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Physical characteristics, blood hormone


concentrations, and plasma lipid concentrations
in obese horses with insulin resistance
Nicholas Frank, DVM, PhD; Sarah B. Elliott, BSc; Laura E. Brandt, BSc; Duane H. Keisler, PhD

ABBREVIATIONS
Objective—To compare obese horses with insulin IR Insulin resistance

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resistance (IR) with nonobese horses and determine PPID Pituitary pars intermedia dysfunction
whether blood resting glucose, insulin, leptin, and CGIT Combined glucose-insulin test
lipid concentrations differed between groups and BCS Body condition score
were correlated with combined glucose-insulin test VLDL Very-low-density lipoprotein
(CGIT) results. PL Phospholipid
Animals—7 obese adult horses with IR (OB-IR group) HDL High-density lipoprotein
and 5 nonobese mares. DEX-TRH Dexamethasone-thyrotropin releasing
hormone
Procedures—Physical measurements were taken, and AUCg Area under the curve for glucose
blood samples were collected after horses had accli- AUCi Area under the curve for insulin
mated to the hospital for 3 days. Response to insulin
NEFA Nonesterified fatty acid
was assessed by use of the CGIT, and maintenance of
plasma glucose concentrations greater than the prein-
jection value for ≥ 45 minutes was used to define IR.
Area under the curve values for glucose (AUCg) and
to develop laminitis and that dietary changes such as
insulin (AUCi) concentrations were calculated. grazing on lush pasture can trigger episodes of
laminitis. Other support is provided by in vitro evi-
Results—Morgan, Paso Fino, Quarter Horse, and
Tennessee Walking Horse breeds were represented in
dence that hoof tissues have a high requirement for
the OB-IR group. Mean neck circumference and BCS glucose.3 Pass et al3 reported that hoof explants
differed significantly between groups and were posi- undergo epidermal-dermal separation readily when
tively correlated with AUC values. Resting insulin and glucose concentrations are low in the medium. It has
leptin concentrations were 6 and 14 times as high, also been reported that ponies with laminitis are less
respectively, in the OB-IR group, compared with the responsive to insulin than healthy ponies4 and that
nonobese group, and were significantly correlated ponies with both obesity and laminitis are more
with AUCg and AUCi. Plasma nonesterified fatty acid, insulin resistant than those that are just obese.5
very low-density lipoprotein, and high-density lipopro- Laminitis has also been associated with PPID, and IR
tein-cholesterol (HDL-C) concentrations were signifi- often accompanies this condition.6,7
cantly higher (86%, 104%, and 29%, respectively) in
OB-IR horses, and HDL-C concentrations were posi- Obesity, hyperinsulinemia, and laminitis some-
tively correlated with AUC values. times develop concurrently in horses without
detectable PPID, and this group of disorders has been
Conclusions and Clinical Relevance—Measurements
of neck circumference and resting insulin and leptin referred to as syndrome X or equine metabolic syn-
concentrations can be used to screen obese horses for drome.6,8,9 The latter term is used most commonly but
IR. Dyslipidemia is associated with IR in obese horses. may not be appropriate because metabolic syndrome is
(J Am Vet Med Assoc 2006;228:1383–1390) defined by the presence of IR or type 2 diabetes melli-
tus as well as at least 2 of 4 additional risk factors,
including obesity or visceral adiposity, dyslipidemia,
microalbuminemia, or arterial hypertension.10 To the
O besity is a major health concern in humans
because of its association with IR, type 2 diabetes
mellitus, and coronary heart disease.1 Insulin resis-
authors’ knowledge, microalbuminemia and hyperten-
sion have not been reported in obese horses, but blood
tance has also been associated with obesity in horses,2 lipid concentrations have been measured in ponies and
and this condition may be of particular concern donkeys with IR, and many of them were obese.11,12
because of the putative link between laminitis and Plasma triglyceride and total cholesterol concentra-
glucose metabolism.3 This link is supported by anec- tions do not differ between clinically normal and
dotal observations that obese horses are more likely hyperinsulinemic ponies,11 but a positive correlation
(r = 0.55; P = 0.002; n = 31) exists between plasma
From the Department of Large Animal Clinical Sciences, College of insulin and triglyceride concentrations in donkeys
Veterinary Medicine, University of Tennessee, Knoxville, TN with naturally occurring hyperlipidemia.12
37996 (Frank, Elliott, Brandt); and the Division of Animal When methods of assessing IR in horses and
Sciences, University of Missouri, Columbia, MO 65211 (Keisler). ponies were recently reviewed, available tests were
Supported by a grant from the University of Tennessee Centers of
Excellence in Livestock Diseases and Animal Health. divided into those that provide nonspecific indications
The authors thank Dr. Mickey Coleman for assistance with this study. of IR and those that give specific quantitative measure-
Address correspondence to Dr. Frank. ments of insulin sensitivity.13 The CGIT used in the

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present study was not discussed, but this test should be normal upright position, and the distance along a straight
included in the first category because it does not pro- line from the poll to the cranial aspect of the withers was
vide values for insulin sensitivity, glucose effectiveness, measured with a measuring tape (Figure 1). Neck circumfer-
or endogenous insulin secretion.13,14 However, delayed ence was measured perpendicular to this line at 0.25, 0.50,
and 0.75 of the distance from the withers to the poll with a
return of blood glucose concentrations to the preinjec- measuring tape (Figure 1). Mean neck circumference values
tion concentrations can be attributed to IR when the were calculated.
CGIT is used because glucose concentrations should
rapidly decrease in response to exogenous insulin CGIT—Horses were provided with ad libitum grass hay
administration.14 The CGIT was also used in the pre- and water before and during the procedure. A 14-gauge
sent study because it is easily interpreted, has low cost, polypropylene catheter was inserted into the left jugular vein,
and an injection cap and infusion setb (length, 30 cm; inter-
and is readily applicable to clinical practice.14 nal diameter, 0.14 cm) were attached. The CGIT was per-
In this study, obese horses were selected on the formed as described.14 Briefly, glucose (150 mg/kg) was
basis of BCS and IR was defined by CGIT results. The infused as a 50% dextrose solutionc through the infusion line
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purpose of the study reported here was to compare and catheter, followed by heparinized saline (0.9% NaCl)
obese horses with IR with nonobese horses to deter- solution and regular insulind (0.1 U/kg). Blood samples were
mine whether blood resting glucose, insulin, leptin, collected via the catheter at 1, 5, 15, 25, 35, 45, 60, 75, 90,
and lipid concentrations differed between groups and 105, 120, 135, and 150 minutes after insulin injection. At
were correlated with CGIT results. We also wanted to each time point, 3 mL of blood was withdrawn from the infu-
identify variables that predicted CGIT responses when sion line and discarded. A 6-mL blood sample was collected,
only a single measurement is available and determine followed by infusion of 5 mL of heparinized saline solution.
Blood was transferred into tubes containing sodium fluoride
whether obesity, IR, and dyslipidemia occur concur- and potassium oxalate that were immediately cooled on ice
rently in horses. and refrigerated or transferred into glass tubes containing no
anticoagulant that were left at 21oC for 1 hour to allow clot
Materials and Methods formation. Serum or plasma was subsequently harvested
Horses—Client-owned horses were admitted to the after low-speed centrifugation (1,000 X g) at 4oC for 20 min-
University of Tennessee Large Animal Hospital and remained utes and stored at –20oC until further analysis.
there for a 7-day evaluation period. Horses were evaluated
between July 2003 and July 2004. Seven obese horses (5 Isolation and quantification of lipoproteins—Plasma
mares, 2 geldings) with CGIT results indicative of IR (OB-IR lipoprotein fractions were isolated by use of the sequential
group) were compared with 5 nonobese mares with a BCS of ultracentrifugation method of Havel et al.17 Briefly, blood was
4/9 to 6/9 and normal insulin sensitivity (NO group). Median centrifuged (1,000 X g) at 4oC for 20 minutes to separate plas-
age for the OB-IR group was 11 years (range, 10 to 16 years), ma from chilled blood samples. Six-milliliter plasma samples
and Morgan Horse (n = 3), Paso Fino (1), Quarter Horse (2), were placed in a fixed-angle rotore for ultracentrifugation at
and Tennessee Walking Horse (1) breeds were represented. 112,000 X g for 18 hours at 10oC. A 1-mL fraction of plasma
Median age of the NO group was 10 years (range, 7 to 16 (density < 1.006 g/mL) was isolated from each tube. This frac-
years), and Quarter Horse (n = 3), Appaloosa (1), and tion was referred to as VLDL. Triglyceride, PL, and total cho-
Mustang (1) breeds were represented. Obesity was defined as lesterol components of VLDL were measured with enzymatic
a BCS ≥ 7 on a 1 to 9 scale.15 Insulin resistance was arbitrari- colorimetric reagentsf in an automated discrete analyzer.g
ly defined by plasma glucose concentrations that remained Lipoprotein lipase, phospholipase D, and cholesterol oxidase,
greater than the preinjection concentration for ≥ 45 minutes respectively, were the principal reagents of the triglyceride,
during the CGIT (ie, attenuation of the negative phase).14
The study protocol was approved by the University of
Tennessee Institutional Animal Care and Use Committee,
and signed owner consent was obtained for each horse.
Experimental design—Horses were admitted on day 0,
and weight, height at the withers, girth, and neck circumfer-
ence were measured. Physical examinations, CBC, and serum
biochemical analyses were performed. An adipose tissue
biopsy specimen was also collected on day 0, and flunixin
megluminea was administered orally for the next 3 days. Each
horse was then housed in a 3.7 X 3.7-m stall within the teach-
ing hospital, provided with mixed timothy-orchard grass hay
and water for ad libitum intake, and acclimated to the envi-
ronment for 3 days.
Blood samples were collected for resting hormone and
blood lipid measurements via jugular venipuncture between
8 AM and 9 AM on the fourth day. An IV catheter was placed,
and a CGIT was performed the next day. Horses were
screened for PPID by use of the combined DEX-TRH sup-
pression test16 on subsequent days, with the exception of 3
horses from the OB-IR group that were excluded at the own-
ers’ request. Resting plasma ACTH concentrations were mea-
sured in these horses instead. The OB-IR horses were dis-
charged 1 week after admission. Figure 1—Illustration of a procedure used to measure mean
neck circumference in horses. a = 0.25 of the distance from poll
Neck circumference measurements—Horses were to withers b = 0.50 of the distance from poll to withers. c = 0.75
restrained so that the head and neck were maintained in a of the distance from poll to withers. Reprinted with permission.

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PL, and total cholesterol assays. Protein content of VLDL was kitm validated for equine insulin.21 Serum samples were ana-
analyzed by use of bovine serum albumin standards and a lyzed for leptin with double-antibody radioimmunoassay
spectrophotometerh in accordance with a modified method of procedures validated for equine leptin.22 Plasma cortisol con-
the Lowry18 procedure.19 Plasma VLDL concentrations were centrations were measured by use of a radioimmunoassay kitn
calculated by summing concentrations of lipid (triglyceride, validated for equine cortisol.21
total cholesterol, PL) and protein components.
Low-density lipoprotein was isolated from the remain- Measurement of plasma ACTH concentrations—Assay
ing plasma by ultracentrifugation under the same conditions of equine ACTH was performed on EDTA plasma by use of a
after addition of potassium bromide to raise the plasma den- commercially available immunoradiometric assay for
sity to 1.063 g/mL. A 1.063 g/mL solution of potassium bro- ACTH.23,o Endogenous plasma ACTH concentrations were
mide prepared in EDTA saline solution was also added to measured in blood collected from 3 OB-IR horses that did not
each sample to increase the volume to 6 mL. After ultracen- undergo combined DEX-TRH testing. For all hormone
trifugation, a 1-mL fraction of plasma (density < 1.063 g/mL) assays, duplicate measurements were performed and an
was isolated from each tube and dialyzed overnight in 0.5M intra-assay coefficient of variance < 10% was required for
ammonium bicarbonate solution to remove dissolved salts. acceptance of results.

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Postdialysis volumes were recorded for each sample, and Statistical analysis—The AUCg and AUCi concentra-
duplicate samples were processed; this fraction was referred tions were calculated from total concentrations by use of the
to as LDL. Compositional analysis of dialyzed LDL samples trapezoidal method and computer software.o Groups were
was performed as described. Plasma LDL concentrations compared by use of the Mann-Whitney U test after distribu-
were adjusted to account for addition of potassium bromide tions were examined and Shapiro-Wilk tests were performed,
solution and volume expansion during dialysis. High-densi- and median, range, mean, and SD values were calculated.
ty lipoprotein was isolated and quantified by use of the same Correlations among variables were examined by calculating
methods as described for LDL, with the exception that the Spearman correlation coefficients. Statistical tests were per-
plasma density was increased to 1.21 g/mL. Equine VLDL, formed with computer software.p Significance was defined at
LDL, and HDL have established density limits of < 1.006, a value of P < 0.05.
1.019 to 1.063, and 1.063 to 1.21 g/mL, respectively.20
Measurement of plasma glucose and lipid concentra- Results
tions—Glucose concentrations were measured by use of a No abnormalities were detected via physical exam-
colorimetric assayi on an automated discrete analyzer.g ination, and results of CBC and serum biochemical
Concentrations of plasma triglyceride and total cholesterol tests were within reference ranges for all horses. Four
were measured by use of enzymatic colorimetric reagentsf
and the automated discrete analyzer.g Plasma NEFA concen- of 7 horses in the OB-IR group had a history of chron-
trations were measured by use of an in vitro enzymatic col- ic laminitis, and abnormal growth rings were seen on
orimetric test kitf on a microtiter plate reader.j For all lipid, hooves, but these horses were not lame at the time of
protein, and glucose measurements, duplicate assays were examination. Groups differed significantly with respect
performed and intra-assay coefficients of variation < 5% were to BCS, girth, and neck circumference, but did not dif-
required for acceptance of results. fer in weight or height (Table 1).
Screening for PPID—A DEX-TRH test was performed in In the OB-IR group, plasma glucose concentra-
4 of 7 horses in the OB-IR group and all of the horses in the tions were greater than baseline concentrations for ≥
NO group according to the method of Eiler at al.16 Briefly, a 45 minutes in 1 horse, 75 minutes in 2 horses, 90 min-
baseline blood sample was collected, and 40 µg of DEXk/kg utes in 1 horse, 120 minutes in 1 horse, 135 minutes in
was injected IV. After collection of another blood sample at 3 1 horse, and > 150 minutes in 1 horse (Figure 2). In
hours, 1.2 mg of TRHl was injected IV and blood samples contrast, plasma glucose concentrations returned to a
were collected at 3.5, 4, and 24 hours after DEX injection. value less than the baseline value within 25 minutes in
Measurement of blood hormone concentrations— 5 horses in the NO group.
Serum or EDTA plasma was obtained via low-speed centrifu- Compared with the NO group, median AUCg and
gation (1,000 X g) at 4oC for 20 minutes. Insulin concentra- AUCi values for CGIT curves were 68% and 81% high-
tions were measured in serum by use of a radioimmunoassay er, respectively, in the OB-IR group (Table 1). Resting

Table 1—Measured variables (median [range]) in 5 nonobese horses and 7 obese


horses with IR (OB-IR).

Nonobese OB-IR
Variable (n = 5) (7) P

Weight (kg) 473 (444–476) 518 (426–610) 0.223


BCS (scale, 1–9) 6 (4–6) 7 (7–9) 0.004
Height (cm) 145 (142–158) 151 (143–155)* 0.927
Girth (cm) 179 (172–183) 189 (182–205)* 0.011
Neck circumference (cm) 87.7 (85.3–95.7) 105.8 (94.3–110.3)* 0.010
Plasma leptin (ng/mL) 0.8 (0.5–11.0) 11.0 (1.0–30.5) 0.028
Resting plasma glucose (mg/dL) 66.9 (49.8–74.1) 83.9 (76.0–97.5) 0.005
Resting serum insulin (µU/mL) 9.1 (6.4–21.1) 50.5 (17.0–93.4) 0.007
Resting glucose-to-insulin ratio 6.4 (3.2–11.6) 1.7 (0.9–5.3) 0.007
AUCg (X 103 mg·dL–1·min–1) 9.2 (7.7–10.1) 15.5 (13.1–22.6) 0.005
AUCi (X 103 µU·mL–1·min–1) 12.8 (10.6–13.2) 23.2 (18.1–86.6) 0.005

*Includes only 6 observations.

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serum insulin and plasma leptin concentrations were 6 Variables significantly correlated with both AUCg
times (P = 0.007) and 14 times (P = 0.028) greater, and AUCi included BCS, neck circumference, leptin
respectively, in the OB-IR group, but ranges over- concentration, glucose concentration, insulin concen-
lapped. These single measurements of insulin (r = 0.76; tration, glucose-to-insulin ratio, and HDL-cholesterol
P = 0.004) and leptin (r = 0.75; P = 0.005) were posi- (Table 3). In this study, AUCg and AUCi were most
tively correlated with BCS. Compared with NO horses, closely correlated with BCS (r = 0.84; P < 0.001) and
plasma NEFA, VLDL, and HDL-cholesterol concentra- neck circumference (r = 0.88; P < 0.001), respectively.
tions were 86%, 104%, and 29% greater, respectively, in Plasma cortisol concentrations were < 10 ng/mL at
OB-IR horses (Table 2). 24 hours after DEX administration in all of the horses
evaluated by use of the DEX-TRH test, with the excep-
tion of 1 OB-IR horse that had a concentration of 10.7
ng/mL. For baseline and 30-minute post-TRH injection
plasma cortisol concentrations, 1 horse from each
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group had responses (74% and 150%) that exceeded


the 66% cutoff established for unaffected horses.16
However, both horses had typical suppression at 24
hours in response to administration of DEX. Median
plasma ACTH concentrations for the 3 OB-IR horses
tested ranged from 2.5 to 3.7 pmol/L, and these values
were less than the 10 pmol/L upper limit of the refer-
ence range provided by the laboratory.q

Discussion
A small population of obese horses with IR was
assembled for this study, and these horses had signifi-
cantly greater resting glucose, insulin, and leptin con-
centrations than healthy nonobese horses. High VLDL,
VLDL-triglyceride, and HDL-cholesterol concentra-
tions were also detected in OB-IR horses, suggesting
that obesity, IR, and dyslipidemia occur concurrently
in certain horses.
Mares predominated in the OB-IR group, but anec-
dotal reports and results of other studies2,5 suggest that
geldings are equally affected by obesity. Although vari-
Figure 2—Mean ± SD serum insulin concentrations (A) and plas- ous breeds were represented in the OB-IR group,
ma glucose concentrations (B) for 5 nonobese horses (squares)
and 7 obese horses with IR (diamonds) during CGITs. Time 0 = clients consistently referred to their horses as “easy
Time of insulin and glucose injection. keepers” because they appeared to require fewer calo-
Table 2—Plasma lipid and lipoprotein concentrations (median [range]) in 5 nonobese
horses and 7 obese horses with IR.

Nonobese OB-IR
Variable (n = 5) (7) P
NEFA (µmol/L) 197.1 (151.5–268.3) 366.5 (187.8–634.0) 0.028
Total cholesterol (mg/dL) 75.3 (73.0–88.1) 78.6 (60.4–86.0) 0.570
Triglyceride (mg/dL) 18.9 (11.0–22.6) 34.6 (13.8–134.8) 0.123
VLDL (mg/dL) 24.4 (19.8–34.6) 49.7 (29.0–162.0) 0.012
VLDL-triglyceride (mg/dL) 15.6 (11.2–23.2) 30.6 (17.4–97.1) 0.019
LDL (mg/dL) 30.3 (13.1–46.8) 30.9 (16.6–54.6) 0.465
LDL-cholesterol (mg/dL) 11.8 (5.2–18.3) 13.5 (6.8–22.4) 0.223
HDL (mg/dL) 139.7 (106.4–169.4) 110.8 (96.4–160.9) 0.168
HDL-cholesterol (mg/dL) 27.5 (24.8–34.2) 35.4 (32.2–52.3) 0.029

Table 3—Variables significantly correlated with plasma AUCg and serum AUCi values
for CGITs in horses.

AUCg AUCi
No. of
Variable horses r P r P
BCS 12 0.84 < 0.001 0.84 < 0.001
Neck circumference 11 0.71 0.015 0.88 < 0.001
Resting leptin concentration 12 0.76 0.004 0.69 0.013
Resting glucose concentration 12 0.83 < 0.001 0.72 0.008
Resting insulin concentration 12 0.66 0.020 0.83 < 0.001
Resting glucose-to-insulin ratio 12 –0.62 0.033 –0.87 < 0.001
HDL-cholesterol concentration 12 0.60 0.039 0.69 0.013

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ries than most horses to maintain body weight. It has technically more challenging and therefore less applic-
been suggested that some horses are genetically predis- able to clinical practice. These tests also require more
posed to obesity because of adaptations to survival on frequent sampling and are therefore more expensive. In
poorer quality forages.8,24 According to this theory, con- this study, the time required for blood glucose concen-
sumption of concentrated feeds or grazing on rich pas- trations to enter the negative phase was used to define
tures might therefore promote obesity in susceptible IR. This definition is easily applied, and only blood
horses. Genetic and environmental factors are likely to glucose measurements are required. Use of the baseline
be important in the development of obesity in horses, measurement as a reference point also allows handheld
and it is interesting that all of the OB-IR horses in this glucometers to be used because variability between
study were 10 years of age or greater, which suggests glucometers does not affect results. This procedure is
that time is required for environmental factors to alter also applicable to ambulatory practice because the
glucose metabolism. CGIT can be abbreviated to 60 minutes.
Because it is likely that diet contributes substantial- Higher resting serum insulin concentrations and

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ly to the development of obesity, the diet consumed by lower resting glucose-to-insulin ratios were detected in
horses prior to admission may have been a confounding horses from the OB-IR group, compared with nonobese
factor in this study. No attempt was made to control the horses, suggesting that these measures may be useful
diet of horses prior to their arrival, so horses had access screening tests for IR. Hyperinsulinemia is a feature of
to pasture and were consuming hay from various IR in humans and occurs when insulin secretion from
sources and sometimes a small quantity of a concentrate the pancreas increases to compensate for reduced
feed as a treat. Hoffman et al2 detected the effects of diet response to insulin.30 In the present study, resting
on response to insulin by determining that horses are serum insulin concentrations and glucose-to-insulin
less responsive to insulin after consuming a diet com- ratios were closely correlated with AUCg and AUCi,
posed primarily of starch and sugar instead of fat and suggesting that the same compensatory responses
fiber. The adipose biopsy procedure performed on day 0 occur in horses. However, it should be noted that a
may have also been a source of variability, but this was resting serum insulin concentration of 17.0 µU/mL was
discounted because all horses underwent the same pro- detected in 1 OB-IR horse, and this value was within
cedure and CGIT results for nonobese horses were con- the range detected for the nonobese group. This find-
sistent with those from another study.14 Effects of flunix- ing therefore underscores the importance of perform-
in meglumine on response to insulin have not been ing challenge tests such as the CGIT when evaluating
investigated, but administration of phenylbutazone for 5 horses with suspected IR.
days did not affect glucose tolerance or insulin secretion Resting glucose concentrations were positively
in geldings.25 correlated with AUCg values, indicating that blood glu-
Obesity was defined by BCS in this study with the cose concentrations increased as IR progressed.
scale developed by Henneke et al.15 Alternative mea- However, it is doubtful that resting blood glucose con-
sures of obesity include cadaver dissection,26 rump-fat centrations will serve as a useful measure of response
thickness,15,26 body mass index,27 and estimation of total to insulin in clinical practice because of the confound-
body water by use of bioelectric impedance analysis or ing effects of stress. Administration of epinephrine31 or
indicator dilution.28 Of these procedures, BCS and dexamethasone11,32 increases blood glucose concentra-
rump-fat thickness are the easiest to perform. A BCS is tions in horses, and stressful procedures such an endo-
assigned after visual appraisal and palpation of fat at 6 scopic examination can markedly alter CGIT results.14
sites on the body, including the neck, withers, back, In this study, horses were acclimated to the hospi-
tailhead, ribs, and area caudal to the shoulder.15 Rump- tal environment for 3 days before the first blood sam-
fat thickness is measured via B-mode ultrasonography ples were collected and mixed timothy-orchard grass
at a site approximately 5 cm lateral from the midline at hay was fed ad libitum before and during the CGIT to
the center of the pelvic bone, and percentage fat is cal- reduce stress. Horses were not tested until 3 days after
culated with an established formula.29 their arrival because transportation increases blood
Mean neck circumference was examined to assess cortisol concentrations and induces hyperglycemia and
regional adiposity because many OB-IR horses have hyperinsulinemia in donkeys.33 A 3-day acclimation
enlargement of adipose tissues at this location, which period was selected for the present study on the basis
is sometimes referred to as a cresty neck. In humans, of previous experiences with the CGIT14 and because
regional adiposity develops within the abdomen and an blood glucose and insulin concentrations returned to
enlarged waist circumference has been identified as a baseline concentrations within 22 hours in donkeys
risk factor for metabolic syndrome.1 Of the physical that were transported.33 Grass hay was fed to horses
measurements collected in this study, mean neck cir- because this feed was assumed to have a low glycemic
cumference was most closely correlated with AUCi, index.34 The glycemic index of the mixed timothy-
which suggested that enlargement of the neck is a risk orchard grass hay fed to horses in this study was not
factor for IR in horses. determined, but Bermuda hay has a mean glycemic
The CGIT was selected for use in this study index of 23%, compared with oats (100%).34 Hay fed to
because of its simplicity and low cost.14 The frequent- horses in the present study was not analyzed, but the
sample IV glucose tolerance test and the euglycemic- mean nonstructural carbohydrate content of hay pur-
hyperinsulinemic clamp test may have been better chased from the same farm was 12.8% of dry matter
choices because both provide specific quantitative when evaluated as part of another study35 performed
measures of insulin sensitivity,13 but these tests are during the same time period (January to October). The

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decision to permit hay consumption during testing was able to an increase in lipoprotein lipase activity. A pos-
also influenced by the observation that obese horses itive correlation between plasma HDL and triglyceride
become particularly agitated when deprived of feed. concentrations has also been detected in Shetland
An association between obesity and IR has been ponies.44
reported in horses2,36,37 and ponies.5,11 When Hoffman et Results of the present study provide evidence of an
al2 studied 3 obese Thoroughbred geldings by use of association between hyperleptinemia and IR in obese
the frequently-sampled IV glucose tolerance test, mean horses. Resting plasma leptin concentrations ranged
insulin sensitivity was < 20% of the value in nonobese from 1.0 to 30.5 ng/mL in OB-IR horses and were pos-
geldings. Obesity and lack of exercise are primary risk itively correlated with AUCg and AUCi values. Leptin is
factors for IR in humans, and the risk of developing a protein hormone synthesized by adipose tissues that
type 2 diabetes mellitus increases with the severity of is secreted in greater quantities when the body is in a
obesity.38 Blood NEFA concentrations increase with positive energy balance.47 Buff et al22 determined that
obesity and lack of physical activity in humans because serum leptin concentrations were positively correlated
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adipose tissues reach their maximum capacity for fat (r = 0.64; P < 0.001) with BCS in a herd of 71 Quarter
storage and the inhibitory effects of insulin on hor- Horses, which indicates that blood leptin concentra-
mone-responsive lipase are reduced.39 As a result, the tions reflect body-fat mass in horses. Plasma leptin
influx of fatty acids into tissues (including those of concentrations and BCS were also positively correlated
skeletal muscle) increases, which causes diacylglyc- in the present study. Hyperleptinemia has also been
erols to accumulate in cells.39 This phenomenon is associated with glucose metabolism disturbances in
sometimes referred to as lipotoxicity because intracel- horses.32 Cartmill et al32 identified obese horses with
lular lipids disrupt insulin receptor signaling in higher serum leptin concentrations and found that glu-
myocytes or β-cell function in the pancreas.39,40 In the cose tolerance test results were abnormal in these hors-
study reported here, plasma NEFA concentrations were es. When obese (BCS ≥ 7.5) horses were divided into
greater in OB-IR horses than in NO horses and were low-leptin (< 5 ng/mL) and high-leptin (7 to 20
significantly correlated with AUCi values. ng/mL) groups, significantly greater insulin responses
Plasma concentrations of VLDL and VLDL-triglyc- to IV glucose infusion were detected in the high-leptin
eride were significantly greater in OB-IR horses than in group.32
NO horses, and the median VLDL concentration (49.7 Results of the present study indicate that resting
mg/dL) was also greater than mean concentrations of serum insulin and leptin concentrations are useful
28.5 and 19.1 mg/dL detected in adult nonobese screening tests for IR in horses, but other factors
mares.41,42 Increased uptake of fatty acids by the liver including time of day and season must be considered
should increase the availability of triglyceride for when interpreting these values. Gordon and
VLDL synthesis and stimulate production of this McKeever48 determined that glucose and insulin
lipoprotein.1 Oversecretion of triglyceride-rich VLDL responses to feeding are higher in the morning, and
particles is associated with abdominal obesity in Fitzgerald and McManus49 determined that serum lep-
humans and attributed to an increase in hepatic fatty tin concentrations were highest in mature mares dur-
acid uptake.1 Increased VLDL production and synthe- ing the late summer and early fall and lowest during
sis of triglyceride-rich VLDLs have also been associat- the winter months in Kentucky. When the same group
ed with feed deprivation and hyperlipemia in horses, measured serum insulin concentrations in obese mares
which are conditions that develop in response to across a 12-month period, mean insulin concentrations
increased mobilization of NEFA from adipose were also higher during the summer and early fall.36
stores.41,43 Because most of the horses included in the present
Hypertriglyceridemia (≥ 150 mg/dL) and low study were evaluated during the summer, concentra-
HDL-cholesterol concentrations are 2 criteria used to tions of these hormones may have been higher than at
define metabolic syndrome in humans.10 These vari- other times of the year.
ables are associated because triglyceride carried by Horses included in this study were screened for
VLDLs and chylomicrons is exchanged for cholesterol PPID because this endocrinopathy has been associated
when these triglyceride-rich lipoproteins interact with with IR in horses.7 Three of 9 horses had abnormal
HDL in the blood.44 Consequently, as VLDL concentra- responses when combined DEX-TRH tests were per-
tions increase, interactions with HDL increase, and formed. However, no clinical signs of PPID were
HDL-cholesterol concentrations decrease in humans.1 observed, with the exception of laminitis, which has
However, VLDL and HDL-cholesterol concentrations been associated with this endocrinopathy in horses,6
were greater in OB-IR horses than in NO horses, which and the 2 components of the test did not agree in any
differs from responses detected in humans.1 This find- of the horses tested. These may have been false-posi-
ing may be explained by the fact that transfer of cho- tive responses, and the time of year that tests were per-
lesterol from HDL to VLDL is catalyzed by cholesterol formed may have influenced results. This issue has not
ester transfer protein in humans,1 whereas there is a been examined extensively, but Donaldson et al50
near absence of cholesterol ester transfer protein activ- recently determined that false-positive results are more
ity in equine blood.45 Higher plasma HDL-cholesterol likely to occur when DEX suppression tests are per-
concentrations have been associated with increased formed in the month of September. Alternatively, hors-
lipoprotein lipase activity in horses fed high-fat diets,46 es with positive test results in the present study may
which suggests that the higher HDL-cholesterol con- have had early or mild PPID that affected only 1 com-
centrations detected in OB-IR horses may be attribut- ponent of the test.

1388 Scientific Reports: Original Study JAVMA, Vol 228, No. 9, May 1, 2006
05-10-0480.qxp 4/11/2006 1:25 PM Page 1389

In the present study, examination of a small popu- 14. Eiler H, Frank N, Andrews FM, et al. Physiologic assess-
lation of OB-IR horses revealed that mean neck cir- ment of blood glucose homeostasis via combined intravenous glu-
cose and insulin testing in horses. Am J Vet Res 2005;66:1598–1604.
cumference should be measured in addition to BCS to 15. Henneke DR, Potter GD, Kreider JL, et al. Relationship
identify horses at risk for IR and that single measure- between condition score, physical measurements and body fat per-
ments of blood insulin and leptin concentrations are centage in mares. Equine Vet J 1983;15:371–372.
useful indicators of IR in obese horses. Horses concur- 16. Eiler H, Oliver JW, Andrews FM, et al. Results of a combined
rently affected by obesity, IR, and dyslipidemia were dexamethasone suppression/thyrotropin-releasing hormone stimula-
identified. tion test in healthy horses and horses suspected to have a pars inter-
media pituitary adenoma. J Am Vet Med Assoc 1997;211:79–81.
17. Havel RJ, Eder HA, Bragdon JH. The distribution and chem-
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f. Wako Chemicals USA, Richmond, Va.
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Selected abstract for JAVMA readers from the


American Journal of Veterinary Research
Correlation of magnetic resonance images with anatomic features of the equine tarsus
Rafael Latorre et al

Objective—To correlate anatomic features of the equine tarsus identified in plastinated sections with
images obtained via magnetic resonance imaging (MRI). May 2006
Animals—4 horses.
Procedures—MRI (1.5-Tesla magnet) of the tarsus was performed on the pelvic limbs of 4 clinically normal
horses following euthanasia. After imaging, tarsocrural joint spaces and vasculature were injected with colored
See the midmonth
latex. Sagittal and transverse sections of the tarsi were plastinated to facilitate interpretation of MR images. issues of JAVMA
Results—Relevant anatomic structures were identified and labeled on the plastinated tissue slices
for the expanded table
and corresponding MRI images. Results indicated high correlations between MRI findings and those of of contents
plastinated sections. for the AJVR
Conclusions and Clinical Relevance—The data obtained provided certain reference standards for or log on to
normal anatomic structure sizes and positions in the equine tarsus. This information may aid future avmajournals.avma.org
physiologic or clinical studies of this joint. (Am J Vet Res 2006;67:756–761) for access
to all the abstracts.

1390 Scientific Reports: Original Study JAVMA, Vol 228, No. 9, May 1, 2006