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ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE) RESEARCH ARTICLE

Effects of Minimum and Maximum Doses of Furosemide on Fractional Shortening Parameter
in Echocardiography of the New Zealand White Rabbit
Roham Vali, Mohammad Nasrollahzadeh Masouleh* and Siamak Mashhady Rafie1
Department of Radiology; 1Department of Small Animal Internal Medicine, Faculty of Specialized veterinary Science,
Science and Research Branch, Islamic Azad University, Tehran, Iran
*Corresponding author:
There is no data on the effect of maximum and minimum doses of furosemide on heart's work performance and amount of fractional shortening (FS) in echocardiography of rabbit. This study was designed to validate probability of the Key words:
mentionable effect. Twenty-four healthy female New Zealand white rabbits were divided into four equal groups. Maximum and minimum doses of furosemide were used for the first and second groups and the injection solution for the third and fourth groups was sodium chloride 0.9% which had the same calculated volumes of furosemide for the first two groups, respectively. The left ventricle FS in statutory times (0, 2, 5, 15, 30 minutes) was determined by echocardiography. Measurements of Mean±SD, maximum and minimum amounts for FS values in all groups before injection and in statutory times were calculated. Statistical analysis revealed non-significant correlation between the means of FS. The results of this study showed that furosemide can be used as a diuretic agent for preparing a window approach in abdominal ultrasonography examination with no harmful effect on cardiac function. To Cite This Article: Vali R, MN Masouleh and SM Rafie, 2013. Effects of minimum and maximum doses of
furosemide on fractional shortening parameter in echocardiography of the New Zealand white rabbit. Pak Vet J, 33(2):
ventricles (Fontes-Sousa et al., 2009). Fractional shortening (end-diastolic diameter minus end-systolic diameter divided Furosemide is one of the most commonly used drugs in by end-diastolic diameter) is a measureable parameter that clinical medicine (Suzuki et al., 2011). It is the most potent can determine ventricular function (Fontes-Sousa et al., and most common diuretic used in heart failure (Pichette and Du-Souich, 1996; Peddle et al., 2012). Congestive heart In routine ultrasonography exams, the sonologist can failure has been reported in pet rabbits (Lord et al., 2011). use the furosemide as a diuretic agent to finding a good Furosemide a loop diuretic, acts on the loop of henle of the window for abdominal investigations, but in the case of nephron by reversibly inhibiting the sodium/potassium/ presence of cardiac disease and probability of the chloride co-transfer. This action increases the amount of furosemide influence on cardiac function, what can the water in the tubule and subsequently increasing the urinary clinician do? As the amount of potassium reduction in volume and decreasing the blood volume (Alván et al., therapeutic doses of furosemide is unclear, this study was 1990). It is used for the treatment of canine pulmonary designed to measure the amount of potassium reduction and edema secondary to left heart failure (Peddle et al., 2012). interaction effects on cardiac function. In addition the Increase of the urinary volume and distention of urinary probability of difference on the effects of the maximum and bladder can be useful in diagnosis of this organ's diseases minimum doses of furosemide on cardiac function was and finding an appropriate window in abdominal determined. ultrasonography. The dose of furosemide in rabbits is 5- 10mg/kg body weight, and the routes of administration are MATERIALS AND METHODS
subcutaneously (SC), intramuscularly (IM) and intravenously IV (Carpenter et al., 2004). Twenty-four healthy female New Zealand white rabbits Echocardiography is a non-invasive technique used for weighing (1.51±0.21kg) in four equal groups were used. the evaluation of cardiac structure and function such as Rabbits were healthy and free from any sign of Pak Vet J, 2013, 33(2): 218-220.
cardiovascular or respiratory tract diseases on the basis of Zealand white rabbits are summarized in Table 1. the physical examination which included careful thoracic Minimum, maximum, mean+SD before injection in 24 auscultation. The animals were housed individually in female New Zealand white rabbits was 32, 53 and cages in a controlled environment, at temperature of 20- 40.8±6.1. Potassium concentrations before injection and 25°C, with 12 hours of light and 12 hours of dark cycle, after injection (min 60) are summarized in Table 2. and were fed a standard pellet diet and adequate water. Blood samples were obtained from saphenous vein, Table 1: Values (mean±SD) fractional shortening in al groups before
(min 0) and after injection of drug at various intervals twenty-four hours before injection of furosemide and echocardiography examination and at Minute 60 after injection. These samples were collected for determining plasma potassium concentration and differences before and after the injection. The normal range of potassium is 3.7-6.8 meq/l (Carpenter et al., 2004). The weight of each rabbit was recorded prior to injection of furosemide. The rabbits of the first and second group received maximum (10mg/kg) and minimum (5mg/kg) dosage of furosemide respectively (SC injection). The rabbits of third and fourth 1=Group1 received maximum dose of furosemide; 2=Group 2 received group received SC injections of sodium chloride 0.9% as minimum dose of furosemide; 3=Group 3 received maximum dose of specified for the first and second group. Feeding was sodium chloride; 4= Group 4 received minimum dose of sodium withheld a few hours before echocardiography chloride. examination to reduce abdominal distension from Table 2: Potassium concentration (Means±SD) before and after drug
intestinal fill, which could mechanically compress the diaphragm and lungs, particularly when the abdomen is compressed during the segment of echocardiography examination in which the images were obtained via the Imaging technique: Rabbits were placed in right lateral
recumbency to obtain right parasternal view, over a gap in There are several reasons that the New Zealand the tabletop through which the ultrasound probe was white rabbits are a good model for cardiovascular brought from below and placed on a shaved area on the research, such as; the ability to perform repeated anterior aspect of the lower portion of the right thoracic measurements in the same animal, low cost to procure and wall. Echocardiography measurements were obtained maintain, easy approach to arterial and venous system, from standard views (Boon, 2011). Transthoracic 2- relatively large heart size in relation to thoracic volume, dimensional and M-mode echocardiography were The possibility of using standard recording equipment and performed by using an ultrasonography system (Acuson, easy management of the rabbits, as a result, these rabbits Cyprus) equipped with a 3-7 MHz (7v3c) neonate are commonly used in cardiac research (Salemi et al., transducer. From the right parasternal short-axis view, 2- 2005; Fontes-Sousa et al., 2006). In addition their dimensional guided m-mode tracing was made just below similarity to humans with regard to cardiac anatomy, the mitral valve at the level of the papillary muscles for physiology and age related changes, are other reasons for measurements of the left ventricular internal diameter using rabbits in these kinds of research (Fontes-Sousa et (LVID), in diastole and systole (Dimitrov et al., 2011). Fractional shortening was calculated from measurements Fractional shortening parameter has been determined for the LVID in systole and diastole by use of the for the mouse, Syrian hamster, rabbit, guinea pig and Chinchilla (Table 3). In the present study, the mean of FS FS (%) = [(LVIDd – LVIDs)/LVIDd] 100 (Fontes- value in all groups before injection is about (40.8±6.1). Sousa et al., 2006; 2009). In our study calculations of the This value compared with the results of other studies on FS were easily obtained in all of the animals, before rabbits has shown an increase (table3). This difference injection (min0) and at 2, 5, 15 and 30 minutes after could be due to the use of anesthetic drugs, since research injection by use of ultrasound system software. All data has shown that anesthetic drugs may reduce fractional about each rabbit was recorded. Measurements of mean, shortening (Yang et al., 1999; Baumgartner et al., 2010; standard deviation of (FS) before injection and in Pelosi et al., 2011). No significant correlation was found statutory times were calculated for all rabbits. Statistical between means of FS values before injection and at 2,5,15 procedures for comparison between mean of FS (min 0) and 30 minutes after injection by using independent- and mean at 2, 5, 15 and 30 minutes after injection were sample t-test, and the p-level for statistical significance done by using independent-sample t-test. Statistical was set at 0.05 (Table 1). differences were considered significant at P<0.05. Normal serum potassium levels are between 3.7 and 6.8 mEq/L (Carpenter et al., 2004; Bielecka-Dabrowa et RESULTS AND DISCUSSION
al., 2012). In our study Means of serum potassium concentration before injection and after injection (min 60) In the present study, none of the animals died during in all groups was compared and no significant correlation or after the study. Values for FS before injection (min 0) was found, (Table 2). Although potassium concentration and after injection (2, 5, 15 and 30 minutes) in 24 New reduced after injection, drug type and dose has no Pak Vet J, 2013, 33(2): 218-220.

Table3: Values for Fractional shortening in rodents as reported by various workers
Nasrol ahzadeh-Masouleh et al. (2010) significant effect on serum potassium before and after Boon JA, 2011. The two dimensional echocardiographic exam. In: injection. This reduction could be due to furosemide Hanbook of Veterinary Echocardiography, 2nd Ed, John Wiley & Sons, Chichester, UK; pp: 37-90. administration, because diuretics like furosemide are Carpenter JW, TY Mashima and DJ Rupiper (eds), 2004. Exotic Animal known to inhibit sodium and chloride co-transport from Formulary, 3rd Ed, WB Saunders, Philadelphia, USA, pp: 428. the luminal side of thick ascending limb of the loop of Cetin N, E Cetin and M Toker, 2005. Echocardiographic variables in Henle. The increased electrolyte excretion will then healthy guineapigs anaesthetized with ketamine-xylazine. Lab Anim, 39:100-106. increase tubular flow and urinary volume. The effect on Dimitrov R, D Vladova, K Stamatova, D Kostov and M Stefanov, 2011. potassium is considered to be transient and mainly Transthoracal two-dimensional ultrasonographic anatomical study
secondary to increase tubular flow (Alván et al., 1990). of the heart in the rabbit (Oryctolagus cuniculus).Trakia J Sci, 9: 45- Drugs such as furosemide may precipitate hypokalemia by Fontes-Sousa AP, C Brás-Silva, C Moura, JC Areias and AF Leite- accelerating renal loss (Bielecka-Dabrowa et al., 2012). Moreira, 2006. M-mode and Doppler echocardiographic reference There are three types of hypokalemia. In mild values for male New Zealand white rabbits. Am J Vet Res, 67: hypokalemia potassium concentration is 3-3.5mEq/L and most often asymptomatic. In moderate hypokalemia Fontes-Sousa AP, C Moura, CS Carneiro, A Teixeira-Pinto, JC Areias potassium concentration is 2.5-3.5mEq/L and non specific and AF Leite-Moreira, 2009. Echocardiographic evaluation including tissue Doppler imaging in New Zealand white rabbits sedated with symptoms like weakness, malaise and constipation appear ketamine and midazolam. Vet J, 181: 326-331. as well. Potassium concentration in severe hypokalemia is Linde A, NJ Summerfield, M Johnston, T Melgarejo, A Keffer and E Ivey, below 2.5 mEq/L and cardiac arrhythmias increase 2004. Echocardiography in the chinchil a. J Vet Intern Med, 18: (Rastegar and Soleimani, 2001). Changes in the potassium Lord B, C Devine and S Smith, 2011. Congestive heart failure in two pet concentration in this study are in normal range of serum potassium concentration (3.7 to 6.8 mEq/L) and it has no Nasrollahzade MM, ORS Amlashi, MS Ghaffari and MR Paryani, 2010. Effect of maximum and minimum dose of atropine on fractional shortening parameter of rabbit echocardiography. J Vet Clin Res, 1: Conclusion: In conclusion, the results of this study
Peddle GD, GE Singletary, CA Reynolds, DJ Trafny, MC Machen and indicate that a single dose of furesemide administration in MA Oyama, 2012. Effect of torsemide and furosemide on clinical, therapeutic doses (minimum or maximum) has no laboratory, radiographic and quality of life variables in dogs with significant effect on reducing the potassium concentration heart failure secondary to mitral valve disease. J Vet Cardiol, 14: 253-259. and changes in the Fractional shortening parameter. The Pelosi A, L St John, L Gaymer, D Ferguson, SK Goyal, GS Abela and J results of this study conclude that furosemide can be used Rubinstein, 2011. Cardiac tissue Doppler and tissue velocity for preparing a window approach in abdominal imaging in anesthetized New Zealand white rabbits. J Am Assoc ultrasonography with no harmful effect on mechanical Pichette V and P Du-Souich, 1996. Role of the kidneys in the metabolism cardiac performance and only prolonged furosemide of furosemide: Its inhibition by probenecid. J Am Soc Nephrol, 7: administration, may induce reduction of serum potassium Rastegar A and M Soleimani, 2001. Hypokalemia and hyperkalemia. Salemi VM, AM Bilate, FJ Ramires, MH Picard, DM Gregio, J Kalil, EC REFERENCES
Neto and C Mady, 2005. Reference values from M-mode and Doppler echocardiography for normal Syrian hamsters. Eur J Alván G, L Helleday, A Lindholm, E Sanz and T Vil én, 1990. Diuretic effect and diuretic efficiency after intravenous dosage of frusemide. Suzuki S, T Ishikawa, L Hamabe, D Aytemiz, H Huai-Che, R Fukushima, Br J Clin Pharmacol, 29: 215-219. N Machida and R Tanaka, 2011. The effects of furosemide on left Baumgartner C, M Bollerhey, J Ebner, L Laacke-Singer, T Schuster and atrial pressure (LAP) in dogs with mitral regurgitation (MR) have W Erhardt, 2010. Effects of ketamine-xylazine intravenous bolus not been documented in a quantitative manner and between injection on cardiovascular function in rabbits. Can J Vet Res, 74: different routes of administration. J Vet Intern Med, 25: 244-250. Yang XP, YH Liu, NE Rhaleb, N Kurihara, HE Kim and OA Carretero, Bielecka-Dabrowa A, DP Mikhailidis, L Jones, J Rysz, WS Aronow and M 1999. Echocardiographic assessment of cardiac function in Banach, 2012. The meaning of hypokalemia in heart failure. Int J conscious and anaesthetized mice. Am J Physiol, 277: H1967-


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