Motion-Mode Echocardiographic Measurement of Cardiac Dimensions in

Muzaffarnagari sheep

Anil Singh*, RP Pandey, G kumar, S Purohit

Department of Veterinary Surgery and Radiology
College of Veterinary Science and Animal Husbandry, DUVASU, Mathura, Uttar
Pradesh, India
*Corresponding author. Email: anil00singh@gmail.com

Abstract: Echocardiographic measurement of cardiac dimensions is a prerequisite for

research and clinical studies of cardiac diseases. These echocardiographic values are
not available in Muzaffarnagari sheep. Therefore, the present study was undertaken
to determine the reference motion (M)-mode echocardiographic values. M-mode
echocardiographic examination was performed in clinically healthy Muzaffarnagari
sheep (n = 12) divided into two groups of age, 3-6 month (n=6) and 12-15 month
respectively. The Mitral valve (MV), left ventricular (LV), left atrial and aortic
dimensions were measured, and parameters for systolic function were calculated and
compared. The negative correlation (p ≥ 0.05) was observed in the values of left
atrium, aortic diameter with age and body weight of the animals while their ratio
(LA/Ao) showed positive correlation. The Mean ± S.E. values of LA, AO and their
ratio LA/AO did not differ significantly between the sheep of group I and group II.
Right and left ventricular diameter measurement at diastole and systole,
Interventricular septum thickness, Posterior wall thickness (PW) at systole and
diastole, E point to septal separation, Ejection fraction, Fractional shortening,
shortening percentage and left ventricular mass.

Keywords: Cardiac dimensions, Motion-mode echocardiography, Muzaffarnagari

sheep

Introduction: Sheep is currently used as experimental animals in human cardiology

studies (Gorman et al. 1998). Similar to human heart, sheep heart also possesses a
limited capacity to develop collateral circulation after myocardial ischemic diseases
(Locatelli et al 2011). Reference values for echocardiographic parameters and indexes
of left ventricular function in healthy, young adult sheep used in translational research:
comparison with standardized values in humans. Despite an increasing interest in sheep
models for cardiovascular research, only few studies have been conducted to determine
motion (M)-mode echocardiographic reference values. Therefore, the present study was
undertaken to determine the reference M-mode echocardiographic values in
Muzaffarnagari sheep.

Material and Methods

Study place: The present study was carried out on12 apparently healthy Muzaffarnagari
sheep maintained at ILFC, DUVASU, Mathura. The study was carried out in the
Teaching Veterinary Clinical Complex, Kothari Veterinary Hospital, College of
Veterinary Science and Animal Husbandry, U.P. Pandit Deen Dayal Upadhyaya Pashu
Chikitsa Vigyan Vishwavidyalaya Evam Go-Anusandhan Sansthan, Mathura (UP).

Group-I : lambs 10-20 kg body weight and 3-6 months of age.

Group-II : Adult sheep 25-40 kg body weight and 12-15 of age.

I) Radiography:

To obtain good quality radiographs, X-ray machine (Heliophos-D, Siemens Healthcare

India) and computed radiography system (Regius Model 110 S with Regius direct
digitiger software, Konica Minolta Healthcare, India) were used. Lateral radiographs
were taken on fixed 95 cm focal film distance (FFD) and 320 mA, 9.5-13 mAs and 58-
70 KVP. The phosphor plates (14 x 17 inches) were horizontally oriented to obtain the
right lateral radiographs for complete visualization of the thorax from spine to sternum
and first rib to diaphragm to rule out cardiomegaly.

Motion-Mode Echocardiographic Examination

Right thoracic area from 2rd to 8th intercostal space just caudal to the triceps muscle,
Hairs/wool were clipped and shaved for echocardiograhic examination. Following this,
the animals were kept quiet and comfortable to bring their heart rate to the normal. All
echocardiographic examination was performed using MyLab40vet ESAOTE INDIA
and probe PA 122 by using 5.0-7.5 mHz. the animals were placed in right lateral
recumbency on a wooden table having a slit. The transducer was placed from the
dependent (right) side at the level of apex beat, through the slit in the table, to obtain
the right parasternal long axis left ventricular inflow-outflow tract view of the heart.
The image was positioned in a way that allowed the M-mode line to transect the LV
wall, IVS, mitral valve leaflets and left atrium-aortic root in a perpendicular fashion in
their respective views.

Mitral valve measurement:

From a right parasternal long-axis image, the cursor line was positioned perpendicular
over the MV leaflets at their tips when they were maximally opened. The anterior MV
leaflet produced an ‘M’ shaped image, whereas, posterior leaflet produced a flattened
‘W’ shaped image on the M-mode(fig).

A) Right ventricular dimensions (RVD):

a) Diastolic measurements: Mean ± S.E. values of right ventricular diameter in
diastole (RVDd) was 10.98 ± 0.87 and 11.15 ± 0.91 cm in the sheep of the group I
and II, respectively. The Mean ± S.E. values of the RVDd were differ significantly
between the sheep of group I and group II. The positive correlation was found in the
values of the RVDd with age and body weight of the animals. (Table 2) (Fig. 2)
In present study, RVDd (mm) was more or less similar to previously recorded values
of 0.99 ± 0.32 in adult sheep (Hallowell et al., 2012) but lower than values recorded
in Saanen goats by leroux et al. (2012), in Swedish goats by Olsson et al. (2001), in
Suffolk-cross sheep by Hallowell et al. (2012), in mixed breed group of Sheep by
Moses and Ross (1987) and in Lori-Bakhtiari sheep by Yadegari (2014). The
difference may be attributed to variations in the size and weight of the animals as
well as due to the variation in physiological status

B) Left ventricular dimensions (LVD):

a) Diastolic measurements (LVDd): Mean ± S.E. values of the left ventricular

diameter in diastole (LVDd) was 24.00 ± 1.50 mm and 26.47 ± 0.91 mm in the animals
of the group I and II, respectively. The positive correlation was observed in the values
of the LVDd with age and body weight of the animals (Table 2). (Fig. 2)
In the animals of the present study, LVDd was recorded lower than the Lori-Bakhtiari
sheep (2.94 ± 0.16 cm, Yadegari, 2014), Suffolk-cross sheep (4.42 ± 0.54 cm, Hallowell
et al., 2012),
Systolic measurements (LVDs):
Mean ± S.E. values of the systolic measurement of the left ventricle (LVDs)
was 12.50 ± 1.02and 13.28 ± 0.96 mm in the sheep of the group-I and II, respectively.
The positive correlation was observed in the values of LVDs with age and body weight
of the animals (Table 2) (Fig. 2). In animals of group II of the present study, LVDs was
recorded higher than for Lori-Bakhtiari sheep (1.28 ± 0.11 cm, Yadegari, 2014) while
lower than the adult Suffolk-cross sheep (2.62 ± 0.35 cm, Hallowell et al., 2012).

C) Interventricular septum thickness (IVS):

a) Diastolic measurements (IVSd):

Mean ± S.E. values of the diastolic measurement of the thickness of

interventricular septum (IVSd) was 8.27 ± 0.31 and 7.87 ± 0.75 mm in the sheep of the
group I and II, respectively. The negative correlation was observed in the values of
IVSd with age and body weight of the animals. (Table 2) (Fig. 2)
In the animals of group I and II of the present study, IVSd was recorded lower
than the Lori-Bakhtiari sheep (0.92 ± 0.11 cm, Yadegari 2014), Suffolk- cross sheep
(1.19 ± 0.15 cm, Hallowell et al., 2012

b) Systolic measurement (IVSs):

Mean ± S.E. values of the systolic measurement of the thickness of

interventricular septum (IVSs) was 11.00 ± 0.41 mm and 10.00 ± 1.06 mm in the sheep
of the group I and II, respectively. The positive correlation was observed in the values
of IVSs with age and body weight of the animals. In the animals of group I and group
II of the present study. (Table 2). IVSs was recorded lower than the Lori-Bakhtiari
sheep (1.45 ± 0.16 cm, Yadegari, 2014), Suffolk-cross sheep (1.55 ± 0.18 mm,
Hallowell et al., 2012).

D) Posterior wall thickness (PW):

a) Diastolic measurements (PWd):

Mean ± S.E. values of the diastolic measurement of the thickness of posterior

wall (PWd) was 8.05 ± 0.52 mm and 7.95 ± 0.58 mm in the sheep of the group I and II,
respectively. The negative correlation was observed in the values of PWd with age and
body weight of the animals. (Table 2) (Fig. 2)

b) Systolic measurements (PWs):

 Mean ± S.E. values of the systolic measurement of the thickness of posterior
wall (PWs) was 10.98 ± 1.06 and 11.08 ± 0.47 mm in the sheep of the group I and II,
respectively. The values of the PWs were negatively correlated with age while
positively correlated with body weight of the animals. (Table 2)

E) E point to septal separation (EPSS):

Mean ± S.E. values of the E point to septal separation (EPSS) was 2.92 ± 0.52
and 2.03 ± 0.31 mm in the sheep of the group I and II, respectively. The negative
correlation was observed in the values of EPSS with age and body weight of the
animals. (Table 2) (Fig. 3).

F) Ventricular functional indices:

a) Ejection fraction (EF):

Mean ± S.E. values of the ejection fraction (EF) was 81.17 ± 1.99 and 82.67 ±
2.30 % in the sheep of the group I and II, respectively. The negative correlation
was observed in the values of EF with age and body weight of the animals.
(Table 2) (Fig. 2)
b) Fractional shortening (FS):

Mean ± S.E. values of the fractional shortening (FS) was 48.17 ± 1.99% and
49.83 ± 2.41% in the sheep of the group I and II, respectively. The positive correlation
was observed in the values of FS with age and body weight of the animals. (Table 2)
(Fig. 2)

c) Shortening percentage (S%):

Mean ± S.E. values of the shortening percentage (S%) was 33.00 ± 6.08% and
46.17 ± 10.63% in the sheep of the group I and II, respectively. The positive
correlation was observed in the values of S% with age and body weight of the
animals. (Table 2)

d) Left ventricular posterior wall thickness percentage (PW%):

Mean ± S.E. values the left ventricular posterior wall thickness percentage
PW% was 38.00 ± 10.97 % and 45.00 ± 4.58 % in the sheep of the group I and II,
respectively. The positive correlation was observed in the values of PW% with age and
body weight of the animals. (Table 2)

e) Left ventricular mass (LVM):

Mean ± S.E. values the left ventricular mass (LVM) g was 40.33 ± 4.01 g and
46.33 ± 6.21 g in the sheep of the group I and II, respectively. The positive correlation
was observed in the values of LVM (g) with age and body weight of the animals. The
mean ± SE of left ventricular mass (LVM) was 1027.14 ± 48.29 g in cattle (Singh,
2017). Table 2.

Left atrium, aortic diameter and their ratio (LA/Ao):

Mean ± S.E. values of the left atrium, aortic diameter and their ratio (LA/Ao)
was 1.495 ± 0.49 mm, 1.462 ± 0.21 mm, 0.77 ± 0.03, and 1.462 ± 0.21 mm, 1.778 ±
0.81 mm, 0.83 ± 0.03 in the sheep of the group I and II, respectively. The negative
correlation (p ≥ 0.05) was observed in the values of left atrium, aortic diameter with
age and body weight of the animals while their ratio (LA/Ao) showed positive
correlation. The Mean ± S.E. values of LA, AO and their ratio LA/AO did not differ
significantly between the sheep of group I and group II. (table 1) (Fig. 1)

Conclusions
Echocardiographic reference values in adult, healthy, Muzaffarnagari sheep have been
established.
Conflict of interest: The authors declare that they have no conflict of interest

Table 1. Mean age (months) and body weight (kg) of animals of group I and II
(Mean ± S.E)

Groups Age (months) Body weight (kg)

Group I (n=6) 4.67 ± 0.33 17.83 ± 0.70

Group II (n=6) 13.83 ± 0.40 37.58 ± 0.66

Table 2. Mean ± S.E. values of left atrium (LA) and aorta (AO) diameters and
their ratio (LA/Ao) in M-mode echocardiography in sheep of group I and II.
 Group Left atrium (LA) Aorta (AO) LA/Ao
diameter (cm) diameter (cm)

Group-I 1.495 ± 0.49 1.462 ± 0.21 0.77 ± 0.03

Group-II 1.462 ± 0.21 1.778 ± 0.81 0.83 ± 0.03

Correlation with
-0.093 -0.459 0.381
Age

Correlation with
-0.098 -0.454 0.374
B.W.

** Correlation is significant at the 0.01 level, *Correlation is significant at the 0.05

level.

Fig. 1: M-Mode echocardiograms obtained from right Fig. 2: M-Mode echocardiograms obtained
parasternal short-axis view showing Ao and LA from right parasternal long-axis view showing
landmarks. RVDd, IVS, IVDd, IVSs, LVDs and PWs
landmarks.
Fig. 3: M-Mode echocardiograms obtained from
right parasternal long-axis view showing E point
to septal separation (EPSS) landmarks.
 3. ECHOCARDIOGRAPHY
Table 3. Mean ± S.E. values of M-mode echocardiographic measurements parameters and indices in sheep of groups I and II.

EPSS RVDd IVSd LVDd PWd IVSs LVDs PWs LVM

Groups EF% FS% S% PW%
(mm) (mm) (mm) (mm) (mm) (mm) (mm) (mm) (gms)

2.92 ± 10.98 ± 8.27 ± 24.00 8.05 11.00 ± 12.50 10.98 81.17 48.17 33.00 40.33 38.00
Group-I
0.52 0.87 0.31 ± 1.50 ± 0.52 0.41 ± 1.02 ± 1.06 ± 1.99 ± 1.99 ± 6.08 ± 4.01 ± 10.97

2.03 ± 11.15 ± 7.87 ± 26.47 7.95 10.00 ± 13.28 ± 11.08 82.67 49.83 46.17 ± 46.33 45.00
Group-II
0.31 0.91 0.75 ± 0.91 ± 0.58 1.06 0.96 ± 0.47 ± 2.30 ± 2.41 10.63 ± 6.21 ± 4.58

Correlation
-0.401 0.043 -0.033 0.393 -0.065 0.156 0.109 -0.003 0.254 0.268 0.239 0.295 0.191
with Age

Correlation
with B.W. -0.411 0.029 -0.054 0.388 -0.033 0.146 0.129 0.029 0.214 0.227 0.252 0.297 0.195

** Correlation is significant at the 0.01 level,*Correlation is significant at the 0.05 level

References

1. Gorman JH, Gorman RC, Plappert T, Jackson BM, Hiramatsu Y, John-Sutton MGS,
Edmunds LH Jr (1998) Infarct size and location determine development of mitral
regurgitation in sheep model. J Thorac Cardiov Surg 115:615–622
2. Hallowell GD, Potter TJ, Bowen IM. 2012. Reliability of quantitative echocardiography in
adult sheep and goats. BMC Veterinary Research, 8: 181.
3. Leroux AA, Moonen ML, Farnir F, Sandersen CF, Deleuze S, Salciccia A, Amory H. 2012.
Two-dimensional and M-mode echocardiographic reference values in healthy adult Saanen
goats. Veterinary Research, 170(6):154.
4. Locatelli P, Olea FD, De Lorenzi A, Salmo F, Vera Janavel GL,Hnatiuk AP, Guevara E,
Crottogini AJ (2011)
5. Olsson K, Hansson A, Hydbring E, von Walter LW, Haggstrom J. 2001. A serial study of
heart function during pregnancy, lactation and the dry period in dairy goats using
echocardiography. Experimental Physiology, 86(1):93-99.
6. Singh CK. 2017. Clinical studies on standardization of echocardiographic indices in
bovine. PhD thesis Submitted to GADVASU, Ludhiana.
7. Yadegari M. 2014. Normal echocardiographic findings in Lori-Bakhtiari sheep. Hum. Vet.
Med. 6, 45-48.