The development of POI is a major complication in horses after colic surgery.1 A factor that may lead to the development of POI is a local inflammatory reaction in the gastrointestinal tract. Mechanical manipulation can induce various degrees of intestinal inflammation in laboratory animals, pigs, horses, and humans.2–7
Local effects of mechanical manipulation of intestines include an inflammatory response in the muscularis externa of the small intestine after surgical manipulation in rodents.2 This results in progressive neutrophil infiltration that peaks on postoperative day 1 and a decrease in circular muscle contractility in vitro.2 Mechanical manipulations can also affect intestine that has not been directly manipulated. Surgical manipulation of jejunum in rodents results in panenteritis.3 Such animals have a decrease in jejunal contractions and gastric emptying time and an increase in gastrointestinal transit time; in addition, colonic circular muscle contractility is decreased in vitro.3 The effects of laparotomy and traditional surgical techniques in pigs have been compared; performance of gastrectomy with intestinal handling results in more severe inflammation than laparoscopically assisted gastrectomy without intestinal manipulation.4 In human patients, traditional gynecologic surgical procedures cause a higher amount of mast cell activation in peritoneal fluid in contrast to minimally invasive surgical procedures.5
In horses, inflammation rapidly develops in colonic mucosa and serosa after direct mechanical manipulation or jejunal ischemia and reperfusion.8 Neutrophil and eosinophil granulocytes are involved in the inflammatory response for both of those experimental conditions.8 Neutrophil granulocytes are inflammatory cells located predominantly in the blood circulation that are recruited to tissues in response to various stimuli. Neutrophil granulocytes are the predominant inflammatory cell type associated with intestinal inflammation after ischemia and reperfusion.9–14 In horses, neutrophil infiltration develops after ischemia and reperfusion injury in the small intestine15 and large colon16 and after distension of the small colon.17 In another study,15 neutrophil infiltration of jejunal serosa and, less severely, of circular muscle developed after mechanical emptying of the jejunum in preparation for experimental induction of ischemia and reperfusion in horses.
Eosinophil granulocytes are part of the resident cell population of equine intestinal submucosa and mucosa.18 Results of other studies indicate mucosal eosinophil granulocytes in horses respond to various stimuli such as parasite infection19,20 and mucosal injury in vitro after mechanical manipulation of the colon.21 Eosinophils in the colon also respond after experimental induction of jejunal ischemia and reperfusion without prior manipulation of the colon.8 Eosinophil granulocytes are capable of initiating and maintaining a local inflammatory reaction.22 Therefore, characteristics of eosinophil and neutrophil granulocyte responses are of interest for improving understanding of the pathogenesis and clinical consequences of intestinal inflammation in horses.
The objective of the study reported here was to determine characteristics of the local jejunal inflammatory reaction in horses after performance of mechanical manipulations, similar to those used during colic surgery. In particular, the objective was to determine the distribution and numbers of neutrophil and eosinophil granulocytes in jejunum after such manipulations. We hypothesized that mechanical manipulations would lead to a measurable local inflammatory reaction in each intestinal wall layer.
Equest Vétoquinol GmbH, Ravensburg, Germany.
Xylapan (Xylazine), CP-Pharma Handelsgesellschaft mbH, Burgdorf, Germany.
Diazepam AbZ, AbZ Pharma GmbH, Blaubeuren, Germany.
Narketan, Vétoquinol GmbH, Ravensburg, Germany.
Isofluran CP, CP-Pharma Handelsgesellschaft mbH, Burgdorf, Germany.
Kardiocap 5 Monitor, Datex-Ohmeda GmbH, Duisburg, Germany.
Dobutamine Ratiopharm, Ratiopharm GmbH, Ulm, Germany.
Ringer's solution, B. Braun Melsungen AG, Melsungen, Germany.
Hemohes, B. Braun Melsungen AG, Melsungen, Germany.
Euthadorm, CP-Pharma Handelsgesellschaft mbH, Burgdorf, Germany
Liquid DAB, Firma BioGenex, San Ramon, Calif.
Anti-calprotectin, clone MAC387, monoclonal, mouse IgG1, DCS Diagnostik Systeme, Hamburg, Germany.
Multilink antibody, Biologo, Steindamm, Germany.
Super-Sensitive IM-Link-Label-Detection, Firma BioGenex, San Ramon, Calif.
Image Pro Express Media Cybernetics Inc. Bethesda, Md.
SAS, version 9.2, SAS Institute Inc, Cary, NC.
Rathmanner M, Staszyk C, Rötting AK, et al. Comparison of the inflammatory reaction in horses with and without development of post operative ileus (POI)(abstr), in Proceedings. 20th Annu Sci Meet Eur Coll Vet Surg 2011;57.
Vente C. Intestinale Entzündungsreaktionen während Ischämie und Reperfusion im Jejunum des Pferdes unter besonderer Berücksichtigung der eosinophilen und neutrophilen granulozyten. Dr med vet dissertation, Tierarztliche Hochschule Hannover, Germany, 2011.
1. Freeman DE, Hammock P & Baker GJ, et al. Short- and long-term survival and prevalence of postoperative ileus after small intestinal surgery in the horse. Equine Vet J 2000; 32:42–51.
2. Kalff JC, Schraut WH & Simmons RL, et al. Surgical manipulation of the gut elicits an intestinal muscularis inflammatory response resulting in postsurgical ileus. Ann Surg 1998; 5:652–663.
3. Schwarz NT, Kalff JC & Türler A, et al. Selective jejunal manipulation causes postoperative pan-enteric inflammation and dysmotility. Gastroenterology 2004; 126:159–169.
4. Hiki N, Simizu N & Yamaguchi H, et al. Manipulation of the small intestine as a cause of the increased inflammatory response after open compared with laparoscopic surgery. Br J Surg 2006; 93:195–204.
5. The FO, Bennink RJ & Ankum WM, et al. Intestinal handling-induced mast cell activation in human postoperative ileus. Gut 2008; 57:33–40.
6. De Jonge WJ, The FO, Ankum WM, et al. Mast cell degranulation during abdominal surgery initiates postoperative ileus in mice. Gastroenterology 2004; 127:535–545.
7. Kalff JC, Turler A & Schwarz NT, et al. Intra-abdominal activation of a local inflammatory response within the human muscularis externa during laparotomy. Ann Surg 2003; 237:301–315.
8. Hopster-Iversen C, Hopster K, Staszyk C, et al. Influence of mechanical manipulations on the local inflammatory reaction in the equine colon. Equine Vet J Suppl 2011; 43:1–7.
9. Grisham MB, Granger DN. Neutrophil-mediated mucosal injury: role of reactive oxygen metabolites. Dig Dis Sci 1988; 33:6S–15S.
10. Grisham MB, Gaginella TS & von Ritter C, et al. Effects of neutrophil-derived oxidants on intestinal permeability, electrolyte transport and epithelial cell viability. Inflammation 1990; 14:531–542.
11. Schoenberg MH, Poch B, Younes M. et al. Involvement of neutrophils in postischaemic damage to the small intestine. Gut 1991; 32:905–912.
12. Kubes P, Hunter J, Granger DN. Ischemia/reperfusion-induced feline intestinal dysfunction: importance of granulocyte recruitment. Gastroenterology 1992; 103:807–812.
13. Moore RM, Bertone AL, Bailey MQ. Neutrophil accumulation in the large colon of horses during low-flow ischemia and reperfusion. Am J Vet Res 1994; 55:1454–1463.
14. Gayle J, Jones SL & Argenzio RA, et al. Neutrophils increase para-cellular permeability of restituted ischemic-injured porcine ileum. Surgery 2002; 132:461–470.
15. Little D, Tomlinson JE, Blikslager AT. Post operative neutrophilic inflammation in equine small intestine after manipulation and ischaemia. Equine Vet J 2005; 37:329–335.
16. Grosche A, Morton AJ & Polyak MMR, et al. Detection of calprotectin and its correlation to the accumulation of neutrophils within equine large colon during ischaemia and reperfusion. Equine Vet J 2008; 40:393–399.
17. Faleiros RR, Macoris DG & Alves GES, et al. Local and remote lesions in horses subjected to small colon distension and decompression. Can J Vet Res 2008; 72:68–76.
18. Rötting AK, Freeman DE, Constable PD, et al. Mucosal distribution of eosinophilic granulocytes within the gastrointestinal tract of horses. Am J Vet Res 2008; 69:874–879.
19. Collobert-Laugier C, Hoste H, Sevin C, et al. Mast cell and eosinophil mucosal responses in the large intestine of horses naturally infected with cyathostomes. Vet Parasitol 2002; 107:251–264.
20. Rötting AK, Freeman DE, Constable PD, et al. The effects of strongylus vulgaris parasitism on eosinophil distribution and accumulation in equine large intestinal mucosa. Equine Vet J 2008; 40:379–384.
21. Rötting AK, Freeman D, Eurell JAC, et al. Effects of acetylcystein and migration of resident eosinophils in an in vitro model of mucosal injury and restitution in equine right dorsal colon. Am J Vet Res 2005; 64:1205–1212.
22. Hogan SP, Rosenberg HF & Moqbel R, et al. Eosinophils: biological properties and role in health and disease. Clin Exp Allergy 2008; 38:709–750.
23. Luna LG. Histopathologic methods and color atlas of special stains and tissue artifacts. Gaithersburg, Md: American Histolab Inc, 1992;255–256.
24. Roussel AJ Jr, Cohen ND, Hooper RN, et al. Risk factors associated with development of postoperative ileus in horses. J Am Vet Med Assoc 2001; 219:72–78.
25. Gerard MP, Blikslager AT & Roberts MC, et al. The characteristics of intestinal injury peripheral to strangulating obstruction lesions in the equine small intestine. Equine Vet J 1999; 31:331–335.
28. Barquist E, Bonaz B & Martinez V, et al. Neuronal pathways involved in abdominal surgery-induced gastric ileus in rats. Am J Physiol 1996; 270:R888–R894.
29. Wehner S, Vilz TO & Stoffels B, et al. Immune mediators of postoperative ileus. Langenbecks Arch Surg 2012; 397:591–601.
32. Köninger J, Gutt CN, Wente MN, et al. Postoperative ileus. Pathophysiology and prevention [in German]. Chirurg 2006; 77:904–912.
33. Shippers E, Tittel A & Öttinger A, et al. Laparoscopy versus laparotomy: comparison of adhesion-formation after bowel resection in a canine model. Dig Surg 1998; 15:145–147.
34. Gorvy DA, Edwards GB, Proudman CJ. Intra-abdomianal adhesions in horses: a retrospective evaluation of repeat laparotomy in 99 horses with acute gastrointestinal disease. Vet J 2008; 175:194–201.
35. Cook VL, Shults JJ & McDowell MR, et al. Anti-inflammatory effects of intravenously administered lidocaine hydrochloride on ischemia-injured jejunum in horses. Am J Vet Res 2009; 70:1259–1268.