Cytokeratins of the stratum medium and stratum internum of the equine hoof wall in acute laminitis

The cytoskeleton of living keratinocytes consists mainly of cytokeratins that have polymerised into intermediate filaments. The aim of this study was to describe the expression of cytokeratins in the living epidermal cells of the weight-bearing parts of the equine hoof wall during acute spontaneous laminitis. A total of 9 hooves from 3 horses subjected to euthanasia within 48 h of the first clinical signs of laminitis were sectioned and examined. The cytokeratins in the stratum medium and stratum internum of the hoof wall were characterized by 1- and 2-dimensional gel electrophoresis, and the tissue distribution of the cytokeratins was studied by immunohistochemical staining. The biochemical results showed the same set of cytokeratins as was seen in 8 normal horses, reported on previously, used as controls. The immunohistochemical results indicated a difference between normal horses and horses with acute laminitis in the content of cytokeratins in the basal cells of the matrix of the stratum medium of the hoof wall and in the basal and suprabasal cells in the stratum internum at the mid level of the hoof wall. However, no conclusion could be drawn as to whether this change in the cytokeratin distribution in laminitis was primary or was caused by the initiation of the local tissue-repairing process.     

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.     

Evaluation of platelet-rich plasma applied in the coronary band of healthy equine hooves

Platelet-rich plasma (PRP) is a widely used hemocomponent that holds great promise in equine medicine due to its feasible production and regenerative therapy potential. Its use has been considered as a treatment for chronic laminitis, mainly in terms of its analgesic properties and because it can induce growth in affected hooves. The aim of this study was to evaluate the effect on hoof growth attributable to PRP applied in the coronary band of clinically healthy horses by comparing the responses to PRP, saline, and trimming alone. After randomization, the forelimbs of 9 horses received specific treatment at intervals of 33 days and were trimmed, measured, and radiographed at the same time. Neither hoof growth nor hoof angles were different between the treatment groups at any time point. The application of PRP in the coronary band of horses may be a safe procedure; however, it had no effect on the growth or conformation of hooves in clinically healthy horses.     

Clinical Outcome of 14 Obese,Laminitic Horses Managed with the Same Rehabilitation Protocol

A specific method of rehabilitation was used to manage obese horses with laminitis, and clinical outcome was evaluated after 5 to 20 months. Clinical data from 14 similar laminitis cases were statistically analyzed to evaluate response to rehabilitation. Data were analyzed using repeated measures or logistic regression methodologies. Each horse presented as obese and laminitic with no history of a systemic inflammatory disease. The rehabilitation method emphasized a mineral-balanced, low nonstructural carbohydrate diet; daily exercise; hoof trimming that minimized hoof wall loading; and sole protection in the form of rubber hoof boots and/or hoof casts. Distal phalanx alignment within the hoof capsule was significantly improved, and hoof wall thickness was significantly decreased (P < .0001) following treatment. Solar depth was significantly increased (P < .0015). Reduction of palmar angle measurements was detected in acutely and chronically affected horses. This treatment effect was statistically greater for horses with chronic laminitis than for horses with acute laminitis (P interaction < .0001). Horses were 5.5 times more likely to be sound post-treatment than before treatment. Daily exercise, dietary modification, and removal of ground reaction force from the hoof wall were foci of the rehabilitation program. Hoof care and husbandry as applied to these horses may be an effective method of rehabilitation of horses from obesity-associated laminitis.     

Equine laminitis model: Lamellar histopathology seven days after induction with oligofructose

Reasons for performing study: The histopathology of laminitis during its transition from the acute to the chronic phase has not been previously documented. Studying hoof lamellar tissues 7 days after induction of laminitis may provide insight into the intractable nature of the chronic phase of the disease. Objectives: To induce laminitis and investigate hoof wall lamellar tissues 7 days after dosing. Methods: Laminitis was induced using oligofructose in 6 normal Standardbred horses. The dorsal hoof lamellar tissues of these and 12 normal horses were processed and examined by light microscopy. Serial sections of a lamellar tip affected by laminitis were used to create a 3 dimensional reconstruction. Results: Transverse sections of dorsal hoof wall lamellae were significantly longer than normal. Many secondary epidermal lamellae were not connected to primary lamellae and existed as spherical or ovoid, discrete islands isolated in the lamellar dermis. The lamellar basement membrane was intact. Conclusions: Lamellar tissue has the ability to reorganise rapidly following an episode of acute laminitis. Although histopathological evidence of ongoing acute laminitis was absent by 7 days, there was marked disruption of lamellar architecture. Potential relevance: The architecture and subsequent strength of the resultant lamellar interface could be greatly influenced for the better by strategies that minimise mechanical displacement during the acute phase of laminitis.     

Evaluation using hoof wall strain gauges of a therapeutic shoe and a hoof cast with a heel wedge as potential supportive therapy for horses with laminitis

OBJECTIVE: To evaluate using strain gauges, a hoof cast with heel wedge, and a therapeutic shoe with unsupported toe for their effectiveness in redistribution of load from the dorsal hoof wall. STUDY DESIGN: In vitro biomechanical study. SAMPLE POPULATION: Twenty forelimb specimens. METHODS: Rosette strain gauges were placed on the dorsal and lateral hoof wall of 20 normal shaped hooves. Limbs were loaded vertically using a tensile testing machine with a 1 Hz sinusoidally cycling load up to 3000 N during 15 seconds. Mean values of principal strain and direction at 2500 N load were calculated for 3 experimental conditions (unshod, therapeutic shoe with unsupported toe, and hoof cast with heel elevation) and tested by ANOVA (P<.05). RESULTS: Vertical limb loading in an unshod hoof leads to a biaxial compression of the dorsal wall with high longitudinal compression (epsilon2 = -1515 microm/m). Principal strain at the dorsal wall (epsilon2) was decreased by 23% with the therapeutic shoe and by 59% with the hoof cast. On the lateral hoof wall principal strain was unchanged with the shoe, but increased by 34% with the cast. CONCLUSIONS: Strain measurements indicate unloading of the dorsal hoof wall by both methods with the cast being more effective than the shoe. CLINICAL RELEVANCE: The hoof cast with wedge offers substantial unloading of the dorsal wall, but increases load on the quarter. Therefore a hoof cast would likely be most helpful in acute laminitis when palmar structures can still bear load. The therapeutic shoe offers rehabilitation and regrowth of the dorsal wall without increased load on the quarter wall.     

Equine laminitis caused by distal displacement of the distal phalanx: 12 cases (1976-1985)

Clinical data from 12 cases of equine laminitis characterized by distal displacement of the distal phalanx (P3) were reviewed. Clinical features of horses that survived the syndrome were compared with the nonsurvivors to obtain prognostic indicators. Animals affected included 8 Quarter Horses, 2 Welsh ponies, 1 Thoroughbred, and 1 Arabian. Eight of the animals were females (67%), 2 were stallions, and 2 were geldings. The mean age of affected animals was 8.6 years (2 to 14 years), and the mean body weight was 442 kg. The survivors weighed less than the nonsurvivors (384 kg vs 473 kg, respectively), suggesting that body weight may be of prognostic value for horses affected with distal displacement of P3. Ten of the 12 animals (83%) were admitted because of a disorder other than laminitis, but subsequently developed laminitis during the treatment period. All affected animals had clinical evidence of endotoxemia and/or sepsis before the onset of laminitis. Cavitation or depression of the dorsal coronary band was detected in all animals and was the most reliable clinical indicator of distal displacement of P3. Five horses had fluid (blood or serum) ooze from their coronary bands and 2 of these sloughed one or more of their hooves. Necropsy findings of the 8 horses that were euthanatized included severe hemorrhagic, congested laminae and complete detachment of P3 from the hoof wall. Histologic examination of affected laminae revealed vascular thrombosis and multifocal areas of hemorrhage and necrosis. Radiography failed to reveal distal displacement of P3 in 8 animals, but the remaining 4 animals had an accentuation of the dorsal proximal hoof wall and cavitation of the coronary band visible on lateral radiographs.     

RADIOGRAPHIC APPEARANCE OF THE FEET OF MAMMOTH DONKEYS AND THE FINDING OF SUBCLINICAL LAMINITIS

All feet of 10 clinically sound mammoth donkeys (Group I) were radiographed to determine the appearance of the distal phalanx. The distal phalanges had blunted to concave-shaped dorsal solar margins which varied in appearance from slight to pronounced. The distal phalanges of the forefeet were wider than those of the hindfeet, and also were positioned a greater distance from the dorsal aspect of the hoof wall. The greater distance between the dorsal aspect of the hoof wall and the distal phalanges seemed related to the presence of a periosteal-like bony proliferation on the dorsum of the distal phalanx. This bony proliferation occurred in those distal phalanges which also had radiographic findings consistent with pedal osteitis. Next, all feet of 5 additional mammoth donkeys (Group II) that were to be necropsied for various reasons, were examined similarly to Group I, necropsied and found to have laminitis. Only 2 of these 5 donkeys had been lame; only one had rotation of the distal phalanges (in the forefeet). Radiographic data from the 4 donkeys without rotation seemed most similar to that found in those Group I donkeys which had periosteal reactions on their distal phalanges. Conclusions from this study were that: 1) feet of mammoth donkeys have some anatomic differences from those of domestic horses, 2) subclinical laminitis and pedal osteitis can occur in mammoth donkeys, 3) rotation of the distal phalanx occurs in some, but not all laminitic donkeys, 4) laminitic changes may be more pronounced in their fore than in their hindfeet, and 5) additional studies of donkeys need to be done, examining both proven normal and confirmed laminitic feet.     

In vitro measurement of internal hoof strain

REASONS FOR PERFORMING STUDY: Strains during stance on the hoof wall surface have been measured by a number of authors in vitro and in vivo. Histological structure and mechanical properties vary through the wall thickness (radially); radial strain measurements may therefore aid the understanding of mechanical function of the capsule and adjacent tissues. OBJECTIVES: To develop instrumentation capable of measuring internal hoof strain, and to carry out a preliminary comparison of normal and laminitic hooves. METHODS: Six forelimbs from 4 horses, including 2 with laminitis from the same horse, were tested using an Instron test rig designed to simulate the walk at impact, midstance and breakover. Internal strains were measured at a dorsal site using strain gauges moulded into a plug made of 007 fast-set structural adhesive. In addition, kinetic and kinematic data were collected from each specimen. RESULTS: When simulating the walk, a significant (P<0.0001) increase in gradient of radial tensile strain was found in a normal hoof wall, from 5.6 +/- 73.9 microepsilon at the outer gauge to 418.5 +/- 170.6 microepsilon at the inner gauge. However, radial strains measured at the inner gauge site in limbs with laminitis were found to be significantly (P<0.0001) compressive, with values of -406.7 +/- 156.3 and -109.9 +/- 72.4 microepsilon for Specimens 1 and 2, respectively. CONCLUSIONS AND POTENTIAL RELEVANCE: These preliminary data indicate that a marked redistribution may well occur in the wall of laminitic hooves. With a larger sample size, the results should have relevance to the treatment and management of laminitis.     

Effect of prolonged water immersion on equine hoof epidermis in vitro

OBJECTIVE: To evaluate the effect of prolonged water exposure on tissue mass and solutes of outer and inner layers of the stratum medium, sole, frog, and the stratum medium (SMZA) zona alba layer of horses' hooves. SPECIMEN POPULATION: 10 hooves from 10 horses without foot abnormalities. PROCEDURES: Hoof wall tissue specimens were obtained and immersed for 10 days in distilled deionized water. Serial changes in mass were recorded during the immersion period. Subsequently, osmolarity and Na+, K, Cl-, and protein concentrations of the immersion solution were quantified. RESULTS: Fully cornified outer hoof wall, sole, and frog epidermal structures increased in mass, whereas the SMZA lost mass when immersed in water. All hoof structures had a variable loss of crystalloids during immersion, but none of the specimens lost proteins. The frog epidermis was distinct in that total solute lost during immersion could not be ascribed to N+, K+, and Cl-. CONCLUSIONS AND CLINICAL RELEVANCE: Data support a 2-compartment model for the fully cornified outer stratum medium, frog, and sole that permits the exchange of crystalloids, but not proteins, across the cell membrane and infers that topical agents containing proteins cannot benefit the hoof. The unique osmotic behavior of the SMZA relative to other hoof structures suggests the hypothesis that it is composed of transitional epithelial cells. The solutes lost from frog epithelium are interpreted to reflect its unique lipid composition.     

Scanning electron microscopy and fungal culture of hoof horn from horses suffering from onychomycosis

Horn samples were taken from the hooves of eight horses with clinical signs of equine onychomycosis in at least one hoof capsule. None of the horses had a documented mycological history. The predominant alterations of the horn capsules were sand cracks, white line disease, brittleness (especially around the nail holes), parakeratosis and bruising. The horn samples were stored in sterile tubes for transportation and transferred onto Sabouraud Dextrose Agar and dermatophyte test agar for mycological examination within 6 h. Fungal cultures were incubated for 30 days at room temperature. Fungal identification was based on colonial morphology and microscopic examination of conidia. Horn samples were also stored at ?80°C until used for scanning electron microscopy (SEM). The fungal culture revealed that the hoof horn from all eight horses was infected with keratinophilic fungi. The keratinopathogenic fungi Trichophyton spp and Scopulariopsis brevicaulis were also detected in six horses. SEM revealed severe alterations of the horn structure in horn samples infected with keratinopathogenic fungi compared to horn samples from a sound hoof. The most evident changes were deterioration of the tubular structure of the horn wall, disruption of the horny layers, superficial lysis of cornified cells and the presence of fungal elements. Samples without dermatophyte or Scopulariopsis infection, in contrast, were similar to healthy hoof horn.     

A Novel Approach to the Treatment and Prevention of Laminitis: Botulinum Toxin Type A for the Treatment of Laminitis

Laminitis is an inflammation of the lamina of hoofed animals. According to the United States Department of Agriculture (USDA), laminitis impacts approximately 2% of the horse population each year. Because of the severity of the inflammation and chronic pain, it is frequently necessary to euthanize these horses. Surviving horses may be left useless, with resultant economic and social impact on the industry. Current interventions for laminitis are based on supportive care and alteration of biomechanical forces on the digit. The transition from laminitis to laminar failure (founder) is believed to be caused by the “weight of the horse and the forces of locomotion driving the bone down into the hoof capsule.” We hypothesize that an important factor is the torsional forces placed on the distal phalanx (coffin bone) by the deep digital flexor muscle and tendon. The paralyzing effect of botulinum toxin will result in a decrease of these torsional forces and therefore will aid in the prevention or treatment of the sequelae of laminitis. Seven horses with varying degrees of laminitis were evaluated. Each underwent pretreatment and posttreatment radiographs as well as pretreatment and posttreatment Obel grading. Each horse received botulinum toxin type A injected into the belly of the deep digital flexor muscle. In all cases there was radiographic stabilization of pedal displacement from the dorsal hoof wall. Obel scores showed improvement of 1 to 2 grades during the time monitored.     

The treatment of laminitis in horses

The structural and vascular anatomy of the healthy equine foot is compared with the pathologic changes in the foot of horses with acute and chronic laminitis. The structural and vascular abnormalities present in the foot of horses with laminitis are demonstrated in order to explain the abnormal manner in which their feet grow. The medical, surgical, dietary, and endocrine management of acute and chronic laminitis is discussed. Various forms of hoof trimming beneficial to the reestablishment of normal digital perfusion, normal hoof growth, and normal spatial orientation among the distal phalanx, hoof wall, and sole are described. Guidelines for the provision of frog support provided by adjustable heart-bar shoes are presented.     

MAGNETIC RESONANCE IMAGING OF THE EQUINE DIGIT WITH CHRONIC LAMINITIS

Chronic laminitis is a severe disease affecting the equine digit. It was hypothesized that magnetic resonance (MR) imaging would improve visualization of structures within the foot and pathology associated with chronic laminitis. This study aimed to describe the MR imaging findings in chronic laminitis, compare different pulse sequences for visualization of pathology, and to compare MR imaging with standard radiography. Twenty (10 forelimb, 10 hindlimb) cadaver limbs from 10 horses clinically diagnosed with chronic laminitis (group L) and 10 limbs without laminitis (group N) were used. Lateromedial radiographs and sagittal and transverse MR images of the foot were obtained. Radiographs and MR images were evaluated for anatomic definition and evidence of pathology. Dorsal hoof wall thickness and angle of rotation and displacement distance of the distal phalanx were measured. Comparisons were made between group L and N, forelimb and hindlimb within each horse, and MR imaging and radiography. Features consistently noted with MR images in group L, but not detected using radiography, included laminar disruption, circumscribed areas of laminar gas, laminar fluid, and bone medullary fluid. Other findings seen only on MR images included increased size and number of vascular channels, alterations in the corium coronae, and distal interphalangeal joint distension. Magnetic resonance imaging allowed better definition of laminar gas lines and P3 surface irregularity observed on radiographs. Based on measurements, group L had a greater angle of rotation, distal displacement, and dorsal hoof wall thickness than group N; forelimb hoof wall thickness was greater than hindlimb; and distal displacement and hoof wall thickness measurements were smaller using MR imaging than radiography, but had a similar pattern. It is concluded that there are features of chronic laminitis consistently observed using MR imaging and that these may be additional to features observed radiographically.     

Laminitis in the pony: conservative treatment vs dorsal hoof wall resection

The clinical and radiographic findings in 21 ponies with laminitis and its treatment and results are described. All ponies received non-steroidal anti-inflammatory drugs. All were fed hay and no concentrates. They were box rested for varying periods depending on clinical improvement. Dorsal hoof wall resection was performed in 11 ponies and all regained complete soundness. To shorten the period of non-activity, working ponies were shod and the hoof wall defect was packed with technovit or a combination of glue with cotton cuttings. Ten were treated conservatively; two recovered completely, four remained lame and four were killed because of recurrent laminitis.     

A Cross-Sectional Survey of Forelimb Hoof Conformation and the Prevalence of Flat Feet in a Cohort of Thoroughbred Racehorses in New Zealand

Racehorses in New Zealand predominantly train counter clockwise. This training pattern has been associated with between forelimb differences in bone mineral density profile and asymmetrical limb loading after training. At present, there is limited data on the hoof conformation of these racehorses. Distal forelimb and digital hoof conformation data were collected from 75 Thoroughbred racehorses (2–5 years old) from two training yards. Digital conformation was subjectively graded, and multiple hoof measurements were made with a modified tire gauge (sole and sulci depth) and from digital photographs. All the horses were shod by two registered master farriers within a median of 15 (interquartile range [IQR], 1–25) days before measurement. There were few distal limb conformation abnormalities scored. Most (62/75) horses presented with some deviation from normal hoof parameters, with 2 (IQR, 1–3) abnormalities reported per horse. The most common hoof abnormality was uneven sulci, which was identified in 43 horses and 59 affected hooves, followed by higher medial hoof wall height in 38 horses and 53 affected hooves. Many of the linear and hoof angle measurements and their ratios were within the bounds reported within the literature and indicative of a balanced foot. The length and width measurements increased with horse age. The dorsal hoof wall (DHW) length:heel length ratios were consistently less than 3:1, and the absolute difference between toe and heel angle was generally greater than 5°. Between limb hoof variation was identified for a number of the morphologic measurements including frog length and sole length and the ratio of sole width:sole length. Flat feet (lack of concave solar surface) were identified in 21/75 (28%) horses and in 28/150 (19%) forelimb hoofs. More horses had a flat left foot (10/75) than right foot (4/75), but seven horses had both feet classified as being flat. Flat feet had 2.4 (1.1–5.6, P = .036) greater odds of presenting with uneven sulci. These data indicate that uneven sulci depth and flatter hooves with may be a typical presentation of Thoroughbred feet. Asymmetry in measurements between limb may reflect the greater loading of the left forelimb when race training counter clockwise.     

Subclinical laminitis in dairy heifers

By causing poorer horn quality, subclinical laminitis is considered to be a major predisposing cause of other hoof problems, particularly sole ulcers in newly calved heifers. In this study the hind hooves of 136 female Friesian/Holstein cattle aged between four months and two years were examined to discover at what age the signs of subclinical laminitis appeared. Sole haemorrhages were found in the hoof horn of calves as young as five months. The consistent finding of these lesions in heifers of all ages indicated that subclinical laminitis of varying degree was a common condition during the early growing period of young dairy heifers.     

Decreased expression of p63, a regulator of epidermal stem cells, in the chronic laminitic equine hoof

Reasons for performing study: Abnormal epidermal stem cell regulation may contribute to the pathogenesis of equine chronic laminitis. Objective: To analyse the involvement of p63, a regulator of epidermal stem cell proliferative potential, in chronic laminitis. Methods: Epidermal tissues from skin, coronet and lamellae of the dorsal foot were harvested from 5 horses with chronic laminitis and 5 control horses. Tissues were analysed using histopathology, immunofluorescence microscopy and quantitative immunoblotting Results: Hoof lamellae of laminitic horses had a lower frequency of p63 positive cells than control lamellae, particularly in the distal region. Quantitative immunoblotting confirmed reduced p63 expression in the laminitic distal lamellar region. The decreased p63 expression in laminitic epidermal lamellae was most apparent in the abaxial region adjacent to the hoof wall and highly associated with the formation of terminally differentiated, dysplastic and hyperkeratotic epidermis in this region, whereas lamellae from control horses maintained high p63 expression throughout the axial‐abaxial axis. Conclusions: Expression of p63 in equine skin resembles that reported in other species, including man and rodents, suggesting that p63 can serve as a marker for the proliferative potential of equine epidermal stem cells. p63 expression was significantly lower in the chronic laminitic hoof than in that of control horses, suggesting laminitic hoof epithelium has more limited proliferative potential with a shift towards differentiation. This may reflect reduced activity of epidermal stem cells in laminitic hoof. It is proposed that p63 contributes to the maintenance of hoof lamellae and that misregulation of p63 expression may lead to epidermal dysplasia during lamellar wedge formation. Potential relevance: This study suggests that loss of epidermal stem cells contributes to the pathogenesis of equine laminitis. Autologous transplantation of p63‐positive epidermal stem cells from unaffected regions may have regenerative therapeutic potential for laminitic horses.     

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.     

Keratinopathogenic mould fungi and dermatophytes in healthy and diseased hooves of horses

Specimens of hoof horn from 187 horses were examined for a possible relationship between clinically affected hooves and the occurrence of pathogenic fungi. Specimens were taken from the coronary band and from the stratum externum and medium of the coronary horn and transferred on to Sabouraud dextrose agar, with and without cycloheximide, and incubated at 28 degrees C. Dermatophytes and mould fungi were identified by their macroscopic and microscopic characteristics. The 732 isolates could be assigned to 26 species of moulds, two different species of the dermatophyte Microsporum and three different species of the dermatophyte Trichophyton. Depending on their pathogenic potential they were assigned to three groups: (i) fungi known to be keratinopathogenic (Acremonium blochii, Alternaria alternata, Alternaria chlamydospora, Geotrichum candidum, Microsporum ferrugineum, Microsporum gypseum, Scopulariopsis brevicaulis, Trichophyton species, Trichophyton mentagrophytes, Trichophyton sch?nleinii, 57 isolates), (ii) a group of uncertain pathogenicity (223 isolates), and (iii) a group of non-pathogenic species (452 isolates). Eighty per cent of the samples from horses with hoof horn lesions and 66.7 per cent of the samples from horses with slightly affected hoof horn contained fungi of the keratinopathogenic group, whereas only 8.9 per cent of the samples from horses with healthy hoof horn contained fungi of this group. There were no significant correlations between the clinical data and the age, sex or breed of the horses or their bedding and hygiene. Twelve species of fungi were isolated from the air in the horses' stables, but none of them belonged to the keratinopathogenic group.