Banding or Burdizzo castration and carprofen administration on peripheral leukocyte inflammatory cytokine transcripts W.Y. Pang a,b , B. Earley a, * , M. Murray a , T. Sweeney b , V. Gath b , M.A. Crowe b a Animal and Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Dunsany, Co. Meath, Ireland b School of Agriculture, Food Science and Veterinary Medicine, and the Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland article info Article history: Received 28 January 2010 Accepted 26 April 2010 Keywords: Castration Cattle Peripheral blood leukocytes Transcripts abstract The objective was to investigate if Banding or Burdizzo castration of bulls would alter the gene expression profile of a range of peripheral leukocyte inflammatory cytokines (IL-1, IL-6, IL-8, IL-10, interferon-c and tumor necrosis factor-a) and to determine if the administration of carprofen (C) before castration would affect the expression of these genes. Thirty Holstein–Friesian bulls (5.5 months; Mean 191 ± (SEM) 3.7 kg) were blocked by weight and randomly assigned to one of five treatments: (1) untreated control (CON); (2) Banding castration at 0 min (BAND); (3) BAND following an i.v. injection of 1.4 mg/kg BW of carprofen (C) at À20 min (BAND + C); (4) Burdizzo castration at 0 min (BURD); or (5) BURD following 1.4 mg/kg BW of carprofen at À20 min (BURD + C). Blood samples were collected at 1 h before castration and 6, 24 and 48 h post-castration for routine hematology and quantitative real-time PCR analysis of cytokine gene expression analysis. Generally, there were no differences (P > 0.05) among treatment groups in hemato- logical variables following castration. Cortisol concentrations were unchanged throughout the experi- mental period in CON bulls. BURD animals had greater cortisol concentrations than BAND and CON animals at 6 h post treatment. Transitory effects were observed only in the expression of IL-6 and TNF- a. The relative expression of IL-6 was greater in the BURD than in the BAND treatment (P < 0.05) at 24 h post-castration and was greater in the BURD + C group than in the BURD group (P < 0.05) at 48 h. The relative expression of TNF-a was greater in BAND than in the BURD group (P < 0.05) at 48 h. In con- clusion, these findings indicate that Banding or Burdizzo castration did not have any major effect on peripheral leukocyte inflammatory cytokine gene expression; Banding castration caused a greater pro- inflammatory cytokine gene expression reaction than Burdizzo castration and carprofen administration can affect IL-6 gene expression levels in BURD castrated animals. Ó 2010 Elsevier Ltd. All rights reserved. 1. Introduction The application of rubber rings or tightened latex bands (re- ferred to as Banding) (Fell et al., 1986; Chase et al., 1995), and use of a Burdizzo instrument to crush the testicular cords (Robert- son et al., 1994) are two techniques that are used to castrate male cattle. Castration by these methods has been shown to elicit phys- iological stress (indicated by an acute cortisol rise in plasma), inflammatory reactions (indicated by acute phase proteins), pain- associated behaviour, suppression of immune function (indicated by in vitro IFN-c production), and a reduction in performance to varying degrees (Molony et al., 1995; Pang et al., 2006). Acute inflammation is initiated and sustained by the participa- tion of many cytokines including tumor necrosis factor a (TNF-a), interleukin-1 (IL-1), IL-6, and interferon-gamma (IFN-c)(Cheville, 1999). Cytokines released by inflammatory leukocytes induce hepatocytes to synthesize and release acute phase proteins (APP’s) (Cheville, 1999). Early signs of clinical inflammation are associated with the appearance of APP’s in blood and tissue fluids that sustain the inflammatory process. APP’s are not normally present in plas- ma or are present in very low amounts and increase markedly immediately after injury and are an indication of tissue injury and inflammation (Cheville, 1999). The acute phase response is the reaction of the animal to disturbances in its homeostasis. It is thought to be beneficial to the injured animals in acting to restore homeostasis and prevent bacterial growth. In later stages of inflammation, cytokines such as IL-10 are produced by infiltrating leukocytes that limit inflammation and counteract hyperalgesia (Cunha and Ferreira, 2003). IL-8 was identified as an agonist for neutrophils (Baggiolini et al., 1994, 1999) and neutrophils are in- volved in many stress models (Murata, 1997; Barton et al., 2000; Weber et al., 2001; Ting et al., 2004). Eicher et al. (2004) reported that the expression of a range of acute phase cytokine genes including IL-1b, IL-1Ra and TNF-a can be altered in dairy calves treated with dexamethasone from days 3 to 56 of age, which 0034-5288/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.rvsc.2010.04.023 * Corresponding author. Tel.: +353 46 9061166; fax: +353 46 9026154. E-mail address: bernadette.earley@teagasc.ie (B. Earley). Research in Veterinary Science 90 (2011) 127–132 Contents lists available at ScienceDirect Research in Veterinary Science journal homepage: www.elsevier.com/locate/rvsc