导尿在反刍动物的肠袢
Summary
我们描述了一种新的手术方法catheterizing在羊回肠“肠袢”。一旦动物有从手术中恢复，并已清除抗生素和止痛药，多治疗，可以直接存入通过导管循环。
Abstract
肠道是一个复杂的结构，不仅在营养物质的吸收，而且还充当之间的个人和外界的障碍。因此，肠道内起着举足轻重的作用，从肠道致病菌的免疫监视和保护。调查肠道生理学和免疫学通常采用“肠袢”作为实验模型。这些循环模型大部分都是非恢复手术过程中，研究短期（<24小时）在肠道内的变化（1-3）。我们先前创建的恢复肠袢模型来具体衡量长期（<6个月）羊暴露后疫苗，佐剂和病毒（4）肠道免疫变化。此过程的本地化处理，以具体的“循环”，让我们品尝这一领域的肠道。这种方法的一个显着的缺点是管理治疗的机会，即在手术时的单一窗口。此外，肠道黏膜和管腔内容的样本，只能采取在项目终止。上述模型中的其他显着的局限性，手术操作和必要的手术后的措施（如抗生素和止痛药的管理），可直接影响治疗本身和/或改变免疫功能，从而混淆的结果。因此，我们修改了我们的肠袢模型，通过长期导管插入循环。羊完全恢复的程序，并exteriorized导管的影响。值得注意的是，建立在循环导管，使我们能够引进在延长的时间间隔多次治疗后，手术和清理手术期间服用过的药物和手术后的时期的恢复。
Protocol
实验过程： 所有仪器蒸压;非autoclaveable材料Germex冷至少10分钟的灭菌水溶液消毒。 羊acepromazine（0.05毫克/公斤），格隆（0.005毫克/公斤），与布托啡诺酒石酸盐（0.2毫克/公斤）IM的前药，并且留15-20分钟。 剃光和颈部皮肤手术擦洗。一旦颈内静脉是可视化的，在静脉注射0.1毫升利多卡因资深大律师，引入静脉导管，并固定在手术过程中使用牲畜识别标签水泥和2“医院磁带皮肤…
Discussion
在此过程中，我们能够成功地建立在羊回肠内建立的肠袢的导管。坚持严格的术后护理，动物迅速恢复从程序和手术后恢复正常活动5-7天。我们决定建立循环内导管不影响肠黏膜免疫功能（数据未显示）和动物可以生活至少8周后手术。此过程允许我们引入不同的免疫车厢多次治疗，在延长的时间间隔，一旦动物有从手术中恢复，并进行实证衡量这些治疗的生理和免疫效果。
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Disclosures
The authors have nothing to disclose.
Acknowledgements
在农业和农业食品加拿大研究中心，莱斯布里奇，阿尔伯达省，加拿大的许多人没有奉献精神和努力，这项研究可能已经完成。在这方面，我们感谢：塔拉谢尔顿前和术后护理的动物，手术准备，麻醉科，样品分析，并与视频影像的援助;兰迪王尔德和ESTELA哥斯达黎加前和术后护理;珍妮Gusse样品分析;拜伦李套管形象;和布赖恩Egland养羊。我们也感谢提供视频设备和意见，以及其他援助（即执行生产）布伦特Selinger（莱斯布里奇大学生物科学系），和课程的再开发中心，莱斯布里奇大学录像专长。这项研究是支持部分由高级食品材料网（GDI），加拿大阿尔伯塔省肉牛业发展基金（GDI），和加拿大农业和农业食品（GDI和JPK）的赠款。
Materials
| Category / Item Number | Company | Catalogue |
| Surgeon supplies | ||
| Surgical gloves | WDDCa | 103091 |
| Surgical mask | WDDC | MAX53013 |
| Surgical gown | CDMVb | 3365 |
| OB Apron | WDDC | 900062 |
| Surgical cap | WDDC | 903271 |
| EZ Scrub brush | WDDC | BDC377479 |
| Anesthesia and drugs | ||
| Anesthetic machine | Boyle | — |
| Sodasorb | WDDC | 108638 |
| Laryngoscope | WDDC | — |
| Laryngoscope blade (Miller blade #4) | WDDC | 902667 |
| Endotracheal tube (7.0 – 8.5 mm) | WDDC | — |
| Lidocaine endotracheal spray | WDDC | 103364 |
| Acepromazine | WDDC | AUS01020 |
| Glycopyrrolate | WDDC | SAB2880 |
| Butorphanol tartrate | WDDC | AYE304541 |
| Diazepam | WDDC | sab790 |
| Thiopental sodium | WDDC | 103508 |
| Epinephrine hydrochloride | WDDC | 103856 |
| Doxapram | Strathcona Pharmacy | — |
| Isoflurane | WDDC | 100339 |
| Hemostam | WDDC | 103857 |
| Flunixin | WDDC | sch140100 |
| Enrofloxacin | WDDC | hll0049 |
| Metronidazole | WDDC | 101653 |
| Ampicillin | WDDC | NOV67185-6 |
| Cefazolin | WDDC | nov69397 |
| Surgical equipment and supplies | ||
| Electroscapel | Macan | MV8 |
| Heating mat | Kane | PHMD-48 |
| Stomach tube (Nalgene tubing 5/16″ ID) | Fisher Scientific | — |
| Needle driver | WDDC | 108338 |
| Addison brown forcep | WDDC | 911075 |
| Rat tooth forcep | CDMV | 5275 |
| Kelly forcep | WDDC | 901133 |
| Mosquito hemostat | WDDC | 108471 |
| Carmalt forcep | WDDC | 901225 |
| Scalpel handle | WDDC | 900285 |
| Scalpel blade #22 | WDDC | 900511 |
| Metzenbaum scissor | WDDC | 901166 |
| Sharp-sharp scissor | WDDC | 108264 |
| Towel clamp | WDDC | 901213 |
| Allis tissue forcep | WDDC | 901201 |
| Doyen’s forcep | WDDC | 904226 |
| Stainless steel bowl | WDDC | 903601 |
| Stainless steel trocar (curved) | Local supplier | — |
| Catheter introducer | WDDC | 11440 |
| Surgical towel | CDMV | 7972 |
| Disposable drape | WDDC | 900035 |
| Steri drape | CDMV | 2323 |
| Drape (40 x 40 cm) | CDMV | 8358 |
| Surgical drape (7″ x 11 ¼” opening) | CDMV | 2694 |
| Vicryl (2-0) | WDDC | JJMJ333H |
| Silk 2-0 | WDDC | 105248 |
| Catgut #3 | WDDC | CSL547-2070 |
| Supramid #1 | WDDC | CSL547-3040 |
| Needles (cutting and non-cutting) | WDDC | — |
| Catheter | ||
| Silastic tubing (0.040″ ID, 0.085″ OD) | Fisher Scientific | 11-189-15D |
| Orbeseal ointment | WDDC | 106344 |
| Clear silicone (732 multi-purpose sealant) | Local supplier | — |
| Blunt end needle | WDDC | 905035 |
| Marker (permanent industrial) | Local supplier | — |
| Catheter pouch (constructed of Elastoplast 7.5 cm) | WDDC | 104094 |
| Bandages | ||
| Hospital tape (2″) | WDDC | 103842 |
| Hospital tape (1″) | WDDC | VLMRENF20 |
| Gauze sponge 4×4 | WDDC | 107551 |
| Gauze cling | WDDC | 107633 |
| Elastoplast 7.5cm | WDDC | 104094 |
| Elastoplast rippable bandage | WDDC | 104093 |
| Needles and syringes | ||
| Syringe (60 ml) | WDDC | 104004 |
| Syringe (10 ml) | WDDC | bdc301604 |
| 25G Needle | WDDC | BDC305122 |
| 20G Needle | WDDC | BDC305175 |
| 18G Needle | WDDC | 903353 |
| Intravenous | ||
| Intracath IV catheter (16G 8″) | WDDC | bdc3831621 |
| IV administration set (10 drop) | WDDC | 101797 |
| 0.9% NaCl solution | WDDC | BAXJB1322P |
| Plasmalyte 148 solution | WDDC | BAXJB2534 |
| Dextrose | WDDC | MCT60190 |
| Peristaltic pump | Cole Parmer | 7524-10 |
| Livestock identification tag cement | Local supplier | — |
| Sanatizers / disinfectants | ||
| Germex solution | WDDC | mtc60270 |
| Isopropyl alcohol | WDDC | 104590 |
| Stanhexidine cleanser | WDDC | 109248 |
| Prepodyne solution | WDDC | 105710 |
| Prepodyne scrub | WDDC | 105711 |
aWestern Drug Distribution Center LTD, Edmonton, AB.
bThe Veterinary Medication Distribution Center Inc., St. Hyacinthe, QE.
References
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- Greibel, P. J., Davis, W. C., Reynolds, J. D. An analysis of the growth and differentiation of b cells isolated from follicles from ileal Peyer’s patch of sheep. Immunology. 75, 601-607 (1992).
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Tags
CatheterizationIntestinal LoopsRuminantsIntestinal PhysiologyImmunologyExperimental ModelSurgical ProceduresShort-term ChangesLong-term ChangesRecovery Intestinal Loop ModelSheepVaccinesAdjuvantsVirusesTreatmentsIntestinal MucosaLuminal ContentsSurgical ManipulationPost-operative MeasuresAntibioticsAnalgesicsCatheters
Cite This Article
Uwiera, R. R. E., Kastelic, J. P., Inglis, G. D. Catheterization of Intestinal Loops in Ruminants. J. Vis. Exp. (28), e1301, doi:10.3791/1301 (2009).