Name: surgical placement of catheters for long-term cardiovascular exercise testing in swine
Uploaded: 2016-02-09T15:00:00
Description: Watch this Scientific Journal Video about Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine at JoVE.com

This study was supported by Netherlands Heart Foundation grant 2000T038 (to D.J. Duncker) grant 2000T042 (to D. Merkus), European Commission FP7-Health-2010 grant MEDIA-261409 (to D.J. Duncker and D. Merkus), Netherlands CardioVascular Research Initiative: the Dutch Heart Foundation, the Dutch Federation for University Medical Centers, the Netherlands Organisation for Health Research and Development and the Royal Netherlands Academy of Sciences &#160;CVON- ARENA CVON 2011-11 (to D.J. Duncker), CVON-PHAEDRA CVON2012-08 ( to D.&#160;Merkus) &#160;and CVON-RECONNECT CVON 2014-11 (to D.J. Duncker and D. Merkus), Sophia Foundation (to D. de Wijs-Meijler, D. Merkus and I.K.M. Reiss).

Procedures involving animal subjects have been approved by the Animal Care Committee at Erasmus Medical Center Rotterdam (NL). Swine with weights between 6 and 80 kg have been successfully instrumented using this protocol.
1. Adaptation of the Animals to Human Handling
<ol>
	<li>After arrival in the facility, house the animals solitarily but enable them to interact with each other.</li>
	<li>Accustomize swine to human handling and transportation from the animal facility to the experimental laboratory, by handlin...

<ol>
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The present study describes the surgery for chronical instrumentation of swine as well as the protocol for exercising the instrumented swine on a motor-driven treadmill while measuring hemodynamics and taking blood samples for measurement of oxygen content in arterial, mixed venous and coronary venous blood.
Critical Steps within the Protocol 
There are several critical steps within the protocol that start already during the intubation procedure. Thiopental (2.1.5) is a respiratory depres...

Adequate cardiopulmonary function is essential to supply the body with oxygen and nutrients, particularly during conditions of increased metabolic demand such as during exercise1. The cardiopulmonary response to exercise is characterized by a number of adaptations in cardiac function, i.e., an increase in heart rate, contractility and stroke volume, and microvascular function, i.e., vasodilation in the vascular beds supplying exercising muscles as well as in the pulmonary vasculature, and vasoconstriction in the vascular beds supplying the gastrointestinal system as well as inactive muscles1. Impaired exercise capacity is an early hallmark of cardiopulmonary dysfunction, and cardiopulmonary exercise testing is used as an effective method to delineate between cardiac dysfunction, vascular dysfunction and/ or pulmonary dysfunction in patients with impaired exercise capacity2. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia, with many anesthetic agents possessing cardiodepressive properties3.
Chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, and when the animals are fully adjusted to the laboratory conditions measurements can be obtained under quiet resting conditions without the effects of anesthesia and acute surgical trauma. Furthermore, when the animals are appropriately trained, measurements can also be obtained during graded treadmill exercise4,5. More specifically, left and right ventricular function can be assessed and related to myocardial perfusion, while regulation of vasomotor tone in the coronary, systemic and pulmonary microcirculation can be determined. The use of fluid-filled catheters allows measurement of pressure as well as taking blood samples without imposing additional stress on the animals. Another advantage of studying chronically instrumented animals is that cardiopulmonary exercise testing can be repeated allowing the use of an animal as its own control, either during disease development or during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.
Cardiopulmonary anatomy of swine closely resembles that of humans and it is possible to induce various forms of cardiopulmonary disease, such as diabetes 6, myocardial infarction 7, pulmonary hypertension 8,9 and pacing-induced heart failure10,11. Moreover, the size of swine allows chronic instrumentation, and repeated blood sampling of sufficient quantity to analyze not only blood gases, but also to perform neurohumoral measurements and/or to search for biomarkers of disease.
This protocol describes the surgery used to chronically instrument swine as well as the protocol for exercising the swine on a motor-driven treadmill.

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			<td height="21" style="height:21px;width:496px;">3-way stopcocks</td>
			<td style="width:212px;">B. Braun</td>
			<td align="right" style="width:179px;">16496</td>
			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;">Perfusor lines PVC (DEHP-free) 150cm/2.6ml&#160;</td>
			<td>B. Braun</td>
			<td align="right" style="width:179px;">8722960</td>
			<td>Used for fluid filled catheters</td>
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			<td height="21" style="height:21px;">&#8220;python &#8220; silicontubing</td>
			<td>Rubber BV</td>
			<td>1757 ID 1mm, OD 2mm</td>
			<td>Used for fluid filled catheters</td>
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			<td height="21" style="height:21px;width:496px;">Sodium Chloride 0.9%</td>
			<td style="width:212px;">Baxter</td>
			<td style="width:179px;">TKF7124</td>
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			<td height="21" style="height:21px;width:496px;">Suction device</td>
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			<td></td>
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			<td height="21" style="height:21px;">Slim-Line electrosurgical pencil with 2 buttons</td>
			<td>ERBE ELEKTROMEDIZIN GMBH</td>
			<td>20190-066</td>
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			<td>Vererinary Technics Int.</td>
			<td>11.02.47</td>
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			<td style="width:212px;">3M</td>
			<td style="width:179px;">1818FS</td>
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			<td height="21" style="height:21px;width:496px;">surgical hat</td>
			<td style="width:212px;">Klinidrape</td>
			<td align="right" style="width:179px;">621301</td>
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			<td>Molnlycke Health Care</td>
			<td align="right" style="width:179px;">97027809</td>
			<td>Surgical drape, gauze pads, syringes, beaker etc</td>
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			<td style="width:212px;">Alcon</td>
			<td style="width:179px;">288-28282-01</td>
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			<td>Meda Pharma BV</td>
			<td>RVG08939</td>
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			<td>Meda Pharma BV</td>
			<td>RVG01331</td>
			<td></td>
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			<td height="21" style="height:21px;width:496px;">Cuffed Endotracheal tube</td>
			<td style="width:212px;">Emdamed</td>
			<td style="width:179px;"></td>
			<td>size depends on animal size</td>
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			<td height="21" style="height:21px;width:496px;">Breathing filter Hyrdo therm 3HME</td>
			<td style="width:212px;">Intersurgical</td>
			<td align="right" style="width:179px;">1560000</td>
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			<td style="width:212px;">Kawe Germany</td>
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			<td height="21" style="height:21px;width:496px;">Manual resuscitator- Combibag</td>
			<td style="width:212px;">Weinmann</td>
			<td>6515-12-313-5596</td>
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			<td height="21" style="height:21px;width:496px;">Perivascular flow probe 3PS</td>
			<td style="width:212px;">Transonic</td>
			<td style="width:179px;"></td>
			<td>For coronary artery; Size 2.5-4 mm depending on animal size</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Confidence flow probe</td>
			<td style="width:212px;">Transonic</td>
			<td style="width:179px;"></td>
			<td>For aorta/pulmonary artery, 16-20 mm; size depends on animal size</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Venflon-Venisystem 20Gx 32 mm</td>
			<td style="width:212px;">BD</td>
			<td align="right" style="width:179px;">393224</td>
			<td>For coronary venous catheter</td>
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			<td height="21" style="height:21px;width:496px;">Blunt Needle 18G</td>
			<td style="width:212px;"></td>
			<td style="width:179px;"></td>
			<td>For coronary venous catheter</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Tygon Tubing</td>
			<td style="width:212px;">Rubber BV</td>
			<td>2802 ID 0.8mm (1/32&#8217;&#8217;), OD 2.4mm (3/32&#8217;&#8217;)</td>
			<td>For coronary venous catheter</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Suction Handle 17 cm 6 6/8 &#34; Coupland 18/8 martinit with tube connector</td>
			<td>KLS Martin Group</td>
			<td>18-575-24</td>
			<td></td>
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			<td height="21" style="height:21px;width:496px;">Scalple blade&#160;</td>
			<td style="width:212px;"></td>
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			<td></td>
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			<td height="21" style="height:21px;">Scalpel Handle 13.5 cm 5 3/8 &#34; Stainless Steel solid</td>
			<td>KLS Martin Group</td>
			<td>10-100-04</td>
			<td></td>
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			<td height="21" style="height:21px;">Vascular Forceps 20.2 cm 8 &#34; De Bakey Stainless Stee</td>
			<td>KLS Martin Group</td>
			<td>24-388-20</td>
			<td>&#177; 14 cm</td>
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			<td height="21" style="height:21px;">Dressing Forceps 17 cm 6 6/8 &#34; Cushing Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>12-189-17</td>
			<td>&#177; 18 cm</td>
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			<td height="21" style="height:21px;">halsted-musquito straight 12.5cm - 5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-3100-13</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">halsted-musquito curved 12.5cm - 5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-3101-12</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">Dissecting and Ligature Forceps 13 cm 5 1/8 &#34; Gemini Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>13-451-13</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">Dissecting and Ligature Forceps 18.5 cm 7 2/8 &#34; Schnidt Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>13-363-18</td>
			<td></td>
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			<td height="21" style="height:21px;">Rib Retractor Finochietto, Baby Aluminium -</td>
			<td>KLS Martin Group</td>
			<td>24-162-01</td>
			<td></td>
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			<td height="21" style="height:21px;">suture forceps Mayo-Hegar 3mm 18cm - 7&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-6050-18</td>
			<td></td>
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			<td height="21" style="height:21px;">Metchenbaum blunt curved 14,5cm - 5(3/4)&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-1311-14M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Retrector farabeuf 12cm - 4 (3/4)&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-4497-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Towel forceps schr&#228;del curved 9cm - 3,5&#34;</td>
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			<td>Rudolf Medical</td>
			<td>RU-1001-13</td>
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			<td height="21" style="height:21px;">Gauzes Cutisoft 10 x 10 cm 4-ply</td>
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			<td>BSN Medical</td>
			<td>45844-00</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:496px;">Flowmeter -CM2 / SF2 - 2gas (O2 and Air)</td>
			<td>UNO BV</td>
			<td align="right" style="width:179px;">180000008</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tec 7 Vaporizer</td>
			<td>Datex-Ohmeda</td>
			<td style="width:179px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Acederm wound spay</td>
			<td style="width:212px;">Ecuphar NV</td>
			<td style="width:179px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Vaseline Album</td>
			<td style="width:212px;">Bufa</td>
			<td align="right" style="width:179px;">165313</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">silkam 3-0 Natural silk, non-absorbable</td>
			<td>B. Braun</td>
			<td>F 1134043</td>
			<td>sutures for placement of catheters</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">silkam 2-0 Natural silk, non-absorbable</td>
			<td>B. Braun</td>
			<td>F 1134051</td>
			<td>sutures for muscular approximation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">dagrofil 3-0 Polyester, non-absorbable</td>
			<td>B. Braun</td>
			<td>C 0842478</td>
			<td>sutures for fluid fille catheters after tunneling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vicryl rapide 3-0, 1x45 cm FS2, V2930G</td>
			<td>Daxtrio medische producten</td>
			<td>15560</td>
			<td>sutures for electrical catheters after tunneling</td>
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			<td>10 ml and 2.5 ml</td>
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			<td>tiletamine / zolazepam</td>
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			<td height="21" style="height:21px;width:496px;">Sedazine</td>
			<td style="width:212px;">AST farma</td>
			<td align="right" style="width:179px;">108855</td>
			<td>xylazine</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Temgesic</td>
			<td style="width:212px;">RB Pharmaceuticals</td>
			<td align="right" style="width:179px;">5429</td>
			<td>buprenorphine</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Tensogrip</td>
			<td style="width:212px;">BSN Medical</td>
			<td style="width:179px;">71522-00</td>
			<td>elastic vest</td>
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	</tbody>
</table>

surgical placement of catheters for long-term cardiovascular exercise testing in swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Exercise up to 5 km/hr resulted in a doubling of cardiac output from 4.3 &#177; 0.3 to 8.5 &#177; 0.7 L/min which was principally accomplished by an increase in heart rate from 137 &#177; 7 to 256 &#177; 8 beats per min in combination with a small increase in stroke volume from 32 &#177; 2 to 36 &#177; 3 ml (Figure 3). The increase in stroke volume was facilitated by an increase in left ventricular contractility, as evidenced by an increase in the maximum of the first der...

Watch this Scientific Journal Video about Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine at JoVE.com

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

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