor. P_aco ₂− ETco ₂ and V_D/V_T during the first 60 min of anaesthesia was significantly less than after 60 min of anaesthesia. Mode of ventilation, position of recumbency, anaesthetic drug protocol, patient age and type of procedure did not have a significant affect on either value. Conclusions P_aco ₂− ETco ₂ in anaesthetised horses can be large, making ETco ₂ unreliable as a predictor of P_aco ₂ and for assessment of pulmonary ventilation. For anaesthesia lasting less than 60 min at least one blood gas analysis of an arterial blood sample is required to assess P_aco ₂− ETco ₂. Arterial blood gas analysis should be repeated after 60 min of general anaesthesia.