Swine dysentery: studies of gnotobiotic pigs inoculated with Treponema hyodysenteriae, Bacteroides vulgatus, and Fusobacterium necrophorum

Transmission experiments were carried out in gnotobiotic pigs to determine whether lesions typical of swine dysentery could be produced by oral inoculation of Treponema hyodysenteriae in combination with Bacteroides vulgatus or Fusobacterium necrophorum, or both. Each of the organisms had been isolated from swine with early lesions of the disease. Lesions were not found in 6 pigs inoculated with T hyodysenteriae alone, in 4 pigs given F necrophorum and T hyodysenteriae, or in 4 pigs given B vulgatus and F necrophorum. Lesions typical of swine dysentery developed in 8 pigs given B vulgatus, F necrophorum, and T hyodysenteriae as well as in 3 of 4 pigs given B vulgatus and T hyodysenteriae. In both of these groups, the inoculated bacteria were recovered from the colon, and T hyodysenteriae was demonstrated in the colonic crypts, epithelium, and lamina propria. The pathogenicity of the T hyodysenteriae was shown by the development of characteristic signs and lesions of swine dysentery in 12 of 14 naturally farrowed pigs inoculated with T hyodysenteriae alone.     

Influence of Diet on Experimental Swine Dysentery. 3. Pathological Changes

Three experiments have been carried out to elucidate the possible influence of the vitamin E and selenium content of the feed on experimental swine dysentery. In most of the pigs given a vitamin E and selenium deficient diet, large and diffuse pseudomembranes appeared in the spiral colon, which also usually displayed a distended appearance and prominent oedematous infiltrations in the mesentery. The histological examination revealed large fibrinous pseudomembranes attached to defects on the mucosal surface. There were also pseudo-membranes containing necrotic mucosal tissue. Fibrinoid thrombi within minute vessels were readily observed in the latter lesions. The vitamin E supplemented pigs had colonic lesions very much like the deficient animals, while half of the selenium supplemented animals developed none or moderate inflammatory changes, the other half displayed, however, prominent pseudomeinbranes in the colon. Ten out of 26 pigs supplemented with both vitamin E and selenium were not affected by swine dysentery. In the remaining pigs catarrhal inflammatory lesions dominated in the colonic mucosa. In some of these animals pseudomembranes occurred, but they were usually small and of limited distribution. The vascular thromboses and tissue necrosis demonstrated within the colonic lesions are found to be compatible with a Shwartzman reaction. Erythrocytic “thrombi” and other phenomena associated with stasis are further believed to be of pathogenetic importance in this respect. It is emphasized that this report also illustrates the enhancing effect of a combined supplement of vitamin E and selenium on resistance to swine dysentery.     

Studies on the pathogenesis of swine dysentery. I. Characterization of the lesions in colons and colonic segments inoculated with pure cultures or colonic content containing Treponema hyodysenteriae.

Swine dysentery was induced in pigs and in ligated colonic segments by inoculation of pure cultures of, or colonic contents containing, Treponema hyodysenteriae. The mildest changes, best seen in ligated segments 48 or 72 hours after inoculation, were congestion and leucocytic margination in mucosal capillaries and depletion of mucigen from goblet cells lining the base of the crypts of Lieberkühn. Superficial mucosal necrosis and crypt cell hyperplasia were later changes. Perfusion studies with India ink did not demonstrate occlusive mucosal ischemia in acute swine dysentery. Mucosa with lesions of swine dysentery contained at least 10(5) colony forming units of T. hyodysenteriae per gram. Mucosa without lesions had 10(5) or fewer T. hyodysenteriae per gram. Segments with acute swine dysentery were distended with clear mucoid fluid with electrolyte composition indicative of net colonic secretion. No increase in the concentration of volatile fatty acids was detected in content from intact colons or colonic segments with lesions of acute swine dysentery.     

Influence of Diet on Experimental Swine Dysentery. 1. Effects of A Vitamin E and Selenium Deficient Diet Supplemented with 6.8 % Cod Liver Oil

Sixteen growing pigs were fed a vitamin E and selenium deficient diet; half of the animals (Group 2) were given a daily supply of vitamin E and selenium. After having been fed these diets for 53 days, the pigs were infected orally with minced colonic material from cases with typical swine dysentery. This exposure resulted in outbreaks of swine dysentery in both groups. The incubation times were, however, distinctly shorter and the clinical symptoms much more pronounced in Group 1 than in Group 2. The patho^morphological lesions in the colon also differed between the 2 groups. In the pigs of Group 1 evident pseudomembraneous lesions were observed in the spiral colon. In Group 2, the colonic alterations consisted predominantly of a catarrhal enteritis; pseudomembranes occurred in a minor part of colon in only 4 pigs. Both the clinical and the chemical observations and the pathological findings indicated a much better vitamin E and selenium balance in the pigs of Group 2. It is concluded that the treatment with vitamin E and selenium in Group 2 greatly increased resistance to swine dysentery.     

Intestinal spirochetosis in a 21-month-old thoroughbred colt

A 21-month-old Thoroughbred colt showed continuous diarrhea and developmental retardation for 7 months, and was thereafter subjected to euthanasia for necropsy and laboratory examinations. At necropsy, the cecal and colonic mucosae were diffusely rough and hyperemic. Histopathologically, the mucosa and submucosa were edematous and were infiltrated by numerous lymphocytes and macrophages. Meanwhile, three morphological types of Brachyspira antigen-containing spirochetes were found to be numerous in the crypts and in the mucus layer over the epithelium in the cecal and colonic lesions. They were frequently observed in intercellular gaps and in the cytoplasm of degenerative epithelial cells, and in the lamina propria, particularly in cavities around blood vessels. These invasive intestinal spirochetes might be one of pathogens inducing colitis and diarrhea in horses.     

Induction of swine dysentery in swine by the intravenous injection of filtered Treponema hyodysenteriae.

Swine dysentery was induced in 18 swine exposed by intravenous injection of a filtrate which contained Treponema hyodysenteriae and was obtained from macerated colonic scrapings of swine dysentery. However, swine dysentery did not develop in swine injected intravenously with a pure culture of T. hyodysenteriae or when combined with a colonic filtrate from normal swine. Diarrheal feces from the swine injected intravenously with the filtered T. hyodysenteriae contained more mucus, and fecal smears contained more T. hyodysenteriae and fewer other bacteria than did swine exposed orally to colon infected with swine dysentery or filtered T. hyodysenteriae. In the colons of the 12 swine injected intravenously with filtered T. hyodysenteriae that died, there was a minimum amount of croupous membrane and, microscopically, the T. hyodysenteriae were located deep in the colonic crypts. Five of the six surviving swine injected intravenously with filtered T. hyodysenteriae developed serum anti-T. hyodysenteriae antibodies using the indirect fluorescent antibody test and four of these swine developed diarrhea when reexposed with swine dysentery infected colon six weeks after initial exposure. None of the swine injected intravenously with cultured T. hyodysenteriae developed serum anti-T. hyodysenteriae antibodies and all were highly susceptible to swine dysentery.     

Immunofluorescence of spirochetes with serum from swine recovered from swine dysentery using an indirect fluorescent antibody test.

Using an indirect fluorescent antibody test, immunofluorescence of large spirochetes was observed with serum from swine that had recovered from swine dysentery. The spirochetes were obtained from scrapings of the colonic mucosa on the first day of diarrhea which was the time when the spirochete population was observed to be the highest. Of 29 exposed nonmedicated swine which developed and recovered from a diarrhea characteristic of swine dysentery 27 had antispirochete serum titers which ranged from 1:2 to 1:16. None of the 50 nonexposes swine developed a titer. Of 19 swine with a serum titer and reexposed with infective swine dysentery inoculum, 18 did not develop a diarrhea and were presumed to be immune. Considering these findings it is possible that this test could be used to detect antispirochete antibody in unknown swine serum.     

Influence of Diet on Experimental Swine Dysentery: 2. Effects of a Vitamin E and Selenium Deficient Diet Supplemented with 3 % Cod Liver Oil,Vitamin E or Selenium

Two experiments (Exps. II and III) were performed with colonic material from swine dysentery as inoculum. The results of Exp. II showed that the inoculation produced less pronounced clinical signs and patho-morphological lesions and also affected fewer pigs in the groups supplemented with vitamin E and selenium than in the group not given a corresponding supplementation. It is therefore concluded that the daily supply of these 2 nutritional factors significantly increased the resistance to swine dysentery. The cod liver oil incorporated in the diet of 2 groups in Exp. II had also a positive effect in this respect. In Exp. III the inoculation of the pigs fed only the basic ration (Group 1) produced relatively moderate clinical signs and patho-morphological lesions in half of the animals, the results in the rest of the group are, however, in accordance with the observations in the corresponding groups of the 2 preceding experiments. The other results of Exp. III indicate an increased resistance to swine dysentery in the group with selenium as the only supplement. However, no equivalent condition could be demonstrated when a similar supplement of vitamin E was used. The pigs in Exp. III given both nutritional factors showed perhaps the best resistance to swine dysentery, especially when the patho-morphological lesions are taken into consideration.     

Probable elimination of swine dysentery after feeding ronidazole, carbadox or lincomycin and verification by feeding sodium arsanilate.

Swine dysentery did not recur during a nine week period after withdrawal of medication in swine fed ronidazole at a level of 60 parts per million of feed for ten weeks or fed either carbadox at 55 ppm or lincomycin at 110 ppm of feed for six weeks. During this period swine dysentery was neither transmitted to accompanying sentinels after the withdrawal of the above medication or was Treponema hyodysenteriae isolated and cultured or observed in stained smears from rectal swabs and feces or from colonic scrapings at necropsy. Beginning three weeks after the withdrawal of medication, all swine were fed sodium arsanilate at a concentration of 220 ppm of feed for three weeks in an attempt to excite the carrier of swine dysentery into developing a swine dysentery diarrhea. A swine dysentery diarrhea did recur during the feeding of sodium arsanilate in swine previously fed ronidazole at a level of 60 ppm of feed for only six weeks. It was concluded: that swine dysentery was probably eliminated with the feeding of ronidazole for the longer duration and with the feeding of carbadox and lincomycin and that sodium arsanilate was of value in identifying the carrier state.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle, Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle,Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

Enterocecocolitis associated with intraepithelial Campylobacter-like bacteria in rabbits (Oryctolagus cuniculus)

We examined 28 suckling, weanling, and young adult rabbits with lethargy, inappetence, and mucinous, semifluid feces. Sixteen of the rabbits had intestinal lesions. In eight of these rabbits, the primary changes were multifocal to diffuse epithelial proliferation and accumulation of lymphocytes, macrophages, or both in the lamina propria of the small intestine, cecum, and sacculated colon. In two of these rabbits, the accumulation of macrophages in the lamina propria was extensive. The other eight rabbits had erosive and suppurative cecocolitis, and four of the rabbits with proliferative lesions also had suppurative cecocolitis. In Warthin-Starry-stained sections of affected intestine, curved or spiral bacteria were visible within degenerated or hyperplastic epithelium, in luminal exudate, or in both. Such organisms were sparse or not found in the other 12 rabbits, which did not have intestinal lesions. The bacteria ultrastructurally resembled intraepithelial Campylobacter-like bacteria previously observed in proliferative enteritis in a variety of species and in acute typhlitis in young rabbits. In immunofluorescence tests, Campylobacter-like bacteria in epithelial cells, crypt lumina, and in luminal exudates in both proliferative and erosive lesions bound monoclonal antibodies and polyclonal antisera prepared against intracellular bacteria found in proliferative enteritis in pigs, hamsters, and ferrets. These observations indicate that a condition similar to proliferative enteritis of swine, hamsters, and other species also occurs in laboratory rabbits.     

Pathogenic Shiga toxin-producing Escherichia coli in the intestine of calves

The purpose of this study was to compare the pathological effects of Shiga toxin-producing Escherichia coli (STEC) that vary in their association with bovine and human disease. Shiga toxin-producing E. coli of serotypes associated with both dysentery in calves and hemolytic uremic syndrome (HUS) in humans (O5:H-, O26:H11, O111:H-, O113:H21) were compared with O157:H7 STEC, which are associated with HUS in humans but not with disease in calves. The STEC were administered orally to 80 day-old chicks and into ligated loops in the ileum and colon of four 2- to 6-day-old calves. Examination of the ceca of the chickens 10 d postchallenge showed no adherence or tissue abnormality for any isolate. The calves were euthanized 8 to 10 h postinoculation, and sections of the intestinal loops were examined by light microscopy, transmission and scanning electron microscopy, and immunohistochemistry. All strains showed consistent focal adherence associated with mild lesions in the colon. Attaching and effacing lesions were observed with the eae-positive strains. Ileal lesions were similar to the colonic ones but were sometimes severe, with marked polymorphonuclear leukocyte proliferation in the lamina propria. It is concluded that chickens were unsuitable for studying interaction of STEC with the intestine and that there was no difference in the interaction of the ligated calf intestine with STEC of serotypes associated with disease in calves compared with O157:H7 STEC.     

Pathophysiologic features of swine dysentery: cyclic nucleotide-independent production of diarrhea

Net electrolyte and water transport and unidirectional Na+ fluxes were examined in ligated colonic loops of clinically normal pigs and in pigs with swine dysentery (etiologic agent Treponema hyodysenteriae) in the presence or absence of theophylline. In normal pigs, theophylline abolished net Na+ absorption via a reduction in the lumen-to-blood flux, decreased Cl- absorption, and increased HCO3- accumulation in the lumen. In infected pigs, all net ion transport was abolished, with the addition of theophylline producing little effect. The absence of net Na+ absorption in infected pigs was also the result of a decreased lumen-to-blood flux. Seemingly, colonic malabsorption may be the primary transport alteration in swine dysentery. Concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were measured in samples of colonic mucosa from normal and infected pigs after in vitro exposure to a Ringer's solution containing 0 or 20 mM theophylline. Basal values of cAMP or cGMP did not increase in infected colonic mucosa. There was a diminished capacity of the infected mucosa to respond to theophylline. Alterations in ion transport in conjunction with measurements of cAMP and cGMP indicated that the pathogenic mechanism(s) in swine dysentery were not similar to those of Salmonella, Shigella, Vibrio cholerae, or Escherichia coli diarrhea.     

Morphology of the non-sensory tissue components in rat aging vomeronasal organ

With 30 figures, 3 histograms and 3 tables SUMMARY: The vomeronasal organ (VNO) is a chemosensory organ that detects environmental pheromones. The morphology of the 'non-sensory' epithelium (NSE) of the VNO and its lamina propria, as well as how it relates to ageing has received little attention. Histological, histochemical, morphometric and ultrastructural techniques were used to study the morphological structure of the rat NSE in five adult (3 months old) and five aged (2-2.5 years old) male albino rats. In adult rats, the NSE contained dark and light columnar cells with predominance of the latter. The surface of the epithelial cells was covered with microvilli and/or cilia. The lamina propria contained serous vomeronasal glands (VNGs), smooth muscles with numerous variable-sized mitochondria, vessels including lymphatic capillaries and nerve bundles. The following changes were detected in aged rats. The NSE exhibited an increase in number of dark columnar cells. Some cells revealed a prominent cell coat, dense aggregation of filaments in the luminal cytoplasm and appearance of multinucleated cells. Their surface revealed malformed configuration. Large mitochondria (2 μm), formed by fusion, were frequently observed in the smooth muscle cells of the lamina propria. Lipid droplets were frequently detected both in the VNGs acini and in the lymphatic endothelium. Ageing affected both the cells of the tissues and the extracellular matrix.     

Studies on the pathogenesis of swine dysentery. II. Search for a cytotoxin in spirochetal broth cultures and colon content.

Broth cultures of Treponema hyodysenteriae and colonic content from pigs with swine dysentery were tested for cytotoxicity in cell cultures, erythrocyte suspensions and in ligated segments of pig colon. Live cells of T. hyodysenteriae attached to the surface of cells in all cultures tested but did not penetrate them nor cause morphologic change detectable by light microscopy. Only live T. hyodysenteriae caused erythrolysis. Broth cultures or colonic content sterilized by filtration or by disruption with ultrasound had no visible effect on the cell cultures, erythrocyte suspensions or the mucosa of ligated colonic segments.     

Uremic gastropathy in the dog.

Stomachs of four dogs with uremia and four normal dogs were examined. Uremic stomachs represented four types of disease: atrophic, amyloidotic, ulcerative and necrotic gastropathy. Pathologic changes common to all uremic stomachs were expansion of the lamina propria, atrophy of gastric glands, and submucosal arteriopathy; lesions were limited to body and fundic zones. Lamina propria was markedly expanded by edema, mastocytosis, deposition of acidic mucosubstances, fibroplasia and mineralization. Capillaries in lamina propria had swollen endothelium and calcium salts were present extracellularly as amorphous granular laminae. Gastric glands were distorted and irregular and had fewer cells per unit of tissue. Parietal cells were swollen and had fragmentation of cytocavitary network and mitochondrial swelling with calcification. Chief cells were shrunken, agranular and atrophic with foci of glycogen and dilation of endoplasmic reticulum. Argentaffin cell content was diminished. Muscular arteries of submucosae had segmental degenerative lesions characterized by myocyte necrosis, calcification, and deposition of acidic mucosubstances and fibrin; thrombosis and obstructive arteriopathy were common. These studies suggest that uremic gastropathy is a disease of mucosal lamina propria and that lesions were due to anoxia caused by diffuse vascular injury and to altered parietal cell function.     

Campylobacter jejuni colitis in gnotobiotic dogs.

Campylobacter jejuni of human and canine origin was inoculated orally into six gnotobiotically reared Beagle puppies and reactions were compared with two controls. Inoculated dogs developed transient lassitude, inappetence, mild diarrhea and tenesmus during the period 36-72 hours after inoculation. Pairs of dogs killed 43 hours, and five and seven days after inoculation had lesions limited to typhlitis and colitis. Congestion of colonic mucosa, associated loss of goblet cells, attenuation and exfoliation of surface epithelium with microerosions, hypertrophy of glands and neutrophil infiltration of lamina propria were seen during the acute phase. Less severe surface and inflammatory lesions were evident at five and seven days, with hyperplasia of the proliferative compartment in mucosal glands. Campylobacter established at over 10(10) organisms per gram of colonic content but did not invade the mucosa. It was concluded that the gnotobiotic dog may be a suitable model for investigation of the pathogenesis of Campylobacter colitis.     

Porcine colonic spirochetosis: a retrospective study of eleven cases.

This retrospective study of porcine colonic spirochetosis was done in order to characterize the clinical signs, the macroscopical and microscopical changes, and the bacteriological results of cases observed in Quebec. Necropsy records of all cases of colitis were reviewed. Eleven cases with filamentous bacteria colonizing the colonic epithelium were selected. This condition was only observed in weaned piglets, and was associated with mild persistent diarrhea and growth retardation. Macroscopic changes were generally limited to the presence of soft to liquid colonic contents. Adherence of filamentous helicoidal bacteria to the apical surface of the colonic epithelium and mild diffuse infiltration of the colonic lamina propria by mononuclear cells were the main histological findings. Weakly beta-hemolytic spirochetes were isolated in 6 cases. This condition seems to be underestimated for various reasons, and it is possible that some cases diagnosed as nonspecific colitis or superficial colitis, in fact, represent later stages of porcine colonic spirochetosis.     

Flow cytometric analysis of canine colonic mucosal lymphocytes from endoscopically obtained biopsy specimens

OBJECTIVE: To validate use of canine colonic biopsy specimens obtained via endoscopy as a source of mucosal lymphocytes (ML) for flow cytometric analysis. SAMPLE POPULATION: Mucosal biopsy specimens from 10 adult dogs. PROCEDURE: Mucosal lymphocyte subsets obtained from excised colon were compared with ML subsets obtained from biopsy specimens obtained by use of an endoscopic forceps (6 dogs). Endoscopic colonic biopsy specimens from 4 other dogs were used to define whether obtained ML were predominantly of intraepithelial or lamina propria origin. Mucosal lymphocytes were isolated and labeled, using commercially available monoclonal antibodies directed against canine cell surface antigens. Lymphocyte subsets (cytotoxic or helper T cells; B cells) were determined by use of flow cytometric analysis. RESULTS: A large number of viable ML was obtained after dissociation of the colonic epithelium from excised colon (45.5 + 21.5 X 10(6)) and endoscopic (7.2+/-3.4 X 10(6)) biopsy specimens. Lymphocyte subsets obtained with both methods were identical for each dog and consisted predominantly of intraepithelial lymphocytes, with some lymphocytes from the lamina propria. Collagenase digestion of excised colon also yielded a large number of viable lymphocytes from the lamina propria (56.7+/-20.4 X 10(6)), but collagenase digestion of endoscopic biopsy specimens was less rewarding. CONCLUSION AND CLINICAL RELEVANCE: A representative sample of viable intraepithelial ML is obtainable from endoscopic biopsy specimens. Flow cytometric analysis, a minimally invasive technique, can be used to study ML of client-owned animals.