Phenotype, intestinal morphology, and survival of homozygous and heterozygous endothelin B receptor--deficient (spotting lethal) rats.

BACKGROUND/PURPOSE: Spotting lethal (sl) rats, a model for Hirschsprung's disease, recently have been found to carry a deletion in the endothelin B (ET(B)) gene, causing functional lack of ET(B) receptors. The ET(B) receptor mediates, together with and in counterbalance to the ET(A) receptor, endothelin actions on vessels, cell proliferation, and migration. The authors investigated the effect of homozygosity (sI/sI) or heterozygosity (+/sl) on phenotype, intestinal morphology, and survival. METHODS: Weight, circumference, and serum albumin were measured. Histological tests of major organs and immunoperoxidase reaction for Peripherin, glial fibrillary acid protein (GFAP), and S-100 in small and large intestine were performed. Peripherin-immunostained sections of colon and jejunum were analyzed morphometrically. Screening for sepsis included search for enterocolitis, bacterial infection, endotoxin, and iNOS mRNA. RESULTS: Sl/sl rats died within 4 weeks of life, showing an early and a later death group. Serum albumin levels were decreased in sl/sl rats, whereas signs of sepsis were rare. Immunostaining uncovered alterations in nerve and glial cells in the whole gut of sl/sl rats, and to a subtle degree also in +/sl rats, which appear clinically normal. Morphometric quantification yielded statistically significant alterations in sl/sl rats only. No obvious abnormalities were found in other organs. CONCLUSIONS: Sl/sl rats die from malnutrition rather than sepsis, too early for ischemic complications to occur. Rats of the later death group are a suitable model for studying the ET8 receptor in vivo. Subtle abnormalities in the enteric nervous system of heterozygous rats underline the critical role of the "gene dose" for functional compensation.

Biodecontamination of animal rooms and heat-sensitive equipment with vaporized hydrogen peroxide.

Common methods used to decontaminate and disinfect laboratory animal areas are difficult to standardize, labor-intensive, and potentially hazardous for staff members and the environment. As an alternative to traditional methods, we tested fumigation with vaporized hydrogen peroxide by using the VHP 1000 Biodecontamination System. The design of our air-conditioning system allowed the connection of the generator to any animal room by using the ventilation piping, thus forming a closed circuit. A 3-h cycle consisting of dehumidification, conditioning, sterilization, and aeration was developed and shown to be effective. The biodecontamination process was monitored during five independent trials using chemical and biological (Bacillus stearothermophilus spores) indicators. Contact plates for testing surfaces and room air for environmental bacteria, yeasts, and fungi consistently showed fewer than 10 colony-forming units per 100 cm2 or per 1 liter air. In addition, this method proved successful with heat-sensitive equipment like the blower units of individually ventilated caging systems. Overall, the system was easy to use and very effective in biodecontaminating animal rooms and equipment in a reproducible manner. There were no signs of corrosion or functional damage after more than 10 fumigation cycles. Work load and potential health risk for staff members and the environment was negligible.

[The laboratory animal management system--an animal housing management data-processing system]. / Das Versuchstierhaltungs-System (VTH)--ein Tierhaus-Verwaltungs-DV-System.

The Laboratory Animal Management System (LAMS) is a flexible, multi-purpose animal house management tool. It has a decentralized structure and was developed using UNIFACE and Oracle-database software. The multiuser LAMS system runs on a Micro-Vax computer and can be accessed from several workstations. LAMS has been designed to manage the following functions: animal study details; animal procurement; book keeping and follow-up; amendments; update of data; inquiries; statistics and numerous additional tasks. LAMS is a user-friendly interactive system which does not allow input of incorrect data and can be operated by staff with very little computer experience. The system fully complies with German legal requirements and is becoming an increasingly important tool for routine management of animal house facilities and animal experimentation.

Acidification of the colonic mucins following polyvalent colonization of the germ-free rat.

Colonic mucins of germ-free (GF) and conventional rats (CV) were compared. After isolation by gel filtration on Sepharose CL-4B and purification by density gradient centrifugation, the content of isolated colonic mucins was estimated by determination of PAS positive carbohydrates. Purified mucins were subjected to carbohydrate and amino acid analysis and separated into mucin subclasses by ion exchange chromatography. While the total amount of colonic mucins was not statistically different in GF and CV animals, analysis of carbohydrate composition demonstrated an increased amount of sialic acid in CV rat mucin. This was in accordance with results of ion exchange chromatography, revealing a significant higher amount of negative charged mucin subclasses in CV mucin, compared to the germ-free counterpart. The results of amino acid analysis were similar in both groups. The compositional differences in carbohydrate moieties are attributed to modulations by the intestinal flora. A selective bacterial degradation of the neutral mucin subclasses and modifications in the mucin composition due to a stimulated synthesis are discussed.

[Physiologic hypothermia of newborn kittens up to 6 weeks old]. / Physiologische Hypothermie der Katzenwelpen bis zur 6. Lebenswoche.

Kittens showed a physiological hypothermia until they were 6 weeks of age. In the first 3 weeks of age rectal temperature was constantly low (37.6 +/- 0.3 degrees C). A linear increase of rectal temperature followed from the fourth to the sixth week of age inclusively and from the seventh week on rectal temperature reached the final temperature level (38.4 +/- 0.3 degrees C). This finding has to be considered in clinical assessment of kittens' body temperature.

Colonic glycoproteins in monozygotic twins with inflammatory bowel disease.

Colonic glycoprotein composition was evaluated in monozygotic twins with inflammatory bowel disease using ion-exchange chromatography. Fifty-three individuals, 12 pairs and 1 single twin with ulcerative colitis and 14 pairs with Crohn's disease, were evaluated. Seven twin pairs were concordant for the presence of ulcerative colitis or Crohn's disease, whereas twin siblings of 10 ulcerative colitis probands and 9 Crohn's disease probands were not known to have inflammatory bowel disease. Content of one chromatographically defined component of colonic mucin, designated HCM species IV, was reduced in both patients with ulcerative colitis (1040 +/- 300 cpm/10,000 cpm total HCM) and their apparently healthy twins (1340 +/- 540 cpm/10,000 cpm total HCM) compared with control subjects (4030 +/- 1,000 cpm/10,000 cpm total HCM). Composition of mucin in Crohn's disease patients and their nonaffected twins was not significantly different than in controls. These observations suggest that altered profiles of mucin glycoprotein may be present before the onset of ulcerative colitis and may be genetically defined. Conversely, it appears that alterations in glycoproteins only are not sufficient to initiate mucosal inflammation.

Pre-epithelial mucus layer in the colon of conventional and germ-free rats.

The pre-epithelial mucus layer (PML) and epithelial mucins were studied by mucin histochemistry in 10 microns-thick celloidinstabilized cryostat sections in the proximal and distal colon of conventional and germ-free rats aged 120 and 350 days. No continuous PML was found in the proximal colon. A continuous mucus blanket, of fairly homogenous thickness, was observed in the distal colon, where the PML-thickness was 40 +/- 24 microns at 120 days of age and 44 +/- 22 microns at 350 days of age in conventional rats, and 25 +/- 17 microns (120 days) and 22 +/- 10 microns (350 days) in germ-free rats. The stainability of the PML by periodic acid-Schiff and Alcian Blue at pH 2.5 and 1.0 was stronger in conventional rats than in germ-free rats, indicating higher concentrations of mucosubstances and of acid and sulphated mucins, respectively. The PML of the conventional rat distal colon showed a stratified structure of up to eight sublayers. In the distal colon of germ-free rats, the whole gut wall thickness was reduced 47% compared to the conventional rat (germ-free; 185 +/- 73 microns, conventional: 350 +/- 115 microns). No stratification of the PML was observed. The presence of intestinal microflora obviously had a strong influence on the thickness, compactness, mucin content, mucin composition and structure of the pre-epithelial mucus layer.