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.     

The development of multiple cutaneous inverted papilloma following ovariohysterectomy in a dog

Abstract

CASE HISTORY: Ovariohysterectomy was performed on an adult Cavalier King Charles Spaniel. The skin that had been clipped for surgery was noticed to be erythematous 8 days later.

CLINICAL AND PATHOLOGICAL FINDINGS: Poorly defined patches containing multiple papules were visible bilaterally within the clipped skin. These became larger over the following 2 weeks, and samples were collected for histology. Seven days later, the lesions were multiple raised masses, up to 5 cm in diameter. Histology revealed numerous cup-shaped epidermal proliferations extending into the dermis. The presence of keratinocytes with increased quantities of blue-grey cytoplasm, and koilocytosis suggested papillomavirul infection; Canis familiaris papillomavirus (CfPV-2) DNA was amplified from two separate samples. Complete regression was observed 8 weeks after the lesions had been initially observed.

DIAGNOSIS: Multiple inverted papilloma confined to skin that had been clipped for surgery.

CLINICAL RELEVANCE: This is the first time that the development of canine cutaneous papillomas has been associated with surgery. The nature of the association between surgery and development of the papillomas is uncertain. However, it is possible that damage to superficial skin could promote the formation of papillomas. This is the first identification of CfPV-2 in New Zealand.     

Increased Risk of Conception Failure in German Holstein Friesian Cows with Chronic Endometritis

The objective of this retrospective field study was to evaluate the effect of chronic clinical endometritis (CCE) on days open in German Holstein Friesian cows. Two hundred and sixty‐four cows diagnosed with CCE from day 14 to 42 postpartum were included in this study. A Cox’s proportional hazards regression model with stepwise forward selection for days open was used and the variables type of vaginal discharge, ovarian cysts and a marked loss of body condition score proved to be significant with hazard ratios of 1.5, 2.0 and 1.9 respectively. Although not remaining in the model, a weak positive effect of treatment for CCE and a negative effect of high milk production could also be found in the univariate analysis. These results suggest that the suppression of fertility of animals diagnosed with CCE varies according to the different symptoms, which could be used to support decisions regarding treatment. Management factors such as a marked loss in body condition and high milk production prolonged the number of days open of animals with CCE.     

Biological control of Phytophthora parasitica var. Nicotianae on tobacco seedlings with non-pathogenic binucleate Rhizoctonia fungi

Nonpathogenic binucleate Rhizoctonia fungi (BNR) controlled black shank caused by Phytophthora parasitica var. nicotianae on greenhouse-grown tobacco seedlings in styrofoam float trays. Three BNR isolates were incorporated into a soil-less mix on colonized, pulverized, sifted rice particles; colonized whole rice grains; or on pelleted tobacco seeds coated with 0.5% methyl cellulose. Five-wk-old seedlings were inoculated with zoospores of P. parasitica var. nicotianae and disease rated over 10 d. The level of protection varied with method of BNR application, ranging from 40 to 70%. Overall, control was better when BNR isolates were applied on rice inocula rather than on BNR-colonized tobacco seeds. From 15% to 80% of individual roots from seedlings grown in soil-less mix amended with BNR-colonized rice grains were colonized while only 0–20% of roots from seedlings grown from BNR-colonized tobacco seeds were colonized. Likewise, 37–100% of soil-less mix amended with BNR-colonized rice grains contained BNR's while less than 3% of soil-less mix was colonized when seedlings emerged from BNR-colonized tobacco seeds. This is the first demonstration of biocontrol of Phytophthora by BNR fungi.     

Landscape-level variation in temperature sensitivity of soil organic carbon decomposition

We examined landscape-level variation in temperature sensitivity of labile SOC across 71 sites at a central North American grassland. The observed range in activation energy of decomposition (E_a), an index of temperature sensitivity, was as great at the landscape scale as has been observed at the continental scale. E_a was lower for soils with more labile C, consistent with the ‘Carbon quality-temperature’ hypothesis. Soil pH explained 67% of the variation in E_a. Although there are strong environmental correlates with the E_a of SOC decomposition at landscape scales, the amount of variation within landscapes could confound regional- to global-scale predictions of the response of soil C to warming.     

Relationship between milk production and somatic cell count in dairy cows in East Gippsland

SUMMARY The analysis of records for 75 821 herd tests on 23 700 cows in the Macalister Irrigation District of East Gippsland, using a model that also included stage of lactation, herd and cow effects, found a strong relationship between somatic cell count and milk production. Unit increases in the natural logarithm of the somatic cell count were associated with decreases in production of up to 2.6% in the range 54 598 to 403 429 cells per mL. This is equivalent to a decrease of 1.9% for a doubling of cell count from 100 000 to 200 000 cells per mL. Subclinical mastitis was assessed to be a significant source of loss in milk production in the study area.