Development of a gut perfusion model as an alternative to the use of live fish

Laboratory of Veterinary Bacteriology and Mycology, Faculty of Veterinary Medicine, Ghent University, B 9820 Merelbeke, Belgium.
Laboratory Animals (Impact Factor: 1.12). 05/2005; 39(2):194-9. DOI: 10.1258/0023677053739710
Source: PubMed


An alternative fish model with the principal aim of studying the interaction between fish pathogens and the intestinal tissue was developed. The preparation consisted of an excised gut tractus from rainbow trout (Oncorhynchus mykiss), perfused through cannulation of the aorta intestinalis ventralis with filtered and heparinized Cortland+dextran 1% as the perfusion fluid. This perfusion fluid was delivered by means of a drip. The perfused gut tractus was suspended in a circular bath filled with Ringer solution, which was aerated and kept at a constant temperature of 12 degrees C. Unperfused gut placed in Ringer solution at the same temperature served as the negative control. Perfusion was effective in maintaining the gut in a healthy condition for at least 60 min with only slight oedema and sloughing of the epithelium. Conversely, the unperfused gut revealed excessive tissue degeneration and severe necrosis.

Download full-text


Available from: Freddy Haesebrouck, Dec 05, 2014
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Flavobacterium psychrophilum is an important pathogen in rainbow trout Oncorhynchus mykiss. The portal of entry for F. psychrophilum is not well known. In this study, the role of the intestine as a colonization site for F. psychrophilum was determined. For this purpose, the ability of a high (Dubois) and a low (99/10A) virulence strain of F. psychrophilum to adhere to intestinal explants of rainbow trout was evaluated. After incubation, samples of the gut were examined bacteriologically, histologically and by electron microscopy. The number of gut-associated F. psychrophilum bacteria was significantly higher for the Dubois than for the 99/10A strain. Histological samples clearly showed numerous bacteria of the high virulence strain associated with the intestinal tissue as opposed to only a few bacteria of the low virulence strain. Additionally, extensive exfoliation of intestinal epithelium was noted after incubation with the high virulence strain, but less with the low virulence strain. These findings were confirmed using scanning electron microscopy and suggest that the intestinal epithelium might represent an important site for colonization of the F. psychrophilum strain and may act as a portal of entry for high virulence F. psychrophilum.
    Full-text · Article · Dec 2005 · Diseases of Aquatic Organisms
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Yersinia ruckeri is the causative agent of enteric redmouth disease leading to significant losses in salmonid aquaculture worldwide. Little information is available on the pathogenesis of this disease. Basic steps in the establishment of an infection include attachment to the epithelium followed by invasion at the portal of entry. In this study, the interactions of Y. ruckeri with the gills and the gut of rainbow trout Oncorhynchus mykiss (Walbaum, 1792) were studied using standardized perfusion models. Virulent and avirulent Y ruckeri isolates appeared to adhere to and invade both tissues without significant differences. For the first time, the gill and gut perfusion models are shown to be suitable to study bacterial invasiveness.
    Full-text · Article · Jul 2010 · Diseases of Aquatic Organisms
  • [Show abstract] [Hide abstract]
    ABSTRACT: Summary The present study describes and visualizes the circulatory system of rainbow trout with emphasis on the heart and main blood vessels, employing corrosion cast methodology. Ten rainbow trout (Oncorhynchus mykiss (Walbaum) of 1000 g average weight were obtained from a commercial fish farm. Fish were anaesthetised using a benzocain solution in ethanol. After 40 min, the fish were killed using an overdose of the benzocain solution. The aorta caudalis and aorta coeliaco-mesentric were cannulated and attempts were made to fill the blood vessels and heart with fluid artificial resin made on the basis of methylmetacrylate. The fish were further prepared by submersion for 12–24 h in a room temperature waterbath until polymerisation and hardening of the methylmetacrylate was complete. This was followed by 24–48 h submersion in a 25% solution of KOH to obtain full maceration of the organic tissues. Various parts of the heart and blood vessels were retained in their natural positions, thereby demonstrating the anatomical details of the main circulatory system. Main elements depicted included the sinus venosus, atrium, ventricle, bulbus arteriosus and related vessels such as the dorsal aorta, subclavian vein, hepatic vein, common cardinal vein, coeliaco-mesenteric artery, gastero-intestinal artery, and dorsal intestinal artery. Related smaller vessels were also determined.
    No preview · Article · Jun 2011 · Journal of Applied Ichthyology
Show more