Cytokeratins of the matrices of the chestnut (torus carpeus) and periople in horses with acute laminitis

OBJECTIVES: To determine whether there is a change in the expression of cytokeratins in the epidermal cells of the non-weight-bearing parts of the limb in horses with acute laminitis and thus determine whether the morphologic changes that develop in the periople and chestnut (torus carpeus) of horses early in acute laminitis are caused by inhibition of keratinocyte differentiation. ANIMALS: 8 horses with acute laminitis. PROCEDURE: Tissue specimens were obtained from the chestnuts of all 8 horses and from the stratum externum of the hoof wall of 3 horses. Tissue specimens were obtained within 48 hours of the first clinical signs of laminitis. The cytokeratins were characterized by 1- and 2-dimensional gel electrophoresis, and the tissue distribution of the cytokeratins was studied by immunohistochemical staining. RESULTS: The biochemical findings indicated that the epidermal cells of tissues from horses affected by laminitis contained the same set of cytokeratins as corresponding tissues from clinically normal horses. Immunohistochemistry on sections from specimens of horses with laminitis versus clinically normal horses indicated a difference in the expression of cytokeratin in the basal cells in the matrix of the stratum externum of the hoof wall and in the matrix of the chestnut of horses with laminitis in which the most severe morphologic changes were observed. CONCLUSIONS: Inhibition of keratinocyte differentiation, as observed by immunohistochemical changes, in cells in parts of the chestnut and periople may indirectly indicate that the observed epidermal changes in horses with laminitis are primary and are unaffected by weight-bearing.     

Detection of calprotectin and apoptotic activity within the equine colon from horses with black walnut extract-induced laminitis

The black walnut extract (BWE) model of equine laminitis is associated with a systemic inflammatory response manifest by increased expression of inflammatory cytokines in the lungs and liver as well as the laminae. The specific role of the gastrointestinal tract in development of this response is unclear and is of utmost importance, as gastrointestinal disease and laminitis are intimately related. We investigated calprotectin expression and epithelial and endothelial apoptosis in the colon of horses exposed to orally administered BWE. Sections of colon from 19 horses including 7 controls not exposed to BWE, 6 horses at the developmental time-point of leukopenia (DTP) and 6 at the onset of Obel grade 1 laminitis (LAM) after BWE-administration were histologically examined. Immunohistochemical evaluation for calprotectin expression with MAC 387 antibody was performed along with assessment of epithelial and endothelial apoptosis with caspase-3 active antibody. Calprotectin expression and percentage of apoptotic cells were compared between controls and the two treatment groups and presence of a correlation between calprotectin expression and apoptosis was evaluated. Histological findings from BWE-treated horses included eosinophil and lymphocyte epitheliotropism. The DTP group had a higher (p<0.01) calprotectin score with respect to the control group, while there was no significant difference in percentage of epithelial and endothelial apoptotic cells between groups (p=0.08 and p=0.48 respectively). No significant correlation was found between calprotectin score and epithelial or endothelial apoptosis (p=0.69 and p=0.29 respectively). There is preliminary evidence that exposure of horses to BWE results in an early inflammatory response in the colon. Further studies are needed to characterize the nature of the colonic injury in BWE-exposed horses and the link to the development of laminitis.     

MAGNETIC RESONANCE IMAGING OF THE INITIAL ACTIVE STAGE OF EQUINE LAMINITIS AT 4.7 T

Equine laminitis is a severely debilitating disease. There is a poor understanding of the underlying pathophysiology, and traditional imaging modalities have limited diagnostic capacity. High field strength magnetic resonance (MR) imaging allows direct visualization of the laminae, which other modalities do not. This would prove useful both in assessment of clinical patients and in further investigation into the pathophysiology of the disease. The objective of this study was to characterize the anatomic changes within the equine foot associated with the initial active stage of laminitis. Images obtained using a 4.7 T magnet were compared with digital radiographs using histologic diagnosis as the reference standard. Objective measurements and subjective evaluation for both modalities were evaluated for the ability to predict the histologic diagnosis in horses with clinical signs of laminitis as well as in clinically normal horses and horses that were in a population at risk for developing laminitis. Signal intensity and architectural changes within the corium and laminae were readily seen at 4.7 T, and there was a strong association with the histologic diagnosis of active laminitis. Measurements obtained with MR imaging were more sensitive and specific predictors of laminitis than those obtained radiographically. Subjective evaluation with MR imaging was more sensitive than with radiography and should become more specific with greater understanding of normal anatomy.     

Circulatory and blood gas changes accompanying the development and treatment of induced laminitis

A peripheral vasodilatory agent, isoxsuprine hydro-chloride, was evaluated in a controlled study for its efficacy in the treatment of acute equine laminitis. Eight healthy, adulthorses of variable age and sex were used in the trial. Acute laminitis was induced in 5 of the horses by oral carbohydrate overload. Intravenous isoxsuprine therapy (1.8 mg/kg) was initiated in 3 of the horses receiving carbohydrate overload at first sign of clinical lameness and repeated at 12-hour intervals. Intravenous saline placebos were administered on a similar schedule to 2 control horses which also received a carbohydrate overload. The remaining 3 horses served as further controls. Local and systemic responses to induction of laminitis and isoxsuprine administration were assessed by subjective evaluation of clinical lameness in a double blind trial; nuclear scintigraphy and radiography of the distal forelimbs; and assessment of physical, hematological and biochemical parameters.

Pronounced tachycardia, hypotension and sweating accompanied the intravenous infusion of isoxsuprine. The 3 horses treated with isoxsuprine following the induction of laminitis showed a more rapid improvement in soundness than horses receiving saline placebos. No horse developed rotation of the third phalanx in response to the diet Nuclear scintigraphy indicated that blood perfusion patterns within the hoof of laminitic horses altered with isoxsuprine therapy, but an overall increase or decrease in perfusion was not apparent. Alterations in serum enzyme and electrolyte profiles with the onset of laminitis generally concurred with findings previously reported for this model of the disease. No change in coagulation profiles accompanied the onset of laminitis or isoxsuprine administration. Blood gas analysis indicated an increase in median palmar vein oxygen partial pressure (PO₂) levels with onset of laminitis. A concurrent decrease in the median palmar arteriovenous oxygen partial pressure difference (AVO₂) was significant at the P<0.01 level. There was no difference in median palmar vein PO₂ values between thoseanimals receiving isoxsuprine and those receiving saline placebo therapy. Results of the trialindicated that isoxsuprine may be beneficial in the treatment of acute laminitis. Further controlled studies are appropriate.     

The prevalence of endocrinopathic laminitis among horses presented for laminitis at a first-opinion/referral equine hospital

Endocrinopathic causes of laminitis may be a common underlying causative pathogenesis in first-opinion or field cases presenting with laminitis, as opposed to laminitis produced in inflammatory research models. This study aimed to determine whether evidence of an underlying endocrinopathy was present in horses presented for laminitis to a first-opinion/referral veterinary teaching hospital. A second aim was to compare the signalment of horses and ponies with laminitis with the equine hospital population during the same period. All horses presenting for laminitis at Helsinki University Equine Teaching Hospital, Finland, over a 16-month period were examined for an underlying endocrinopathy. Horses presenting for laminitis were compared with the hospitalized population over the same period. There were 36 horses presented for laminitis, and evidence of endocrinopathy was present in 89%. Of the horses showing an underlying endocrinopathy, one-third had a diagnosis of pituitary pars intermedia dysfunction, and two-thirds showed basal hyperinsulinemia indicative of insulin resistance, without evidence of hirsutism. Phenotypic indicators of obesity were present in 95% of horses with basal hyperinsulinemia without hirsutism. Compared with the hospital population during the same period, horses with laminitis associated with an underlying endocrinopathy were significantly older and more likely to be pony breeds. Our data support that endocrine testing should be performed on all cases of laminitis that do not have a clear inflammatory or gastrointestinal origin.     

Equine laminitis model: Cryotherapy reduces the severity of lesions evaluated seven days after induction with oligofructose

Reasons for performing study: A previous preliminary study demonstrated the potential of distal limb cryotherapy (DLC) for preventing laminitis. Clinically, DLC must be effective for periods longer than 48 h and the preventive effect must extend beyond its discontinuation. Objectives: To evaluate the effect of DLC, applied during the developmental phase of induced laminitis, on the severity of clinical laminitis and lamellar histopathology 7 days after dosing. Methods: Eighteen normal Standardbred horses were divided into 3 groups of 6. Continuous cryotherapy was applied for 72 h to the distal limbs of the first group. The second and third groups were administered laminitis inducing doses of oligofructose and 72 h of cryotherapy applied (immediately after dosing) to the second group. After clinical assessment all horses were subjected to euthanasia 7 days after dosing and hoof lamellar tissues were harvested and analysed. Results: In the laminitis induced horses clinical lameness and laminitis histopathology was significantly reduced in horses that underwent 72 h of DLC compared with untreated controls. Cryotherapy alone produced no significant lameness or other ill effect. Conclusions: Continuous, medium‐ to long‐term (72 h) cryotherapy applied to the distal limbs of horses safely and effectively ameliorates the clinical signs and pathology of acute laminitis. Potential relevance: Pre‐emptive distal limb cryotherapy is a practical method of ameliorating laminitis in ill horses at risk of developing the disease.     

Glucocorticoids and laminitis in the horse.

The administration of exogenously administered GCs and syndromes associated with GC excess are both attended by increased risk for the development of laminitis in adult horses. However, there exists substantial controversy as to whether excess GCs cause laminitis de novo. If true, the pathogenesis of laminitis arising from the effects of GC excess is probably different from that associated with diseases of the gastrointestinal tract and endotoxemia. Although a satisfactory explanation for the development of laminitis as a consequence of GC action is currently lacking, numerous possible and plausible theoretical mechanisms do exist. Veterinarians must exert caution with respect to the use of GCs in adult horses. The extent to which individual horses are predisposed to laminitis as a result of GC effect cannot be predicted based on current information. However, the administration of systemic GCs to horses that have been previously affected by laminitis should be used only with extreme caution, and should be accompanied by careful monitoring for further signs of laminitis. The risk of laminitis appears to be greater during treatment using some GCs (especially dexamethasone and triamcinalone) compared with others (prednisone and prednisolone). Whenever possible, to reduce the risk of laminitis, GCs should be administered locally. For example, the risk of GC-associated laminitis is evidently considerably reduced in horses affected with chronic obstructive pulmonary disease (COPD) if GC treatment is administered via inhalation. We have hypothesized that structural changes in the equine hoof that resemble laminitis may arise as a consequence of excess GC effect. Although these changes are not painful per se, and are not associated with inflammation, they could likely predispose affected horses to the development of bona fide laminitis for other reasons. Moreover, the gross morphological appearance of the chronically GC-affected hoof resembles that of a chronically foundered hoof in some respects. Further investigation into the effect of GC on the hoof lamellar interface is clearly needed.     

Dynamic changes in circulating leukocytes during the induction of equine laminitis with black walnut extract

Administration of black walnut heartwood extract (BWHE) via nasogastric tube induces acute laminitis in horses. However, the processes responsible for the development of laminitis, including laminitis induced with BWHE, remain unclear. The results of recent studies indicate that administration of BWHE initiates an inflammatory response in the laminar tissues and that this response may be due to extravasation of activated leukocytes from the circulation. This study examines the effects of BWHE administration on the dynamics of circulating neutrophils and monocytes, and the capacity of blood leukocytes to produce radical oxygen species (ROS) over the time period from administration of BWHE to the development of lameness consistent with Obel grade I laminitis.

Individual horses, free of pre-existing musculoskeletal disease, were administered either 6 l of BWHE or an equal volume of water at time 0 (T = 0). Blood samples were collected prior to dosing and at 1, 2, 3, 4, 6, 8, 10 and 12 h after dosing, or until the onset of Obel grade I laminitis. For each sample, total leukocyte counts were determined followed by collection of buffy coats and removal of erythrocytes by hypotonic lysis. Leukocytes were either fixed for flow cytometric assessment of differential counts or maintained in culture to measure endogenous and phorbol ester-induced production of ROS. At each sample time, the number of cells recovered and the flow cytometric differential counts were compared with corresponding total leukocyte counts determined by the Clinical Pathology laboratory.

Horses administered BWHE had a significant reduction in circulating leukocytes at 3–4 h relative to values for horses administered the same volume of water. Horses that developed Obel grade I laminitis had a significant reduction in circulating leukocytes when compared to values for horses administered BWHE that did not become lame. Flow cytometric analysis revealed a consistent decrease in the total number of monocytes obtained from horses that developed laminitis. In these same horses, the endogenous level of ROS production was significantly higher at T = 0 than for horses that did not become lame. Furthermore, production of ROS by leukocytes from horses that developed laminitis increased significantly and coincided with the decrease in circulating leukocytes.

Collectively, these findings support a role for systemic activation of leukocytes and induction of inflammation by BWHE as a factor in the early pathogenesis of acute laminitis. Because laminitis often develops as a sequel to diseases characterized by systemic inflammatory events, activation and emigration of neutrophils and monocytes may be important factors in the early pathogenesis of laminitis in clinical cases.     

Evaluation of suspected pituitary pars intermedia dysfunction in horses with laminitis

OBJECTIVE: To determine prevalence and clinical features of pituitary pars intermedia dysfunction (PPID) in horses with laminitis. DESIGN: Case series. ANIMALS: 40 horses with laminitis. PROCEDURES: Horses with laminitis that survived an initial episode of pain and were not receiving medications known to alter the hypothalamic-pituitary-adrenal axis were tested for PPID by evaluation of endogenous plasma ACTH concentration. Signalment, suspected cause, month of onset and duration of laminitis, Obel grade of lameness, pedal bone rotation, physical examination findings, results of endocrine function tests, treatment, outcome, and postmortem examination findings were recorded. RESULTS: Prevalence of PPID as defined by a single high plasma ACTH concentration was 70%. Median age of horses suspected of having PPID (n = 28) was 15.5 years, and median age of horses without PPID (12) was 14.5 years. Laminitis occurred most frequently in horses with and without suspected PPID during September and May, respectively. Chronic laminitis was significantly more common in horses suspected of having PPID. In horses suspected of having PPID, the most common physical examination findings included abnormal body fat distribution, bulging supraorbital fossae, and hirsutism. Five horses suspected of having PPID had no clinical abnormalities other than laminitis. Seventeen horses suspected of having PPID that were treated with pergolide survived, and 3 horses that were not treated survived. CONCLUSIONS AND CLINICAL RELEVANCE: Evidence of PPID is common among horses with laminitis in a primary-care ambulatory setting. Horses with laminitis may have PPID without other clinical signs commonly associated with the disease.     

Assessment of apoptosis in epidermal lamellar cells in clinically normal horses and those with laminitis

OBJECTIVE: To determine and compare the number, type, location, and distribution of apoptotic epidermal cells in the laminae of clinically normal horses and horses with laminitis. SAMPLE POPULATION: Formalin-fixed samples of digital lamellar tissue from 47 horses (including clinically normal horses [controls; n = 7], horses with acute [4] and chronic [7] naturally acquired laminitis, and horses with black walnut extract-induced [11] or carbohydrate overload-induced [18] laminitis). PROCEDURE: Blocks of paraffin-embedded lamellar tissues were stained for DNA fragmentation with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Differential immunohistochemical staining for caspases 3 and 14 were used to confirm apoptosis. RESULTS: The number of TUNEL-positive epidermal cells per 0.1 mm of primary laminae was significantly greater in the acute laminitis group than in the other groups. In the acute laminitis group, there were 17 and 1,025 times as many TUNEL-positive basal layer cells and keratinocytes, respectively, compared with the control group. Apoptosis of TUNEL-positive basal layer cells was confirmed by results of caspase 3 immunohistochemical staining. The TUNEL-positive keratinocytes did not stain for caspases 3 or 14. CONCLUSIONS AND CLINICAL RELEVANCE: The large number of apoptotic basal layer cells detected in the lamellar tissue of horses with acute naturally acquired laminitis suggests that apoptosis may be important in the development of acute laminitis. The role of the large number of TUNEL-positive keratinocytes detected in the interface of primary and secondary epidermal laminae of horses with acute laminitis remains to be elucidated.     

Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses

Equine metabolic syndrome is characterized by obesity and regional adiposity coupled with evidence of recurrent laminitis. Although inflammation has been well characterized in several experimental models of acute laminitis, the inflammatory events associated with endocrinopathic laminitis are not well documented. The aim of this study was to characterize selected markers of inflammation in horses with clinical evidence of equine metabolic syndrome (EMS). Neutrophil phagocytosis and oxidative burst, as well as endogenous and stimulated cytokine expression were evaluated. A marked increase in neutrophil reactive oxygen species production upon phagocytosis was observed in horses with EMS that was strongly correlated to the blood insulin concentration. Increased oxidative burst activity of neutrophils in hyperinsulinemic horses may predispose horses with metabolic syndrome to laminitis. In contrast, peripheral blood cells of obese hyperinsulinemic horses showed decreased endogenous proinflammatory cytokine gene expression (IL-1 and IL-6) and similar cytokine response following immune stimulation compared to that of control horses. This may suggest that, unlike in people, cytokine-mediated inflammation does not increase in direct response to obesity or insulin resistance in horses. This species-specific disparity may explain the difference in clinical outcomes observed in obese horses compared to obese people.     

Equine laminitis: Induced by 48 h hyperinsulinaemia in Standardbred horses

Reasons for performing study: Hyperinsulinaemia is known to induce laminitis experimentally in healthy ponies with no history of the condition. Horses are more insulin sensitive than ponies and whether prolonged hyperinsulinaemia and euglycaemia would have a similar laminitogenic effect requires study. Objectives: To determine if laminitis results when the prolonged euglycaemic hyperinsulinaemic clamp technique (p‐EHC) is applied to clinically normal Standardbred horses, and to monitor hoof wall temperature seeking an association between vascular activity and laminitis development. Methods: Eight young, clinically normal Standardbred horses were assigned into 4 pairs and within each pair, one was assigned randomly to either treatment (n = 4) or control (n = 4) groups. Treated horses received continuous infusions of insulin and glucose until clinical signs of laminitis developed, at which point the horses were subjected to euthanasia. Control horses received an equivalent volume of a balanced electrolyte infusion for the same period. Hoof wall surface temperature (HWST) was monitored continuously throughout the experimental period. Results: All horses in the treatment group were calculated to have normal insulin sensitivity. All treated horses, and none in the control group, developed laminitis (P = 0.01). Pronounced digital pulses were a feature of the treatment group, while insignificant digital pulses occurred in control horses. HWST was higher and less variable in treated horses once hyperinsulinaemia was established. Conclusions: Healthy Standardbred horses subjected to prolonged hyperinsulinaemia develop laminitis within 48 h, demonstrating that laminitis in horses can be triggered by insulin. Potential relevance: Insulin resistance and the associated hyperinsulinaemia place horses and ponies at risk of developing laminitis. This study demonstrates a need for prompt management of the persistent hyperinsulinaemia seen in some endocrinopathies.     

Voluntary limb-load distribution in horses with acute and chronic laminitis

OBJECTIVES: To compare limb-load distribution between horses with and without acute or chronic laminitis. ANIMALS: 10 horses with carbohydrate-induced acute laminitis, 20 horses with naturally occurring chronic laminitis, and 20 horses without foot abnormalities (controls). PROCEDURES: Limb-load distribution was determined, using a custom-designed system that allowed simultaneous quantification of the mean percentage of body weight voluntarily placed on each limb (ie, mean limb load) and the SD of the mean load over a 5-minute period (ie, load distribution profile [LDP]). Load distribution profile was used as an index of frequency of load redistribution. RESULTS: Mean loads on fore- and hind limbs in control horses were 58 and 42%, respectively, and loads were equally and normally distributed between left and right limbs. In addition, forelimb LDP was greater, compared with hind limbs, and was affected by head and neck movement. In comparison, limb-load distribution in horses with chronic laminitis was characterized by an increase in the preferential loading of a forelimb, a decrease in total forelimb load, and an increase in LDP that was correlated with severity of lameness. In horses with carbohydrate-induced acute laminitis, mean limb loads after onset of lameness were not different from those prior to lameness; however, LDP was significantly decreased after onset of lameness. CONCLUSION AND CLINICAL RELEVANCE: Quantification of limb-load distribution may be an applicable screening method for detecting acute laminitis, grading severity of lameness, and monitoring rehabilitation of horses with chronic laminitis.     

Initial Experience Using Contrast Magnetic Resonance Imaging in Laminitic Horses: 18 Studies

Plain radiographic imaging inadequately identifies soft-tissue pathology and only distinguishes chronic laminitis after the development of notable displacement of the distal phalanx. The window of opportunity for maximum response to treatment occurs before biomechanical failure of the lamellar attachment. Radiographic and magnetic resonance venograms allow vascular assessment of patients affected with acute laminitis. When vascular findings are interpreted with additional magnetic resonance information, the degree of damage to the foot in patients affected with laminitis is understood more thoroughly. This article describes the technique, findings, and outcome in a group of laminitic horses that underwent contrast magnetic resonance imaging.     

The Role of Insulin in Endocrinopathic Laminitis

Laminitis is a devastating disease of horses that usually arises as a consequence of major systemic disease or endocrine disturbances. Research has been confounded by apparently disparate results and theories on pathogenesis. Models of laminitis have greatly advanced our understanding of the disease, yet have mostly involved perturbations of the gastrointestinal tract or inflammatory models. A major trend in research on laminitis in the past few years has been the increasing interest in endocrine dysfunction resulting in laminitis. A new model of laminitis associated with hyperinsulinemia has recently been discovered and the central role of high insulin in triggering endocrinopathic laminitis highlighted. This review discusses the pathophysiology of insulin resistance and hyperinsulinemia in horses and possible mechanisms of insulin-induced laminitis.     

The developmental and acute phases of insulin-induced laminitis involve minimal metalloproteinase activity

Metalloproteinases have been implicated in the pathogenesis of equine laminitis and other inflammatory conditions, through their role in the degradation and remodelling of the extracellular matrix environment. Matrix metalloproteinases (MMPs) and their inhibitors are present in normal equine lamellae, with increased secretion and activation of some metalloproteinases reported in horses with laminitis associated with systemic inflammation. It is unknown whether these enzymes are involved in insulin-induced laminitis, which occurs without overt systemic inflammation. In this study, gene expression of MMP-2, MMP-9, MT1-MMP, ADAMTS-4 and TIMP-3 was determined in the lamellar tissue of normal control horses (n=4) and horses that developed laminitis after 48 h of induced hyperinsulinaemia (n=4), using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Protein concentrations of MMP-2 and MMP-9 were also examined using gelatin zymography in horses subject to prolonged hyperinsulinaemia for 6h (n=4), 12h (n=4), 24h (n=4) and 48 h (n=4), and in normal control horses (n=4). The only change in gene expression observed was an upregulation of MMP-9 (p<0.05) in horses that developed insulin-induced laminitis (48 h). Zymographical analysis showed an increase (p<0.05) in pro MMP-9 during the acute phase of laminitis (48 h), whereas pro MMP-2 was present in similar concentration in the tissue of all horses. Thus, MMP-2, MT1-MMP, TIMP-3 and ADAMTS-4 do not appear to play a significant role in the pathogenesis of insulin-induced laminitis. The increased expression of MMP-9 may be associated with the infiltration of inflammatory leukocytes, or may be a direct result of hyperinsulinaemia. The exact role of MMP-9 in basement membrane degradation in laminitis is uncertain as it appears to be present largely in the inactive form.     

Submural histopathologic changes attributable to peracute laminitis in horses

OBJECTIVE: To describe submural histopathologic changes attributable to peracute laminitis in horses. ANIMALS: 20 adult horses. PROCEDURE: A concurrent-control design was used to compare laminar lesions in 10 horses subjected to carbohydrate-induced laminitis with laminar characteristics of 10 sex- and aged-matched control horses with normal feet. Horses in the treatment group were administered an overload of carbohydrate. Tissues were obtained by biopsy 4 to 8 hours after onset of lameness or 72 hours after administration of the carbohydrate overload when lameness did not develop. Sections were stained with H&E, Masson's trichrome, and periodic acid-Schiff stains. Histopathologic changes were analyzed to detect differences between groups and to correlate epidermal changes with severity and duration of lameness. RESULTS: Analysis indicated that dermal and epidermal lesions were evident despite lack of visible separation of the epidermal basement membrane, can be found in horses without detectable lameness, and were nonspecific and progressive following onset of lameness. Furthermore, severity and location of lesions were associated with severity and duration of lameness. CONCLUSION AND CLINICAL RELEVANCE: These observations are consistent with the concept that separation of the laminar epithelial basement membrane is a delayed step in the pathogenesis of acute laminitis, digital vascular hypoperfusion is an underlying cause for laminitis, and the potential for repeated episodes of subclinical laminitis may underlie the development of structural and mechanical changes consistent with chronic laminitis despite lack of clinical signs of acute laminitis.     

Serum markers of lamellar basement membrane degradation and lamellar histopathological changes in horses affected with laminitis

In order better to evaluate the extent to which degradation of the lamellar basement membrane (LBM) by matrix metalloproteinases (MMP) occurs in equine laminitis, we determined the concentration of type IV collagen and laminin in normal and laminitic horses, using specific immunoassays. Blood samples were obtained from both the jugular and the cephalic veins of horses (n = 10) before and after the induction of acute alimentary laminitis by carbohydrate overload. Jugular and cephalic venous blood samples were also obtained from horses affected with naturally occurring laminitis (n = 16) and nonlaminitic controls (n = 8). The serum collagen IV concentration was not changed following the induction of laminitis in the experimental group. Serum collagen IV concentration was increased in jugular venous blood obtained from cases of naturally occurring laminitis (mean +/- s.e. 218.04 +/- 18.59 ng/ml) compared with nonlaminitic controls (157.50 +/- 10.93 ng/ml) (P<0.05). Serum collagen IV concentration was also increased in jugular venous blood obtained from severely laminitic horses (219.50 +/- 18.18 ng/ml) compared with nonlaminitic controls (157.50 +/- 10.93 ng/ml) (P<0.05). A difference in serum concentration of collagen IV was not identified based on chronicity of naturally occurring laminitis. Serum laminin concentration did not differ between laminitic and nonlaminitic horses. Differences in serum laminin concentration were not identified based on sampling location (jugular or cephalic vein), severity of laminitic pain, or chronicity of spontaneous laminitis. In conclusion, the circulating concentration of collagen IV was increased in horses affected with naturally occurring laminitis. The potential role for serum collagen IV assay for characterisation of equine laminitis warrants further investigation.     

Chronic laminitis is associated with potential bacterial pathogens in the laminae

A common sequella of chronic laminitis in horses is repeated abscesses with variable lameness and drainage. It is unclear whether the exudate represents the debridement phase of a non-septic inflammatory process involving clearance of laminar tissue damaged during the acute episode of laminitis, or a response to a microbial infection developed by ascent of microbes from the environment to the tissue via the white line. The objective of this study was to evaluate the possibility that an undiagnosed microbial infection in laminar tissue is present in laminar tissue collected from chronically laminitic horses without an active hoof abscess. Methods to collect laminar tissue, aseptically, from control (non-laminitic) horses and those with chronic/recurrent laminitis are described. Laminae homogenates were evaluated for the presence of bacteria. Bacteria were identified using biochemical tests and sequencing of 16S rRNA and virulence genes. Laminae from chronically laminitic horses revealed 100-fold higher levels (P=0.002) of bacteria compared to control, non-laminitic horses. Although environmental organisms were identified, potential pathogens were identified. Included were Gram positive bacteria, Brevibacterium luteolum, coagulase-negative Staphylococcus spp. as well as Gram negative bacteria, enterohemorrhagic Escherichia coli and Alcaligenes faecalis. Further research is warranted to evaluate the role of bacteria in equine chronic laminitis.     

Evaluation of heparin for prophylaxis of equine laminitis: 71 cases (1980-1986)

The records of 71 horses with small intestinal disorders requiring surgical correction were disorders requiring surgical correction were reviewed to compare the prevalence of laminitis in those horses treated prophylactically with heparin and the prevalence of horses not treated with heparin. The prevalence of laminitis was 13% (9/71), and there was no significant difference (P less than 0.05) in the prevalence of laminitis between the 2 groups. The lack of significant benefit after treatment with heparin indicates that further work needs to be done on the equine coagulation system before heparin can be advocated for prevention of laminitis.