OBJECTIVE To compare initial leak pressure (ILP) between cadaveric canine and synthetic small intestinal segments that did and did not undergo enterotomy. SAMPLE Eight 8-cm grossly normal jejunal segments from 1 canine cadaver and eight 8-cm synthetic small intestinal segments. PROCEDURES Intestinal segments were randomly assigned to undergo enterotomy (6 cadaveric and 6 synthetic segments) or serve as untreated controls (2 cadaveric and 2 synthetic segments). For segments designated for enterotomy, a 2-cm full-thickness incision was created along the antimesenteric border. The incision was closed in a single layer with 4-0 suture in a simple continuous pattern. Leak testing was performed with intestinal segments occluded at both ends and infused with dilute dye solution (999 mL/h) until the solution was observed leaking from the suture line or serosal tearing occurred. Intraluminal pressure was continuously monitored. The ILP at construct failure was compared between cadaveric and synthetic control segments and between cadaveric and synthetic enterotomy segments. RESULTS Mean ± SD ILP did not differ significantly between cadaveric (345.11 ± 2.15 mm Hg) and synthetic (329.04 ± 24.69 mm Hg) control segments but was significantly greater for cadaveric enterotomy segments (60.77 ± 15.81 mm Hg), compared with synthetic enterotomy segments (15.03 ± 6.41 mm Hg). CONCLUSIONS AND CLINICAL RELEVANCE Leak testing should not be used to assess the accuracy or security of enterotomy suture lines in synthetic intestinal tissue. Synthetic intestinal tissue is best used for students to gain confidence and proficiency in performing enterotomies before performing the procedure on live animals.

OBJECTIVE To evaluate the effect of kernel and window settings on the assessment of small and complicated vasculature in CT angiographic (CTA) images of kidneys, jejunum with mesentery, and tumors in dogs. ANIMALS 20 healthy dogs and 20 dogs with tumors. PROCEDURES Images from CTA performed previously in dogs were reconstructed with 3 different combinations of kernel and window settings (soft kernel with soft tissue window, soft kernel with bone window, and sharp kernel with bone window), and reconstructed images of the left kidney and the jejunum with the mesentery in healthy dogs and tumors in affected dogs were evaluated by reviewers blinded to the settings. RESULTS For images of kidney and jejunum with mesentery, reviewers’ scores for the conspicuity of vascularity in the arterial phase and the differentiation of the organs from the adjacent structures were significantly higher when viewed in bone window (vs soft tissue window) regardless of kernel setting. For images of head and gastrointestinal tumors, reviewers’ scores for differentiation of intratumoral vasculature were higher when viewed in sharp kernel with bone window versus other setting combinations. However, the conspicuity of gastrointestinal, hepatic, or splenic tumoral vessels from the adjacent structures had higher reviewer scores for images in soft kernel with soft tissue window, compared with other setting combinations. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that reconstruction of CTA images with sharp kernel combined with bone window settings might have clinical utility in evaluating and planning treatments for dogs with various tumors; however, additional research is warranted to further identify effects of various kernel and window setting combinations on assessments of small and complicated vasculature in larger and more diverse populations of dogs with and without tumors.

OBJECTIVE To evaluate the effect of presurgical storage conditions on leakage pressures of enterotomy sites closed with unidirectional barbed suture material in fresh, chilled, and frozen-thawed cadaveric canine jejunal specimens. SAMPLE 36 grossly normal jejunal segments obtained from 4 dog cadavers. PROCEDURES 9 jejunal segments were harvested immediately from each euthanized dog and randomly assigned to be tested within 4 hours after collection (fresh segments), stored at 4°C for 24 hours before testing (chilled segments), or stored at −20°C for 7 days and thawed at 21°C for 6 hours before testing (frozen-thawed segments). For leakage pressure testing, a 3-cm-long antimesenteric enterotomy was performed and repaired with 3-0 unidirectional barbed suture material in a simple continuous pattern in each segment. Time to complete the enterotomy, initial leakage pressure, maximum intraluminal pressure, and leakage location were recorded for each segment. RESULTS Mean ± SD initial leakage pressure for fresh, chilled, and frozen-thawed segments was 52.8 ± 14.9 mm Hg, 51.8 ± 11.9 mm Hg, and 33.3 ± 7.7 mm Hg, respectively. Frozen-thawed segments had significantly lower mean initial leakage pressure, compared with findings for fresh or chilled segments. Time to complete the enterotomy, maximum intraluminal pressure, and leakage location did not differ among groups. CONCLUSIONS AND CLINICAL RELEVANCE Leak pressure testing of cadaveric jejunal segments that are fresh or chilled at 4°C for 24 hours is recommended for enterotomy studies involving barbed suture material in dogs. Freezing and thawing of cadaveric jejunal tissues prior to investigative use is not recommended because leak pressure data may be falsely low.

OBJECTIVE To determine whether stored (cooled or frozen-thawed) jejunal segments can be used to obtain dependable leak pressure data after enterotomy closure. SAMPLE 36 jejunal segments from 3 juvenile pigs. PROCEDURES Jejunal segments were harvested from euthanized pigs and assigned to 1 of 3 treatment groups (n = 12 segments/group) as follows: fresh (used within 4 hours after collection), cooled (stored overnight at 5°C before use), and frozen-thawed (frozen at −12°C for 8 days and thawed at room temperature [23°C] for 1 hour before use). Jejunal segments were suspended and 2-cm enterotomy incisions were made on the antimesenteric border. Enterotomies were closed with a simple continuous suture pattern. Lactated Ringer solution was infused into each segment until failure at the suture line was detected. Leak pressure was measured by use of a digital transducer. RESULTS Mean ± SD leak pressure for fresh, cooled, and frozen-thawed segments was 68.3 ± 23.7 mm Hg, 55.3 ± 28.1 mm Hg, and 14.4 ± 14.8 mm Hg, respectively. Overall, there were no significant differences in mean leak pressure among pigs, but a significant difference in mean leak pressure was detected among treatment groups. Mean leak pressure was significantly lower for frozen-thawed segments than for fresh or cooled segments, but mean leak pressure did not differ significantly between fresh and cooled segments. CONCLUSIONS AND CLINICAL RELEVANCE Fresh porcine jejunal segments or segments cooled overnight may be used for determining intestinal leak pressure, but frozen-thawed segments should not be used.

OBJECTIVE To evaluate a method for identifying intact and degranulated eosinophils in the small intestine of dogs with inflammatory bowel disease (IBD) by use of a monoclonal antibody (mAb) against eosinophil peroxidase (EPX). ANIMALS 11 untreated dogs with IBD, 5 dogs with IBD treated with prednisolone, and 8 control dogs with no clinical evidence of gastrointestinal tract disease and no immunosuppressive treatment. PROCEDURES 4-μm-thick sections of paraffin-embedded tissues from necropsy specimens were immunostained with EPX mAb. Stained intact and degranulated eosinophils in consecutive microscopic fields (400X magnification) of the upper (villus tips) and lower (between the muscularis mucosae and crypts) regions of the lamina propria of the jejunum were manually counted. RESULTS Compared with control and treated IBD dogs, untreated IBD dogs had a significantly higher number of degranulated eosinophils in the lower region of the lamina propria. However, no significant differences were detected in the number of intact eosinophils in this region among groups. In the upper region of the lamina propria, untreated IBD dogs had a significantly higher number of degranulated and intact eosinophils, compared with control and treated IBD dogs. Number of degranulated and intact eosinophils did not differ significantly between control and treated IBD dogs. CONCLUSIONS AND CLINICAL RELEVANCE Immunohistologic analysis with EPX mAb yielded prominent granule staining that allowed reliable morphological identification of degranulated and intact eosinophils, which may provide a strategy for quantitative and selective evaluation of eosinophils in gastrointestinal biopsy specimens and a potential method to diagnose IBD and evaluate treatment outcome.

OBJECTIVE To examine effects of continuous rate infusion of lidocaine on transmural neutrophil infiltration in equine intestine subjected to manipulation only and remote to ischemic intestine. ANIMALS 14 healthy horses. PROCEDURES Ventral midline celiotomy was performed (time 0). Mild ischemia was induced in segments of jejunum and large colon. A 1-m segment of jejunum was manipulated by massaging the jejunal wall 10 times. Horses received lidocaine (n = 7) or saline (0.9% NaCl) solution (7) throughout anesthesia. Biopsy specimens were collected and used to assess tissue injury, neutrophil influx, cyclooxygenase expression, and hypoxia-inducible factor 1α (HIF-1α) expression at 0, 1, and 4 hours after manipulation and ischemia. Transepithelial resistance (TER) and mannitol flux were measured by use of Ussing chambers. RESULTS Lidocaine did not consistently decrease neutrophil infiltration in ischemic, manipulated, or control tissues at 4 hours. Lidocaine significantly reduced circular muscle and overall scores for cyclooxygenase-2 expression in manipulated tissues. Manipulated tissues had significantly less HIF-1α expression at 4 hours than did control tissues. Mucosa from manipulated and control segments obtained at 4 hours had lower TER and greater mannitol flux than did control tissues at 0 hours. Lidocaine did not significantly decrease calprotectin expression. Severity of neutrophil infiltration was similar in control, ischemic, and manipulated tissues at 4 hours. CONCLUSIONS AND CLINICAL RELEVANCE Manipulated jejunum did not have a significantly greater increase in neutrophil infiltration, compared with 4-hour control (nonmanipulated) jejunum remote to sites of manipulation, ischemia, and reperfusion. Lidocaine did not consistently reduce neutrophil infiltration in jejunum.

OBJECTIVE To evaluate the eosinophilic response in intestinal mucosa of horses with intestinal ischemia and reperfusion or with strangulation of the jejunum or colon. SAMPLE Mucosal samples from horses with naturally occurring strangulation (n = 24 horses) or distention (n = 6) of the jejunum or colon (11), with experimentally induced ischemia and reperfusion of the jejunum (6) or colon (15), or that were euthanized for reasons other than gastrointestinal tract disease (13). PROCEDURES Mucosal samples were collected and grouped by type of intestinal injury. Slides were stained with Luna eosinophil stain and histologically examined to determine eosinophil accumulation and distribution. Number of eosinophils per mm2 of mucosa was calculated as a measure of eosinophil accumulation. Additionally, mucosa was categorized into 5 regions; the percentage of eosinophils in each of the 5 regions, relative to the total eosinophil count in all regions, was determined. RESULTS Eosinophil migration toward and onto the luminal surface was evident in tissues after ischemia and reperfusion and after naturally occurring strangulating disease of the jejunum and colon, as indicated by a decrease in the number of eosinophils near the muscularis mucosa and an increase in the number of eosinophils on or near the luminal surface. Ischemia alone did not change eosinophil distribution in the jejunum or colon. CONCLUSIONS AND CLINICAL RELEVANCE Eosinophils responded to mucosal damage evoked by ischemia and reperfusion by migration toward and onto the luminal surface. This migration could represent an important component of the inflammatory response to injury in equine gastrointestinal mucosa.

This study examined known or possible virulence-associated genes in type A Clostridium perfringens from cases of both bovine clostridial abomasitis (BCA) and jejunal hemorrhage syndrome (JHS) and compared these to isolates from calves that were healthy or had undifferentiated diarrheal illness. A real-time polymerase chain reaction (PCR) assay was used to genotype the 218 C. perfringens isolates. Isolates were sourced from healthy and diarrheic young and mature cattle (n = 191), from calves with confirmed or suspected BCA (n = 22), and from mature cattle with JHS (n = 5). Of 216 isolates (96%), 208 were positive for the cpa gene and 13% (29/218) were positive for atypical cpb2. Three of 8 (37.5%) confirmed BCA isolates, 2 of 13 (15.4%) suspected BCA isolates, and no JHS isolates tested positive for atypical cpb2. As all isolates were negative for cpb, cpb2, cpe, etx, netB, and tpeL, the results of the present study do not support a role for these genes in BCA or JHS. A subset of unique genes identified in 1 bovine clostridial abomasitis isolate (F262), for which a genome sequence is available, was searched for in 8 BCA isolates by PCR. None of the 10 genes was consistently present in all or even in a majority of BCA isolates. Many of these genes were also variably and inconsistently present in type A isolates from calves that did not have BCA. Although a virulence signature to aid in the diagnosis of BCA caused by C. perfringens type A was not identified, further work may discover a gene or group of genes that would constitute such a signature.