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 other assistance (i.e. executive production), and the Curriculum Re-Development Centre, University of Lethbridge for videography expertise. This study was supported in part by grants from the Advanced Foods and Materials Network (G.D.I.), the Canada-Alberta Beef Industry Development Fund (G.D.I.), and Agriculture and Agri-Food Canada (G.D.I. and J.P.K.).

Experimental procedure:
<ol>
<li>All instruments are autoclaved; non-autoclaveable materials are sterilized in Germex cold sterilization solution for a minimum of 10 min.</li>
<li>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.</li>
<li>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...

<ol>
	<li>Vidal, J. E., McClane, B. A., Saputo, J., Parker, J., Uzal, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Clostridium+perfringens+beta-toxin+on+the+rabbit+small+intestine+and+colon.">Effects of Clostridium perfringens beta-toxin on the rabbit small intestine and colon.</a> Infect. Immun. 76, 4396-4404 (2008).</li><li>Santos, R. L., Zhang, S., Tsolis, R. M., Baumler, A. J., Adams, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphologic+and+molecular+characterization+of+Salmonella+typhimurium+infection+in+neonatal+calves.">Morphologic and molecular characterization of Salmonella typhimurium infection in neonatal calves.</a> Vet. Pathol. 39, 200-215 (2002).</li><li>Uzzau, S., Leori, G. S., Petruzzi, V., Watson, P. R., Schianchi, G., Bacciu, D., Mazzarello, V., Wallis, T. S., Rubino, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salmonella+enterica+serovar-host+specificity+does+not+correlate+with+the+magnitude+of+intestinal+invasion+in+sheep.">Salmonella enterica serovar-host specificity does not correlate with the magnitude of intestinal invasion in sheep.</a> Infect. Immun. 69, 3092-3099 (2001).</li><li>Gerdts, V., Uwiera, R. R. E., Mutwiri, G. K., Wilson, D. J., Bowersock, T., Kindane, A., Babiuk, L. A., Griebel, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+intestinal+'loops'+provide+an+in+vivo+model+to+analyse+multiple+mucosal+immune+responses.">Multiple intestinal 'loops' provide an in vivo model to analyse multiple mucosal immune responses.</a> J. Immunol. Methods. 256, 19-33 (2001).</li><li>Greibel, P. J., Davis, W. C., Reynolds, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+analysis+of+the+growth+and+differentiation+of+b+cells+isolated+from+follicles+from+ileal+Peyer's+patch+of+sheep.">An analysis of the growth and differentiation of b cells isolated from follicles from ileal Peyer's patch of sheep.</a> Immunology. 75, 601-607 (1992).</li><li>Motyka, G., Greibel, P. J., Reynolds, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Agents+that+activate+protein+kinase+C+resue+sheep+ileal+Peyer's+patch+B+cells+from+apoptosis.">Agents that activate protein kinase C resue sheep ileal Peyer's patch B cells from apoptosis.</a> Eur. J. Immunol. 23, 1314-1321 (1993).</li></ol>

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...

<table border="1">
<tr><td>Category / Item	Number</td><td>Company</td><td>Catalogue</td></tr> 
<tr><td colspan="3">Surgeon supplies</td></tr>
<tr><td>Surgical gloves</td><td>WDDCa</td><td>103091</td></tr>
<tr><td>Surgical mask</td><td>WDDC</td><td>MAX53013</td></tr>
<tr><td>Surgical gown</td><td>CDMVb</td><td>3365</td></tr>
<tr><td>OB Apron</td><td>WDDC</td><td>900062</td></tr>
<tr><td>Surgical cap</td><td>WDDC</td><td>903271</td></tr>
<tr><td>EZ Scrub brush</td><td>WDDC</td><td>BDC377479</td></tr>

<tr><td colspan="3">Anesthesia and drugs</td></tr>
<tr><td>Anesthetic machine</td><td>Boyle</td><td>&#8212;</td></tr>
<tr><td>Sodasorb</td><td>WDDC</td><td>108638</td></tr>
<tr><td>Laryngoscope</td><td>WDDC</td><td>&#8212;</td></tr>
<tr><td>Laryngoscope blade (Miller blade #4)</td><td>WDDC</td><td>902667</td></tr>
<tr><td>Endotracheal tube (7.0 - 8.5 mm)</td><td>WDDC</td><td>&#8212;</td></tr>
<tr><td>Lidocaine endotracheal spray</td><td>WDDC</td><td>103364</td></tr>
<tr><td>Acepromazine</td><td>WDDC</td><td>AUS01020</td></tr>
<tr><td>Glycopyrrolate</td><td>WDDC</td><td>SAB2880</td></tr>
<tr><td>Butorphanol tartrate</td><td>WDDC</td><td>AYE304541</td></tr>
<tr><td>Diazepam</td><td>WDDC</td><td>sab790</td></tr>
<tr><td>Thiopental sodium</td><td>WDDC</td><td>103508</td></tr>
<tr><td>Epinephrine hydrochloride</td><td>WDDC</td><td>103856</td></tr>
<tr><td>Doxapram</td><td>Strathcona Pharmacy</td><td>&#8212;</td></tr>
<tr><td>Isoflurane</td><td>WDDC</td><td>100339</td></tr>
<tr><td>Hemostam</td><td>WDDC</td><td>103857</td></tr>
<tr><td>Flunixin</td><td>WDDC</td><td>sch140100</td></tr>
<tr><td>Enrofloxacin</td><td>WDDC</td><td>hll0049</td></tr>
<tr><td>Metronidazole</td><td>WDDC</td><td>101653</td></tr>
<tr><td>Ampicillin</td><td>WDDC</td><td>NOV67185-6</td></tr>
<tr><td>Cefazolin</td><td>WDDC</td><td>nov69397</td></tr>

<tr><td colspan="3">Surgical equipment and supplies</td></tr>		
<tr><td>Electroscapel</td><td>Macan</td><td>MV8</td></tr>
<tr><td>Heating mat</td><td>Kane</td><td>PHMD-48</td></tr>
<tr><td>Stomach tube (Nalgene tubing 5/16" ID)</td><td>Fisher Scientific</td><td>&#8212;</td></tr>
<tr><td>Needle driver</td><td>WDDC</td><td>108338</td></tr>
<tr><td>Addison brown forcep</td><td>WDDC</td><td>911075</td></tr>
<tr><td>Rat tooth forcep</td><td>CDMV</td><td>5275</td></tr>
<tr><td>Kelly forcep</td><td>WDDC</td><td>901133</td></tr>
<tr><td>Mosquito hemostat</td><td>WDDC</td><td>108471</td></tr>
<tr><td>Carmalt forcep</td><td>WDDC</td><td>901225</td></tr>
<tr><td>Scalpel handle</td><td>WDDC</td><td>900285</td></tr>
<tr><td>Scalpel blade #22</td><td>WDDC</td><td>900511</td></tr>
<tr><td>Metzenbaum scissor</td><td>WDDC</td><td>901166</td></tr>
<tr><td>Sharp-sharp scissor</td><td>WDDC</td><td>108264</td></tr>
<tr><td>Towel clamp</td><td>WDDC</td><td>901213</td></tr>
<tr><td>Allis tissue forcep</td><td>WDDC</td><td>901201</td></tr>
<tr><td>Doyen's forcep</td><td>WDDC</td><td>904226</td></tr>
<tr><td>Stainless steel bowl</td><td>WDDC</td><td>903601</td></tr>	
<tr><td>Stainless steel trocar (curved)</td><td>Local supplier</td><td>&#8212;</td></tr>
<tr><td>Catheter introducer</td><td>WDDC</td><td>11440</td></tr>
<tr><td>Surgical towel</td><td>CDMV</td><td>7972</td></tr>
<tr><td>Disposable drape</td><td>WDDC</td><td>900035</td></tr>
<tr><td>Steri drape</td><td>CDMV</td><td>2323</td></tr>
<tr><td>Drape (40 x 40 cm)</td><td>CDMV</td><td>8358</td></tr>
<tr><td>Surgical drape (7" x 11 &#188;" opening)</td><td>CDMV</td><td>2694</td></tr>
<tr><td>Vicryl (2-0)</td><td>WDDC</td><td>JJMJ333H</td></tr>
<tr><td>Silk 2-0</td><td>WDDC</td><td>105248</td></tr>
<tr><td>Catgut #3</td><td>WDDC</td><td>CSL547-2070</td></tr>
<tr><td>Supramid #1</td><td>WDDC</td><td>CSL547-3040</td></tr>
<tr><td>Needles (cutting and non-cutting)</td><td>WDDC</td><td>&#8212;</td></tr>

<tr><td colspan="3">Catheter</td></tr>
<tr><td>Silastic tubing (0.040" ID, 0.085" OD)</td><td>Fisher Scientific</td><td>11-189-15D</td></tr>
<tr><td>Orbeseal ointment</td><td>WDDC</td><td>106344</td></tr>
<tr><td>Clear silicone (732 multi-purpose sealant)</td><td>Local supplier</td><td>&#8212;</td></tr>
<tr><td>Blunt end needle</td><td>WDDC</td><td>905035</td></tr>
<tr><td>Marker (permanent industrial)</td><td>Local supplier</td><td>&#8212;</td></tr>
<tr><td>Catheter pouch (constructed of Elastoplast 7.5 cm)</td><td>WDDC</td><td>104094</td></tr>

<tr><td colspan="3">Bandages</td></tr>
<tr><td>Hospital tape (2")</td><td>WDDC</td><td>103842</td></tr>
<tr><td>Hospital tape (1")</td><td>WDDC</td><td>VLMRENF20</td></tr>
<tr><td>Gauze sponge 4x4</td><td>WDDC</td><td>107551</td></tr>
<tr><td>Gauze cling</td><td>WDDC</td><td>107633</td></tr>
<tr><td>Elastoplast 7.5cm</td><td>WDDC</td><td>104094</td></tr>
<tr><td>Elastoplast rippable bandage</td><td>WDDC</td><td>104093</td></tr>
		
<tr><td colspan="3">Needles and syringes</td></tr>		
<tr><td>Syringe (60 ml)</td><td>WDDC</td><td>104004</td></tr>
<tr><td>Syringe (10 ml)</td><td>WDDC</td><td>bdc301604</td></tr>
<tr><td>25G Needle</td><td>WDDC</td><td>BDC305122</td></tr>
<tr><td>20G Needle</td><td>WDDC</td><td>BDC305175</td></tr>
<tr><td>18G Needle</td><td>WDDC</td><td>903353</td></tr>

<tr><td colspan="3">Intravenous</td></tr> 
<tr><td>Intracath IV catheter (16G 8")</td><td>WDDC</td><td>bdc3831621</td></tr>
<tr><td>IV administration set (10 drop)</td><td>WDDC</td><td>101797</td></tr>
<tr><td>0.9% NaCl solution</td><td>WDDC</td><td>BAXJB1322P</td></tr>
<tr><td>Plasmalyte 148 solution</td><td>WDDC</td><td>BAXJB2534</td></tr>
<tr><td>Dextrose</td><td>WDDC</td><td>MCT60190</td></tr>
<tr><td>Peristaltic pump</td><td>Cole Parmer</td><td>7524-10</td></tr>
<tr><td>Livestock identification tag cement</td><td>Local supplier</td><td>&#8212;</td></tr>

<tr><td colspan="3">Sanatizers / disinfectants</td></tr>		
<tr><td>Germex solution</td><td>WDDC</td><td>mtc60270</td></tr>
<tr><td>Isopropyl alcohol</td><td>WDDC</td><td>104590</td></tr>
<tr><td>Stanhexidine cleanser</td><td>WDDC</td><td>109248</td></tr>
<tr><td>Prepodyne solution</td><td>WDDC</td><td>105710</td></tr>
<tr><td>Prepodyne scrub</td><td>WDDC</td><td>105711</td></tr>
</table>
aWestern Drug Distribution Center LTD, Edmonton, AB.
bThe Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE.

catheterization of intestinal loops in ruminants

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 (&lt;24 hr) changes in the intestine (1-3). We previously created a recovery intestinal loop model to specifically measure long-term (&lt;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.

Watch this Scientific Journal Video about Catheterization of Intestinal Loops in Ruminants at JoVE.com

Catheterization of Intestinal Loops in Ruminants

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.

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 ...

catheterization-of-intestinal-loops-in-ruminants

Research

JoVE Journal

Biology

13.6K visualizações.  University of Alberta. Descrevemos um novo método cirúrgico para cateterizar "alças intestinais" dentro do íleo de ovelhas. Uma vez que os animais têm recuperado de uma cirurgia e que tenha limpado antibióticos e analgésicos, múltiplos tratamentos podem ser depositados diretamente em loops através dos cateteres.