We describe a novel surgical method for catheterizing 'intestinal loops' within the ileum of sheep. Once animals have recovered from surgery and have cleared antibiotics and analgesics, multiple treatments can be deposited directly in loops via the catheters.
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Method Article
Catheterization of Intestinal Loops in Ruminants
In This Article
Summary
Abstract
The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the outside world. As such, the intestine plays a pivotal role in immunosurveillance and protection from enteric pathogens. Investigating intestinal physiology and immunology commonly employs 'intestinal loops' as an experimental model. The majority of these loop models are non-recovery surgical procedures that study short-term (<24 hr) changes in the intestine (1-3). We previously created a recovery intestinal loop model to specifically measure long-term (<6 mo) immunological changes in the intestine of sheep following exposure to vaccines, adjuvants, and viruses (4). This procedure localized treatments to a specific 'loop', allowing us to sample this area of the intestine. A significant drawback of this method is the single window of opportunity to administer treatments (i.e. at the time of surgery). Furthermore, samples of both the intestinal mucosa and luminal contents can only be taken at the termination of the project. Other salient limitations of the above model are that the surgical manipulation and requisite post-operative measures (e.g. administration of antibiotics and analgesics) can directly affect the treatment itself and/or alter immune function, thereby confounding results. Therefore, we modified our intestinal loop model by inserting long-term catheters into the loops. Sheep recover fully from the procedure, and are unaffected by the exteriorized catheters. Notably, the establishment of catheters in loops allows us to introduce multiple treatments over an extended interval, following recovery from surgery and clearance of drugs administered during surgery and the post-operative period.
Protocol
Experimental procedure:
- All instruments are autoclaved; non-autoclaveable materials are sterilized in Germex cold sterilization solution for a minimum of 10 min.
- Sheep are pre-medicated with acepromazine (0.05 mg/kg), glycopyrrolate (0.005 mg/kg), and butorphanol tartrate (0.2 mg/kg) IM, and left for 15-20 min.
- The neck is shaved and the skin surgically scrubbed. Once the jugular vein is visualized, 0.1 ml of lidocaine is injected SC over the vein, the catheter is introduced into the vein and secured to the skin using livestock identification tag cement and 2" hospital tape. During surgery, the animals are given 10 ml/kg/hr of Plasmalyte 1....
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Discussion
In this procedure, we were able to successfully establish catheters in intestinal loops established within the ileum of sheep. By adhering to stringent post-operative care, animals rapidly recovered from the procedure and resumed normal activity 5-7 days after surgery. We determined that the establishment of catheters within loops does not affect intestinal mucosal or immune function (data not shown) and animals can live for at least 8 wk after surgery. This procedure allows us to introduce multiple treatments into disti.......
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Disclosures
All procedures on experimental animals were approved by the Institutional Animal Care Committee (Animal Use Protocol #0609) and followed the guidelines set by the Canadian Council of Animal Care.
We have nothing to disclose.
Acknowledgements
The research could not have been completed without the dedication and efforts of many persons at the Agriculture and Agri-Food Canada Research Centre, Lethbridge, Alberta, Canada. In that regard, we thank: Tara Shelton for pre- and post-operative care of animals, surgical preparation, and anesthesiology, analysis of samples, and assistance with video imaging; Randy Wilde and Estela Costa for pre- and post-operative care; Jenny Gusse for sample analysis; Byron Lee for the trocar image; and Brian Egland for sheep husbandry. We also thank Brent Selinger (Department of Biological Sciences, University of Lethbridge) for providing video equipment and advice, as well as othe....
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Surgical gloves | Western Drug Distribution Center, Ltd. | 103091 | |
| Surgical mask | Western Drug Distribution Center, Ltd. | MAX53013 | |
| Surgical gown | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 3365 | |
| OB Apron | Western Drug Distribution Center, Ltd. | 900062 | |
| Surgical cap | Western Drug Distribution Center, Ltd. | 903271 | |
| EZ Scrub brush | Western Drug Distribution Center, Ltd. | BDC377479 | |
| Anesthetic machine | Boyle | - | |
| Sodasorb | Western Drug Distribution Center, Ltd. | 108638 | |
| Laryngoscope | Western Drug Distribution Center, Ltd. | - | |
| Laryngoscope blade (Miller blade #4) | Western Drug Distribution Center, Ltd. | 902667 | |
| Endotracheal tube (7.0 - 8.5 mm) | Western Drug Distribution Center, Ltd. | - | |
| Lidocaine endotracheal spray | Western Drug Distribution Center, Ltd. | 103364 | |
| Acepromazine | Western Drug Distribution Center, Ltd. | AUS01020 | |
| Glycopyrrolate | Western Drug Distribution Center, Ltd. | SAB2880 | |
| Butorphanol tartrate | Western Drug Distribution Center, Ltd. | AYE304541 | |
| Diazepam | Western Drug Distribution Center, Ltd. | sab790 | |
| Thiopental sodium | Western Drug Distribution Center, Ltd. | 103508 | |
| Epinephrine hydrochloride | Western Drug Distribution Center, Ltd. | 103856 | |
| Doxapram | Strathcona Pharmacy | - | |
| Isoflurane | Western Drug Distribution Center, Ltd. | 100339 | |
| Hemostam | Western Drug Distribution Center, Ltd. | 103857 | |
| Flunixin | Western Drug Distribution Center, Ltd. | sch140100 | |
| Enrofloxacin | Western Drug Distribution Center, Ltd. | hll0049 | |
| Metronidazole | Western Drug Distribution Center, Ltd. | 101653 | |
| Ampicillin | Western Drug Distribution Center, Ltd. | NOV67185-6 | |
| Cefazolin | Western Drug Distribution Center, Ltd. | nov69397 | |
| Electroscapel | Macan | MV8 | |
| Heating mat | Kane | PHMD-48 | |
| Stomach tube (Nalgene tubing 5/16" ID) | Fisher Scientific | - | |
| Needle driver | Western Drug Distribution Center, Ltd. | 108338 | |
| Addison brown forcep | Western Drug Distribution Center, Ltd. | 911075 | |
| Rat tooth forcep | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 5275 | |
| Kelly forcep | Western Drug Distribution Center, Ltd. | 901133 | |
| Mosquito hemostat | Western Drug Distribution Center, Ltd. | 108471 | |
| Carmalt forcep | Western Drug Distribution Center, Ltd. | 901225 | |
| Scalpel handle | Western Drug Distribution Center, Ltd. | 900285 | |
| Scalpel blade #22 | Western Drug Distribution Center, Ltd. | 900511 | |
| Metzenbaum scissor | Western Drug Distribution Center, Ltd. | 901166 | |
| Sharp-sharp scissor | Western Drug Distribution Center, Ltd. | 108264 | |
| Towel clamp | Western Drug Distribution Center, Ltd. | 901213 | |
| Allis tissue forcep | Western Drug Distribution Center, Ltd. | 901201 | |
| Doyen’s forcep | Western Drug Distribution Center, Ltd. | 904226 | |
| Stainless steel bowl | Western Drug Distribution Center, Ltd. | 903601 | |
| Stainless steel trocar (curved) | Local Supplier | - | |
| Catheter introducer | Western Drug Distribution Center, Ltd. | 11440 | |
| Surgical towel | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 7972 | |
| Disposable drape | Western Drug Distribution Center, Ltd. | 900035 | |
| Steri drape | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 2323 | |
| Drape (40 x 40 cm) | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 8358 | |
| Surgical drape (7" x 11 ¼" opening) | The Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE | 2694 | |
| Vicryl (2-0) | Western Drug Distribution Center, Ltd. | JJMJ333H | |
| Silk 2-0 | Western Drug Distribution Center, Ltd. | 105248 | |
| Catgut #3 | Western Drug Distribution Center, Ltd. | CSL547-2070 | |
| Supramid #1 | Western Drug Distribution Center, Ltd. | CSL547-3040 | |
| Needles (cutting and non-cutting) | Western Drug Distribution Center, Ltd. | - | |
| Silastic tubing (0.040" ID, 0.085" OD) | Fisher Scientific | 11-189-15D | |
| Orbeseal ointment | Western Drug Distribution Center, Ltd. | 106344 | |
| Clear silicone (732 multi-purpose sealant) | Local Supplier | - | |
| Blunt end needle | Western Drug Distribution Center, Ltd. | 905035 | |
| Marker (permanent industrial) | Local Supplier | - | |
| Catheter pouch (constructed of Elastoplast 7.5 cm) | Western Drug Distribution Center, Ltd. | 104094 | |
| Hospital tape (2") | Western Drug Distribution Center, Ltd. | 103842 | |
| Hospital tape (1") | Western Drug Distribution Center, Ltd. | VLMRENF20 | |
| Gauze sponge 4x4 | Western Drug Distribution Center, Ltd. | 107551 | |
| Gauze cling | Western Drug Distribution Center, Ltd. | 107633 | |
| Elastoplast 7.5cm | Western Drug Distribution Center, Ltd. | 104094 | |
| Elastoplast rippable bandage | Western Drug Distribution Center, Ltd. | 104093 | |
| Syringe (60 ml) | Western Drug Distribution Center, Ltd. | 104004 | |
| Syringe (10 ml) | Western Drug Distribution Center, Ltd. | bdc301604 | |
| 25G Needle | Western Drug Distribution Center, Ltd. | BDC305122 | |
| 20G Needle | Western Drug Distribution Center, Ltd. | BDC305175 | |
| 18G Needle | Western Drug Distribution Center, Ltd. | 903353 | |
| Intracath IV catheter (16G 8") | Western Drug Distribution Center, Ltd. | bdc3831621 | |
| IV administration set (10 drop) | Western Drug Distribution Center, Ltd. | 101797 | |
| 0.9% NaCl solution | Western Drug Distribution Center, Ltd. | BAXJB1322P | |
| Plasmalyte 148 solution | Western Drug Distribution Center, Ltd. | BAXJB2534 | |
| Dextrose | Western Drug Distribution Center, Ltd. | MCT60190 | |
| Peristaltic pump | Cole-Parmer | 7524-10 | |
| Livestock identification tag cement | Local Supplier | - | |
| Germex solution | Western Drug Distribution Center, Ltd. | mtc60270 | |
| Isopropyl alcohol | Western Drug Distribution Center, Ltd. | 104590 | |
| Stanhexidine cleanser | Western Drug Distribution Center, Ltd. | 109248 | |
| Prepodyne solution | Western Drug Distribution Center, Ltd. | 105710 | |
| Prepodyne scrub | Western Drug Distribution Center, Ltd. | 105711 |
References
- Vidal, J. E., McClane, B. A., Saputo, J., Parker, J., Uzal, F. A. Effects of Clostridium perfringens beta-toxin on the rabbit small intestine and colon. Infect. Immun. 76, 4396-4404 (2008).
- Santos, R. L., Zhang, S., Tsolis, R. M., Baumler, A. J., Adams, L. G.
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