AL SC R IEN TU C A E N F D O N U A N D D A E I T L I Journal of Applied and Natural Science 8 (3): 1253 - 1259 (2016) O P N P JANS A ANSF 2008 Pathomorphological changes of flunixin meglumine toxicity in layer chicks * R. A. Patel, K. B. Kapadiya and D. J. Ghodasara Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand-388110 (Gujarat), INDIA *Corresponding author. E-mail: [email protected] Received: October 04, 2015; Revised received: April 29, 2016; Accepted: July 18, 2016 Abstract: The aim of the 21 day toxicity study was to evaluate the pathomorphological effect of flunixin meglumine in layer chicks. The chicks of Group I were kept as control while groups II, III and IV were fed with diet containing flunixin meglumine @ 10 ppm, 25 ppm and 50 ppm respectively for 21 days. Clinical signs viz. anorexia, dullness, lethargy, lameness and uneven growth were noticed in chicks of treatment groups III and IV only. Maximum mortal- ity was observed in group IV (12%) followed by group III (4%). A dose dependant reduction in body weight was ob- served in all the treatment groups. The mean values of Kidney: Body weight ratio was significantly increased in group IV. The plasma uric acid, creatinine and BUN values were significantly increased in group III whereas in- crease in group IV was highly significant. Grossly, there was deposition of chalky white urates on serosal surface of kidney, heart and liver in chicks of group IV which died during experiment. Microscopically, lesions were character- ized by varying degrees of congestion, haemorrhages, degeneration, necrosis and deposition of urate crystals in visceral organs of group III and group IV chicks. The intensity and distribution of pathological lesions were more severe in chicks of group IV, followed by chicks of group III. The overall lesions gave an impression that flunixin me- glumine was nephrotoxic in nature. Keywords: Flunixin meglumine, Nephrotoxicity, Urate crystals, Visceral gout INTRODUCTION properties, it has been used extensively alone or in combination with antimicrobial agents to treat a num- Diclofenac, a non-steroidal anti-inflammatory drug ber of conditions in domestic animals like lameness, (NSAID) was the only cause of rapid decline in vulture colic, endotoximia, mastitis, metritis and respiratory population across the Indian subcontinent (Oaks et al., diseases. This suggests that flunixin meglumine may 2004). Surveys of livestock carcasses across India indi- be potential alternative of diclofenac for veterinary use. cate that over 10% contained diclofenac residue that This highlights need for robust safety testing before occur at sufficient concentrations, in relation to dose recommending any NSAIDs as a safe replacement for dependent mortality of Oriental white backed vultures diclofenac (Swan et al., 2006). Poultry feed also con- (Gyps bengalensis) (Naidoo et al., 2009). Scientific tain many animal source byproducts like meat and evidences following the observation of waste disposal bone meal, blood meal, bone based DCP, mutton tal- practices of carcasses confirmed without any doubt low etc. which are likely to have flunixin meglumine that the veterinary use of diclofenac is the main cause residue and probably responsible for visceral gout in of decline in vulture population throughout the Indian layers. Therefore, the aim of this study was to evaluate subcontinent (Green et al., 2004). Thus, Government whether the flunixin meglumine can be used in animals of India banned diclofenac in veterinary use from May, as an alternative to diclofenac, without serious toxicity 2006 due to vulture crisis. After the ban of diclofenac in vultures and poultry. in veterinary field other NSAIDs such as ketoprofen, meloxicam, flunixin meglumine, carprofen, phenylbu- MATERIALS AND METHODS tazone etc. are now commonly used in veterinary prac- Experimental birds: A total of 100 apparently healthy tice. Diclofenac, and now ketoprofen, have been clearly day old BV-300 layer chicks (Gallus domesticus) were shown to be toxic to Gyps vultures, and carprofen and procured from a local commercial hatchery (Shakti flunixin are also likely to be toxic (Cuthbert et al., 2006). hatcheries, Sarsa, Anand, Gujarat, India) and were Flunixin, 2-(2-methyl-3-trifluoromethylenilino) nico- maintained at Experimental unit, Department of Vet- tinic acid, is a non-steroidal anti-inflammatory agent erinary Pathology, College of Veterinary science and that is a highly substituted derivative of nicotinic acid. Animal husbandry, Anand under standard managemen- In veterinary medicine, it is used with meglumine as a tal conditions. The chicks were given feed and fresh solubilizer as flunixin meglumine. As flunixin meglu- water ad libitum. mine has anti-inflammatory, analgesic and antipyretic ISSN : 0974-9411 (Print), 2231-5209 (Online) All Rights Reserved © Applied and Natural Science Foundation www.jans.ansfoundation.org R. A. Patel et al. / J. Appl. & Nat. Sci. 8 (3): 1253 - 1259 (2016) Experimental design: Chicks were randomly divided Table 2. Weekly body weight (Mean ± S.E., g) in layer into 4 equal groups. Chicks were fed graded dose of chicks of different experimental groups. FM (Virbac animal health India Pvt. Ltd., Mumbai, Da Group - Group - Group – Group - III Batch No. VB107) through dietary inclusion for 21 y I II VI consecutive days at the dose rate of 10, 25 and 50 mg/ 36.16±0. 0 36.4±0.31 35.08±0.43 35.68±0.31 kg (ppm) of feed in group II, III and IV respectively, 28 60.46±0. 59.52±0.8 while group I was offered feed free of FM and served 7 58.82±0.55 56.22±0.55 as control. All the birds were observed daily for any 70 0 100.2±0. 83.84±1.54 abnormal physical or behavioural changes and mortal- 14 98±0.75 97.3±0.88* 71 ** ity throughout the period of 21 days of experiment. 163.33±2 160.97±1. 159.43±1.3 143.73±1.6 21 Weight of chicks was recorded at day 1 and at the end .41 53 4* 9** of every week. At the end of experiment i.e. on 22 nd day, about 2 ml blood was collected from jugular vein *: Significant (p < 0.05); **: Highly significant (p < 0.01) in vaccutainer having K 3EDTA (1-2 mg/ml) as an anti- Table 3. Average feed conversion ratio (FCR), feed con- coagulant for plasma separation. Plasma samples were sumption (g) and body weight (g) in different experimental stored in deep freeze at -20 0C for uric acid, creatinine groups at end of experiment. and BUN estimation. After the blood collection all the surviving birds in all four groups were subjected to a Experimen- Feed con- Body weight FCR terminal sacrifice. Detailed post mortem examination tal Group sumption (g) (g) was performed on sacrificed and dead birds. The gross I 290.62 163.33 1.77 pathological lesions were recorded and for histopathologi- II 289.67 160.97 1.79 III 288.00 159.43 1.80 cal examination, tissue from kidney, liver, heart, lung, IV 263.65 143.73 1.83 spleen, proventriculus and intestine were collected in 10% formalin. Pieces of kidney, liver and heart tissues were Table 4. Mean values of plasma uric acid (mg/dl), creatinine also fixed in absolute alcohol for De-Galantha’s special (mg/dl) and BUN (mg/dl) in layer chicks of different experi- staining for demonstration of urate crystals. mental groups. RESULTS AND DISCUSSION Experimen- Uric acid Creatinine BUN (mg/ tal Group (mg/dl) (mg/dl) dl) Clinical signs: Chicks under group I and II did not I 6.105±0.17 0.28±0.02 1.74±0.07 reveal any observable clinical signs during the entire II 7.005±0.18 0.38±0.05 1.88±0.07 experimental period. Birds in the group III and IV III 8.55±0.12* 0.39±0.06* 1.92±0.09* 10.67±0.56* 0.91±0.06* showed similar clinical signs with gradually more pro- IV 2.68±0.08** * * nounced in group IV. Birds exhibited a tendency to remain standing at one place with apathy, dullness and *: Significant (p < 0.05); **: Highly significant (p < 0.01) drooping of the wings. Birds appeared emaciated, de- present experiment indicated that the compound FM hydrated, depressed and lethargic with shrunken eyes. could produce clinical signs like dullness, dehydration, They also exhibited clinical signs like anorexia and anorexia, uneven growth, ruffled feather, lethargy and uneven growth. shifting leg lameness were in agreement with the find- Muhammad et al., (2012) noticed signs of depression, ings of Muhammad et al., 2012. anorexia, lethargy and reluctance to move in FM Mortality : Maximum mortality was observed in group treated broiler birds. They showed stiff gait and felt IV (12%) followed by group III (4%) (Table 1). Mortality difficulty in walking and standing. Affected birds in group IV was observed on 15 th and 16 th day whereas in perched aside with ruffled feathers and eyes closed. group III it was on 16 th day of experiment. Prior to death affected birds became comatose. The Cuthbert et al., (2007) documented that carprofen and FM were associated with mortality of Gyps vultures and other Table 1. Mortality and Kidney: Body weight ratio (Mean ± species, with a reported mortality of 13% (5/40 cases) and SE, ×10 -3) of layer chicks of different experimental groups at 30% (7/23 cases), respectively as a result of renal failure the end of treatment period. and/or visceral gout, in a surveys of veterinarians and zoos document on treatment with NSAIDs. Muhammad No. of Experimen- Mortal- Kidney: Body et al., (2012) reported 20%, 40% and 60% mortality in layer birds tal Group ity % weight ratio five weeks old broiler chickens treated with FM at the died I 0 0 7.763 ± 0.104 dose rate of 2.5, 5 and 10 mg/kg body weight for four II 0 0 8.017 ± 0.133 consecutive days, intramuscularly.