Airway inflammation is associated with mucous cell metaplasia and increased intraepithelial stored mucosubstances in horses

This study was performed to determine if a peripheral sample of lung from the site where biopsy is conducted is representative of the rest of the lung and to investigate the relationship between airway inflammation and intraepithelial mucous production in the peripheral airways. Lung parenchyma samples were collected from five different regions of the lung in five control and five heaves-affected horses. Horse groups were defined by clinical response to stabling. Tissue sections were used for semi-quantitative scoring of lesions, to count the number of airways, to quantify the amount of stored mucosubstances (Vs) within the epithelium, and to count the number of epithelial cells in terminal airways. No significant differences were found between lung regions or between groups of horses. Lack of regional differences in airway structures means that a biopsy sample can be used for diagnosis and investigation of diffusely distributed diseases. Airway inflammation was correlated with mucous cell metaplasia and Vs. Therefore, in horses, mucus accumulation is partly caused by increased number of mucous cells and is associated with airway inflammation. Therapy targeted to reduce airway inflammation will help reduce the excessive mucous accumulation in horses.     

Characterisation of three novel canine osteosarcoma cell lines producing high levels of matrix metalloproteinases

Three canine osteosarcoma cell lines were established from spontaneous pelvic and radial osteosarcomas. The cell populations cultured exhibited characteristics of malignancy and consisted of adherent, pleomorphic, mostly large spindle-shaped or polyhedral cells, characterised by the presence of numerous cytoplasmic granules and vacuoles. The main ultrastructural features included the presence of abundant rough endoplasmic reticulum and numerous cytoplasmic vesicles, deposit vacuoles and small cytoplasmic protrusions. Zymography showed that the cell lines produce high levels of MMP-2 and MMP-9, enzymes directly involved in crucial aspects of the metastatic process. Consistent with their osteoblastic lineage and malignant phenotype, all cell lines were immunoreactive to vimentin, osteopontin, PCNA, p53, MMP-2 and MMP-9, while they were negative for cytokeratin, desmin, SMA, Factor VIII, NSE, GFAP, Rb and p21 protein. No retroviral particles or RNA were detected ultrastructurally or with RT-PCR, although the possibility of viral involvement in osteosarcoma cannot be excluded. The new cell lines provide excellent in vitro models that may allow further studies on the pathobiology of canine osteosarcoma to be undertaken.     

Adrenergic and peptidergic innervation of the trachealis muscle in the normal horse: a preliminary report

The tone of respiratory smooth muscle is largely determined by the input from autonomic nerves. The distribution of adrenergic and selected nonadrenergic, non-cholinergic (NANC) nerves in the normal equine trachealis muscle was investigated using immunohistochemistry. The smooth muscle of the trachealis was found to contain numerous nerves immunoreactive for an enzymatic marker of adrenergic nerves, as well as many nerves immunoreactive for a putative NANC neurotransmitter, peptide histidine isoleucine, a potent bronchodilator. The tissues surrounding the respiratory smooth muscle contained numerous nerves immunoreactive for the neuropeptides substance P and calcitonin gene-related peptide, which can cause marked vasodilation and bronchoconstriction. The complex innervation of the equine trachea should be kept in mind when interpreting the results of physiological experiments.     

The in vitro diagnosis of anthelmintic resistance in cyathostomins

Cyathostomins are the primary parasitic pathogens of equids. For over 40 years, these nematodes have been controlled using broad spectrum anthelmintics. Three classes of anthelmintic are currently available for this use but, unfortunately, resistance to each of these has now been recorded in cyathostomin populations. As part of an optimal strategy to control cyathostomin infections in the field, it will be important to identify drug-resistant worms at as early a stage as possible. This objective needs to be supported by methodologies that will allow the accurate comparison of anthelmintic resistance in different nematode populations. At present, the faecal egg count reduction test is considered the most suitable method for initial screening for anthelmintic resistance in equine nematode populations. However, in its current state, this test lacks sensitivity. It is also costly and time-consuming to perform. Laboratory-based techniques, such as the egg hatch assay, larval development assay, larval migration inhibition assay and the larval feeding inhibition assay offer alternative options for assessing anthelmintic resistance in nematode populations. All of these tests have been investigated for their utility in measuring drug resistance in sheep nematode populations and some have proven useful. The egg hatch assay, larval development assay and larval migration inhibition assay have been investigated for use in measuring levels of drug resistance in equine nematode populations. However, at best, the results obtained thus far indicate that these tests require further refinement.     

Letters to the editor

Sir;- Outbreaks of ovine abortions caused by Campylobacter fetus fetus (previously C. fetus intestinalis) occur sporadically in mimy areas of New Zealand. The devastating effects on some flocks have stimulated recent studies of the disease,^(1)(3)(7) and have also led to the commercial production of a single strain vaccine (Campylovexin; Wellcome N.Z. Ltd, Auckland). However, it has been shown that single strain C. Fetus vaccines do not protect against serologically distinct strains.⁽²⁾ Since at least two different serotypes of C. fetus fetus are present in New ZealandY) it is not surprising that the commercially available vaccine is only partially effective in some outbreaks.⁽⁷⁾     

Chronic subclinical ovine fascioliasis: plasma glutamate dehydrogenase, gamma-glutamyl transpeptidase and aspartate aminotransferase activities and their significance as diagnostic aids

The effect of low grade chronic Fasciola hepatica infection on the concentration of plasma glutamate dehydrogenase (GD), gamma-glutamyl transpeptidase (gamma-GT) and aspartate aminotransferase (AST) was investigated in sheep dosed daily with three (AL3), eight (AL8) or 14 (AL14) metacercariae over 22 weeks or given a single dose of 200 metacercariae. Significant increases in plasma GD activity first occurred after nine, 12 and 23 weeks and in gamma-GT activity after 12, 24 and 32 weeks for groups AL14, AL8 and AL3 respectively. Changes in AST activity were not as clearly related to dose level. In sheep with single infection, both GD and gamma-GT were capable of detecting liver damage resulting from the migration of 10 or more flukes. Plasma GD and gamma-GT activities are more sensitive indicators of liver cell damage in chronic subclinical fascioliasis than AST activity and gamma-GT may be more suitable as a diagnostic aid on account of its greater stability.     

SEASONAL AVAILABILITY OF NEMATODE LARVAE ON PASTURES GRAZED BY CATTLE IN NEW SOUTH WALES

Trends in the numbers of infective nematode larvae on pasture plots contaminated by cattle at different seasons of the year were defined in 3 different climatic regions. The main nematodes were Ostertagia ostertagi, Trichostrongylus spp, Haemonchus spp and Cooperia spp.
On the North Coast of New South Wales with a sub-tropical climate, the numbers of infective larvae of all 4 nematodes rose rapidly to peak levels soon after each seasonal period of contamination began, then fell quickly within a few months. On the Central Coast of New South Wales, the trends were similar to those on the North Coast, except that the larvae persisted on the pasture for a much longer time. On the Northern Tablelands of New South Wales, where temperatures were much colder than on the coast, larval development was slower and major peaks of larval availability did not occur until early spring. These different seasonal trends in each region were considered to be related to the climatic differences between the regions.
On pastures which were contaminated continuously, larval numbers reached maximum levels in mid-winter on the Central Coast and in early spring on the Northern Tablelands. It was concluded that the majority of these larvae were derived from the contamination of pastures in autumn and winter. Subsequently in summer, a rapid dying out of larvae was observed in all the regions, probably due to the effect of hotter weather.
The studies suggest that a reduction in the contamination of pasture with nematode eggs in autumn and winter could result in pastures carrying fewer larvae and thus form the basis of effective worm control programs for cattle.     

Effect of colostrum deprivation of goat kids on the natural transmission of caprine retrovirus infection

An investigation was conducted to determine if depriving goat kids of colostrum and rearing them away from the herd would prevent transmission of caprine retrovirus infection. Twenty-four newborn goat kids were deprived of colostrum and reared on cow's milk away from their dams from an endemically infected goat herd. Twenty-three colostrum-deprived kids had no evidence of retrovirus infection at birth. One kid had sucked briefly and obtained some colostrum resulting in passive transfer of antibody but it did not develop evidence of infection. Nineteen showed no serological evidence of infection over the 370 days of the study. One colostrum-deprived, segregated goat was subsequently found to be infected and developed arthritis-synovitis. Three had doubtful positive response in one or 2 serological tests during the period but no evidence of infection in leucocyte co-cultures. Cells centrifuged from colostrum of infected goats were co-cultivated with foetal goat synovial membrane cultures. Caprine retrovirus was isolated from cells in the colostrum from the 3 goats examined.     

Validation of candidate bovine reference genes for use with real-time PCR

Accurate quantification with real-time PCR requires the use of stable endogenous controls. Recently, there has been much debate concerning the stability of commonly used reference or housekeeping genes. To address this concern, a number of statistical approaches have been designed to analyse data and assist in determining the most appropriate reference genes for experimental comparisons. In this study, three programs, BestKeeper, Norm Finder, and geNorm were used to assess four candidate reference genes: 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), acidic ribosomal protein large (RPLP0) and beta-actin, for use in expression profiling of individuals from divergent cattle genotypes subject to parasitic challenge with the cattle tick Boophilus microplus. Results demonstrated beta-actin and GAPDH were the most suitable reference genes in blood and could be used either individually or combined as an index to normalise data. RPLP0 was identified as the least stable gene, while 18S rRNA was omitted as being too highly expressed. As the recommendations on the most suitable reference genes varied between the programs, it is recommended that more than one should be utilised, to ensure the most robust experimental tools are selected.     

Isoflurane to prolong medetomidine/ ketamine anaesthesia in six adult female chimpanzees (Pan troglodytes)

Six adult female chimpanzees (Pan troglodytes) were anaesthetised for the placement of intrauterine contraceptive devices, microchips for identification, routine blood sampling, and physical measurements. Anaesthesia was induced with medetomidine in combination with ketamine administered by intramuscular injection with a projectile syringe. Induction was smooth and rapid, but five of the animals were insufficiently relaxed for orotracheal intubation. The plane of anaesthesia was deepened by administering isoflurane delivered in oxygen and nitrous oxide, and general anaesthesia was maintained for up to 74 minutes. The action of medetomidine was reversed at the end of each procedure with atipamezole, and the animals recovered smoothly and uneventfully.