Detection of Clostridium botulinum types C and D toxin by ELISA

An ELISA to detect Clostridium botulinum type D toxin was developed using polyclonal antibodies to a semi-purified toxic complex of the neurotoxin. Sensitivity of the ELISA for detecting type C and type D toxin compared with mouse inoculation was 70% and specificity 96% on samples from animals with botulism diagnosed on clinical signs and herd history. However, both mouse inoculation and the ELISA failed to detect toxin in many animals with a presumptive diagnosis of botulism. Some cross-reaction was seen with Clostridium novyi type A, but not with other clostridial species. While the ELISA described here cannot replace mouse inoculation for the diagnosis of botulism, it is a useful additional test.     

Skin concentrations of thromboxane synthetase inhibitor after topical application with bioelastic membrane

Elevated thromboxane levels are associated with a number of disease states, including dermal pressure ulcers. When dazmegrel was orally administered to greyhound dogs wearing leg casts, it resulted in a sparring effect on the skin areas of potential pressure ulcer development. The objective of this research was to determine if bioelastic matrices could provide controlled release of thromboxane A2 synthetase inhibitor (dazmegrel) at tissue concentrations sufficient for inhibition of thromboxane synthesis. The animal used for these studies was the greyhound, which has thin skin, angular conformation, limited body fat and is predisposed to pressure ulcers similar to those occurring in humans. In vivo skin penetration studies showed that epidermal exposure to bioelastic thromboxane synthetase inhibitor (TSI) matrix resulted in local tissue concentrations of TSI sufficient for thromboxane synthetase inhibition. There were no significant differences between dazmegrel in the skin layers (epidermis, dermis and subcutaneous layers) on 1, 7 and 14-day exposures.     

Increased expression of MAIL, a cytokine-associated nuclear protein, in the prodromal stage of black walnut-induced laminitis

REASONS FOR PERFORMING STUDY: The mediators and signalling cascades important in the initiation of laminitis remain unclear. We therefore wanted to explore the genes and overall signalling mechanisms that play an important role in the developmental stage of laminitis. OBJECTIVE: To use a broad genomic screening technique to identify novel genes that are differentially regulated in the equine lamellae during the developmental period of laminitis. METHODS: Differential mRNA display (DRD) was performed to discover regulated genes, and real-time quantitative polymerase chain reaction (RT-qPCR) was then used to evaluate lamellar mRNA levels of a regulated gene (MAIL) and mediators related to that gene (IL-1beta and IL-6) in control horses (n = 5) and horses administered black walnut extract (BWE; n = 5). RESULTS: Using DRD, MAIL was identified as a regulated gene. RT-qPCR indicated a 4-fold increase in expression of the MAIL mRNA in BWE lamellae compared to controls. A 30-fold increase in IL-1beta, and a 160-fold difference in IL-6 mRNA expression was present in BWE lamellae. Differences in MAIL, IL-1beta and IL-6 mRNA expression were statistically significant between groups (P < 0.05). CONCLUSIONS AND POTENTIAL RELEVANCE: The data strongly support a role for inflammatory cytokines in the developmental stages of laminitis, possibly inducing the vascular and metabolic alterations reported to occur in the affected digit. These results potentially support the use of anti-inflammatory drugs in horses at risk of laminitis, and warrant further investigation of the link between systemic disease processes associated with laminitis and the reported digital inflammation.     

Seasonal changes in plasma adrenocorticotropic hormone and α-melanocyte-stimulating hormone in response to thyrotropin-releasing hormone in normal, aged horses

Background: Results of diagnostic tests for equine pituitary pars intermedia dysfunction (PPID), including endogenous ACTH concentration and the overnight dexamethasone suppression test (DST), are affected by season. New and potentially more sensitive diagnostic tests for equine PPID, such as thyrotropin‐releasing hormone (TRH)‐stimulated ACTH response, have been developed, but have had limited evaluation of seasonality. Objective: Our purpose was to evaluate seasonal changes in plasma ACTH and alpha‐melanocyte‐stimulating hormone (α‐MSH) responses to TRH administration. Animals: Nine, healthy, aged horses with normal DST results. Methods: Synthetic TRH (1 mg) was administered IV. Plasma ACTH and α‐MSH concentrations were measured at 0, 5, 10, 15, 20, 25, 30, 45, 60, and 180 minutes. Testing was performed in February, July, August, September, October, and November. Mean TRH‐stimulated ACTH and α‐MSH concentrations were compared across months and time by repeated measures analysis of variance. Significance was set at the P < .05 level. Results: Concentrations of ACTH and α‐MSH significantly increased after TRH administration. Endogenous and TRH‐stimulated ACTH and α‐MSH concentrations were significantly different across months with higher concentrations in the summer and fall compared with February. Conclusions and Clinical Importance: Plasma ACTH and α‐MSH responses to TRH administration experience seasonal variation, with TRH‐stimulated ACTH and α‐MSH concentrations increasing from summer through fall. These results support previous evidence of a seasonal influence on the equine pituitary‐adrenal axis. More research is warranted with a larger number of horses to determine if seasonal reference ranges for TRH stimulation testing need to be defined.     

Developmental alterations in the regulation of glucagon and insulin secretion in Holstein calves

Twenty-one Holstein bull calves were randomly assigned at birth to 3 groups. Two groups (each of 7 calves) were raised as follows: fed a milk diet alone or fed milk with grain supplementation after 2 weeks of age; studies were done when calves reached 4 weeks of age. The 3rd group was fed on milk with grain supplementation until weaning after which the calves were maintained on grain and pasture. These calves (older calves) were studied at 12 weeks of age. Either propionate (0.28 mmol/kg) or glucose (0.56 mmol/kg) was injected IV in a random order. Samples of blood were obtained from the calves before and immediately after injections were done and at 2, 5, 10, 15, 30, 45, and 60 minutes after secretagogue injection. Plasma was examined for glucose by a glucose oxidase procedure and for immunoreactive insulin (IRI) and glucagon (IRG) by radioimmunoassay. The IRI response to the injection of glucose was greater in older calves (P less than 0.02). Patterns of IRI secretion, as determined by heterogeneity of regression, showed age differences for both secretagogues (P less than 0.05). Base-line IRG was greater in milk/grain-fed calves than in milk-fed calves (P less than 0.05). Mean IRG response to propionate injection was higher (P less than 0.05) in milk/grain-fed calves than in milk-fed calves. Plasma glucose concentration increased in older calves, but decreased in milk-fed calves after propionate injection. The data indicate that maturation in the ruminant is accompanied by altered regulation of insulin and glucagon secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Observations on the indirect transmission of virulent ovine footrot in sheep yards and its spread in sheep on unimproved pasture

SUMMARY: Virulent ovine footrot was transmitted accidentally to a group of 23 adult Merino sheep (flock B) after holding for 1 hour in sheep yards, which earlier the same day had contained another flock (flock A) with < 1% prevalence of sheep with footrot lesions. Sheep in flock B were rendered susceptible to virulent footrot by grazing 600 mm high unimproved pasture dominated by paspalum ( Paspalum dilatatum ) and kangaroo grass ( Themeda australis ) during warm, humid and wet weather. In addition to moisture, interdigital abrasions caused by the pasture might have predisposed the interdigital skin to infection with Dichelobacter nodosus .     

Restriction-endonuclease analysis of Australian isolates of Leptospira interrogans serovar hardjo from cattle with agalactia and abortion

SUMMARY Polyacrylamide gel electrophoresis and agarose gel electrophoresis were used to resolve restriction endonuclease digests of 20 Australian isolates of Leptospira interrogans cultured from urine samples of cattle with agalactia and abortion. The restriction endonuclease profiles of 19 isolates closely matched the profiles of L interrogans serovar hardjo subtype hardjobovis reference strains. The remaining isolate had a different restriction profile from subtype hardjobovis and subtype hardjoprajitno reference strains and was serologically identified as serovar pomona. Silver staining of polyacrylamide gels gave enhanced resolution of restriction fragments compared with the traditional method of ethidium bromide staining of agarose gels.     

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.