Echocardiographic variables in healthy

guineapigs anaesthetized with

ketamine–xylazine

Nazmi Çetin1, Ebru Çetin1 & Melahat Toker2

1
Department of Physiology, Faculty of Veterinary Medicine, University of Erciyes, Kayseri, Turkey and
2
Social Security Organization, Ankara Training and Research Hospital, Department of Cardiovascular
Surgery, Ankara, Turkey

Summary
Echocardiographic parameters were recorded, measured and statistically analysed on a
population of 12 male Hartley albino guineapigs under ketamine–xylazine anaesthesia.
Additionally, the effect of body weight on these parameters and the correlation between the
parameters were assessed. The mean values of left ventricular internal diameter in end
diastole (LVIDD), left ventricular internal diameter in end systole (LVIDS), interventricular
septum thickness in diastole (IVSD), interventricular septum thickness in systole (IVSS), left
ventricular posterior wall thickness in diastole (LVPWD), left ventricular posterior wall thick-
ness in systole (LVPWS), left atrial diameter (LA), aortic diameter (AO), left ventricular frac-
tional shortening (FS) and left ventricular ejection fraction (EF) were measured or calculated as
6.85 ⫾0.36, 4.35 ⫾0.17, 1.75 ⫾0.31, 2.26 ⫾0.35, 2.28 ⫾0.40, 2.80 ⫾0.58, 4.95 ⫾0.34,
4.65 ⫾0.25 mm, 35.62 ⫾2.62 and 70.87 ⫾3.01%, respectively. A significant (P ⬍0.01) positive
correlation to body weight was found with LVIDD, LVPWD, IVSD, aortic root diameter and
LA. Significant correlation was also found between a number of echocardiographic parameters.

Keywords Echocardiography; guineapig

Transthoracic echocardiography is an The guineapig is frequently used as an

established non-invasive procedure that has
frequently been used to assess cardiac
morphology and physiology in dogs (Boon
et al. 1983), cats (Jacobs & Knight 1985)
and rabbits (Çetin et al. 2001). Recently,
several investigators have shown that
echocardiography is a valid approach to the
non-invasive evaluation of cardiac function
in small rodents, mice (Yang et al. 1999,
Chaves et al. 2001) and rats (Doursout et al.
2001).
experimental animal in cardiovascular
research. To our knowledge, echocardio-
graphic evaluation of cardiac morphology
and physiology in anaesthetized guineapigs
has not been studied previously.
The purpose of the present study was to
determine echocardiographic values in
anaesthetized guineapigs and to investigate
the effect of body weight on these parameters
and the correlation between parameters.

Material and methods

Correspondence to: Dr Nazmi Çetin, University of Erciyes,
Faculty of Veterinary Medicine, Department of Physiology, Animals
Barış Manço C. Sümer M., Kocasinan-Kayseri, 38090 Turkey
E-mail: cetin@erciyes.edu.tr
Twelve male albino guineapigs of the
Hartley strain weighing 500–650 g were used
Accepted 15 June 2004 © Laboratory Animals Ltd. Laboratory Animals (2005) 39, 100–106
Echocardiography in guineapigs
in this study. They were obtained from the
Experimental Animals Breeding Department
(Refik Saydam Hygiene Center, Ankara,
Turkey). The guineapigs were considered
healthy after successful physical, electrocar-
diographic and echocardiographic examina-
tions. All guineapigs were housed separately
in wire mesh cages (50 ⫻50 ⫻30 cm).
Guineapig chow, fresh vegetables, and water
were available ad libitum. Animals were
maintained at 22 ⫾2°C with a 12 h/12 h
light/dark cycle. Electrocardiographic
tracing was recorded on all awake
guineapigs the week before the echocardio-
graphic investigation and all guineapigs
were within the normal range (Yılmaz
1988).
Echocardiography
Twelve guineapigs were anaesthetized with
a combination of xylazine (5 mg/kg body
weight, i.m) and ketamine (30 mg/kg body
weight, i.m.) to perform echocardiographic
examination. The dosages used were approx-
imately the average of previously published
reports (Harkness & Wagner 1989), provid-
ing approximately 30–50 min of anaesthesia
in guineapigs.
Following the induction of anaesthesia,
the chest of each animal was carefully
shaved. The animals were positioned in the
supine position with spread front legs, and
an ultrasound transmission gel was applied
to the precordium. Transthoracic echocar-
diography was performed using a Toshiba
SSH 160 A ultrasonography and 7.5 MHz.
transducer. The heart was first imaged in
the two-dimensional (2-D) mode in the
parasternal long axis view. From this view,
the M-mode line was positioned perpendi-
cular to the interventricular septum and
passed through the left ventricle (LV) struc-
tures, at the level of the chordae tendinea,
just below the mitral valve, and M-mode
images were obtained. The aortic dimension
(AO) and left atrial dimension (LA) were
obtained after the cursor was moved to the
aortic root and left atrium (Sahn et al. 1978).
Measurements of ventricular septal
thickness (IVS), left ventricular internal
dimension (LVIDD), and left ventricular
101
posterior wall thickness (LVPW) were made
from two-dimensionally directed M-mode
images of the LV in both the systole and the
diastole by use of the leading edge-to-edge
convention adopted by the American
Society of Echocardiography (Sahn et al.
1978). LV per cent fractional shortening (FS)
was calculated from the M-mode using the
following equation
FS (%) : [(LVIDD ⫺LVIDS)/LVIDD] ⫻100.
Left ventricular ejection fraction (EF) was
automatically calculated by the echocardiog-
raphy machine according to the Teicholz
formula. Each measurement was obtained
by averaging results from three consecutive
heart beats.
Statistics
Mean values, standard deviation (SD),
maximum and minimum values (range) for
body weight and the echocardiographically
measured or calculated parameters were
determined with the help of a statistics
program (SPSS 10.0 for Windows). Pearson’s
coefficient of correlation was computed for
all pairs of variables. When the correlation
was found to be statistically significant at
the 5% level (P ⬍0.05), the equation of the
linear regression of body weight on the
echocardiographic parameter was calculated.
The purpose of regression analysis was to
find a formula permitting prediction of the
expected echocardiographic value when
body weight was known. A scatter plot was
created as a test of the linearity assumption.
The graphic representations show the
individual values, the regression line and
the 95% confidence interval. All statistical
calculations were computed using the
statistic program.
Results
Imaging
Two-dimensional echocardiogram, performed
in left parasternal long axis view, revealed
a number of left heart structures; in
particular, left atrial and ventricular cavities
and walls, the interventricular septum, the
Laboratory Animals (2005) 39
102
aortic root and the mitral leaflets (anterior
and posterior mitral leaflet) (Fig 1). The apical
four chamber view could not be achieved in
the animals. The right ventricular (RV) wall
was not delineated clearly enough to allow
an accurate measurement of RV internal
dimension. The arithmetic mean, SD,
extreme values (min and max) for body
weight, and the echocardiographically mea-
sured or calculated parameters are given in
Table 1.
Effect of body weight on echocardiographic
parameters
A significant (P ⬍0.01) positive correlation
to body weight was found with LVIDD,
IVSD, LVPWD, AO and LA (Figs 2–6). On
the other hand, there was no significant
correlation between body weight and LVIDS,
IVSS, LVPWS, EF and FS. The regression
equation and coefficient correlation are
indicated in Table 2.
Correlations between echocardiographic
parameters
There were several statistically significant
positive correlations (Table 3).
There were positive correlations (P ⬍0.01)
between EF and FS, IVSD and LVPWD, AO
Fig 1 M-mode (left) and two-dimensional (right) parasternal long axis view of the left heart obtained
from a guineapig weighing 610 g. LA: left atrium; AO: aortic root; IVS: interventricular septum; PW: left ven-
tricular posterior wall; LV: left ventricle; AML and PML: mitral valve leaflets
Laboratory Animals (2005) 39
Çetin, Çetin & Toker
and EF, AO and LVIDS. EF and FS were
significantly correlated (P ⬍0.05) to LVPWS.
Left ventricular posterior wall thickness
in systole, AO, EF and FS were significantly
correlated (P ⬍0.01) to LVIDD.
Discussion
These data demonstrate that good-quality
2-D directed M-mode echocardiograms can
be obtained in anaesthetized guineapigs.
Echocardiographic examination of the RV
dimension was difficult, and measurement
was less precise than with other cardiac
structures. We were unable to determine
right ventricular wall thickness in this
study as the endo- and epicardium of the
right heart could not be visualized on the
echocardiograms. The apical four-chamber
view could not be achieved in the animals
because of the small size and the position
of the guineapig’s heart.
In this study, anaesthesia (a mixture
of ketamine and xylazine) was used to
facilitate echocardiographic examination.
Ketamine can profoundly alter heart rate
and thereby obscure any intrinsic relation-
ship between echocardiographic parameters
and heart rate (Jacobs & Knight 1985, Yang
et al. 1999). Therefore, correlation between
Echocardiography in guineapigs 103

Table 1 Echocardiographic parameters of anaes-

thetized guineapigs

Parameters Mean ⫾SD Range

LVIDD (mm) 6.85 ⫾0.36 6.30–7.50

LVIDS (mm) 4.35 ⫾0.17 4.20–4.60
IVSD (mm) 1.75 ⫾0.31 1.30–2.10
IVSS (mm) 2.26 ⫾0.35 1.70–2.70
LVPWD (mm) 2.28 ⫾0.40 1.50–2.70
LVPWS (mm) 2.80 ⫾0.58 1.90–3.50
AO (mm) 4.65 ⫾0.25 4.40–5.00
LA (mm) 4.95 ⫾0.34 4.60–5.40
EF (%) 70.87 ⫾3.01 67.00–78.00
FS (%) 35.62 ⫾2.62 32.00–41.00
BW (g) 570.13 ⫾31.09 500.00–650.00

Values represent the mean ⫾SD and range for 12 animals.

LVIDD and LVIDS: left ventricular internal diameter in dias-
tole and systole; IVSD and IVSS: interventricular septum
thickness in diastole and systole; LVPWD and LVPWS: left ven-
tricular posterior wall thickness in diastole and systole; AO:
aortic root diameter; LA: left atrial diameter; EF: ejection
fraction; FS: fractional shortening; BW: body weight
Fig 3 Interventricular septal thickness in end dias-
tole (IVSD) and corresponding weights. The solid
line represents the calculated regression equation
and the broken lines represent 95% confidence
interval. See Table 3 for the regression equation

Fig 2 Left ventricular internal dimension in end
diastole (LVIDD) and corresponding weights. The
solid line represents the calculated regression equa-
tion and the broken lines represent 95% confidence
interval. See Table 3 for the regression equation
Fig 4 Left ventricular posterior wall thickness in end
diastole (LVPWD) and corresponding weights. The
solid line represents the calculated regression equa-
tion and the broken lines represent 95% confidence
interval. See Table 3 for the regression equation

echocardiographic parameters and heart rate
was not determined in this work.
In veterinary medicine, most researchers
have used body weight rather than surface
area or age for regression analyses (Lombard
1984, Amory & Lekeux 1991, Hanton et al.
1998). This seems to be useful, because body
weight is a variable that can be determined
more easily and more accurately than age or
surface area in animals (Schille & Skrodzki
1999). The results of the present study demon-
strate the significant linear relationship
between increase in body weight and
increase in cardiac dimension for several
Laboratory Animals (2005) 39
104
Fig 5 Aortic root dimension (AO) and correspon-
ding weights. The solid line represents the calculat-
ed regression equation and the broken lines
represent 95% confidence interval. See Table 3 for
the regression equation
Fig 6 Left atrial dimension (LA) and corresponding
weights. The solid line represents the calculated
regression equation and the broken lines represent
95% confidence interval. See Table 3 for the regres-
sion equation
echocardiographic values. When body
weight was evaluated with each variable,
LVIDD, IVSD, LVPWD, AO and LA were
significantly correlated. Similar relation-
ships have been previously described
between body weight and LVIDD, IVSD,
IVSS and LVPWD (Hanton et al. 1998), and
Laboratory Animals (2005) 39
Çetin, Çetin & Toker
Table 2 Effect of body weight on echocardiographic
parameters
Regression Coefficient of
Parameters equation correlation (r)
LVIDD (mm) 1.88 ⫹0.010 BW 0.89**
IVSD (mm) 1.51 ⫹0.010 BW 0.77**
LVPWD (mm) 2.92 ⫹0.010 BW 0.82**
AO (mm) 2.18 ⫹0.001 BW 0.76**
LA (mm) 2.17 ⫹0.010 BW 0.83**
BW: body weight in g. Significance of the coefficient of cor-
relation: **P ⬍0.01. See Table 1 for other abbreviations
LVID, LA, AO and IVS in dogs (Lombard
1984), and IVST and LVPW (Philip et al.
1985), and LA in cats (Allen 1982). Also,
a positive correlation between body weight
and LVIDD, IVS, LVPWT, LA and AO was
reported by Bradley and Ross (1987) in
sheep.
The size of the left ventricle cavity and
walls were higher in our guineapigs than
values in mice and rats (Table 4). These
differences can be explained by the higher
body weight in guineapigs and the correla-
tion we have found between body weight
and these parameters.
Fractional shortening is an important
parameter of left ventricular function
(Bonagura 1983, Feigenbaum 1994). In the
present study, FS does not depend on body
weight. This is in agreement with the
findings of others (Stewart et al. 1984, De
Madron et al. 1985, O’Grady et al. 1986,
Amory & Lekeux 1991) who also studied
young animals of various species in the
growing phase in order to determine the
influence of body weight on echocardio-
graphic parameters. Fractional shortening in
our guineapigs had a mean 35.62 ⫾3.62. The
FS of the left ventricular values observed in
this study was comparable with the data
obtained by Yang et al. (1999) in mice, rats
(Akula et al. 2003) (Table 4) and rabbits
(33.35±0.8%) (Çetin et al. 2001).
The EF, which can be calculated from the
left ventricular dimensions via M-mode
echocardiography, is another parameter of
cardiac function. Our echocardiograph
calculated the EF automatically using the
Teicholz formula. The accuracy of the
Echocardiography in guineapigs 105

Table 3 Correlations between echocardiographic parameters in the anaesthetized guineapigs

LVIDD LVIDS IVSD IVSS LVPWD LVPWS AO LA EF FS

LVIDD 1.000 0.542 0.098 ⫺0.316 0.286 0.729** 0.815** 0.472 0.957** 0.747**
LVIDS 1.000 ⫺0.085 0.083 0.158 ⫺0.092 0.806** 0.453 0.484 0.124
IVSD 1.000 0.573 0.737** 0.356 0.112 0.051 0.202 0.271
IVSS 1.000 0.273 ⫺0.460 ⫺0.210 0.275 ⫺0.302 ⫺0.308
LVPWD 1.000 0.597* 0.518 0.007 0.279 0.142
LVPWS 1.000 0.454 ⫺0.147 0.652* 0.669*
AO 1.000 0.389 0.736** 0.322
LA 1.000 0.412 0.364
EF 1.000 0.820**
FS 1.000

Significance of the coefficient of correlation: *P ⬍0.05; **P ⬍0.01. See Table 1 for other abbreviations

Table 4 A comparison of the values of the echocardiographic variables observed

in healthy guineapigs with the corresponding values in other species

Mouse Rat Guineapig

Parameters (Yang et al. 1999)* (Akula et al. 2003)* Present study*

LVIDD (mm) 3.57 ⫾0.09 6.18 ⫾0.25 6.85 ⫾0.11

LVIDS (mm) 2.34 ⫾0.08 4.25 ⫾0.17 4.35 ⫾0.05
IVSD (mm) 0.43 ⫾0.01 1.21 ⫾0.15 1.75 ⫾0.08
IVSS (mm) 0.91 ⫾0.02 1.65 ⫾0.25 2.26 ⫾0.10
LVPWD (mm) 0.42 ⫾0.01 1.41 ⫾0.29 2.28 ⫾0.10
LWPVS (mm) 0.89 ⫾0.03 2.36 ⫾0.34 2.80 ⫾0.13
AO (mm) NR NR 4.65 ⫾0.06
LA (mm) NR NR 4.95 ⫾0.12
EF (%) 55.00 ⫾1.7 68.18 ⫾2.8 70.87 ⫾1.20
FS (%) 34.6 ⫾1.1 31.22 ⫾1.7 35.62 ⫾1.04
BW (g) 24.3 ⫾0.6 225.00 ⫾3.4 570.13 ⫾11.09

Values are given as the mean ⫾SE. See text for definition of echocardiographic variables
*Values measured in anaesthetized subjects. NR: not reported

Teicholz formula has not been evaluated in
small rodents, but has been shown to
predict EF in human beings more accurately
than other proposed formulas that use
M-mode measurements (Teicholz et al.
1976, Kronik et al. 1979). The values of EF
in the present study are comparable to the
values in other species (Table 4). Like the
FS, it was not dependent of the guineapigs’
body weight (Table 2). In agreement with
other findings (Lombard 1984, Schille &
Skrodzki 1999), the EF did not correlate
with body weight in the present study.
Further studies are necessary to validate the
significance and validity of EF measure-
ments in guineapigs.
To conclude, we have generated reference
values for a number of M-mode echocar
diographic parameters in guineapigs and we
have demonstrated the effect of body weight
on some of these parameters and a
number of correlations between echo-
cardiographic parameters. By use of
transthoracic echocardiography, it is
possible to quantitatively assess left
ventricular dimension and wall thickness
in guineapigs. These findings give a better
understanding of the functional changes
investigated by echocardiography
in guineapigs and will be helpful when
interpreting echocardiographic data in small
experimental animals.
Laboratory Animals (2005) 39
106
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