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.     

The sympatho-adrenal system and plasma levels of adrenocorticotropic hormone, cortisol and catecholamines in equine grass sickness

Plasma levels of adrenocorticotropic hormone (ACTH), cortisol and catecholamines were used to study the role of the sympatho-adrenal system in equine grass sickness. Statistical evaluation determined differences of hormone levels between seven horses with grass sickness (one acute, five subacute and one chronic), six horses with colic (one with laminitis) and 16 control horses before and after mild stress. Plasma levels of the hormones were higher in horses with acute and subacute grass sickness than in the other groups. No differences were detected between horses with colic and stressed control horses but some hormone levels differed between control and colic horses and control horses before and after stress. It is possible that hyperactivation of the sympatho-adrenal system is caused by stress but it is uncertain whether the stress is only a result of the severity of the disease or also plays a role in its aetiology.     

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.     

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.     

Changes in nasal mucosal innervation in horses with grass sickness

REASONS FOR PERFORMING STUDY: Equine grass sickness is a dysautonomia characterised by widespread destruction of autonomic ganglia, resulting in the clinical signs of dysphagia, constipation, profuse sweating, tachycardia, rhinitis sicca and high mortality rate. Rhinitis sicca is a common finding in horses with the chronic form and we have postulated that alterations in autonomic innervation of the nasal mucosa might underlie this clinical presentation. OBJECTIVES: In this study, the expression and distribution of nerve fibres immunoreactive for calcitonin gene-related peptide (CGRP), substance P (SP), the general neuronal marker protein gene-product 9.5 (PGP 9.5; ubiquitin) and the intermediate neurofilaments (PAN-N; neurorfilaments L, M and H) in the nasal mucosa of normal horses (n = 10) and horses with EGS (n = 18; acute n = 8, subacute n = 3, chronic n = 7) was assessed. METHODS: Innervation density and distribution was investigated in the different groups using standard immunohistochemical techniques. RESULTS: Significant differences were noted when comparing the density and distribution of nerve fibres immunoreactive for PGP 95 and PAN-N, with PGP 95 consistently giving better staining in all groups and at all sites in the nasal mucosa. An apparent increase in the density of innervation was noted for acute vs. normal cases. A significant reduction in the density of innervation was noted only with PAN-N when comparing normal horses and acute cases with the chronic group (P < 0.05). CGRP and SP immunoreactive nerve fibres were typically most abundant in the epithelial and subepithelial layers, but the quality of staining and nerve fibre density was greater for SP, achieving significant difference in several comparisons. The density of innervation for SP was significantly reduced in the chronic group compared to the normal and acute groups (P < 0.01). A significant decrease was noted for CGRP only for the acute and chronic groups (P<0.05). CONCLUSIONS: These results demonstrate a reduction in the expression of the sensory neuropeptides in nasal mucosal innervation as a consequence of equine dysautonomia, and may underlie the clinical presentation of rhinitis sicca noted with this disease. POTENTIAL RELEVANCE: Nasal biopsy may be of use in antemortem diagnosis of grass sickness and identification of mucosal denervation; and might also be useful in the treatment of rhinitis in EGS cases.     

Serum protein changes in grass sickness

Polyacrylamide gel electrophoresis was used to compare serum taken from ponies before and during clinical illness confirmed as grass sickness. A consistent rise in the level of haptoglobin was seen in serum from animals which had shown symptoms for more than two days. Serum albumin was also shown to have altered mobility at the onset of clinical disease. Estimation of the haemoglobin-binding capacity confirmed the haptoglobin increase. This haptoglobin has been purified and some of its properties determined. In contrast to the situation in acute inflammatory conditions no other acute-phase proteins increased to a significant extent in grass sickness. It is concluded that the neurotoxin known to be present in sera of animals experiencing acute grass sickness cannot itself be detected by polyacrylamide gel electrophoresis but may be bound to serum albumin.     

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.     

Clinical equine dysautonomia and autonomic neuron damage.

Damage to the neurons of selected autonomic ganglia was quantified in relation to the severity of the clinical signs shown in acute, subacute and chronic cases of dysautonomia (grass sickness). No connection between the clinical severity of acute or subacute dysautonomia and the amount of neuronal damage in the superior cervical, stellate and coeliaco-mesenteric ganglia could be demonstrated. However, a higher proportion of normal neurons were found in chronic cases. Jejunal submucosal neuronal damage was correlated with clinical severity but further work is required to confirm this finding and to establish how widespread the alimentary neuronal lesions are in dysautonomia of different severities.     

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.     

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     

Chronic lung inflammation affects plasma amino acid concentrations in pigs

Metabolic changes associated with inflammatory processes and immune response can modify protein and AA requirements. Improved knowledge of these processes will provide opportunities for nutritional intervention to sustain growth and animal defense at the same time. The objective of the study was to identify AA whose metabolism is affected by chronic lung inflammation. Six pairs of littermate piglets were selected at 28 d of age on the basis of their BW. After catheterization of the jugular vein, one littermate received complete Freund's adjuvant (CFA) intravenously, whereas its littermate was injected with a sterile saline solution (CON). Piglets within a litter were pair-fed in order to avoid confounding effects of feed intake and inflammation on plasma AA concentrations. Blood samples were taken after an overnight fast and 2 h after the morning meal for 9 d. Rectal temperature, food consumption, weight gain, plasma haptoglobin, and AA concentrations were measured. The CFA injection decreased food intake, and increased body temperature and plasma haptoglobin concentration. Plasma tryptophan, glutamine, proline, glycine, tyrosine, ornithine, total AA concentrations, and the ratio of tryptophan to large neutral AA were less in CFA than in CON (P < 0.05), independent of time and meal. In contrast, plasma histidine concentration was higher (P < 0.05) in CFA than in CON pigs. Plasma serine, arginine, alanine, asparagine, and total AA concentrations were lower in CFA than in CON pigs only in the fed state (P < 0.05). Among essential AA, only plasma tryptophan concentration was lower (P < 0.01) in CFA than in CON pigs in both fasted and fed state. These results show that chronic lung inflammation affects individual AA differently and suggest that the utilization of some AA increased during chronic lung inflammation in pigs. Activation of tryptophan catabolism enzyme indoleamine 2,3-dioxygenase seems a relevant hypothesis to explain the increased tryptophan utilization, although its incorporation in acute-phase proteins and the existence of other catabolic pathways cannot be excluded.     

Acute phase protein responses to uterine bacterial contamination in cattle after calving

Repeated ultrasonographic examinations and collections of blood samples and uterine lumenal swabs between seven and 28 days after calving were used to examine the relative effects of bacterial contamination and involution of the uterus on the concentrations of acute phase proteins in the blood of 26 dairy cows. The severity of bacterial contamination, as determined by the total bacterial growth score, was a significant variable for the concentrations of the acute phase proteins alpha1-acid glycoprotein (P < 0.0001), haptoglobin (P < 0.05) and ceruloplasmin (P < 0.0001). In addition, the concentrations of alpha1-acid glycoprotein and ceruloplasmin were increased in the cows from which Escherichia coli (P < 0.0001) and Arcanobacterium pyogenes (P < 0.05), respectively, were isolated from the uterine lumen. Uterine involution, as determined by the decreasing diameter of the previously gravid uterine horn, was associated with a decrease in the concentrations of alpha1-acid glycoprotein (P < 0.005), haptoglobin (P < 0.05) and ceruloplasmin (P < 0.01). However, the response of the acute phase proteins to bacterial contamination was independent of the day on which the samples were collected, indicating that their concentrations were increased by bacterial contamination in addition to the changes associated with uterine involution.     

Comparison of IgG antibody levels to Clostridium botulinum antigens between euthanased and surviving cases of chronic grass sickness

Serum from 12 horses suffering from chronic grass sickness (CGS) were assayed for IgG antibodies against botulinum neurotoxins C and D (BoNT/C and BoNT/D) and to a surface antigen extract of a neurotoxin negative strain of Clostridium botulinum type C. Collectively, the six surviving CGS cases demonstrated significantly higher initial IgG levels (P=0.05) against surface antigens than the six that were subsequently euthanased. The surviving animals also demonstrated higher initial IgG levels against the BoNT/C but not reaching significance (P=0.06). The two groups demonstrated no difference between IgG levels against BoNT/D. This study supports existing evidence of the involvement of C. botulinum type C in the aetiology of grass sickness.     

Acute phase proteins in cattle: discrimination between acute and chronic inflammation.

Acute phase proteins such as serum amyloid A, haptoglobin, and alpha 1-acid glycoprotein have been identified as markers of inflammation in cattle because they are produced by the liver in response to pro-inflammatory cytokines. This study was designed to assess whether they could be used to discriminate between acute and chronic inflammation. Their concentrations were measured in serum samples from 81 cattle in which inflammation was classified by thorough clinical examination, supported by postmortem findings, as being acute in severity in 31 and chronic in 50. The classical haematological markers of inflammation were also determined in blood from the animals. Serum amyloid A had a maximum (100 per cent) clinical sensitivity in discriminating between the acute and chronic cases, and haptoglobin had the highest clinical specificity of 76 per cent; counts of neutrophils and band neutrophils had sensitivities of 71 per cent and 42 per cent and specificities of 30 per cent and 72 per cent, respectively. It was concluded that serum amyloid A and haptoglobin may be used to discriminate between acute and chronic inflammatory conditions.     

Re: The safety of oral calcium formate

A Short Communication was published in the October 2002 issue of the New Zealand Veterinary Journal entitled An investigation of the safety of oral calcium formate in dairy cows using clinical, biochemical and histopathological parameters (McIntyre and Weston 2002). It appeared to be comparing results directly with those of a prior communication (Scott and Van Wijk 2000). The following should be considered when comparing the two papers. Haptoglobins were propounded by McIntyre and Weston as definitive indicators of acute inflammation, yet current literature does not give consistent viewpoints as to their value. The authors cited instances where the haptoglobin concentration was measured in cases of infective disease. However, Skinner et al (1991) believed that haptoglobin concentrations increase as a result of the infection rather than the trauma. To further complicate matters Alsemgeest et al (1994) found that haptoglobin concentrations were elevated in cattle with chronic rather than acute conditions whereas Horadagoda et al (1999) believed the concentrations of acute-phase proteins were higher in acute inflammation. However, they found serum amyloid A was more reliable as an indicator of acute, rather than chronic, inflammation than was haptoglobin. Therefore, histopathology would appear to give much more conclusive evidence of mucosal inflammation than the determination of haptoglobin...     

Comparison of duodenitis/proximal jejunitis and small intestinal obstruction in horses: 68 cases (1977-1985)

Sixty-eight horses with colic caused by small intestinal disease were allotted into 2 groups of 34 on the basis of recorded findings during exploratory celiotomy, necropsy, or response to medical treatment alone. Signalment, history, physical examination findings, and laboratory findings were compared between the group of horses with small intestinal obstruction and the group with duodenitis/proximal jejunitis. A significantly greater proportion of horses with duodenitis/proximal jejunitis were older than 2 years old (P less than 0.05). Differences in sex or breed distribution, or in seasonality of the 2 disease syndromes were not observed. Horses with duodenitis/proximal jejunitis had significantly greater signs of depression than those with small intestinal obstruction (P less than 0.01), and horses with small intestinal obstruction had significantly greater signs of abdominal pain (P less than 0.05). The mean heart and respiratory rates were significantly lower (P less than 0.01) and the volume of nasogastric reflux was significantly greater (P less than 0.05) in the group of horses with duodenitis/proximal jejunitis. Sections of small intestine that were palpable per rectum were less distended and there were more auscultable borborygmi in horses with duodenitis/proximal jejunitis, compared with those with small intestinal obstruction (P less than 0.05 and P less than 0.01). The group of horses with duodenitis/proximal jejunitis had lower mean plasma potassium and higher mean plasma bicarbonate concentrations (P less than 0.05) than the group with small intestinal obstruction. The mean nucleated cell count and total protein concentration of peritoneal fluid specimens were significantly less in the group with duodenitis/proximal jejunitis (P less than 0.01); however, these values were greater than normal.(ABSTRACT TRUNCATED AT 250 WORDS)     

Behavioural and EEG changes in sheep in response to painful acute electrical stimuli

Objective To measure acute pain in sheep, based on a human pain model, and examined changes in both electroencephalogram frequency spectrum and behavioural responses to increased electrical stimulation in sheep. Design Analysis of variance (treatment and animal effects) for stimulus intensity where each animal received each electric shock treatment given in the order 0, 5,10 and 20 mA. Procedure Eight sheep with electrodes implanted over the surface of the brain were examined for escape-avoidance and electroencephalogram responses to four levels of electrical stimulation from 0–20 mA. Results With increasing stimulus intensity at the time of feeding, the sheep were more hesitant to return to the feeder or remain near the feeder following stimulation. There was little difference between the 0 and 5 mA stimuli for any of the behaviour variables (P > 0.05). However, there were marked increases in the time taken to re-approach the feeder after receiving an electric shock of 5 mA and of 20 mA (P < 0.05; mean values 3 and 119 s, respectively) and remaining near the feeder for 5 s (P < 0.001; mean values 10 and 167 s, respectively). Following the stimulus, there was an overall increase in the electroencephalogram power spectrum in the first four seconds, which then rapidly returned to normal. In particular, the 20 mA stimulus resulted in higher absolute power values than in the control (0 mA) treatment for delta 2 (P < 0.001), theta 1 (P < 0.05), theta 2 (P < 0.05), alpha 1 (P < 0.001), alpha 2 (P < 0.001) and beta 1 (P < 0.01) band-widths. Similarly, for the 10 mA stimulus, the absolute power values were greater than the control treatment for delta 2 (P < 0.05), alpha 1 (P < 0.01), alpha 2 (P < 0.001) and beta 1 (P < 0.01) bandwidths. Conclusion The experiment suggests that a human acute pain model is applicable to sheep and that these electroencephalogram changes may provide a good measure of acute pain in sheep.     

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.     

Clinical relevance of monocyte procoagulant activity in horses with colic

Endotoxin-activated monocytes express a thromboplastin-like procoagulant activity on the cell surface that may serve as a focal point for formation of microvascular thrombi. Because coagulopathy is a common sequela to endotoxemia in the equine species, we investigated the ability of monocytes, isolated from horses with colic, to express procoagulant activity. On the day of admission, and on the third and fifth day of hospitalization, monocytes were isolated from 30 adult horses with colic. A coagulation profile, including prothrombin time, activated partial thromboplastin time, thrombin time, and plasma fibrinogen and serum fibrin degradation products concentrations, was determined at each sample collection. The concentration of endotoxin in the plasma was quantitated at the time of admission. Ten clinically normal adult horses served as controls. The procoagulant activity of monocytes isolated from horses with colic was significantly (P less than 0.05) greater than that of the monocytes isolated from clinically normal horses. On the first and third day of hospitalization, the mean prothrombin time was significantly (P less than 0.05) longer in horses with colic, compared with clinically normal horses, and was the most common abnormality in the coagulation profile on the day of admission (25/30; 83%). Mean fibrin degradation products concentration was significantly (P less than 0.05) greater in horses with colic on the day of admission and was the second most common abnormality in the coagulation profile on day 1 (23/30; 77%). In horses with colic, the mean prothrombin and activated partial thromboplastin times were significantly (P less than 0.05) longer in horses that did not survive, compared with horses that survived.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of type of forage fed to dairy heifers: associations between claw characteristics, clinical lameness, environment and behaviour

Heifers were reared separately on wet fermented (WF) (grass silage based) or dry unfermented (DU) (straw based) diets. Clinical lameness was recorded and physical claw attributes were measured regularly, up to six months after calving at approximately two years of age. Two months before calving, some aspects of animal behaviour and the properties of slurry were studied. The incidence of lameness due to claw horn lesions was significantly higher (P < 0.01) in WF than in DU. WF suffered more serious heel erosion prior to calving (P < 0.05) and had softer claw horn at two (heel and axial sole) out of five sites throughout (P < 0.01 and P < 0.05). The slurry produced by WF had lower total solids content (P < 0.05) and viscosity (P < 0.001). WF spent longer standing and feeding (P < 0.001). It is proposed that standing for longer in less viscous slurry contributed to softer claw horn and more severe heel erosion, predisposing WF to claw horn lesions causing lameness. Reducing contact with low dry matter slurry is recommended for improving claw health.