Lesions in the small intestine of newborn pigs inoculated with porcine, feline, and canine coronaviruses

The infectivity and pathogenicity to newborn pigs of antigenically related coronaviruses from pigs (transmissible gastroenteritis virus; TGEV), cats (feline infectious peritonitis virus; FIPV), and dogs (canine gastroenteritis virus; CGEV) were studied by light, scanning electron, and immunofluorescence microscopy. Hysterectomy-derived, 12-hour-old pigs were orally given tissue culture or frozen preparations of 6 coronavirus strains (3 porcine, 2 feline, and 1 canine). The pigs were killed at regular intervals between 24 and 144 hours after exposure. Virulent TGEV and virulent FIPV produced necrosis of villous epithelium, resulting in villous atrophy in the jejunum and the ileum. Similar, but less extensive and severe lesions, were produced by the 4 other viruses. Coronaviral antigens were identified by immunofluorescence in villous epithelial cells of pigs that had been inoculated with virulent TGEV, attenuated TGEV, virulent FIPV, and tissue culture-adapted FIPV. In contrast, coronaviral antigens were not induced by the small plaque variant TGEV and virulent CGEV in the villous epithelium, but rather in cells of the lamina propria and crypt epithelium.     

Functional significance of histologic alterations induced by Escherichia coli pig-specific,mouse-negative,heat-stable enterotoxin (STb)

In contrast to cholera enterotoxin and other Escherichia coli enterotoxins, a pig-specific, heat-stable E. coli enterotoxin (ST_b) causes morphologic lesions (loss of villous epithelial cells and partial villous atrophy). These lesions reflect a loss of absorptive cells and thus suggest that ST_b causes impaired absorption as well as inducing net secretion. The present studies assess functional significance of morphologic changes induced by ST_b. Net fluid movement, mucosal surface area, sucrase activity and the electrical response induced by alanine were measured in swine jejunal loops exposed to E. coli culture filtrates with and without ST_b. Net fluid secretion (-11.1±1.1 ml) occurred in some ST_b loops (secretors) and net absorption (2.7±0.3 ml) in others (nonsecretors), but net absorption occurred in all control loops (4.9±0.2 ml). The mucosal surface area of St_b loops was about 20% less than that of controls (P<0.01). Sucrase activity was also lower (about 15%) in ST_b loops than in control loops (P<0.01). The electrical response induced by alanine in mucosa from nonsecreting ST_b loops did not differ from that induced in mucosa from control loops. However, the response to alanine in mucosa from secreting ST_b loops was reduced about 70% from that in mucosa from nonsecreting ST_b loops or from control loops (P<0.05). It is concluded that reduced sucrase activity is a functional correlate to villous atrophy induced by ST_b, that ST_b impairs alanine absorption in some loops (secretors), and that the impaired alanine absorption is independent of the decreased surface area caused by St_b. Because the impaired alanine absorption occurred independent of the decreases in surface area, it is suggested that the secretory response to ST_b is associated with an impairment of active absorption of alanine.     

Microscopic alterations in jejunal epithelium of 3-week-old pigs induced by pig-specific, mouse-negative, heat-stable Escherichia coli enterotoxin

The objective of this study was to determine whether exposure of swine jejunum to crude culture filtrates containing Escherichia coli pig-specific, mouse-negative, heat-stable enterotoxin (STb) induces structural alterations in the jejunal mucosa of pigs. Two ligated intestinal loops in each of twelve 3-week-old pigs were exposed for 2 hours to sterile E coli culture filtrates from each of the following strains: 431 (STa-producing), 1261 (STa and STb-producing), and 1790 (STb-producing); recombinant strain HB101-pRAS-1 (STb-producing); the nontoxigenic K-12 variant HB101; or trypticase soy broth. Formalin-fixed sections from these loops were examined for sloughed cells around villi, and a lesion score was determined, indicating a change in villous epithelium from columnar to cuboidal or squamous cell types or to discontinuous epithelium. Villous lengths and crypt depths also were determined. For loops exposed to culture filtrates containing STa and STb or containing only STb, lesion scores and numbers of sloughed cells were greater (P less than 0.05) and villous length was shorter (P less than 0.01) than in loops not exposed to toxin. For loops exposed to culture filtrates containing STa, lesion scores, villus lengths, and numbers of sloughed cells were not different from those of loops not exposed to toxin. Therefore, exposure of swine jejunum to STb induced structural alterations in intestinal mucosa (ie, loss of villous absorptive cells and partial atrophy of villi) that were consistent with those causing compromised absorptive capacity.     

Pathogenesis of porcine enteric calicivirus-like virus in four-day-old gnotobiotic pigs

Eighteen 4-day-old gnotobiotic pigs were orally inoculated with porcine enteric calicivirus-like virus (C strain). Seven additional gnotobiotic pigs served as noninoculated controls. Mild diarrhea developed in all inoculated pigs by postinoculation day (PID) 3 and persisted for 3 to 7 days. Severe diarrhea developed in 2 inoculated pigs between PID 4 and 5. Twelve inoculated and 7 control pigs were euthanatized over a 7-day period. Small intestinal mucosal smears were stained with a fluorescein-conjugated anti-porcine enteric calicivirus-like virus serum. Immunofluorescence was observed in villous epithelial cells (primarily in the duodenum or jejunum) of all inoculated pigs, except for 1 pig euthanatized at PID 7. Villus length was determined in histologic sections of the small intestinal specimens from control and inoculated pigs. Statistically significant (P less than 0.01) villus atrophy was found in the duodenum and/or jejunum of inoculated pigs at PID 3 to 7. These observations were confirmed by scanning electron microscopy, which revealed shortening, blunting, fusion, or absence of villi in the duodenum and jejunum of inoculated pigs at PID 3 to 7. Lesions were not seen in control pigs. Calicivirus-like particles were detected by immune electron microscopy in the large intestinal contents and feces of inoculated pigs from PID 1 to 7.     

Campylobacter sputorum subsp mucosalis and Campylobacter hyointestinalis infections in the intestine of gnotobiotic pigs

At 4 days of age, 7 gnotobiotic pigs were orally inoculated with broth cultures of both Campylobacter sputorum subsp mucosalis (CSM) and Campylobacter hyointestinalis (CH). One pig was killed and evaluated each week for 7 weeks. Forty-eight hours after inoculation, CH and CSM were recovered from the feces of the pigs; thereafter, only CH was recovered. Organisms morphologically typical of Campylobacter sp were observed on the mucosal surface and on the crypt epithelial cells of the ileum, cecum, and colon from post-inoculation weeks (PIW) 2 through 7. Bacteria were clustered around the surface opening of goblet cells in pigs at PIW 6 and 7. Crypt epithelial cell proliferation and intracellular bacteria were not seen, except in 1 pig (killed at PIW 7) in which intracellular bacteria were seen only in the cecum. Therefore, CSM and CH did not induce porcine proliferative enteritis in gnotobiotic pigs.     

The pathogenesis of an enteric infection in pigs,experimentally induced by the coronavirus-like agent,CV 777

Sixteen 2–3-days-old caesarean-derived, colostrum-deprived piglets were each dosed oro-nasally with 2 ml of a bacteria-free filtrate containing 10⁴ pig-infectious-doses of CV 777. The piglets were killed at intervals of 12 to 120 h after infection. The coronvirus-like agent caused a local infection of the intestinal tract which resulted in villous atrophy, malabsorption and diarrhea. The pathogenesis of this infection was similar to that of transmissible gastroenteritis (TGE), a known coronaviral infection of pigs. However, where were some differences. By immunofluorescent staining, CV 777 antigens were not only detected in the epithelial cells covering the small intestinal villi, but also in the cells of the colonic surface epithelium. Occasional fluorescence was also seen in the small intestinal crypt epithelium, but the regenerative capacity of the crypts was not affected. The progress of intestinal epithelial cell infection by CV 777 was much slower than that in TGE, resulting in a longer incubation period and in less drastic epithelial cell destruction. The infection of regenerating cells occurred to a much higher degree during the late stage of a CV 777 infection than has been observed in TGE.     

Age-dependent resistance to transmissible gastroenteritis of swine. III. Effects of epithelial cell kinetics on coronavirus production and on atrophy of intestinal villi.

Coronavirus titers in small intestine, degree of villous atrophy and apparent rates of regeneration of intestinal villi were compared in newborn, 3-week-old and adult pigs for 1 week after they were exposed to the transmissible gastroenteritis virus of swine. The response within the newborn group was homogeneous, resulting in high virus titers, maximal villous atrophy and comparatively slow regeneration. In general, virus titers were lower, villous atrophy was less severe and regeneration more rapid in both older groups than in the newborn pigs. However, the response varied greatly in the older groups. The 3-week-old group was divided into two populations. The major population had low virus titers and developed partial villous atrophy, whereas the minor population had marked villous atrophy and virus titers comparable to those of the newborn pigs. These observations support the hyposthesis that the accelerated replacement of villous epithelium in the small intestine of pigs during the first 3 weeks contributes to the innate age-dependent resistance to transmissible gastroenteritis. The accelerated replacement of villous epithelial cells in older pigs contributes to resistance in two ways. The increased proliferative capacity of crypt epithelium results in a more rapid regeneration of atrophic villi; and the comparatively young villous absorptive cells resulting from accelerated replacement produce less virus per cell than the older ones of the newborn pig.     

Immunolocalization of epidermal growth factor (EGF) and EGF receptors in the porcine upper gastrointestinal tract.

The factors controlling growth and maturation in the porcine gastrointestinal tract are not well understood. Epidermal growth factor (EGF) is a polypeptide that has been implicated in the control of gastrointestinal tract growth, maturation, and protection in other species. Immunoreactive EGF (IR-EGF) and EGF receptors (EGF-R) were histochemically identified in formalin-fixed tissues of the upper digestive tract of 1-, 3-, 7-, 14-, 21-, and 28-day-old pigs. The ductal epithelium consistently contained IR-EGF in the parotid salivary gland of pigs of all ages and in the mandibular salivary gland in pigs greater than or equal to 7 days old. Immunoreactive EGF was detected in the mucosal epithelium of the esophagus and nonglandular portion of the stomach, and in the pancreas and liver in all pigs. Gastric gland IR-EGF was inconsistently detected in pigs less than 14 days old and was consistently observed in all older pigs. Enterocyte EGF immunoreactivity was usually weak and was variably detected in the duodenum of pigs less than or equal to 7 days old and in the jejunum of pigs less than or equal to 14 days old, but was consistently observed in older pigs. Ileal immunoreactivity was erratic. Immunoreactive EGF-R were identified in the esophageal epithelium of all pigs, and in the nonglandular gastric and glandular gastric mucosa of all pigs, except for two 7-day-old pigs and one 7-day-old pig, respectively. Immunoreactive EGF-R were detected in the duodenal, jejunal, and ileal enterocytes of pigs of all ages examined.     

Morphologic studies of intrapulmonary airway mucosa-associated lymphoid tissues in swine

Morphologic structures of the lymphoid tissues associated with the mucosa of the respiratory tract in swine aged 11 to 13 weeks were studied by dissecting, light, and transmission electron microscopy. Tissues along the respiratory tract were collected for further examination. Light microscopic examination showed that the lymphoid tissues were associated with bronchi (8.38%), bronchioles (81.63%), and respiratory bronchioles (9.98%). The lymphoid tissues were identified at different levels of the airways and were designated intrapulmonary airway mucosa-associated lymphoid tissues. Anatomically, the lymphoid tissues were found mostly in the bronchiolar bifurcations, with an arteriole frequently nearby. Histologically, the lymphoid tissues contained a single dome-shaped lymphoid follicle, which bulged out from the mucosal surface into the airway, where it was covered along the mucosal side by a layer of specialized epithelium. Ultrastructurally, the mucosal surface of the specialized epithelial cells was equipped with microfolds or cytoplasmic projections instead of cilia. The cytoplasm was elongated, and therefore much thinner than that of ordinary epithelial cells. Many lymphocytes were found scattered in the mucosal epithelium between or beneath the specialized epithelial cells. This structure was similar to that seen in the Peyer's patches of the small intestine in swine and other animals.     

Scanning electron microscopy and fluorescent in situ hybridization of experimental Brachyspira (Serpulina) pilosicoli infection in growing pigs

Two groups of six 8-week-old pigs were challenged with 1x10(9) cfu Brachyspira (Serpulina) pilosicoli or Serpulina intermedia daily for 3 consecutive days to study the pathology of porcine colonic spirochetosis by scanning electron microscopy (SEM) and fluorescent in situ hybridization (FISH) with oligonucleotide probes targeting ribosomal RNA specific for B. pilosicoli and the genus Brachyspira/Serpulina. Six pigs served as noninoculated controls. The animals were euthanatized successively between postinoculation days 14 and 24. B. pilosicoli was reisolated in feces from all of the inoculated pigs; however, only two pigs developed transient watery diarrhea. S. intermedia was reisolated from four of the inoculated pigs, but clinical signs were not observed. Gross examination of the B. pilosicoli-infected pigs revealed dilated large intestines with a hyperemic mucosa, whereas the large intestines of the S. intermedia-inoculated pigs and the control pigs appeared normal. SEM examination of B. pilosicoli-infected pigs revealed degenerated epithelial cells and spirochetal colonization of the colonic mucosa in four pigs. By FISH, B. pilosicoli cells were found colonizing and invading the surface epithelium and the crypts in all the pigs. Spirochetal crypt colonization markedly exceeded the occurrence of spirochetes on the mucosal surface. SEM examination of S. intermedia-inoculated pigs revealed no abnormalities, and Serpulina cells were detected only sporadically in the otherwise normal-appearing mucosa of four pigs by FISH. The results provide further evidence that B. pilosicoli is associated with colitis in pigs, although the gross lesions are mild. The spirochete is capable of colonizing the large intestine, inducing mucosal damage, invasion of the crypt and surface epithelium, and focal infiltration of the lamina propria. In addition, the study shows the applicability of FISH for specific identification of B. pilosicoli in formalin-fixed tissue.     

Pathogenicity of Escherichia coli O115:K"V165" strains isolated from pigs with diarrhea

Eighteen strains of Escherichia coli serogroup O115:K"V165" isolated from 1- to 8-week-old pigs with diarrhea were tested for toxigenicity, pathogenicity in pigs and mice, serum resistance, mannose-resistant hemagglutination (MRHA), F165 and other surface antigens, colicin V (Col V), aerobactin, and biotype. Twelve strains were positive for heat-stable enterotoxin (STb), MRHA-negative, and F165-negative; 5 strains were enterotoxin-negative, MRHA-positive, and F165-positive; and 1 strain was MRHA-positive, but F165- and enterotoxin-negative. Six of the 12 STb-positive strains moderately colonized the ileum of newborn colostrum-deprived pigs within 24 hours after inoculation. Two of the colonizing strains were able to induce watery diarrhea. All 12 STb-positive strains were nonpathogenic for adult mice and were serum-sensitive; 11 of 12 were Col V-negative, 9 of 12 did not produce aerobactin, and 10 of 12 belonged to biotypes other than 1 or 2. All 6 enterotoxin-negative strains colonized the small and large intestines, associated with peritoneal serosal surfaces, and induced septicemia and polyserositis in newborn colostrum-deprived pigs 1 to 2 days after inoculation. In contrast, 3 STb-positive strains poorly colonized the intestines and did not induce septicemia in pigs at 3 days after inoculation. All 6 enterotoxin-negative strains were Col V-positive, produced aerobactin, and belonged to biotype 1 or 2. Of the 5 enterotoxin-negative, F165-positive strains, only 4 were pathogenic for intraperitoneally inoculated adult mice and were serum-resistant. The enterotoxin-negative, F165-negative strain was neither serum-resistant nor mouse-pathogenic.(ABSTRACT TRUNCATED AT 250 WORDS)     

Decreased activity of brush border membrane-bound digestive enzymes in small intestines from pigs experimentally infected with porcine epidemic diarrhea virus

Brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase were measured in jejunum from pigs experimentally infected with porcine epidemic diarrhea virus (PEDV). Three piglets from the infected and control groups were euthanized by electrocution and subjected to necropsy at 24, 36, 48, 60, and 72 hours post-inoculation (hpi). The infection of PEDV to jejunum resulted in significant decreases in brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase. PEDV replication results in massive destruction of villous enterocytes leading to a marked reduction of intestinal epithelial surface and brush border membrane-bound digestive enzyme activity. Reduced enzymatic activity and villous atrophy in the small intestine is thought to result in a maldigestive and malabsorptive diarrhea.     

Evidence of porcine circovirus infection in pigs with wasting disease syndrome from 1985 to 1999 in Hokkaido, Japan

An epizootiological survey with histopathological methods was conducted for porcine circovirus in 220 diseased pigs (1-200 days old) in 49 farms from 1985 to 1999. Histopathological lesions containing PCV antigen were detected mainly in the lymphoid tissues from 42 of 189 diseased pigs (22.2%) in 4 of 45 farms (8.9%) from 1990 to 1999. The rate of positive pigs gradually increased from 1997 onward and PCV infection was found in 50% of diseased pigs in 1999. Histopathologically, the lesions in the lymphoid tissues (including lymph nodes, Peyer's patches, tonsil and spleen) were highly correlated with the presence of numerous spherical basophilic intracytoplasmic inclusion bodies with PCV antigen, and consisted of lymphocellular depletion and infiltration of macrophages. Although most affected cells showed cytoplasmic reactivity for PCV, intranuclear antigen was also seen in the lymphocytes, macrophages and ileal epithelial cells. Ultrastructurally, macrophages and giant cells contained electron-dense, round to ovoid lysosomal bodies, in which there were concentric circle or paracrystalline arrays of small nonenveloped icosahedral viral particles, approximately 15-17 nm in diameter. Other consistent infectious agents were present in 90.5% of cases, and porcine reproductive and respiratory syndrome virus infection was in 52.4% of the cases with PCV. The histopathological findings suggested that PCV induced systemic immunosuppression in the infected pigs and made them more susceptible to infection of the organisms. Because of the presence of PCV antigens in the intestinal epithelium, feces may play a significant role in dissemination of PCV.     

Effect of virus-induced destruction of villous epithelium on intestinal secretion induced by heat-stable Escherichia coli enterotoxins and prostaglandin E1 in swine

Villous atrophy and crypt hyperplasia were induced in the jejunal epithelium of thirteen 3-week-old pigs by inoculation with transmissible gastroenteritis virus. The responses (changes in net fluid movement) induced in ligated intestinal loops of these pigs by intraloop injections of prostaglandin E1 (PGE1) or Escherichia coli broth culture filtrates containing either or both E coli heat-stable enterotoxins (STa and STb) were compared with the responses induced by these preparations in littermates not inoculated with virus. Villous atrophy was associated with a marked decrease in response to preparations containing STa, STb, or STa + STb, but the response to PGE1 was undiminished. These results were consistent with the reports of others that the response to cyclic adenosine monophosphate-mediated secretogogues (PGE1) is a function of crypt epithelium; however, the present results also suggest that the secretory response to STa and to STb is dependent on the integrity of the villous epithelium. In the present study, loss of villous epithelium was associated with loss of response to STa and STb, but not to PGE1.     

Expression of secreted mucins (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound mucin (MUC4) in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae

The expression patterns of different secreted (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound (MUC4) mucins were determined immunohistochemically in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae. Forty-seven-week-old colostrum-deprived pigs were randomly allocated to infected (n=20) or control groups (n=20). Five infected and uninfected pigs were euthanized at 0, 6, 12, and 48 h post-inoculation (hpi). In the infected pigs, the expression of both types of mucins, which were invariably observed, was associated with bronchiolar and respiratory bronchiolar lesions. Strong positive mucin signals were seen on the surface of bronchiolar and respiratory bronchiolar epithelium with neutrophil infiltration. The mean mucin-positive area peaked at 6 hpi and decreased significantly to control levels by 48 hpi on the surface of the bronchiolar and respiratory bronchiolar epithelium. Further studies are needed to establish the functional relationship between mucin expression and the host defense mechanism against A. pleuropneumoniae in the lungs of infected pigs.     

Histology, immunohistochemistry and ultrastructure of the equine tubal tonsil

The tubal tonsil of the horse surrounds the pharyngeal opening of the eustachian tube and is lined by pseudostratified columnar ciliated epithelium interspersed with areas of follicle-associated epithelium (FAE) heavily infiltrated by lymphocytes but devoid of goblet and ciliated cells. Scanning and transmission electron microscopy revealed microvillous cells and cells with features characteristic of M cells such as reduced microvilli or depressed bare surface, more numerous mitochondria, small vesicles and lysosomes, as well as vimentin filaments and epitopes specific for GS 1-B4 as previously seen in the nasopharyngeal tonsil. M cells were also identified in areas of respiratory epithelium not associated with lymphoid follicles and appeared to be the nasal mucosal counterparts of recently described intestinal villous M cells in the mouse. The underlying lymphoid tissue of the FAE was generally organized as solitary lymphoid follicles without germinal centres in contrast to the diffuse and large amount of organized lymphoid follicles with germinal centres that characterize the nasopharyngeal tonsil. CD8+ T and B-lymphocytes were much fewer than in the nasopharyngeal tonsil. High endothelial venules were mainly oriented towards the parafollicular area and contained much fewer endothelial pores and vesiculo-vacuolar organelles. Finally, scattered small clusters of mucus acini and striated muscles were other features that differentiated the tubal and nasopharyngeal tonsils.     

Transcytosis of F4 fimbriae by villous and dome epithelia in F4-receptor positive pigs supports importance of receptor-dependent endocytosis in oral immunization strategies

Very few antigens have been described that induce an intestinal immunity when given orally. Our laboratory demonstrated that oral administration of isolated F4 (K88) fimbriae of Escherichia coli to F4-receptor positive (F4R(+)) pigs induces protective mucosal immunity against challenge infection. However, presence of F4-receptors (F4R) on villous enterocytes is a prerequisite for inducing the immune response, as no F4-specific antibody-secreting cells (ASC) can be induced in F4R(-) pigs. In this study, the in vivo binding of isolated F4 fimbriae (F4) to the gut epithelium was examined in F4R(+) and F4R(-) pigs. It was further investigated whether binding of F4 to the F4R results in endocytosis in and translocation across the gut epithelium using microscopy. F4 did not adhere to the intestinal epithelium of F4R(-) pigs, whereas it strongly adhered to the villous epithelium and the follicle-associated epithelium (FAE) of the jejunum and ileum of F4R(+) pigs. Following binding to F4R, F4 was endocytosed by villous enterocytes, follicle-associated enterocytes and M cells. Transcytosis of F4 across the epithelium resulted in the appearance of F4 in the lamina propria and dome region of the jejunal and ileal PP. This is the first study showing transcytosis of fimbriae across the gut epithelium. This receptor-dependent transcytosis can explain the success of F4 fimbriae as oral immunogen for inducing protective immunity in F4R(+) pigs strengthening the importance of receptor-dependent endocytosis and translocation in oral vaccine strategies. Further identification of the receptor responsible for this transport is in progress.     

A comparison of histopathological changes in calves associated with K99- and K99+ strains of enterotoxigenic Escherichia coli.

Enterotoxigenic colibacillosis was experimentally produced in four colostrum-deprived calves given 10(10) Escherichia coli strain 210 (serotype 09+:K30+:K99-:F41-:H-) orally and the histopathological changes compared to those seen in colostrum-fed calves infected in an earlier study with strain B44 (serotype 09+:K30+:K99+:F41+:H-). Escherichia coli strain 210 caused diarrhea, atrophic villi with cuboidal epithelium, and focal accumulations of a few neutrophils in the dome villi above Peyer's patches but neither the clinical nor the histopathological changes were as pronounced as with strain B44. The extent and distribution of adherence to the mucosal surface differed between the two strains. Strain B44 adhered as a continuous layer over most of the absorptive epithelial surface of both the jejunum and ileum. Adherence of strain 210 was restricted to the ileum and the bacteria often adhered focally in "clumps" rather than as a continuous layer, especially on the distal half of the villous surface.     

Intestinal trichomonads (Tritrichomonas mobilensis) in the natural host Saimiri sciureus and Saimiri boliviensis

A retrospective study of cecal and colonic tissues from 28 squirrel monkeys (Saimiri sciureus and Saimiri boliviensis) demonstrated enteric trichomonads within luminal crypts. Twenty-one of 28 (75%) had trichomonads in the mucosal epithelium either in cup-like depressions or intraepithelial vacuoles. Organisms were also beneath the superficial luminal mucosal epithelium and between the basement membrane and crypt epithelial cells. Immunoperoxidase staining also identified organisms within the lamina propria and submucosa. Additional histologic changes included mucosal ulceration, multifocal cryptitis, and focal epithelial necrosis. Most areas containing trichomonads did not have an associated inflammatory response.     

Pathogenesis of porcine rotaviral infection in experimentally inoculated gnotobiotic pigs.

Porcine rotavirus was shown to infect gnotobiotic pigs and induce an acute enteric disease clinically characterized by diarrhea, anorexia, depression, and occasional vomition. Onset of clinical signs correlated closely with the appearance of lesions within the small intestinal mucosa, and recovery from infection was associated with the regeneration of normal, functional villous epithelium. Villous atrophy, especially in the caudal two-thirds of the small intestine, was the consistent lesion observed in pigs with clinical signs of rotaviral infection. Villi were often short, blunt, and covered with cuboidal epithelial cells. Immunofluorescent microscopy methods demonstrated that the principal site of rotaviral replication was the villous columnar epithelial cells in the small intestine.