Intestine Of Sheep Research Articles

Signal amplification by combining two advanced immunohistochemical techniques

The immunohistochemical techniques known as EnVision trade mark + System (EVS) and Mirror Image Complementary Antibody (MICA) were recently introduced into laboratory practice because of their high sensitivity. In this paper these techniques were compared and their sequences combined to obtain a new method possibly more sensitive than the original ones. The immunohistochemical staining employing the avidin-biotin complex (ABC), largely used as routine, was adopted as a term of comparison. Samples from the small and large intestine of pigs and sheep were fixed in Bouin and embedded in Paraplast. The primary antibodies utilized were directed against the neuronal nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP) and chromogranin A (Cr A). Targets of these antibodies were nerve structures of the intestinal wall, as well as endocrine cells scattered in the mucosa of the bowel, defined neuroendocrine cells or paraneurons. The EVS method appeared as slightly superior to the MICA method regarding sensitivity of detection. The EVS/MICA (combined) method resulted four/eight times more effective than the original techniques regarding sensitivity of detection and staining intensity, both at low and high dilutions of the primary antibodies. Of these latter, immunopositive structures were still clearly identifiable, at a dilution of 1:256,000. Such efficiency could be explained by the high number of revealing molecules of peroxidase contained in the new sequence. The application of the combined method is recommended when a small quantity of tissue antigens needs to be detected immunohistochemically.

Catheterization of Intestinal Loops in Ruminants

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the outside world. As such, the intestine plays a pivotal role in immunosurveillance and protection from enteric pathogens. Investigating intestinal physiology and immunology commonly employs 'intestinal loops' as an experimental model. The majority of these loop models are non-recovery surgical procedures that study short-term (<24 hr) changes in the intestine (1-3). We previously created a recovery intestinal loop model to specifically measure long-term (<6 mo) immunological changes in the intestine of sheep following exposure to vaccines, adjuvants, and viruses (4). This procedure localized treatments to a specific 'loop', allowing us to sample this area of the intestine. A significant drawback of this method is the single window of opportunity to administer treatments (i.e. at the time of surgery). Furthermore, samples of both the intestinal mucosa and luminal contents can only be taken at the termination of the project. Other salient limitations of the above model are that the surgical manipulation and requisite post-operative measures (e.g. administration of antibiotics and analgesics) can directly affect the treatment itself and/or alter immune function, thereby confounding results. Therefore, we modified our intestinal loop model by inserting long-term catheters into the loops. Sheep recover fully from the procedure, and are unaffected by the exteriorized catheters. Notably, the establishment of catheters in loops allows us to introduce multiple treatments over an extended interval, following recovery from surgery and clearance of drugs administered during surgery and the post-operative period.

Development of a Gas Empowered Drug Delivery system for peptide delivery in the small intestine

The aim of this investigation was to design a novel Gas Empowered Drug Delivery (GEDD) system for CO 2 forced transport of peptide drugs together with mucoadhesive polymers to the surface of the small intestine. The GEDD effect of the core tablet was achieved using CO 2 gas to push insulin together with the mucoadhesive excipients poly(ethyleneoxide) (PEO) and the permeation enhancer trimethyl chitosan (TMC) to the surface of the small intestine. The in-vitro insulin release showed that almost 100% of the insulin was released from enterically coated tablets within 30 min at pH 6.8. The designed GEDD system was shown to increase the insulin transport by approximately 7 times in comparison with the free insulin across sheep's intestine ex-vivo. Three different peroral formulations were administered to male rabbits: F1 containing no TMC or PEO, F2 containing PEO but no TMC and F3 containing both PEO and TMC. The administrations of insulin using the formulation F1 resulted in a low FR value of 0.2% ± 0.1%, while the formulations F2 and F3 resulted in a much higher FR values of 0.6 ± 0.2% and 1.1% ± 0.4%, respectively. Hence, the insulin permeation achieved by the GEDD system is primarily due to the enhancing effect of TMC and the mucoadhesive properties of PEO both of which synergistically increase the bioavailability of insulin.

TSE risk assessment from carcasses of ovine and caprine animals below 6 months of age from TSE infected flocks intended for human consumption - Scientific Opinion of the Panel on Biological Hazards.

TSE risk assessment from carcasses of ovine and caprine animals below 6 months of age from TSE infected flocks intended for human consumption - Scientific Opinion of the Panel on Biological Hazards.

Sutureless Nerve Repair with Laser-Activated Chitosan Adhesive: A Pilot in Vivo Study

The anastomosis of peripheral nerves is a demanding procedure that has potential complications due to foreign body reactions elicited by sutures. In this study, the sutureless in vivo anastomosis of rat tibial nerves was successfully performed, using for the first time a chitosan-based laser-activated adhesive. The nerve thermal damage caused by the laser irradiation was quantitatively assessed. A novel adhesive composed of chitosan, indocyanine green, acetic acid, and water, was fabricated in thin sheets. Its adhesive strength was tested in vitro by bonding strips (surface area approximately 20 mm2, thickness approximately 20 microm) onto rat sciatic nerves and sheep intestine by laser activation with low fluence ( approximately 50 J/cm2), using a fiber-coupled diode laser (n = 13). The tensile strength of the adhesive/tissue bonds was measured after tissue repair. The chitosan adhesive was then used to perform sutureless anastomosis of tibial nerves in vivo (n = 6). Adhesive strips were also bonded in vivo onto intact rat sciatic nerves (n = 6) in order to quantitatively assess, by counting myelinated axons, the thermal damage induced by the laser. The adhesive bonded well to tissue with a tensile strength of 12.5 +/- 2.6 KPa (mean +/- SD; n = 13). The in vivo anastomosed nerves were in continuity 3 d after surgery. Axon counting showed the number and morphology of myelinated axons were normal proximally ( approximately 96%) compared with intact nerves (100%). Axon demyelination was observed at the operation site ( approximately 49%) and distally ( approximately 27%), and was attributed to laser-induced thermal damage. Nerve anastomosis, performed by the laser-adhesive procedure, was successful 3 d postoperatively. Proximal myelinated axons were not significantly damaged by the low laser fluence.

Detection and Frequency of Stx2 Gene in Escherichia coli O157 and O157:H7 Strains Isolated from Sheep Carcasses in Shiraz-Iran

Enterohaemorrhagic Escherichia coli constitute a subset of serotypes (E. coli O157 and some other serogroups) of Shiga toxin-producing E. coli firmly associated with severe human illnesses like bloody diarrhoea and haemolytic uraemic syndrome. Escherichia coli O157:H7 is a zoonotic pathogen. They rarely cause disease in animals, live in the intestines of healthy sheep and ruminants are recognized as their main natural reservoir, so they can contaminate meat during slaughtering practices. The purpose of this study was epidemiological survey on the occurrence of E. coli O157:H7 in healthy sheep in Shiraz-Iran. Polymerase Chain Reaction (PCR) assay was developed to detect the Stx2 gene the only bacterial factor that has been associated with more severe disease. During a period of 7 months (December 2005 to June 2006), 153 slaughtered sheep at Shiraz slaughterhouse, were randomly selected and examined for surface carriage of E. coli O157:H7 by conventional plating and Stx2 gene detection by PCR technique. E. coli O157:H7 was found in 6(3.92%) of 153 sheep. The bacteria were isolated from 5(3.34%) of 114 and 1(2.63%) of 38 sheep two or under two and more than 2 years old, respectively (p = 0.5). The contamination rate might vary depending on season, age and infection time. The higher frequency for younger animals may be due to differences in the composition of the gastrointestinal flora resulting from differences in diet. This is the first report of the presence of E. coli O157:H7 in sheep from Iran.

Localization and Control of Expression of VEGF-A and the VEGFR-2 Receptor in Fetal Sheep Intestines

We studied expression of vascular endothelial growth factor A (VEGF-A) and its main receptor, VEGFR-2, in the small intestine from five groups of fetal sheep (each n = 5): 1) preterm controls, 2) term controls, 3) preterm animals where the fetus was infused with cortisol, or 4) saline, and 5) term animals where adrenalectomy had been performed preterm. The main transcript expressed in fetal small intestine was VEGF-A165. Comparing term with preterm animals, there were significantly higher levels of expression of VEGF-A protein (p = 0.005). Levels of VEGF-A protein expression were lower in term adrenalectomized animals (p = 0.01) and were higher in preterm animals infused with cortisol (p = 0.01), compared with their respective control groups. Immunohistochemistry demonstrated strongest expression of VEGF-A protein in the epithelial cells and lamina propria of the villi. Intestinal expression of mRNA encoding the VEGFR-2 receptor did not significantly vary with gestational age. In situ hybridization localized VEGFR-2 to the lamina propria of the villous core and receptor autoradiography using 125I VEGF-A demonstrated binding in the same site. These data show that intestinal VEGF-A is up-regulated with advancing gestation in a glucocorticoid-dependent manner--novel findings consistent with a role for VEGF-A stimulated angiogenesis in preparing the fetal gut for birth.

Surgical instruments, sutures and suturing techniques

Anthropological studies suggest that prehistoric man used various techniques to close skin wounds (Ellis, 2001). The jaws of termites or beetles were used to bite across wounds and hold skin edges together in India and South America, and in East Africa acacia thorns were stuck along the two wound edges and then plaited together (Ellis, 2001). Suture materials used included horsehair, fibres from tree bark, cotton and, in ancient Rome, human hair and catgut (made from sheep's intestine) (Burnand and Young, 1998), which was used until recently in the UK. The sutures used in modern surgical practice are almost exclusively synthetic materials.

Clostridium Perfringens type A Toxin Production in 3 Commonly Used Culture Media

In vitro toxin production is an important tool not only for diagnostic purposes but also for the study of pathogenesis of Clostridium perfringens infections. The present study was carried out to compare the level of toxin production by several strains of C. perfringens type A, isolated from the intestine of animals, when cultured in 3 different conventional culture media. Six strains of C. perfringens type A isolated from the small intestine of healthy sheep were cultured in commercial cooked meat medium (CMM), brain heart infusion (BHI), and tryptone glucose yeast (TGY). Intravenous lethality in mice and phospholipase C (PLC) activity were measured in filtered culture supernatants. Lethality of culture supernatants was highest for all isolates when grown in BHI, followed by CMM. No supernatants from any isolates grown in TGY produced lethality in mice. Phospholipase C activity was highest when the isolates were grown in BHI and CMM and significantly lower when grown in TGY.

Bovine venereal campylobacteriosis: an overview.

Abstract Campylobacter fetus is a well-recognized pathogen of animals and humans. The organism has been divided into two subspecies, C. fetus subsp. venerealis and C. fetus subsp. fetus , that are highly related phenotypically and genotypically. Due to this relatedness, it has become imperative to differentiate these two subspecies in order to avoid misdiagnosis and highlight clonal origins geographically. The principal phenotypic differentiating test at the moment is the growth of C. fetus subsp. fetus in the presence of 1% glycine, but glycine-tolerant variants of C. fetus subsp. venerealis have also been described. Molecular tools such as polymerase chain reaction (PCR), amplified fragment length polymorphism (AFLP), numerical analysis of pulsed-field gel electrophoresis (PFGE)-DNA profiles and gene sequence analysis amongst others have been used to differentiate C. fetus subspecies; and they seem to hold promise for the future. C. fetus subsp. fetus colonize the intestines of cattle and sheep and is associated with sporadic abortions and enteritis, while, C. fetus subsp. venerealis is highly adapted to the genital tracts of cattle and is the causative agent of bovine venereal campylobacteriosis (BVC). This disease has a worldwide distribution and is of major economic concern to cattle industries in various parts of the world. The prevalence of the disease is highest in the developing countries where natural breeding in cattle is widely practiced. BVC is associated with lowered fertility, embryo mortality and abortion; other clinical features of the disease include many services per conception, poor pregnancy rates, long calving intervals, stillbirths and birth of weak calves that may eventually die. Vaccines have been shown to be useful in the protection of cattle against the menace of BVC, but commercial vaccines are only available in some parts of the world where advanced agricultural practices have limited its occurrence. The continued presence of this disease in the developing world poses a threat to sustainable cattle production efforts in these countries. Previous reviews highlighted the epidemiology, pathogenesis and diagnosis of the disease; in this review the clinical features of BVC, advances in diagnosis, treatment and prevention, as well as the geographic distribution of the disease have been reviewed. To achieve faster results worldwide, the need for collaborative research on BVC between investigators in developing countries and their counterparts in the developed countries has been recommended.

In vitro and in vivo tissue repair with laser‐activated chitosan adhesive

Sutures are currently the gold standard for wound closure but they are still unable to seal tissue and may induce scarring or inflammation. Biocompatible glues, based on polysaccharides such as chitosan, are a possible alternative to conventional wound closure. In this study, the adhesion of laser-activated chitosan films is investigated in vitro and in vivo. In particular we examine the effect of varying the laser power, as well as adding a natural cross-linker (genipin) to the adhesive composition. Flexible and insoluble strips of chitosan films (surface area approximately 34 mm(2), thickness approximately 20 microm) were bonded to sheep intestine using several laser powers (0, 80, 120, and 160 mW) at 808-nm wavelength. The strength of repaired tissue was tested by a calibrated tensiometer to select the best power. A natural cross-linker (genipin) was also added to the film and the tissue repair strength compared with the strength of plain films. The adhesive was also bonded in vivo to the sciatic nerve of rats and the thermal damage induced by the laser assessed 4 days post-operatively. Chitosan adhesives successfully repaired intestine tissue, attaining a maximum repair strength of 14.7+/-4.3 kPa (n = 30) at the laser power of 120 mW. The chitosan-genipin films achieved lower repair strength (9.1+/-2.9 kPa). The laser caused partial demyelination of axons at the site of operation, but the myelinated axons retained a normal morphology proximally and distally. The chitosan adhesive effectively bonded to tissue causing only localized thermal damage in vivo, when the appropriate laser parameters were selected.

Detection of Transgenic and Endogenous Plant DNA in Digesta and Tissues of Sheep and Pigs Fed Roundup Ready Canola Meal

The persistence of plant-derived recombinant DNA in sheep and pigs fed genetically modified (Roundup Ready) canola was assessed by PCR and Southern hybridization analysis of DNA extracted from digesta, gastrointestinal (GI) tract tissues, and visceral organs. Sheep (n = 11) and pigs (n = 36) were fed to slaughter on diets containing 6.5 or 15% Roundup Ready canola. Native plant DNA (high- and low-copy-number gene fragments) and the cp4 epsps transgene that encodes 5-enolpyruvyl shikimate-3-phosphate synthase were tracked in ruminal, abomasal, and large intestinal digesta and in tissue from the esophagus, rumen, abomasum, small and large intestine, liver, and kidney of sheep and in cecal content and tissue from the duodenum, cecum, liver, spleen, and kidney of pigs. High-copy chloroplast-specific DNA (a 520-bp fragment) was detected in all digesta samples, the majority (89-100%) of intestinal tissues, and at least one of each visceral organ sample (frequencies of 3-27%) from sheep and swine. Low-copy rubisco fragments (186- and 540-bp sequences from the small subunit) were present at slightly lower, variable frequencies in digesta (18-82%) and intestinal tissues (9-27% of ovine and 17-25% of porcine samples) and infrequently in visceral organs (1 of 88 ovine samples; 3 of 216 porcine samples). Each of the five cp4 epsps transgene fragments (179-527 bp) surveyed was present in at least 27% of ovine large intestinal content samples (maximum = 64%) and at least 33% of porcine cecal content samples (maximum = 75%). In sheep, transgene fragments were more common in intestinal digesta than in ruminal or abomasal content. Transgene fragments were detected in 0 (esophagus) to 3 (large intestine) GI tract tissues from the 11 sheep and in 0-10 of the duodenal and cecal tissues collected from 36 pigs. The feed-ingested recombinant DNA was not detected in visceral tissues (liver, kidney) of lambs or in the spleen from pigs. Of note, however, one liver and one kidney sample from the pigs (different animals) were positive for a 278-bp fragment of the transgenic cp4 epsps (denoted F3). Examination of genomic libraries from these tissues yielded no conclusive information regarding integration of the fragment into porcine DNA. This study confirms that feed-ingested DNA fragments (endogenous and transgenic) do survive to the terminal GI tract and that uptake into gut epithelial tissues does occur. A very low frequency of transmittance to visceral tissue was confirmed in pigs, but not in sheep. It is recognized that the low copy number of transgenes in GM feeds is a challenge to their detection in tissues, but there was no evidence to suggest that recombinant DNA would be processed in the gut in any manner different from endogenous feed-ingested genetic material.

Monocarboxylate transporter 1 gene expression in the ovine gastrointestinal tract

In this study, we investigated the tissue distribution and expression of monocarboxylate transporter 1 (MCT1) along the gastrointestinal tract of sheep. Western blot analysis suggested the presence of MCT1 as a 43-kDa protein in immunoblots of membranes from the various tissues examined. The results of Western blotting were further confirmed by immunohistochemical studies, which revealed intense immunoreactivity for the MCT1 protein in the forestomach (rumen, reticulum and omasum) and large intestine (caecum, proximal and distal colon). Moderate reactivity, however, was detected in the abomasum, while no immunoreactivity could be seen in any regions of the small intestine examined. Furthermore, MCT1 was expressed at the mRNA level as determined by reverse transcriptase polymerase chain reaction (RT-PCR), which showed a band of the expected size (300 bp) in all tissues examined. From these results we concluded that MCT1 protein is highly expressed and distributed in the stomach and large intestine of sheep suggesting that MCT1 may play a significant role in the transport of short chain fatty acids and their metabolites in the gastrointestinal tract of ruminants.

Morphologic and physiologic changes induced by Clostridium perfringens type A α toxin in the intestine of sheep

To evaluate the morphologic and physiologic changes induced by Clostridium perfringens type A (alpha toxin in the ileum and colon of sheep. 16 ligated intestinal loops in 4 Merino lambs and 18 explants of ileum and colon from slaughtered lambs. alpha Toxin-induced fluid accumulation was evaluated in ligated ileal and colonic loops of sheep. Tissues were evaluated morphologically by use of gross and histologic examination. Effects of toxin on in vitro intestinal net water transport were tested in modified Ussing chambers. Ovine ileal and colonic loops incubated with C perfringens type A alpha toxin retained more fluid than control loops. Histologically, in the ileum of lambs inoculated with 300 LD50 of alpha toxin/mL, there was a mild to moderate multifocal infiltration of neutrophils in the lamina propria and submucosa. The colonic loops of lambs inoculated with 30 or 300 LD50 of alpha toxin/mL had excessive mucus in the lumen, a moderate amount of neutrophils mixed with mucus in the intestinal lumen, and moderate multifocal infiltration of the lamina propria and submucosa with neutrophils; the blood vessels of these layers were engorged with neutrophils. In vitro measurements of water transport also revealed inhibition of net epithelial water absorption in ileum and colon incubated with alpha toxin on the mucosal side. These results indicate that alpha toxin induces alterations in sheep intestine. Clostridium perfringens type A organisms that produce alpha toxin could be responsible for diseases of intestinal origin in some ruminants.

Chitosan adhesive for laser tissue repair: In vitro characterization

Laser tissue repair usually relies on hemoderivate protein solders, based on serum albumin. These solders have intrinsic limitations that impair their widespread use, such as limited tensile strength of repaired tissue, poor solder solubility, and brittleness prior to laser denaturation. Furthermore, the required activation temperature of albumin solders (between 65 and 70 degrees C) can induce significant thermal damage to tissue. In this study, we report on the design of a new polysaccharide adhesive for tissue repair that overcomes some of the shortcomings of traditional solders. Flexible and insoluble strips of chitosan adhesive (elastic modulus approximately 6.8 Mpa, surface area approximately 34 mm2, thickness approximately 20 microm) were bonded onto rectangular sections of sheep intestine using a diode laser (continuous mode, 120 +/- 10 mW, lambda = 808 nm) through a multimode optical fiber with an irradiance of approximately 15 W/cm2. The adhesive was based on chitosan and also included indocyanin green dye (IG). The temperature between tissue and adhesive was measured using a small thermocouple (diameter approximately 0.25 mm) during laser irradiation. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were cultured in extracted media from chitosan adhesive to assess cytotoxicity via cell growth inhibition in a 48 hours period. Chitosan adhesive successfully repaired intestine tissue, achieving a tensile strength of 14.7 +/- 4.7 kPa (mean +/- SD, n = 30) at a temperature of 60-65 degrees C. Media extracted from chitosan adhesive showed negligible toxicity to fibroblast cells under the culture conditions examined here. A novel chitosan-based adhesive has been developed, which is insoluble, flexible, and adheres firmly to tissue upon infrared laser activation.

Comparative Histological Studies of Mechanically Versus Manually Processed Sheep Intestines Used To Make Natural Sausage Casings

The natural sausage casings industry is large and worldwide, and casings prepared from the small intestine of sheep form a large part of it. Food safety authorities in several countries have been concerned about the risk to consumers from the bovine spongiform encephalopathy (BSE) agent. Although this agent could enter the European small ruminant population via infected feed, there is no evidence that it has. Because the BSE agent introduced experimentally into sheep and goats has a tissue distribution very similar to that found in animals with natural cases of scrapie, the agent would likely be found in the intestine and lymph nodes of some infected sheep from an early age. When natural casings are prepared from the intestine, the ileum (known to be infected in animals with natural cases of scrapie) is removed and the intestine is cleaned such that the inner (tunica mucosa) and outer (tunica serosa and tunica muscularis) layers are removed, leaving only the submucosa. There are two main methods for cleaning the intestine: manual and mechanical. The cleaning efficiency of these two methods was examined in the commercial environment as practiced on healthy sheep considered fit for human consumption in Turkey and Great Britain. The investigation involved a qualitative and quantitative histological approach. There was no significant difference in cleaning efficiency between the two methods, although there was some variation. No Peyer's patches or residues of them were found in any part of the cleaned casings. This finding is important because in sheep infected with transmissible spongiform encephalopathies (TSEs) Peyer's patches are likely to contain a major part of the intestinal infectivity. No serosa was found in any casing, but some residual mucosa and muscularis was retained, with more of the former than the latter. The results indicate that the cleaning efficiency of the two methods was broadly equivalent, that there was significant removal of tissue that could promote TSE infection, and that TSE risk reduction likely would be achieved by both methods, although this probability could not be quantified by the methods used in this study.

Distribution and estimation of anaerobic zoosporic fungi along the digestive tracts of sheep

The status of anaerobic zoosporic (Chytridiomycota) fungi along the entire digestive tract of sheep was assessed both analytically and microscopically. Digest samples were taken from different segments of the digestive tracts of three newly killed sheep that previously had been used in experimental dietary studies. These digest samples were tested for the presence of rumen fungi by assessing the recovery of live fungi from the samples, direct observation of digested plant remains under the scanning electron microscope (SEM), and using a chitin assay as an estimation of fungal biomass. Live anaerobic fungi were recovered from the abomasum, small and large intestine, caecum and faeces of sheep, but not from the digest samples of rumen and omasum. However, SEM examination of the samples confirmed the presence of fungal structures from all of these organs. In the large intestine and caecum samples the observed sporangial structures were rounded and showed conspicuous surface pitting. Results of the chitin assay indicated that the anaerobic fungi might account for up to 20% of the total microbial biomass in the rumen of sheep. The results of this study support the view that anaerobic fungi may be present as a resistant stage in the lower reaches of the digestive tract.

Long-term collection and characterization of afferent lymph from the ovine small intestine

Reliable methods for long-term collection of afferent lymph draining from the small intestine of sheep are described and validated. The procedure was used successfully in normal sheep, in animals infected experimentally with the parasitic intestinal nematode Trichostrongylus colubriformis and in animals infected naturally with Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease. Our approach enabled afferent lymph draining from the small intestine to be collected continuously for up to 4 months, without any detrimental effects on the animals. Based on cytokine gene expression profiles of afferent intestinal lymph cells, the two infections induced contrasting regional immune responses, namely, Th2-type immunity in the case of T. colubriformis infection and Th1-type immunity in natural cases of Johne's disease. Some immune parameters differed markedly between the two disease models, highlighting the potential value of this approach to gain real-time insights into distinctive host–pathogen interactions as they occur in vivo within the regional immune system of the gastrointestinal tract.

Albumin-genipin solder for laser tissue repair.

Laser tissue soldering (LTS) is an alternative technique to suturing for tissue repair that avoids foreign body reaction and provides immediate sealing of the wound. One of the major drawbacks of LTS, however, is the weak tensile strength of the solder welds when compared to sutures. In this study, a crosslinking agent of low cytotoxicity was investigated for its ability to enhance the bond strength of albumin solders with sheep intestine. Solder strips were welded onto rectangular sections of sheep small intestine using a diode laser. The laser delivered in continuous mode a power of 170 +/- 10 mW at lambda = 808 nm, through a multimode optical fiber (core size = 200 microm) to achieve a dose of 10.8 +/- 0.5 J/mg. The solder thickness and surface area were kept constant throughout the experiment (thickness = 0.15 +/- 0.01 mm, area = 12 +/- 1.2 mm2). The solder was composed of 62% bovine serum albumin (BSA), 0.38% genipin, 0.25% indocyanin green dye (IG), and water. Tissue welding was also performed with a BSA solder without genipin, as a control group. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were also cultured in extracted media from heat-denatured genipin solder to assess cell growth inhibition in a 48 hours period. The tensile strength of the genipin solder was doubled that of the BSA solder (0.21 +/- 0.04 N and 0.11 +/- 0.04 N, respectively; P = 10(-15) unpaired t-test, N = 30). Media extracted from crosslinked genipin solder showed negligible toxicity to fibroblast cells under the culture conditions examined here. Addition of a chemical crosslinking agent, such as genipin, significantly increased the tensile strength of adhesive-tissue bonds. A proposed mechanism for this enhanced bond strength is the synergistic action of mechanical adhesion with chemical crosslinking by genipin.

Involvement of gut-associated lymphoid tissue of ruminants in the spread of transmissible spongiform encephalopathies

Scrapie is a transmissible spongiform encephalopathy (TSE) and its spread across the intestine of sheep is linked to the biology of intestinal Peyer's patches (PPs). Specialized epithelial cells, M cells, would appear to be the portal of entry for the scrapie agent, PrP Sc, while lymphoid nodules of PPs become major sites of accumulation of PrP Sc as the infection becomes established. Furthermore, evidence suggests that the enteric nervous system supplying the PPs is important for neuroinvasion. The gut-associated lymphoid tissue (GALT) of ruminants shows morphological and functional differences to the GALT of mice and humans. Recent investigations of aging scrapie-affected sheep revealed a substantial network of nerve fibres in the lymphoid nodules of PPs, contradicting the widely held notion that lymphoid nodules are poorly innervated. Advances in the understanding of the pathogenesis of scrapie may be achieved by a deeper appreciation of the development, morphology and function of GALT in small ruminants.