Serum lipids and lipoproteins in equine colic and grass sickness

Serum total lipids, lipoprotein fractions, triglycerides, cholesterol, phospholipids and free fatty acids were measured in horses with acute, subacute and chronic grass sickness (equine dysautonomia) and in colic cases. The values were compared with those of normal grazing and stabled horses. A marked individual variation occurred, but total lipids, triglycerides and free fatty acids were significantly higher than normal in grass sickness and colic cases with cholesterol was significantly higher than normal in grass sickness cases only. Pre-beta lipoprotein was significantly increased in colic and subacute grass sickness although all abnormal groups showed this fraction which was absent from normal horses. The percentage of alpha 2b lipoprotein was significantly higher in colic and grass sickness. The changes described are typical of those occurring in fat mobilisation in the horse and are considered to be due to a number of factors including decreased food intake, cortisol and catecholamine release and insulin resistance.     

Acute phase proteins in grass sickness (equine dysautonomia)

Four acute phase proteins were assayed in the serum of normal horses and those with acute, subacute and chronic grass sickness, colic and inflammatory conditions, in order to investigate their diagnostic value in grass sickness. The grass sickness and inflammation group had a significantly increased haptoglobin concentration (P less than 0.01-P less than 0.001). Orosomucoid was elevated in acute, subacute and chronic grass sickness and inflammation (P less than 0.001, P less than 0.001, P less than 0.05 and P less than 0.05, respectively). Highest concentrations of haptoglobin and orosomucoid were recorded in subacute grass sickness. Ceruloplasmin was significantly higher in acute grass sickness cases than all other groups except the colic group (P less than 0.05-P less than 0.01). alpha 2-macroglobulin was significantly higher in acute grass sickness than normal, colic and chronic grass sickness cases (P less than 0.01, P less than 0.05 and P less than 0.05). The time scale of changes suggests that the stimulus to haptoglobin and orosomucoid synthesis occurs at the onset of clinical signs whereas the increase in ceruloplasmin and alpha 2-macroglobulin is more likely to reflect haemoconcentration.     

Biomarkers of exposure to cyanogens in horses with grass sickness

To test the hypothesis that equine grass sickness may be associated with the ingestion of cyanogenic glycosides from white clover (Trifolium repens), the concentrations of whole blood cyanide, and plasma and urinary thiocyanate, the main metabolite of cyanide, were measured in 12 horses with acute grass sickness and 10 horses with subacute grass sickness, and in 43 control horses, of which 21 were co-grazing with cases of acute grass sickness, 12 grazed pastures where grass sickness had not been reported, and 10 were stabled horses. The healthy horses which grazed with cases of acute grass sickness had higher concentrations of blood cyanide, and plasma and urinary thiocyanate than the other control horses, consistent with an increased exposure to cyanogens. The horses with grass sickness had no evidence of a recent intake of cyanogens, but may have been exposed to increased levels of cyanogens before they became anorexic.     

Evaluation of urinalysis as an aid in the diagnosis of equine grass sickness

To determine whether urinalysis can aid the diagnosis of equine grass sickness, samples of urine from 15 horses with acute grass sickness, eight horses with subacute grass sickness, 17 co-grazing horses and 17 stabled control horses were analysed. The samples from all of the horses with grass sickness had a significantly higher specific gravity, higher protein and creatinine concentrations and a significantly lower pH; the samples from the horses with acute grass sickness also had significantly higher glucose concentrations. These differences may support a diagnosis of grass sickness but they are not pathognomonic for the disease.     

Analysis of peritoneal fluid as a diagnostic aid in grass sickness (equine dysautonomia)

The analysis of peritoneal fluid is of value in the differential diagnosis of equine colic but its characteristics have not been evaluated in grass sickness. Peritoneal fluid was collected from 15 normal horses and from 11 cases of medical colic, 11 cases of surgical colic, 20 cases of acute grass sickness and 13 cases of subacute grass sickness. The fluid was analysed for its appearance, total and differential white cell count, specific gravity, total protein concentration and total and intestinal alkaline phosphatase activity. Fluid from cases of medical colic was normal in these respects. Surgical cases were unique in having bloodstained fluid with a high alkaline phosphatase activity. Grass sickness cases had a higher specific gravity and protein content than the cases of medical colic although the appearance of the fluid was similar. Grass sickness cases were distinguishable from cases of surgical colic on the basis of the appearance of the fluid and its lower alkaline phosphatase activity.     

Clinical and biochemical features of grass sickness (equine dysautonomia)

An attempt has been made to assess the diagnostic value of clinical features seen at initial examination of horses with grass sickness, colic cases and cases submitted as possible grass sickness but diagnosed subsequently as some other condition. There appears to be no single pathognomonic sign for grass sickness. A range of signs has been associated with grass sickness but these are of value only when related to the length of illness and the history. Biochemical tests related to intestinal tissue damage, stress and dehydration were evaluated and most were found to be of value in diagnosing acute grass sickness. No evidence was found relating selected mineral or vitamin deficiencies to grass sickness, nor were protein changes specific. A marked fall in glutathione peroxidase values was associated with grazing rather than disease.     

Functional and histopathological evidence of cardiac parasympathetic dysautonomia in equine grass sickness

The parasympathetic terminal cardiac ganglia were examined in three normal horses and in five horses with grass sickness. Histopathological changes, consistent with those observed in other ganglia of horses with grass sickness, were identified in the terminal cardiac ganglia of the affected horses. A functional analysis of cardiac autonomic control by time domain analysis (TDA) of heart rate variability was applied to eight horses with grass sickness, and double-paired to 16 normal horses on the basis of their age, sex and breed, and the time of day at which the electrocardiographs were obtained. There were statistically significant differences between the normal and the affected horses in terms of the triangular index (P=0.01), a geometric method of TDA, the sNN100 index (P=0.009), a statistical method of TDA, and the R to R interval (P<0.0005). These results indicate that there is a pathological and functional parasympathetic decentralisation to the heart in equine grass sickness.     

Systemic concentrations of antioxidants and biomarkers of macromolecular oxidative damage in horses with grass sickness

REASONS FOR PERFORMING STUDY: The aetiopathogenesis of equine grass sickness (EGS) is unknown. The role of free radical-mediated neuronal damage has not previously been investigated in this condition. OBJECTIVES: To investigate the potential contribution of oxidative damage and antioxidant status to neurodegeneration in EGS. METHODS: Systemic levels of surrogate biomarkers were determined in 10 horses with acute EGS and in 2 control populations; 10 healthy horses co-grazing with the 10 EGS horses at the onset of clinical disease, and 10 healthy mares grazing where EGS has not been reported. RESULTS: EGS horses had alterations in levels of several antioxidants, consistent with oxidative stress, the acute phase response and/or the secondary metabolic complications of EGS. EGS horses had elevated plasma dihydroxyphenylalanine (DOPA) levels. CONCLUSIONS: The elevated DOPA levels probably reflected a generalised disturbance of catecholamine metabolism rather than increased DOPA production via free radical-mediated oxidation of tyrosine. However, there was no evidence of systemic macromolecular oxidative damage. POTENTIAL CLINICAL RELEVANCE: Further work is required to determine whether macromolecular oxidative damage occurring at the neuronal level contributes to EGS.     

Unusual compound of small molecular weight in the serum of horses with acute grass sickness

An unusual compound of small molecular weight has been detected in serum from horses with acute grass sickness by solvent extractions and thin-layer chromatography. The substance has not been detected in the serum of normal horses or cases of equine colic and apparently disappears from grass sickness serum after two to three days of clinical illness. Although this compound is found in sera which are known to possess in vivo neurotoxicity, this property could not be demonstrated in either the total chloroform extract which contains the compound or in the hydrophilic serum components remaining after extraction. These findings suggest that, although the compound may be related to the neurotoxin present in the original serum, the toxicity is inactivated during the extraction procedure or that the emulsion necessary for administration precludes absorption of an active toxin.     

Cholinergic activity of intestinal musclein vitro taken from horses with and without equine grass sickness

Equine grass sickness (EGS) is a pan-dysautonomia of horses that involves central and peripheral neuronal degeneration and ultimately depletion. This is the first reported functional study on the motility of equine intestine taken immediatelypost mortem from horses with EGS. Strips of smooth muscle from the small intestine of healthy and EGS-affected horses were suspended in an organ bath and their motility was measured isometrically. The activity of the cholinergic system was studied. Physostigmine enhanced the motility of all muscle strips. Tissues taken from horses suffering from acute grass sickness (AGS) had the longest latency before a measurable response could be obtained (p<0.05). The ileum appeared to be damaged by EGS to a greater extent than the duodenum. For the duodenal strips the enhanced rate of spontaneous contractions was significant (p<0.05) for both normal tissue and that affected by grass sickness but this was not the case for the ileal strips. Muscarinic receptor sensitivity investigation using bethanecol suggested a hypersensitivity of receptors with AGS material,Abbreviations AGS acute grass sickness - CGS chronic grass sickness - ED₅₀ median effective dose - EGS equine grass sickness - VIP vasoactive intestinal peptide     

Plasma arginine vasopressin concentration in horses undergoing surgery for colic

Objective – To determine if horses before undergoing anesthesia for surgical correction of colic would have lower plasma arginine vasopressin (AVP) concentrations than healthy horses undergoing anesthesia for arthroscopic surgery, and would not increase their plasma AVP concentrations in response to anesthesia and surgery. Design – Prospective clinical study. Setting – University teaching hospital. Animals – Fourteen horses with colic and 8 healthy horses. Interventions – Horses with colic underwent anesthesia and surgery for alleviation of colic, and healthy horses underwent anesthesia and surgery for arthroscopy. Measurements and Main Results – Plasma AVP was measured perioperatively in horses with colic and in healthy horses. Before anesthesia, and 30 and 60 minutes after induction, horses with colic had greater median plasma AVP concentrations than control horses (P≤0.001); thereafter during anesthesia differences in AVP concentrations between the 2 groups were not significant. In the control group, plasma AVP concentration increased during 120 minutes of anesthesia; no such increase occurred in colic horses. Conclusions – Compared with healthy horses, horses with colic had higher preanesthesia plasma AVP concentrations that did not increase further in response to anesthesia and surgery. Exogenous AVP is associated with decreased splanchnic perfusion in a variety of animal species and, therefore, could be detrimental to horses with colic. Thus, it may be inappropriate to use exogenous AVP in support of blood pressure in anesthetized horses with colic. Further studies are warranted to define appropriate indications for the use of AVP in horses with colic.     

Comparison of the effects of fructose and glucose supplementation on metabolic responses in resting and exercising horses

The aim of this study was to compare the effects of two different carbohydrate sources (fructose and glucose) on the metabolic responses in resting and exercising horses. The following regimes were fed in randomized order to five trained horses at rest and immediately before or during exercise. The resting regime comprised 0.6 kg grass meal pellets (control) or 0.6 kg grass meal pellets supplemented with either 50% glucose or 50% fructose. The exercise regime comprised 0.3 kg grass meal (control) or 0.6 kg grass meal pellets supplemented with either 50% glucose or 50% fructose fed immediately before or during simulated endurance exercise on a treadmill (30 km, total running time 120 min; 15 min rest after 60 min running time). Blood samples were collected for the analysis of glucose, insulin, free fatty acids (FFA) and lactate. In resting horses, glucose supplementation resulted in a significantly higher glycaemic and insulinaemic response than the control or fructose feedings (treatment P < 0.05). Plasma glucose levels were significantly higher after glucose supplementation given before or during exercise. Similar plasma glucose concentrations were observed after fructose was fed before exercise, whereas fructose supplementation during exercise resulted in a lower glycaemic response than glucose feeding (P < 0.05). Plasma insulin, FFA and lactate levels showed exercise-related changes (time P < 0.05), but treatment did not effect these results. Plasma glucose concentrations were elevated during the simulated endurance exercise after glucose feeding, and no counter-regulation by insulin occurred. The higher glycaemic response might be beneficial as exogenous glucose can be used as an energetic substrate during prolonged exercise. Fructose exhibited no clear advantages in comparison with glucose as a carbohydrate source for exercising horses.     

Studies of oesophageal function

Lesions of the sympathetic nervous system have been associated with grass sickness for many years (Obel, 1955). Dysphagia is also an accepted clinical feature of subacute or chronic cases. Megaoesophagus has been reported in horses with grass sickness, but it was uncertain whether oesophageal dilation was a primary condition, or a sequel to gastric distension (Greet, 1982). Robertson and others, (1948) suggested that dysphagia was alleviated in affected horses after drainage of gastric contents. It is interesting to note that dysphagia was present in less than half the cases in this study although radiological abnormalities of the oesophagus were similar in all cases. Consistent radiological abnormalities of the oesophagus occurred in 12 out of the 14 horses examined (this consisted of incoordination or atony of the thoracic oesophagus). In the two suspected cases of grass sickness which made a clinical recovery, the radiological abnormalities were slightly different. Contrast material was transferred slowly through the cervical oesophagus and pooled at the thoracic inlet as well as at several sites in the cervical oesophagus. Eventually the contrast material passed through the distal oesophagus into the stomach. Although gastric distension was present in many of the cases examined it was not possible to identify this by radiographic means. It is unlikely that the signs of oesophageal incoordination and dilation were related only to gastric distension, as in most of the cases, contrast material pooled in the oesophagus at the thoracic inlet rather than just cranial to the diaphragm. It seems reasonable to suggest that these radiological abnormalities resulted from neurological impairment of the oesophagus. It would be of particular value to examine horses with obstructive lesions of the stomach or duodenum to evaluate the effect of upper gastro-intestinal obstruction on oesophageal motility. A number of horses with other conditions of the upper alimentary tract have been examined radiographically at these clinics. Two horses suspected of suffering from grass sickness were found to have primary oesophageal abnormalities. One had a diverticulum and the other a localized area of dilation; both showed regurgitation of food and weight loss. Barium swallows in both horses defined the sites of their localized lesions but there was no evidence of either Type I or Type II oesophageal malfunction, and at post-mortem examination the ganglionic changes associated with grass sickness were absent. Examination of one horse with ileocaecal intussusception did not show radiological features of the oesophagus typical of those shown by cases of grass sickness. However, contrast material passed slowly through the upper oesophagus of a foal with congenital megaoesophagus in a manner similar to the Type II oesophageal malfunction described above. It also demonstrated dilation and gross inco-ordination of the thoracic oesophagus with pooling of contrast material at the thoracic inlet and oscillation between the thoracic inlet and diaphragm as seen in Type I malfunction in proven grass sickness. It is recognized that the radiological findings of oesophageal dilation and inco-ordination merely demonstrate the presence of neuromuscular impairment of oesophageal movement. Until more cases of upper gastro-intestinal disease can be examined, the specificity of these functional abnormalities for grass sickness cannot be accurately assessed. However the results assume considerable diagnostic significance when demonstrated in an adult horse which is exhibiting signs of grass sickness, particularly those of colonic impaction. The need for sophisticated equipment and thus the necessity of transport to centres so equipped, is clearly a drawback to the technique but this is often outweighed by the advantage of being able to avoid unnecessary laparotomy. As there is at present no cure for grass sickness, the improved diagnostic capability will allow severely ill horses to be destroyed without delay. It may also identify horses which on rare occasions appear to be making a slow recovery.     

Long-term survival of equine surgical colic cases. Part 1: patterns of mortality and morbidity

Postoperative complications and mortality can occur many weeks or months after colic surgery. We are interested in the long-term outcome of these cases. This study documents patterns of mortality and morbidity among 341 horses that recovered from colic surgery March 1998-August 2000. The progress of each horse was rigorously followed by periodic telephone and postal questionnaires. Event time data were recorded for each animal and a total of 321 horse years of survival, together with death from all causes, colic-related death and various postoperative complications. Postoperative survival (of all horses excluding grass sickness cases) was triphasic over the first 600 days and there was marked mortality in the first 10 days postoperatively. The probability of survival postoperatively decreased to 0.87 by 10 days, 0.82 by 100 days and declined slowly to 0.75 at 600 days. Horses suffering from epiploic foramen entrapment had a significantly reduced probability of postoperative survival (RR = 2.1, P = 0.033). The causes of death for 104 horses that died postoperatively and the prevalence of postoperative complications are recorded for the study population. Postoperative colic was the most prevalent complication with 100 horses (29%) suffering one or more episodes. However, only 16 horses (4.6%) suffered 3 or more episodes. The incidence of postoperative colic was 0.55 episodes/horse year at risk. This study provides data that will inform the prognosis for postoperative colic cases and identifies epiploic foramen entrapment as carrying a worse prognosis for survival than other strangulating lesions.     

Survival and complication rates in 300 horses undergoing surgical treatment of colic. Part 1: Short-term survival following a single laparotomy

REASONS FOR PERFORMING STUDY: A minority of equine colic cases prove fatal unless treated surgically; however, few studies have considered long-term survival and complication rates, and few have attempted to identify factors that might affect outcomes. Such information is required for owners and veterinary surgeons to make informed decisions about the most appropriate treatment for individual cases. OBJECTIVES: To document short-term survival rates of 300 horses undergoing colic surgery and analyse factors that might have predisposed to short-term death. METHODS: History, clinical and surgical findings, treatments and outcomes of 300 surgical colic cases (1994-2001) were reviewed. Comparisons among groups of discrete data were made using chi-squared or Student's t tests as appropriate. Significance was set at P < 0.05, and 95% confidence intervals were calculated for percentages. RESULTS: The short-term survival rate (to discharge) was 70.3% for all horses and 83.1% for those recovering from anaesthesia; for horses that had a single laparotomy it was 87.2%. The most common reasons for death/euthanasia in the post operative period after a single laparotomy were persistent pain/colic, post operative ileus and grass sickness. Horses with lesions involving the small intestine and caecum had lower survival rates (75.2 and 66.7%, respectively) than those with large colon or small colon lesions (89.9 and 100%, respectively). The survival rate for ischaemic/strangulating lesions (68.9%) was lower than for simple obstructions (90.5%). CONCLUSIONS: Short-term survival of horses undergoing exploratory laparotomy for acute colic is dependent on many factors, including the nature of the underlying disease, cardiovascular status and post operative complications. POTENTIAL RELEVANCE: These retrospective studies may be used as a basis for prospective studies assessing treatments that could ultimately improve survival and decrease complication rates.     

A comparison of traditional and quantitative analysis of acid-base and electrolyte imbalances in horses with gastrointestinal disorders

The purpose of this study was to compare traditional and quantitative approaches in analysis of the acid-base and electrolyte imbalances in horses with acute gastrointestinal disorders. Venous blood samples were collected from 115 colic horses, and from 45 control animals. Horses with colic were grouped according to the clinical diagnosis into 4 categories: obstructive, ischemic, inflammatory, and diarrheic problems. Plasma electrolytes, total protein, albumin, pH, pCO2, tCO2, HCO3-, base excess, anion gap, measured strong ion difference (SIDm), nonvolatile weak buffers (A(tot)), and strong ion gap were determined in all samples. All colic horses revealed a mild but statistically significant decrease in iCa2+ concentration. Potassium levels were mildly but significantly decreased in horses with colic, except in those within the inflammatory group. Additionally, the diarrheic group revealed a mild but significant decrease in Na+, tCa, tMg, total protein, albumin, SIDm, and A(tot). Although pH was not severely altered in any colic group, 26% of the horses in the obstructive group, 74% in the ischemic group, 87% in the inflammatory group, and 22% in the diarrheic group had a metabolic imbalance. In contrast, when using the quantitative approach, 78% of the diarrheic horses revealed a metabolic imbalance consisting mainly of a strong ion acidosis and nonvolatile buffer ion alkalosis. In conclusion, mild acid-base and electrolyte disturbances were observed in horses with gastrointestinal disorders. However, the quantitative approach should be used in these animals, especially when strong ion imbalances and hypoproteinemia are detected, so that abnormalities in acid-base status are evident.     

The role of quantitative electromyography (EMG) in horses suspected of acute and chronic grass sickness

REASONS FOR PERFORMING THE STUDY: Clinical evidence of motor neuron involvement in equine grass sickness (EGS) has not been reported. HYPOTHESIS: Quantitative electromyography (EMG) analysis can elucidate subtle changes of the lower motor neuron system present in horses with EGS, performed ante mortem. METHODS: Fourteen horses diagnosed clinically with acute, subacute or chronic EGS were examined and quantitative EMG performed. Previously published data on healthy horses and horses with proven lower motor neuron disease (LMND) were used as controls. In 8 horses post mortem examination was performed, and in 7 muscle biopsies of the lateral vastus muscle underwent histopathology and morphometry. RESULTS: Clinical electrophysiological evidence of neuropathy was present in 12 horses. Analysis of data from the first 4 horses resulted in 95% confidence intervals (CI) of nontransformed data for motor unit action potential (MUP) duration in subclavian, triceps and lateral vastus muscle of 11.0-13.7, 14.8-20.3 and 12.2-17.2 msecs, respectively, and for MUP amplitude 291-453, 1026-1892 and 957-1736 microV, respectively. For number of phases the 95% CI was 3.6-4.4, 2.9-3.6 and 2.9-3.4, respectively, and for number of turns 5.0-6.5, 4.3-5.3 and 3.7-4.6, respectively. No changes in duration of insertional activity were measured. Pathological spontaneous activity was observed in all horses. EGS as evidenced by degenerative changes in the autonomic ganglia in combination with minor degenerative changes of the spinal lower motor neurons was observed on post mortem examination in all 8 available autopsies. In muscle biopsies of 4 out of 7 horses changes consistent with slight neurogenic atrophy were found. CONCLUSIONS AND POTENTIAL RELEVANCE: EMG results demonstrated the presence of a neuropathy of skeletal muscles in all horses suspected to have EGS. The combination of clinical and electrophysiological evidence may aid differential diagnosis of neurogenic disease in cases of weight loss and colic.     

Prevalence of Gastric Ulcerations in Horses with Colic

The goal of this study was to determine the prevalence of gastric ulcers in horses with acute abdominal crisis (colic) and to examine the temporal effect of hospitalization on ulcer development in equine patients treated for colic. In addition, other factors that may be associated with gastric ulceration were also explored. The study design was a prospective original study incorporating 169 horses that presented to the George D. Widener Hospital for examination. One hundred and twelve horses presenting with the chief complaint of colic were included in the study group, and 57 horses that presented for non-colic or nonemergency complaints were evaluated and included as case controls. Gastroscopy was performed on equine patients presenting with the chief complaint of colic or horses presenting for reasons other than colic (control); mucosal changes were scored 0 to 3. Additionally, horses presenting for colic were gastroscopically evaluated twice during a 5-day period. Medical records were reviewed for history, clinical findings, laboratory results, and treatment. Seventy-six of 112 horses presenting with the chief complaint of colic had gastric ulceration compared with 41 of 57 horses in the control group. There was a significant association between age of the patient and chief complaint (ie, colic vs control) and between breed and chief complaint. There was no association between gastric ulcer score and chief complaint (colic vs control). Thirty-eight of the 112 horses presenting with colic deteriorated in ulcer score while hospitalized. Using a Pearson chi-squared test, there was no statistically significant association between gastric ulceration with age, breed, or sex. Horses with gastric ulceration in the colic group had lower packed cell volumes compared with horses presenting with colic without gastric ulcers, and this was statistically significant (P < .001). The high incidence of gastric ulceration in the study and control groups supports the reports of other investigators that gastric ulcers in horses, no matter the presenting complaint, are widespread. There was a significant association between breed and chief complaint (P = .005); however, breed and outcome of gastric ulceration were not related (Thoroughbreds were the least likely breed to present for colic). Although a trend in increasing gastric ulceration was seen in hospitalized colic patients, it was not statistically significant. This suggests that horses that are hospitalized may be at increased risk for developing gastric ulcers because of stress, feed deprivation, and administration of treatment. Thus, horses that present for colic should be gastroscopically evaluated if clinical signs raise the index of suspicion for gastric ulceration.     

Systemic antibodies to Clostridium botulinum type C: do they protect horses from grass sickness (dysautonomia)?

The aetiology of equine grass sickness (EGS) is still unknown. There is increasing evidence that toxicoinfection with Clostridium botulinum type C is involved. Epidemiological evidence shows that resistance to EGS can occur in older horses and those that have been on a particular pasture for longer or have been in prior contact with the disease. This resistance may be in the form of an immune response to the aetiological agent. Levels of systemic antibodies to the surface antigens of C. botulinum type C (using the closely related and safe C. novyi type A as a phenotypic marker) and to the botulinum type C neurotoxin (BoNT/C) were investigated in horses with and without EGS. Horses with grass sickness were found to have significantly lower levels of systemic IgG to both surface antigens and BoNT/C. Horses with low levels of systemic immunity to these antigens may be more susceptible to developing EGS. There were no significant differences in antibody levels between the different categories of EGS, suggesting systemic immunity to C. botulinum type C does not play a significant role in influencing the severity of the disease. However, horses that had been in contact with EGS or that were grazing land where it had occurred frequently in the past had significantly higher antibody levels to these antigens. These horses may have been exposed to subclinical doses of C. botulinum type C and BoNT/C, resulting in the production of a protective immune response against the putative aetiological agent. This finding is of potential significance for the prospect of prevention of EGS by vaccination against C. botulinum type C.     

A negative serological relationship between cases of grass sickness in Scotland and Clostridium perfringens type A enterotoxin

In an attempt to compare the equine grass sickness as reported in Europe with that described in the Republic of Colombia, sera from horses experiencing grass sickness in Scotland were used in neutralisation tests with Clostridium perfringens type A enterotoxin. The sera, from acute and chronic cases of the disease, failed to neutralise either crude or partially-purified enterotoxin. Neither were precipitin lines formed when the sera were treated against the toxin in immunoelectrophoresis. These results suggest that grass sickness in Europe and the equine disease in Colombia have a different aetiology.