Effect of morphine and ibuprofen on nociceptive behavior, preening and motor activity following tonic chemical pain in the Japanese quail (Coturnix japonica)



      To establish a tonic chemical model of pain in quail and evaluate the efficacy of opioid and non-steroidal anti-inflammatory drugs.

      Study design

      A randomized, blinded, experimental study design.


      A total of 120 male Japanese quail, aged 7 weeks.


      A formalin solution (0.3%, 0.6% and 0.9%; total volume of 40 μL) was injected subcutaneously (SC) into the medial aspect of the right metatarsus (shank). Foot lift and preening activities were recorded for 45 minutes following injection of formalin and scored by an investigator blinded to the treatment. An open field test was used to evaluate motor activity. Treatments included SC saline, SC morphine (1.25, 2.5 and 5 mg kg–1) and oral ibuprofen (5 and 10 mg kg–1). The treatment effect was analyzed by one-way anova and the time course effect analyzed using repeated measures anova, both followed by Dunnett’s post hoc test (p < 0.05).


      All formalin concentrations induced significant foot lifting activity in the first phase (0–5 minutes), whereas only 0.6% and 0.9% formalin elicited responses in both the first and second (25–45 minutes) phases. Neither morphine nor ibuprofen affected phase 1 of the formalin test. Morphine (2.5 and 5 mg kg–1) and ibuprofen (5 and 10 mg kg–1) significantly reduced foot lift responses. Preening activity was significantly decreased following injection of 0.6% and 0.9% formalin. Preening was normalized with ibuprofen, but not with morphine. Morphine, but not ibuprofen, reduced quail activity.

      Conclusion and clinical relevance

      These results suggest that the formalin test was a reliable method for assessing tonic pain behavior in quail. The acute phase of the formalin test was not affected by morphine or ibuprofen. Although ibuprofen reduced the pain response in phase 2, the analgesic effects of morphine were not conclusive because morphine appeared to induce sedation.


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        • Barrot M.
        Tests and models of nociception and pain in rodents.
        Neuroscience. 2012; 211: 39-50
        • Clayton D.H.
        • Cotgreave P.
        Relationship of bill morphology to grooming behaviour in birds.
        Anim Behav. 1994; 47: 195-201
        • Delius J.
        Preening and associated comfort behavior in birds.
        Ann N Y Acad Sci. 1988; 525: 40-55
        • Desmarchelier M.
        • Troncy E.
        • Beauchamp G.
        • et al.
        Evaluation of a fracture pain model in domestic pigeons (Columba livia).
        Am J Vet Res. 2012; 73: 353-360
        • Duncan I.J.
        • Slee G.S.
        • Seawright E.
        • Breward J.
        Behavioural consequences of partial beak amputation (beak trimming) in poultry.
        Br Poult Sci. 1989; 30: 479-488
        • Evrard H.C.
        • Balthazart J.
        The assessment of nociceptive and non-nociceptive skin sensitivity in the Japanese quail (Coturnix japonica).
        J Neurosci Methods. 2002; 116: 135-146
        • Gentle M.J.
        Cutaneous sensory afferents recorded from the nervus intramandibularis of Gallus gallus var domesticus.
        J Comp Physiol A. 1989; 164: 763-774
        • Gentle M.J.
        The acute effects of amputation on peripheral trigeminal afferents in Gallus gallus var domesticus.
        Pain. 1991; 46: 97-103
        • Gentle M.J.
        Sodium urate arthritis: effects on the sensory properties of articular afferents in the chicken.
        Pain. 1997; 70: 245-251
        • Gentle M.J.
        Pain-related behaviour following sodium urate arthritis is expressed in decerebrate chickens.
        Physiol Behav. 1997; 62: 581-584
        • Gentle M.J.
        Pain issues in poultry.
        Appl Anim Behav Sci. 2011; 135: 252-258
        • Gentle M.J.
        • Hunter L.N.
        Physiological and behavioural responses associated with feather removal in Gallus gallus var domesticus.
        Res Vet Sci. 1991; 50: 95-101
        • Gunaydin C.
        • Bilge S.S.
        Effects of nonsteroidal anti-inflammatory drugs at the molecular level.
        Eurasian J Med. 2018; 50: 116-121
        • Guzman D.S.
        • Beaufrère H.
        Avian pain management and anesthesia.
        in: Graham J.E. Doss G.A. Beaufrère H. Exotic Animal Emergency and Critical Care Medicine. John Wiley & Sons Inc., USA2021: 488-502
        • Hawkins M.G.
        • Paul-Murphy J.
        Avian analgesia.
        Vet Clin North Am Exot Anim Pract. 2011; 14: 61-80
        • Hocking P.M.
        • Robertson G.W.
        • Gentle M.J.
        Effects of non-steroidal anti-inflammatory drugs on pain-related behaviour in a model of articular pain in the domestic fowl.
        Res Vet Sci. 2005; 78: 69-75
        • Hoppes S.
        • Flammer K.
        • Hoersch K.
        • et al.
        Disposition and analgesic effects of fentanyl in white cockatoos (Cacatua alba).
        J Avian Med Surg. 2003; 17: 124-130
        • Hughes R.A.
        Strain-dependent morphine-induced analgesic and hyperalgesic effects on thermal nociception in domestic fowl (Gallus gallus).
        Behav Neurosci. 1990; 104: 619-624
        • Hughes R.A.
        • Sufka K.J.
        Morphine hyperalgesic effects on the formalin test in domestic fowl (Gallus gallus).
        Pharmacol Biochem Behav. 1991; 38: 247-251
        • Jones R.B.
        Avian open-field research and related effects of environmental novelty: an annotated bibliography, 1960–1988.
        Psychol Rec. 1989; 39: 397-420
        • Khalilzadeh E.
        • Saiah G.V.
        • Hasannejad H.
        • et al.
        Antinociceptive effects, acute toxicity and chemical composition of Vitex agnus-castus essential oil.
        Avicenna J Phytomedicine. 2015; 5: 218-230
        • Landoni M.F.
        • Lees P.
        Comparison of the anti-inflammatory actions of flunixin and ketoprofen in horses applying PK/PD modelling.
        Equine Vet J. 1995; 27: 247-256
        • Lees P.
        • Landoni M.F.
        • Giraudel J.
        • Toutain P.L.
        Pharmacodynamics and pharmacokinetics of nonsteroidal anti-inflammatory drugs in species of veterinary interest.
        J Vet Pharmacol Ther. 2004; 27: 479-490
        • Lierz M.
        • Korbel R.
        Anesthesia and analgesia in birds.
        J Exot Pet Med. 2012; 21: 44-58
        • Machin K.L.
        Controlling avian pain.
        Compend Cont Educ Pract Vet. 2005; 27: 299-307
        • McNamara C.R.
        • Mandel-Brehm J.
        • Bautista D.M.
        • et al.
        TRPA1 mediates formalin-induced pain.
        Proc Natl Acad Sci USA. 2007; 104: 13525-13530
        • Nilius B.
        • Owsianik G.
        The transient receptor potential family of ion channels.
        Genome Biol. 2011; 12: 218
        • Omote K.
        • Kawamata T.
        • Kawamata M.
        • Namiki A.
        Formalin-induced release of excitatory amino acids in the skin of the rat hindpaw.
        Brain Res. 1998; 787: 161-164
        • Owens J.G.
        • Kamerling S.G.
        • Barker S.A.
        Pharmacokinetics of ketoprofen in healthy horses and horses with acute synovitis.
        J Vet Pharmacol Ther. 1995; 18: 187-195
        • Palocz O.
        • Gal J.
        • Csiko G.
        Application of non-steroidal anti-inflammatory drugs in birds: literature review.
        Magyar Állatorvosok Lapja. 2015; 137: 671-678
        • Paul-Murphy J.R.
        • Brunson D.B.
        • Miletic V.
        A technique for evaluating analgesia in conscious perching birds.
        Am J Vet Res. 1999; 60: 1213-1217
        • Pinkernell S.
        • Becker K.
        • Lindauer U.
        Severity assessment and scoring for neurosurgical models in rodents.
        Lab Anim. 2016; 50: 442-452
        • Roder J.D.
        • Chen C.L.
        • Chen H.
        • Sangiah S.
        Bioavailability and pharmacokinetics of ibuprofen in the broiler chicken.
        J Vet Pharmacol Ther. 1996; 19: 200-204
        • Shafi M.
        • Shabir S.
        Immunosuppressive effect of ibuprofen in Japanese quails.
        J Entomol Zool Stud. 2019; 7: 253-258
        • Shields S.D.
        • Cavanaugh D.J.
        • Lee H.
        • et al.
        Pain behavior in the formalin test persists after ablation of the great majority of C-fiber nociceptors.
        Pain. 2010; 151: 422-429
        • Siegel H.S.
        Physiological stress in birds.
        Bioscience. 1980; 30: 529-534
        • Singh P.M.
        • Johnson C.B.
        • Gartrell B.
        • et al.
        Analgesic effects of morphine and butorphanol in broiler chickens.
        Vet Anaesth Analg. 2017; 44: 538-545
        • Vermeulen B.
        • Remon J.P.
        The oral bioavailability of ibuprofen enantiomers in broiler chickens.
        J Vet Pharmacol Ther. 2001; 24: 105-109
        • Woolley S.C.
        • Gentle M.J.
        Physiological and behavioural responses in the hen (Gallus domesticus) to nociceptive stimulation.
        Comp Biochem Physiol A Comp Physiol. 1987; 88: 27-31
        • Zimmermann M.
        Ethical guidelines for investigations of experimental pain in conscious animals.
        Pain. 1983; 16: 109-110