Influence of Temperature during Glycerol Addition and Post-thaw Dilution on the Quality of Canine Frozen Semen

Contents The aim of the present study was to compare the influence of room temperature (27 degrees C) and 4 degrees C during glycerol addition on canine semen cryopreservation and verify the effect of different post-thawing dilutions on canine semen. Ten ejaculates from five stud dogs were collected by digital manipulation. Semen samples were evaluated and further divided into two aliquots. The first aliquot was extended in Tris-egg yolk-glycerol at 27 degrees C and the second one received glycerol at 4 degrees C. Samples were frozen and stored in liquid nitrogen. After 1 week, samples were thawed and submitted to evaluations of progressive sperm motility, morphology, acrosomal integrity, hypo-osmotic swelling (HOST) and thermoresistance tests. For thermoresistance test, aliquots were divided in two portions: one portion was kept undiluted (1 : 0) and the other one was diluted in a 1 : 4 ratio (one part semen to four parts extender). No differences were observed between temperatures for glycerol addition regarding seminal parameters evaluated. Furthermore, post-thawing dilutions demonstrated similar effect on canine semen longevity. Correlations among post-thaw sperm motility and HOST and results from thermoresistance test were observed for both temperatures for glycerol addition. In conclusion, glycerol could be added to canine semen at room temperature (27 degrees C) or at 4 degrees C. Moreover, there is no need to extend canine semen after thawing for the thermoresistance test, but if we need to increase the inseminating volume for artificial inseminations, the addition of extender will not damage the semen.     

The effect of antibody to caseous lymphadenitis in ewes on the efficacy of vaccination in lambs

The effect of maternal antibody to the toxin of Corynebacterium pseudotuberculosis, produced by caseous lymphadenitis (CLA) in ewes or by vaccinating ewes before lambing, on the efficacy of vaccination against CLA in their lambs was examined. Lambs were allocated to treatments according to either the vaccination history of their dam or level of CLA toxin antibody of their dam. They were vaccinated twice using 2 different inoculation regimes and then artificially infected with CLA organisms. The number of lambs with CLA lesions was assessed at slaughter. In one experiment high levels of CLA toxin antibody activity in ewes were associated with decreased efficacy of CLA vaccination in their lambs, when lambs were vaccinated at 2 and 8 weeks or 8 and 14 weeks of age. In a second experiment the efficacy of lamb vaccination at 8 and 12 weeks, but not at 14 and 18 weeks of age, was decreased. In sheep flocks with a high prevalence of CLA, vaccinating lambs against CLA at less than 10 weeks of age may not produce optimum protection against CLA in lambs. There was no difference in infection rate between lambs from vaccinated and unvaccinated ewes. However, vaccination of lambs at 2 and 8 wks was less effective that vaccination at 8 and 14 weeks, probably due to reduced immunocompetence in young lambs. In sheep flocks where significant numbers of lambs receive their primary vaccination at less than 3 weeks of age vaccination programmes to control CLA in lambs may be less effective.     

Investigations into the toxicity of Echium plantagineum in sheep: 1. Field grazing experiments

A field grazing trial was undertaken to monitor the health and production of crossbred sheep grazing pasture where Echium plantagineum constituted a considerable proportion of the available forage. The trial, conducted for 19 months over successive grazing seasons, demonstrated a significant difference in production, with sheep on the E. plantagineum pasture being lighter and growing less wool compared with sheep on Echium-free pasture. No mortalities involving pyrrolizidine alkaloid poisoning were recorded in sheep grazing E. plantagineum, although there was histological evidence of moderately severe liver damage associated with high liver copper concentrations in at least one sheep following the grazing of large quantities of the plant.     

Factors affecting the natural transmission of bovine leukaemia virus infection in Queensland dairy herds

Natural transmission of bovine leukaemia virus (BLV) infection in south-eastern Queensland dairy herds was slow in 2 herds with a low to moderate (13 to 22%) prevalence of infection. Infection spread much more rapidly in a herd that had a higher prevalence (42%) when first tested. In a 13 month study of this herd, the cumulative incidence of infection was 24%. In one herd new infections were confined almost entirely to calves of uninfected dams. Following the end of feeding bulk milk to calves, a common practice in dairy herds, no more calves in this herd became infected. In laboratory experiments, neither prolonged housing of susceptible calves with infected cattle, consumption of drinking water contaminated with infected blood, nor inoculation of sheep with saliva from infected cattle resulted in transmission of BLV infection. Sheep were infected by subcutaneous inoculation of a suspension of purified lymphocytes from an infected heifer. The minimum infective dose was 10(3) lymphocytes, equivalent to the number of lymphocytes in approximately 0.1 microliter blood. Thus, procedures involving the transfer of a very small volume of blood from animal-to-animal have the potential to transmit infection.     

Use of phenytoin to treat horses with Australian stringhalt

Five horses with Australian stringhalt were treated with 15 mg/kg phenytoin orally for 2 weeks. During the second week of the trial, 3 of the horses were given an additional dose of 10 mg/kg phenytoin. The response to treatment was clinically assessed by grading the severity of the gait abnormality at the walk, trot, turning and backing twice daily. There was a significant (P less than 0.05) improvement in the gait abnormality when pre-treatment values were compared with the mean of the last 3 assessments before treatment stopped. When reassessed 2 weeks after treatment ceased, there remained a significant (P less than 0.05) improvement compared with pre-treatment values at the trot and on backing, but not at the walk or turning. Surface electromyographic recordings were made weekly from the long digital extensor muscle, and there was a change to a near normal recording by the end of treatment. Plasma phenytoin concentrations were monitored during the trial, and the dose rates used achieved a steady state with a mean plasma level of 37 +/- 7 mumol/l. There was wide variability between plasma concentrations in different horses, although there was no difference in absorption between administration of the phenytoin as a paste, or when it was mixed in the feed. Although mild tranquilization was seen after treatment, there were no clinical, haematological or biochemical signs of toxicity from the phenytoin therapy.     

Purification of secretory immunoglobulin A from milk of the brushtail possum (Trichosurus vulpecula)

AIM: To identify and purify secretory immunoglobulin A (sIgA), a key effecter molecule in mucosal immune responses, from milk of the brushtail possum (Trichosurus vulpecula).

METHODS: Milk samples were collected from female possums with pouch young, and clarified by centrifugation and precipitation methods. The clarified fraction was purified by gel filtration and affinity chromatography to yield sIgA. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting techniques were used to assess the purity of the final product, and to identify the heavy (H) chain, light (L) chain and secretory component (SC) of possum sIgA.

RESULTS: Immunoblotting, using antibodies raised against cloned possum sIgA SC and H-chain, and a synthetic peptide fragment of the H-chain, confirmed the identity of the purified protein. The N-terminal amino acid sequence of purified possum sIgA showed strong homology to reported sequences of H-chain variable regions of marsupial immunoglobulins.

CONCLUSIONS: Milk was shown to be a convenient source of mucosal secretion containing sIgA, and a process involving 2 precipitation and 2 chromatography steps produced purified sIgA. This IgA preparation will prove useful for the generation of sIgA-specific immunological reagents for measurement of immune responses in the development of mucosal-based vaccines for biological control of possums.     

Interaction between nutrition and gastrointestinal parasitism in sheep

Effects of gastrointestinal nematode infection on metabolism and nutrient utilisation in sheep are reviewed. Infection induces protein deficiency by increasing the demand for amino acids in the alimentary tract while reducing supply through depression of appetite. Mechanisms through which improved protein nutrition could improve the performance of the host are then discussed. Opportunities for capitalising on such effects are limited by our rudimentary understanding of the cell-mediated immune response in gastrointestinal epithelial tissue. Both resistance of the animal to larval establishment and performance in the face of larval challenge can be enhanced by improved protein nutrition. However, enhanced immune responses may not necessarily be synonymous with improved productivity except at luxurious levels of protein intake, because of apparently competing demands for protein. Such levels of protein nutrition are difficult to achieve in pasture-based systems, because of the protein limiting role of the rumen. Work with protein-protecting tannins to overcome this limitation is discussed. The much more limited evidence for effect of mineral nutrition, particularly copper (Cu), molybdenum (Mo), cobalt (Co) and phosphorus (P), on outcome of larval challenge is also reviewed.     

Comparison of Different Glycerol and Egg Yolk Concentrations Added to Tris‐based Extender for the Collared Peccaries (Tayassu tajacu) Semen Freezing

This study aimed to evaluate various concentrations of egg yolk (5, 10, or 20%) in combination with different concentrations of glycerol (3% or 6%) added to a Tris‐based extender on the post‐thaw characteristics of sperm obtained from Tayassu tajacu. For this purpose, semen from 10 sexually male mature collared peccaries was collected by electroejaculation and evaluated for sperm motility, vigour, viability, morphology and functional membrane integrity. The ejaculates were initially extended in Tris‐fructose plus egg yolk (5%, 10% or 20%). After cooling, the semen was added to Tris‐egg yolk plus glycerol (6% or 12%), resulting in a final concentration of 3% or 6% glycerol of the extender. Straws were frozen using liquid nitrogen and thawed in a water bath at 37°C for 30 s. The frozen–thawed semen was evaluated as reported for fresh semen. After thawing, a significant decrease was verified for sperm motility and vigour, for all the samples in comparison with fresh semen. However, no differences were evidenced among treatments for any sperm characteristics evaluated (p > 0.05), except for the combination between 10% egg yolk and 6% glycerol, which provided the worst preservation of functional membrane integrity (p < 0.05). The interactions between higher concentrations of egg yolk (20%) and glycerol (6%) and also between lower concentrations of the same substances (5% egg yolk and 3% glycerol) added to the Tris‐based extender negatively affected the preservation of the normal sperm morphology after thawing (p < 0.05). In conclusion, the use of Tris‐based extender added to 10% or 20% egg yolk plus 3% glycerol is recommended for effective sperm cryopreservation in collared peccaries.