Induction of laminitis by prolonged hyperinsulinaemia in clinically normal ponies

The purpose of this study was to determine the effects of prolonged administration of insulin, whilst maintaining normal glucose concentrations, on hoof lamellar integrity in vivo on healthy ponies with no known history of laminitis or insulin resistance. Nine clinically healthy, unrelated ponies were randomly allocated to either a treatment group (n =5; 5.9+/-1.7 years) or control group (n =4; 7.0+/-2.8 years). The treatment group received insulin via a euglycaemic hyperinsulinaemic clamp technique modified and prolonged for up to 72 h. Control ponies were infused with an equivalent volume of 0.9% saline. Ponies were euthanized at the Obel grade 2 stage of clinical laminitis and hoof lamellar tissues were harvested and examined for histopathological evidence of laminitis. Basal serum insulin and blood glucose concentrations were 15.7+/-1.8 microU/mL and 5.2+/-0.1 mmol/L, respectively (mean+/-SE) and were not significantly different between groups. Mean serum insulin concentration in treatment ponies was 1036+/-55 microU/mL vs. 14.6 microU/mL in controls. All ponies in the treatment group developed clinical and histological laminitis (Obel grade 2) in all four feet within 72 h (55.4+/-5.5h), whereas none of the control ponies developed laminitis. There was no clinical evidence of gastrointestinal involvement and the ponies showed no signs of systemic illness throughout the experiment. The data show that laminitis can be induced in healthy young ponies, with no prior history of laminitis, by maintaining prolonged hyperinsulinaemia with euglycaemia. This suggests a role for insulin in the pathogenesis of laminitis, independent of hyperglycaemia, or alterations in hind-gut fermentation. For the clinician, early detection and control of hyperinsulinaemia may facilitate management of endocrinopathic laminitis.     

Histopathology of insulin-induced laminitis in ponies

Reasons for performing study: Ponies with laminitis associated with insulin resistance and hyperinsulinaemia lack systemic and/or intestinal inflammatory signs, suggesting a different pathogenesis potentially reflected in differing histopathology. Objectives: To describe the histological appearance and quantify morphological changes in primary and secondary epidermal lamellae (PEL and SEL) of laminitis lesions from ponies with insulin‐induced laminitis. Methods: Equine hoof lamellar tissue was obtained from 4 control ponies and 5 ponies with laminitis induced following infusion of insulin (1036 ± 55 µU/ml) while maintaining euglycaemia for 55.4 ± 5.5 h. Sections from all 4 hooves were stained and examined by a veterinary pathologist. Measurements of lamellar length (PEL and SEL) were made in mid‐dorsal sections of the right forefeet by 2 blinded observers. Immunolabelling for calprotectin was performed using a monoclonal antibody. Results: No lesions were detected in normal ponies. Lesions detected in ponies with laminitis were variable in severity between ponies. Within ponies, SEL lesions were more severe along the axial region of PEL. Lesions included swelling, disorganisation and abnormal keratinisation of epidermal cells, increased mitotic activity and apoptosis. Separation of basement membranes was minimal. Immunostaining revealed inflammatory cells within the lamellar dermis. SEL were significantly elongated in laminitic hooves relative to controls, with the greatest elongation in those attached to abaxial and middle regions of PEL. Conclusions: Laminitis induced by prolonged infusion of insulin lacked widespread basement membrane disintegration, and increases in epidermal cellular proliferation at axial aspects were marked for this acute stage of disease. Potential relevance: Defining equine laminitis entirely in terms of separation of the basement membrane may not be appropriate for laminitis associated with hyperinsulinaemia.     

Seasonal changes in plasma concentrations of cecum-derived amines in clinically normal ponies and ponies predisposed to laminitis

OBJECTIVE: To measure concentrations of amines formed in the cecum of clinically normal ponies, determine amine concentrations in plasma samples collected in spring and winter, and compare concentrations of amines and serotonin in plasma samples obtained from clinically normal ponies and ponies predisposed to laminitis. SAMPLE POPULATION: Cecal contents obtained from 10 ponies euthanatized at an abattoir and blood samples obtained from 42 adult ponies. PROCEDURE: Cecal contents were assayed for amines by high-performance liquid chromatography (HPLC). Blood samples were collected at various times of the year from 20 ponies predisposed to acute laminitis and 22 clinically normal ponies. Plasma serotonin concentration was measured by HPLC, and tryptamine (TRP), tyramine (TYR), phenylethylamine (PEA), and isoamylamine (IAA) were measured by liquid chromatography-mass spectrometry. RESULTS: 15 amines were identified in cecal contents. Plasma TRP, TYR, PEA, and IAA concentrations ranged from 10pM to 100nM in both groups of ponies. Plasma concentrations of serotonin or other amines did not differ between clinically normal ponies and those predisposed to laminitis; however, significantly higher concentrations of TRP, PEA, and IAA were found in samples obtained in the spring, compared with winter samples. CONCLUSIONS AND CLINICAL RELEVANCE: Various amines are found in the cecum of ponies, several of which can be detected in the plasma. Concentrations increase significantly in the spring and may reach concentrations close to the threshold for causing vasoconstriction. Release of amines from the cecum into the systemic circulation may contribute to hemodynamic disturbances in horses and ponies with acute laminitis.     

Gastrointestinal ultrasonography in normal adult ponies

The objective of this study was to determine the characteristics based on ultrasonographic examination of the stomach, duodenum, jejunum, cecum, and peritoneal fluid in normal adult ponies. Abdominal ultrasonographic examination was performed in nine unsedated standing ponies. The duodenum was examined at three sites and the jejunum in 12 regions. Wall thickness, contractility, distention, and luminal contents were recorded. Stomach wall thickness and location, cecal wall thickness, and peritoneal fluid location and character were recorded. Statistical analysis was performed. Wall thicknesses (in cm) were 0.431 +/- 0.069 for the stomach, 0.188 +/- 0.033 for the duodenum (at all sites), 0.195 +/- 0.031 for the jejunum (at all regions), and 0.179 +/- 0.031 for the cecum. Duodenal contractions per minute were 3.78 +/- 1.10. The stomach spanned 5.14 +/- 0.9 intercostal spaces, with the 8th intercostal space being the most cranial and the 15th intercostal space being the most caudal space through which the stomach was identified. It was possible to identify the jejunum in all ponies dorsal to the left dorsal colon and from the ventral abdominal wall. Peritoneal fluid was identified in six ponies. Peritoneal fluid was usually seen transiently and most commonly in the ventral aspect of the abdominal cavity or around the duodenum. Overall, the ponie's abdominal ultrasonographic examinations revealed wall thicknesses that were less than the published normal ranges for horses. It appears that ponies may have increased duodenal contractility than horses and that the conformation of ponies may change the locations for imaging the stomach.     

Kinetics and kinematics of the equine hind limb: in vivo tendon loads and force plate measurements in ponies

Loads on the suspensory ligament, deep digital flexor tendon, superficial digital flexor tendon, and long digital extensor tendon of the equine hind limb were determined in ponies by use of implanted strain gauges consisting of silicone rubber tubes filled with mercury. Recordings were made simultaneously with force plate measurements and high-speed film recordings while the ponies were walking. The relationship between strain gauge signals and tendon loads was obtained from tension-strain tests performed after death of the ponies. The suspensory ligament and the 2 digital flexor tendons were loaded during the stance phase, and the extensor tendon was loaded mainly during the swing phase. The loading pattern of the suspensory ligament, with peak loads of 4.6 N/kg of body weight, correlated well with the vertical component of the ground reaction force. Maximal loading of the deep digital flexor tendon was observed during the second half of the stance phase, with peak values of 6.7 N/kg. The superficial digital flexor tendon was loaded maximally at the beginning of the stance phase, with a peak load of 4.1 N/kg, and the long digital extensor tendon was loaded maximally during the swing phase, with a peak load of 0.3 N/kg. Recordings made from this procedure for calibration of the strain gauge signals to tendon load and tendon strain, in combination with the force plate measurements, enabled verification of the results by torque analysis of the lower portion of the hind limb, using the vector of the ground reaction force, limb conformation, and limb geometric configuration. Torque analysis of the lower extremity indicated that the determined tendon loads were in agreement with the recorded ground reaction forces.     

Glucose tolerance and insulin sensitivity in ponies and Standardbred horses

The existence of an innate insulin insensitivity in ponies was investigated and compared with the situation in larger breeds of horse. Ponies that were fat or had previously suffered laminitis were found to be far more intolerant to oral glucose loading (1 g/kg bodyweight [bwt]) than normal ponies or Standardbreds. These ponies also exhibited a far greater response in plasma insulin levels after glucose loading. Insulin response tests (0.4 iu/kg bwt insulin intravenously) showed only a minimal and very protracted response in both the fat and laminitic groups. The relevance of these findings in regulation of carbohydrate and lipid metabolism, and their role in the pathogenesis of hyperlipaemia, are discussed.     

Histopathology in horses with chronic palmar foot pain and age-matched controls. Part 2: The deep digital flexor tendon

REASONS FOR PERFORMING STUDY: Causes of palmar foot pain and the aetiopathogenesis of navicular disease remain poorly understood, despite the high incidence of foot-related lameness. HYPOTHESES: Abnormalities of the deep digital flexor tendon (DDFT) may contribute to palmar foot pain; ageing degenerative changes may be seen in horses free from lameness; and horses with lameness are likely to have a greater severity of abnormalities than age-matched horses with no history of foot pain. METHODS: Feet were selected from horses with a history of uni- or bilateral forelimb lameness of at least 2 months' duration. Histology of the DDFT from the level of the proximal interphalangeal joint to its insertion were examined and the severity of lesions for each site graded. Associations between lesions of the navicular bone, collateral sesamoidean ligaments (CSL), distal sesamoidean impar ligament, navicular bursa, distal interphalangeal (DIP) joint synovium and collateral ligaments of the DIP joint and DDFT were assessed. RESULTS: There was no relationship between age and grade of histological abnormality of the DDFT. There were significant histological differences between groups for lesions of the dorsal layers of the DDFT, but not for lesions of the palmar aspect. There were significant associations between histological grades for the superficial dorsal layer of the DDFT and flexor aspect of the navicular bone; and between the deep dorsal layer of the DDFT and the proximal border and medulla of the navicular bone. The navicular bursa grade was correlated with grades for the superficial dorsal, deep dorsal and deep palmar layers of the DDFT. The histological grades for the CSL and the superficial dorsal layer of the DDFT were also associated. CONCLUSIONS: Pathological abnormalities in lame horses often involved the DDFT in addition to the navicular bone. Vascular and matrix changes may precede changes in the fibrocartilage of the navicular bone. POTENTIAL RELEVANCE: Identification of factors leading to vascular changes within the interstitium of the DDFT and changes in matrix composition, may help in future management of palmar foot pain.     

Evaluation of genetic and metabolic predispositions and nutritional risk factors for pasture-associated laminitis in ponies

OBJECTIVE: To evaluate genetic and metabolic predispositions and nutritional risk factors for development of pasture-associated laminitis in ponies. DESIGN: Observational cohort study. ANIMALS: 160 ponies. PROCEDURES: A previous diagnosis of laminitis was used to differentiate 54 ponies (PL group) from 106 nonlaminitic ponies (NL group). Pedigree analysis was used to determine a mode of inheritance for ponies with a previous diagnosis of laminitis. In early March, ponies were weighed and scored for body condition and basal venous blood samples were obtained. Plasma was analyzed for glucose, insulin, triglycerides, nonesterified fatty acids, and cortisol concentrations. Basal proxies for insulin sensitivity (reciprocal of the square root of insulin [RISQI]) and insulin secretory response (modified insulin-to-glucose ratio [MIRG]) were calculated. Observations were repeated in May, when some ponies had signs of clinical laminitis. RESULTS: A previous diagnosis of laminitis was consistent with the expected inheritance of a dominant major gene or genes with reduced penetrance. A prelaminitic metabolic profile was defined on the basis of body condition, plasma triglyceride concentration, RISQI, and MIRG. Meeting > or = 3 of these criteria differentiated PL- from NL-group ponies with a total predictive power of 78%. Determination of prelaminitic metabolic syndrome in March predicted 11 of 13 cases of clinical laminitis observed in May when pasture starch concentration was high. CONCLUSIONS AND CLINICAL RELEVANCE: Prelaminitic metabolic syndrome in apparently healthy ponies is comparable to metabolic syndromes in humans and is the first such set of risk factors to be supported by data in equids. Prelaminitic metabolic syndrome identifies ponies requiring special management, such as avoiding high starch intake that exacerbates insulin resistance.     

A comparison of three horseshoeing styles on the kinetics of breakover in sound horses

A variety of horseshoe designs are believed to 'ease' breakover, or the unloading of the foot once the heels leave the ground. In this study, conventional toe-clip shoes, quarter-clip shoes, fitted to the white line at the toe, and Natural Balance horseshoes were fitted to the front feet of 9 sound Irish Draught-cross type horses. Forceplate and video motion analyses were undertaken during trot locomotion to determine the moment arm of the ground reaction force on the distal interphalangeal (DIP) joint, the peak DIP joint moment and the peak compressive force on the navicular bone. DIP joint moment arm during breakover was reduced with both Natural Balance (mean +/- s.d. 77 +/- 7 mm) and quarter-clip shoes (78 +/- 9 mm) compared to the toe-clip shoes (86 +/- 6 mm) (P<0.01). Peak DIP joint moment was not significantly different (175 +/- 37,171 +/- 38 and 175 +/- 31 Nmm/kg, in Natural Balance, quarter-clip and toe-clip shoes, respectively) and neither was peak force on the navicular bone (5.52 +/- 1.52, 5.79 +/- 1.53 and 6.14 +/- 1.47 N/kg, respectively). Breakover duration (heel off to toe off) was not significantly reduced by the Natural Balance shoes (39 +/- 6 ms) or the quarter-clip shoes (40 +/- 6 ms) compared to toe-clip shoes (42 +/- 9 ms). This study has demonstrated that the use of Natural Balance shoes reduces the moment arm of the ground reaction force (GRF) during breakover but does not reduce the peak DIP joint moment or the force on the navicular bone.     

Lameness in 46 horses associated with deep digital flexor tendonitis in the digit: diagnosis confirmed with magnetic resonance imaging

REASONS FOR PERFORMING STUDY: The differential diagnosis of foot pain has long proved difficult and the use of magnetic resonance imaging (MRI) offers the opportunity to further the clinical understanding of the subject. OBJECTIVES: To determine the incidence of deep digital flexor tendon (DDFT) injuries in a series of 75 horses with lameness associated with pain localised to the digit, with no significant detectable radiographic or ultrasonographic abnormalities, using MRI; and to describe a variety of lesion types and relate DDF tendonitis with anamnesis, clinical features, response to local analgesic techniques and nuclear scintigraphic and ultrasonographic findings. METHODS: All horses undergoing MRI of the front feet between January 2001 and October 2002 were reviewed and those with DDFT injuries categorised according to lesion type; horses with primary tendonitis (Group I) and those with concurrent abnormalities of the navicular bone considered to be an important component of the lameness (Group II). The response to perineural analgesia of the palmar digital nerves and palmar (abaxial sesamoid) nerves, intra-articular analgesia of the distal interphalangeal (DIP) joint and analgesia of the navicular bursa were reviewed. The result of ultrasonography of the pastern and foot was recorded. Lateral, dorsal and solar pool and bone phase nuclear scintigraphic images were assessed subjectively and objectively using region of interest (ROI) analysis. RESULTS: Forty-six (61%) of 75 horses examined using MRI had lesions of the DDFT considered to be a major contributor to lameness. Thirty-two horses (43%) had primary DDFT injuries and 14 (19%) a combination of DDF tendonitis and navicular bone pathology. Lesions involved the insertional region of the tendon alone (n = 3), were proximal to the navicular bone (n = 23) or were at a combination of sites (n = 20). Lesion types included core lesions, focal and diffuse dorsal border lesions, sagittal plane splits, insertional injuries and lesions combined with other soft tissue injuries. Many horses had a combination of lesion types. Lameness was abolished by palmar digital analgesia in only 11 of 46 horses (24%). Twenty-one of 31 horses (68%) in Group I showed > 50% improvement in lameness after intra-articular analgesia of the DIP joint, whereas 11 of 12 horses (92%) in Group II had a positive response. Twelve of 18 horses (67%) in Group I had a positive response to analgesia of the navicular bursa. Nineteen horses had lesions of the DDFT extending proximal to the proximal interphalangeal joint seen using MRI, but these were identified ultrasonographically in only 2 horses. Scintigraphic abnormalities suggestive of DDFT injury were seen in 16 of 41 horses (41%), 8 in pool phase images and 8 in bone phase images. CONCLUSIONS AND POTENTIAL RELEVANCE: DDFT injuries are an important cause of lameness associated with pain arising from the digit in horses without detectable radiographic abnormalities. Lameness is not reliably improved by palmar digital analgesia, but may be improved by intra-articular analgesia of the DIP joint in at least 68% of horses. Ultrasonography is not sensitive in detecting lesions of the DDFT in the distal pastern region, but a combination of pool and bone phase scintigraphic images of the digit is helpful in some horses. Further follow-up information is required to determine the prognosis for horses with lesions of the DDFT in the digit and to establish whether this is related to lesion severity and/or location.     

The effects of vasoactive amines found in the equine hindgut on digital blood flow in the normal horse

REASONS FOR PERFORMING STUDY: Disturbances of digital blood flow are thought to be fundamental to the pathophysiology of acute laminitis. However, factors linking the initiating events in the equine hindgut with these disturbances in the foot remain to be determined. HYPOTHESIS: Amine compounds, formed by bacteria in the equine hindgut, have digital vasoconstrictor effects in vivo. METHODS: Tryptamine (1.6 microg/kg/min) and phenylethylamine (2.13 microg/kg/min) were infused i.v. into standing nonsedated horses. Digital blood flow was measured by Doppler ultrasound and foot surface temperature was monitored. Plasma 5-hydroxytryptamine (5-HT) concentrations were measured by HPLC. RESULTS: Tryptamine and phenylethylamine infusions had no effect on systemic arterial blood pressure or heart rate, but caused significant decreases in digital arterial blood flow (mean +/- s.e. 29.2 +/- 8.5 and 18.4 +/- 6.8%, respectively). Both amines also caused decreases in dorsal hoof wall temperature (0.6 +/- 0.1 and 0.5 +/- 0.1 degrees C for tryptamine and phenylethylamine, respectively) and concomitant increases in plasma 5-HT concentration. CONCLUSIONS: Tryptamine and phenylethylamine caused reduction of digital blood flow, effects which may have been mediated, in part, via displacement of 5-HT from platelets. POTENTIAL RELEVANCE: Amine compounds occurring in the equine hindgut, if released into the circulation following carbohydrate overload, could contribute to selective digital vasoconstriction. Further work in ponies and horses, with naturally occurring laminitis, is necessary to determine whether amines represent a therapeutic target in this disease.     

Cardiovascular, acid-base, electrolyte, and plasma volume changes in ponies developing alimentary laminitis.

Twelve Shetland ponies were fed a high-starch ration. Seven ponies which had a transitory metabolic acidosis developed laminitis 56 hours (+/- 3.5, SEM) after overfeeding. These ponies also developed persistent hypokalemia, hyperthermia, and increased heart rate 24 hours before the onset of lameness. Serum sodium, serum chloride, hematocrit, plasma volume, and blood volume were unchanged. At the onset of clinical signs of laminitis, cardiac output and blood pressure increased, but total peripheral resistance was unchanged. None of the measured or calculated values predicted the onset of laminitis. Hypertension appeared to be a response to, rather than a cause of, lameness. Three of the remaining ponies apparently died of shock 29.3 +/- 2.7 hours after overfeeding. All 3 had severe metabolic acidosis; decreased cardiac output, systemic arterial pressure, and plasma volume; and increased hematocrit, total peripheral resistance, and pulmonary vascular resistance. The 11th pony was unaffected and the 12th pony was euthanatized.     

Equine laminitis: Ultrastructural lesions detected in ponies following hyperinsulinaemia

Reasons for performing study: Anatomical changes in the hoof lamellar tissue induced by prolonged hyperinsulinaemia have not been described previously. Analysis of the induced lesions may promote understanding of hyperinsulinaemic laminitis pathogenesis and produce clinical benefit. Objectives: To use light and transmission electron microscopy (TEM) to document hoof lamellar lesions in ponies clinically lame after prolonged hyperinsulinaemia. Methods: Nine clinically normal, mature ponies were allocated randomly to either a treatment group (n = 5) or control group (n = 4). The treatment group received insulin via a modified, prolonged euglycaemic hyperinsulinaemic clamp technique (EHCT) and were subjected to euthanasia when clinical signs of Obel grade II laminitis occurred. The control group was sham treated with an equivalent volume of 0.9% saline and killed at 72 h. Lamellar tissues of the right front feet were harvested and processed for TEM. Results: Lamellae from insulin treated ponies were attenuated and elongated with many epidermal basal cells (EBC) in mitosis. Unlike carbohydrate induced laminitis in horses there was no global separation at the lamellar dermal/epidermal interface among ponies. Sporadic EBC basement membrane (BM) separation was associated with the proximity of infiltrating leucocytes. In 2 ponies, the lamellar BM was thickened. The number of hemidesmosomes/μm of BM was decreased in all insulin treated ponies. Conclusions: Prolonged hyperinsulinaemia causes unique lamellar lesions normally characteristic of acute and chronic laminitis. Lamellar proliferation may be an insulin effect through its mitogenic pathway. Aberrant lamellar mitosis may lengthen and weaken the lamellar, distal phalanx attachment apparatus and contribute to the clinical signs that developed. Potential relevance: The study shows that insulin alone, in higher than normal circulating concentrations, induces profound, changes in lamellar anatomy. Medical control of insulin resistance and hyperinsulinaemia may ameliorate lesions and produce clinical benefit.     

Hypertension and insulin resistance in a mixed-breed population of ponies predisposed to laminitis

OBJECTIVE: To determine the metabolic phenotype of a group of laminitis-prone ponies when at pasture in summer, compared with when at pasture in winter. ANIMALS: 40 ponies of various breeds predisposed to recurrent pasture-associated laminitis and 40 unaffected control ponies. PROCEDURES: Body condition score and size of the crest of the neck were assessed, blood samples obtained, and blood pressure measured by use of an indirect oscillometric technique, while ponies were kept on winter pasture (last week of November or beginning of December) and again on summer pasture (June). Serum insulin concentration and plasma glucose, triglyceride, uric acid, and ACTH concentrations were measured. Insulin sensitivity was calculated with proxies derived from basal serum insulin and plasma glucose concentrations. RESULTS: No significant differences were apparent between ponies predisposed to laminitis and control ponies during winter. However, in June, laminitis-prone ponies had increased serum insulin concentration and plasma triglyceride and uric acid concentrations, compared with control ponies. Also, laminitis-prone ponies were relatively insulin resistant, compared with control ponies. Mean blood pressure was significantly higher during summer in laminitis-prone ponies (median [interquartile range], 89.6 mm Hg [78.3 to 96.9 mm Hg]), compared with control ponies (76.8 mm Hg [69.4 to 85.2 mm Hg]). CONCLUSIONS AND CLINICAL RELEVANCE: Summer pastures appear to induce metabolic responses in some ponies, leading to expression of the prelaminitic phenotype, which includes hypertension as well as insulin resistance. Signs of this metabolic syndrome may not be apparent in affected ponies during periods of grazing winter pasture. Understanding this syndrome may enable improved countermeasures to be devised to prevent laminitis.     

Physiological stimuli of thirst and drinking patterns in ponies

The stimuli that elicit thirst were studied in four ponies. Nineteen hours of water deprivation produced an increase in plasma protein from 67 +/- 0.1 g/litre to 72 +/- 2 g/litre, a mean (+/- se) increase in plasma sodium from 139 +/- 3 to 145 +/- 2 mmol/litre and an increase in plasma osmolality from 297 +/- 1 to 306 +/- 2 mosmol/litre. Undeprived ponies drank 1.5 +/- 0.9 kg/30 mins; 19 h deprived ponies drank 10.2 +/- 2.5 kg/30 mins and corrected the deficits in plasma protein, plasma sodium and plasma osmolality as well as compensating for the water they would have drunk during the deprivation period. In order to determine if an increase in plasma osmolality would stimulate thirst, 250 ml of 15 per cent sodium chloride was infused intravenously. The ponies drank when osmolality increased 3 per cent and when plasma sodium rose from 136 +/- 3 mmol/litre to 143 +/- 3 mmol/litre. Ponies infused with 15 per cent sodium chloride drank 2.9 +/- 0.7 kg; those infused with 0.9 per cent sodium chloride drank 0.7 +/- 0.5 kg. In order to determine if a decrease in plasma volume would stimulate thirst, ponies were injected with 1 or 2 mg/kg bodyweight (bwt) frusemide. Plasma protein rose from 68 +/- 2 g/litre pre-injection to 75 +/- 2 g/litre 1 h after 1 mg/kg bwt frusemide and to 81 +/- 1 g/litre 1 h after 2 mg/kg bwt frusemide.(ABSTRACT TRUNCATED AT 250 WORDS)     

An evaluation of two autologous tendon grafting techniques in ponies

OBJECTIVE: To compare the healing and mechanical strength of a multiple split autologous tendon graft (MG) to a whole autologous tendon graft (WG) in the deep digital flexor tendon of ponies. STUDY DESIGN: In vitro evaluation of two different tendon-grafting techniques. ANIMALS OR SAMPLE POPULATION: Six ponies of mixed gender and age. METHODS: Tenotomies performed in forelimb deep digital flexor tendons (DDFT) distal to the insertion of the accessory ligament (AL-DDFT) were repaired with free autologous grafts from the hindlimb lateral digital extensor tendon (LDET). Grafts were either whole (WG) or split into three longitudinal strips (multiple graft, MG). Tendons and graft sites were collected and loaded (2.54 cm/s) to failure at either 4 or 8 weeks after surgery. Cross-sectional area was determined by both impression cast (IC) and an inkblot (IB) method. Tissue maturity and inflammation were evaluated by microscopy. RESULTS: Gap formation was a consistent finding in all repair sites. No statistical differences were found in healing or mechanical variables between MG and WG techniques. The failure stress for the 8-week repairs (15.51 +/- 3.1 MPa IB and 11.73 +/- .77 MPa IC, 16.13 +/- 2.2 MPa IB and 10.22 +/- .76 MPa IC for MG and WG, respectively) were significantly greater (P <.0005) than for 4-week repairs (3.71 +/- 1.7 MPa IB and 2.68 +/- 1.44 MPa IC, 2.81 +/- 1.46 MPa IB and 2.3 +/- 1.7 MPa IC for MG and WG, respectively). The repair tissue was more mature (P <.05) at 8 weeks than at 4 weeks, but there was no significant difference in inflammatory responses at 4 and 8 weeks. CONCLUSIONS: There was a sixfold increase in strength between 4 and 8 weeks of healing, but no significant difference in healing or strength between the MG and WG techniques. CLINICAL RELEVANCE: In ponies, autologous tendon grafting contributes to a strong repair during the early convalescent period, but splitting a tendon graft seemingly offers no appreciable advantage over use of a whole graft.     

Effect of dietary fructans and dexamethasone administration on the insulin response of ponies predisposed to laminitis

OBJECTIVE: To determine whether pasture, and specifically the addition of fructan carbohydrate to the diet, induces exaggerated changes in serum insulin concentration in laminitispredisposed (LP) ponies, compared with ponies with no history of the condition, and also to determine insulin responses to the dexamethasone suppression test. DESIGN: Prospective study. ANIMALS: 10 LP and 11 control adult nonobese mixed-breed ponies. PROCEDURES: Insulin-modified IV glucose tolerance tests were performed (5 ponies/group). In diet studies, ponies were kept on pasture and then changed to a hay diet (10 ponies/group). Second, ponies were maintained on a basal hay diet (4 weeks) before being fed a hay diet supplemented with inulin (3 g/kg/d [1.4 g/lb/d]). Serum insulin and plasma glucose concentrations were analyzed before and after dietary changes. Serum cortisol and insulin concentrations were also measured in a standard dexamethasone suppression test. RESULTS: The LP ponies were insulin resistant (median insulin sensitivity of 0.27 x 10(4) L min(-1) mU(-1) in LP ponies, compared with 0.64 x 10(4) L min(-1) mU(-1) in control ponies). Median insulin concentration in LP ponies was significantly greater than that in control ponies at pasture, decreased in response to feeding hay, and was markedly increased (5.5-fold) following the feeding of inulin with hay. The LP ponies had a greater increase in serum insulin concentration at 19 hours after dexamethasone administration (median, 222.9 mU/L), compared with control ponies (45.6 mU/L). CONCLUSIONS AND CLINICAL RELEVANCE: Nonobese ponies predisposed to develop laminitis had compensated insulin resistance, and this phenotype was revealed by feeding plant fructan carbohydrate or by dexamethasone administration.     

Effects of training on maximum oxygen consumption of ponies

OBJECTIVES: To establish maximum oxygen consumption VO2max) in ponies of different body weights, characterize the effects of training of short duration on VO2max, and compare these effects to those of similarly trained Thoroughbreds. ANIMALS: 5 small ponies, 4 mid-sized ponies, and 6 Thoroughbreds. PROCEDURE: All horses were trained for 4 weeks. Horses were trained every other day for 10 minutes on a 10% incline at a combination of speeds equated with 40, 60, 80, and 100% of VO2max. At the beginning and end of the training program, each horse performed a standard incremental exercise test in which VO2max was determined. Cardiac output (Q), stroke volume (SV), and arteriovenous oxygen content difference (C [a-v] O2) were measured in the 2 groups of ponies but not in the Thoroughbreds. RESULTS: Prior to training, mean VO2max for each group was 82.6 = 2.9, 97.4 +/- 13.2, and 130.6 +/- 10.4 ml/kg/min, respectively. Following training, mean VO2max increased to 92.3 +/- 6.0, 107.8 +/- 12.8, and 142.9 +/- 10.7 ml/kg/min. Improvement in VO2max was significant in all 3 groups. For the 2 groups of ponies, this improvement was mediated by an increase in Q; this variable was not measured in the Thoroughbreds. Body weight decreased significantly in the Thoroughbreds but not in the ponies. CONCLUSIONS AND CLINICAL RELEVANCE: Ponies have a lower VO2max than Thoroughbreds, and larger ponies have a greater VO2max than smaller ponies. Although mass-specific VO2max changed similarly in all groups, response to training may have differed between Thoroughbreds and ponies, because there were different effects on body weight.     

Determination of reference range values indicative of glucose metabolism and insulin resistance by use of glucose clamp techniques in horses and ponies

OBJECTIVES: To acquire reference range values indicative of glucose metabolism by use of the hyperglycemic clamp technique in healthy horses and evaluate the usefulness of the euglycemic hyperinsulinemic clamp technique in healthy horses and ponies. ANIMALS: Dutch Warmblood horses and 4 Shetland ponies. PROCEDURE: The hyperglycemic clamp technique was used for quantification of the sensitivity of beta cells to exogenous glucose infusion in horses. The euglycemic hyperinsulinemic clamp technique was used to determine the sensitivity and responsiveness of tissues to exogenous insulin in horses and ponies. RESULTS: During the hyperglycemic clamp technique, the mean amount of glucose metabolized (M) in horses was 0.011 +/- 0.0045 mmol/kg x min(-1) (95% confidence interval [CI], 0.0018 to 0.020 mmol/kg x min(-1); range, 0.000035 to 0.021 mmol/kg x min(-1)) and the mean M value-to-plasma insulin concentration (I) ratio (ie, mmol of glucose/kg x min(-1) per pmol of insulin/L x 100) was 0.017 +/- 0.016 (95% CI, -0.014 to 0.049; range, 0.000025 to 0.055). During the euglycemic hyperinsulinemic clamp technique, the mean M value was 0.014 +/- 0.0055 mmol/kg x min(-1) (95% CI, 0.0026 to 0.025 mmol/kg x min(-1); range, 0.0042 to 0.023 mmol/kg x min(-1)) in horses and 0.0073 +/- 0.0020 mmol/kg x min(-1) (95% CI, 0.0034 to 0.011 mmol/kg x min(-1); range, 0.0049 to 0.011 mmol/kg x min(-1)) in ponies. The M value was significantly lower in ponies than in horses, whereas the M:I ratios were not significantly different between horses and ponies. CONCLUSION AND CLINICAL RELEVANCE: Glucose clamp techniques offer good methods to investigate glucose metabolism in horses and ponies. A higher degree of insulin resistance was found in ponies, compared with Dutch Warmblood horses.