CD4+ T-cell responses and distribution at the colonic mucosa during Brachyspira hyodysenteriae-induced colitis in pigs

The spirochaete Brachyspira hyodysenteriae causes swine dysentery, a severe colitis characterized by mucosal enlargement as a result of crypt elongation and epithelial necrosis. Most efforts to understand the pathogenesis of this disease have focused on the aetiological agent and its virulence factors. However, the host immune response has been considered an important factor in disease development. Previous research has shown that B. hyodysenteriae induces systemic CD4(+) and gammadelta T-cell responses after intramuscular immunization. Here, we have evaluated changes in the CD4(+) and gammadelta T-cell composition and distribution the different compartments of the colonic mucosa of pigs challenged with B. hyodysenteriae. We report that, in infected pigs, gammadelta T cells were significantly depleted from the epithelial layer, although their numbers were maintained in the lamina propria. In addition, CD4(+) T cells aggregated in clusters located in the lamina propria and submucosa. Ex vivo analyses of CD4(+) T-cell responses to B. hyodysenteriae antigens correlated with the changes in the mucosal CD4(+) T-cell distribution observed in infected pigs; CD4(+) T cells recovered from peripheral blood and colonic lymph nodes of infected pigs proliferated to B. hyodysenteriae antigens, whereas no differences were found in the gammadelta T-cell responses between challenged and control groups. In addition, colonic lymph node CD4(+) T cells had a predominant memory/activated phenotype. These results indicate that infection with B. hyodysenteriae induces a mucosal CD4(+) T-cell response and points to CD4(+) T cells being important contributors to the immunopathogenesis of swine dysentery.     

Comparison of stained smears and culturing for identification of Treponema hyodysenteriae.

A comparative study was made of stained fecal smears and cultured fecal swabs for identification of the large spirochetes Treponema hyodysenteriae and Treponema innocens. Feces were obtained by swabbing rectums, colons, and stools of nonexposed swine and swine experimentally exposed to swine dysentery. In this study there was a significant (P less than 0.001) correlation between the observation of one or more large spirochetes on stained slides and obtaining either a strong or a weak beta-hemolytic reaction in culture. A significant (P less than 0.001) correlation was also found between the observation of one or more large spirochetes on stained smears or obtaining either a strong or a weak beta-hemolytic reaction in culture and the occurrence of either nonhemorrhagic or hemorrhagic diarrhea in the swine. In the diarrheic swine at the time of swabbing, 157 of 393 samples (40%) were negative for both the presence of large spirochetes on stained smears and the production of either a strong or a weak beta-hemolytic reaction; in nondiarrheic swine, 42 of 278 samples (15.1%) were positive in stained smears and 32 of 268 samples (11.9%) were positive by culturing. In swine infected with swine dysentery, 17 of 1,011 samples produced weak beta-hemolytic reactions, and in swine infected with nonpathogenic large spirochetes of T. innocens, three of 34 samples produced strong beta-hemolytic reactions. It was concluded from this study that neither staining rectal smears nor culturing rectal swabs is sufficient, either together or alone, for the diagnosis of swine dysentery; however, these laboratory methods could be highly supportive of a diagnosis of swine dysentery in swine with clinical signs and lesions of the disease.     

Pathogenic synergism between Treponema hyodysenteriae and other selected anaerobes in gnotobiotic pigs.

Gnotobiotic pigs were orally exposed to various anaerobes at 6 to 9 days of age and similarly inoculated with Treponema hyodysenteriae B204 3 to 6 days later. Watery diarrhea and fecal excretion of large quantities of mucus and some fibrin clots were observed 4 to 20 days after inoculation with B204 if other anaerobes were present. Colonic lesions characteristic of swine dysentery were observed when B204 was present with Fusobacterium necrophorum, three strains of Bacteroides vulgatus, a Clostridium species, and Listeria denitrificans individually and when some of these microbes were present in various combinations, but not when B204 was present alone. These results are consistent with the conclusion that T. hyodysenteriae is the primary pathogen in the etiology of swine dysentery and that the presence of one or more other anaerobes is a prerequisite for expression of pathogenicity of T. hyodysenteriae. This prerequisite can be met by a variety of anaerobes.     

Interaction between Mycoplasma hyopneumoniae and Swine Influenza Virus

An experimental respiratory model was used to investigate the interaction between Mycoplasma hyopneumoniae and swine influenza virus (SIV) in the induction of pneumonia in susceptible swine. Previous studies demonstrated that M. hyopneumoniae, which produces a chronic bronchopneumonia in swine, potentiates a viral pneumonia induced by the porcine reproductive and respiratory syndrome virus (PRRSV). In this study, pigs were inoculated with M. hyopneumoniae 21 days prior to inoculation with SIV. Clinical disease as characterized by the severity of cough and fever was evaluated daily. Percentages of lung tissue with visual lesions and microscopic lesions were assessed upon necropsy at 3, 7, 14, and 21 days following SIV inoculation. Clinical observations revealed that pigs infected with both SIV and M. hyopneumoniae coughed significantly more than pigs inoculated with a single agent. Macroscopic pneumonia on necropsy at days 3 and 7 was greatest in both SIV-infected groups, with minimal levels of pneumonia in the M. hyopneumoniae-only-infected pigs. At 14 days post-SIV inoculation, pneumonia was significantly more severe in pigs infected with both pathogens. However, by 21 days postinoculation, the level of pneumonia in the dual-infected pigs was similar to that of the M. hyopneumoniae-only-infected group, and the pneumonia in the pigs inoculated with only SIV was nearly resolved. Microscopically, there was no apparent increase in the severity of pneumonia in pigs infected with both agents compared to that of single-agent-challenged pigs. The results of this study found that while pigs infected with both agents exhibited more severe clinical disease, the relationship between the two pathogens lacked the profound potentiation found with dual infection with M. hyopneumoniae and PRRSV. These findings demonstrate that the relationship between mycoplasmas and viruses varies with the individual agent.     

A humoral immune response confers protection against Haemophilus ducreyi infection

Haemophilus ducreyi is the etiologic agent of the sexually transmitted genital ulcer disease chancroid. Neither naturally occurring chancroid nor experimental infection with H. ducreyi results in protective immunity. Likewise, a single inoculation of H. ducreyi does not protect pigs against subsequent infection. Accordingly, we used the swine model of chancroid infection to examine the impact of multiple inoculations on a host's immune response. After three successive inoculations with H. ducreyi, pigs developed a modestly protective immune response evidenced by the decreased recovery of viable bacteria from lesions. All lesions biopsied 2 days after the first and second inoculations contained viable H. ducreyi cells, yet only 55% of the lesions biopsied 2 days after the third inoculation did. Nearly 90% of the lesions biopsied 7 days after the first inoculation contained viable H. ducreyi cells, but this percentage dropped to only 16% after the third inoculation. Between the first and third inoculations, the average recovery of CFU from lesions decreased approximately 100-fold. The reduced recovery of bacteria corresponded directly with a fivefold increase in H. ducreyi-specific antibody titers and the emergence of bactericidal activity. These immune sera were protective when administered to na?ve pigs prior to challenge with H. ducreyi. These data suggest that pigs mount an effective humoral immune response to H. ducreyi after multiple exposures to the organism.     

Effect of Treponema hyodysenteriae infection on mucosal mast cells and T cells in the murine cecum.

The pathogenic mechanisms responsible for the development of lesions in swine and mice after infection with Treponema hyodysenteriae have not been fully characterized. The release of inflammatory mediators from mast cells has been postulated to play a role in lesion development during swine dysentery. Therefore, C3H/HeN mice were infected with T. hyodysenteriae, and mucosal mast cell (MMC) numbers were examined in cecal sections. An initial increase in MMC numbers from 13 to 22 per 50 crypt villus units was observed, but at 20 days postinfection the numbers significantly decreased (P less than 0.05) to 5 MMC per 50 crypt villus units. Immunohistochemical analysis performed on cecal sections failed to show a significant change in lamina proprial T-lymphocyte subsets. Numbers of T. hyodysenteriae CFU recovered from the cecum were stable throughout the experimental time period. Mast cell-deficient W/Wv mice and their mast cell-sufficient littermates were also infected to determine whether MMCs were necessary for the occurrence of T. hyodysenteriae-induced lesions. W/Wv mice were as susceptible to infection as their normal littermates and developed similar macroscopic and microscopic lesions. These results indicate that changes in MMC numbers can be detected after an infection with T. hyodysenteriae; however, on the basis of observations of infected W/Wv mice, mast cells are not required for lesion development in the murine model.     

Swine model of Haemophilus ducreyi infection.

Haemophilus ducreyi is a strict human pathogen that causes sexually transmitted genital ulcer disease. We infected domestic swine with H. ducreyi 35000, resulting in the development of cutaneous ulcers histologically resembling human chancroid lesions. Intraepidermal lesions progressed from pustules to ulcers containing polymorphonuclear leukocytes and were accompanied by a dermal inflammatory infiltrate containing T cells and macrophages. H. ducreyi was recovered from lesions up to 17 days after inoculation, and pigs did not develop immunity to reinfection with the challenge strain. Features of the model include inoculation through abrasions in the epidermis, ambient housing temperatures for infected pigs, the ability to deliver multiple different inocula to a single host, and the availability of monoclonal antibodies against porcine immune cells permitting immunohistochemical characterization of the host immune response to H. ducreyi infection.     

Invasion and spread of equine herpesvirus 9 in the olfactory pathway of pigs after intranasal inoculation.

The neuropathogenesis of equine herpesvirus 9 (EHV-9) in pigs was investigated by intranasal inoculation of the virus together with intramuscular administration of dexamethasone (DM). All infected pigs developed characteristic meningo-encephalitis, accompanied by basophilic intranuclear inclusion bodies in the neuronal cells. One non-DM-treated and two DM-treated pigs had prominent malacic lesions in the rhinencephalon. Associated with the encephalitic lesions, there was invariably an increase in the number of nucleated cells in the cerebrospinal fluid (CSF). EHV-9 antigen was first detected in the nasal and olfactory epithelial cells in the nasal cavity, and in the neuroglial cells in the olfactory bulb. Subsequently it was demonstrated in the amygdaloid and caudate nuclei, and putamen. The virus was not isolated from the CSF. These results suggest that, after intranasal inoculation, EHV-9 replicates in the olfactory epithelial cells, spreading to the central nervous system via the olfactory pathway.     

Experimental infection of rabbit ligated ileal loops with Treponema hyodysenteriae

An in vivo animal model was used to assess the enteropathogenicity of the etiological agent (Treponema hyodysenteriae) of swine dysentery. Multiple ligated ileal loops, prepared in New Zealand white rabbits, were challenged with either pathogenic (B78 and B204) or nonpathogenic (Pu) isolates of the organism. The pathogenic isolates induced the onset of intestinal fluid accumulation as early as 4 h, with maximal fluid induction at 18 h postchallenge. Gross lesions of the intestinal mucosa, observed in ileal loops of rabbits sacrificed 24 h postchallenge, were characteristic of swine dysentery. Both pathogenic isolates colonized the epithelial surface and eroded the mucosal barrier, as determined by histological and scanning electron microscopic observations. Intestinal fluid accumulation and erosion of the mucosal barrier were not observed in ileal loops exposed to the nonpathogenic isolate (Pu) or to either of the nonviable pathogenic (B78 and B204) isolates. The ability of pathogenic isolates to initiate and produce infection in rabbit ligated ileal loops, which closely resembles the disease in swine, provides a system with which to study experimental swine dysentery.     

Ultrastructural morphometric investigation of early lesions in the pulmonary alveolar region of pigs during experimental swine influenza infection.

Experimental infection of specific-pathogen-free pigs with swine influenza virus by the intratracheal route resulted in a severe respiratory disease that closely resembled natural swine influenza in clinical course and pathologic lesions. Alveolar epithelial necrosis with sloughing of necrotic cells occurred from 24 to 96 hours after inoculation (p.i.) and was associated with alveolar edema and diffuse interstitial pneumonitis. The latter, initially of neutrophilic character, became histiocytic 48 hours p.i. Ultrastructural analysis of alveolar parenchyma disclosed viral replication in epithelial cells beginning at 5 hours p.i. and lasting to 96 hours. Budding of pleomorphic virus particles from the surface of alveolar epithelial cells and accumulation of viral proteins within the nucleus and cytoplasm of epithelial cells were seen. The extent of parenchymal lesions as quantified by stereologic morphometry within the whole lung was characterized by a marked relative and absolute volume increase of interalveolar septa and increased air-blood tissue barrier thickness. The volume increase of interalveolar septa was due to an increase of interstitial tissue volume by 85% in pigs at 96 hours p.i., compared with control pigs with similar lung volumes.     

Experimental infection with Treponema hyodysenteriae in guinea pigs.

Outbred and inbred (Hartley strain) guinea pigs (GP) were inoculated intragastrically with pathogenic and nonpathogenic Treponema hyodysenteriae. GP 3 to 16 weeks old received T. hyodysenteriae after a fasting period of 36 to 72 h. Infected GP with pathogenic T. hyodysenteriae developed a diarrheal and/or depressive condition, with mucus but not blood in the feces. Of 88 GP, 40 had gross lesions resembling those of swine dysentery. Lesions were limited mainly to the large intestine. TP used as controls or inoculated with nonpathogenic T. hyodysenteriae did not develop these lesions in the large intestine. These studies suggest that the GP may be used as an animal model for swine dysentery.     

Pathogenicity of porcine intestinal spirochetes in gnotobiotic pigs.

Twelve intestinal spirochete strains of porcine origin were characterized on the basis of their phenotypic properties, by multilocus enzyme electrophoresis, and by pathogenicity testing in gnotobiotic pigs. The spirochetes used included two strains of Serpulina hyodysenteriae (B204 and P18A), two strains of Serpulina innocens (B256 and 4/71), one strain from the proposed new genus and species "Anguillina coli" (P43/6/78), and seven non-S. hyodysenteriae strains recently isolated from United Kingdom pig herds with a history of nonspecific diarrhea and typhlocolitis. By multilocus enzyme electrophoresis, five of these were identified as S. innocens, one was identified as an unspecified Serpulina sp., and one was identified as "A. coli." S. hyodysenteriae B204 and P18A, "A. coli" P43/6/78 and 2/7, and three (22/7, P280/1, and 14/5) of the five S. innocens field isolates induced mucoid feces and typhlocolitis in gnotobiotic pigs. None of the other spirochetes produced clinical signs or large intestinal pathology in this model. The "A. coli" strains induced a more watery diarrhea, with lesions present more proximally in the large intestine, than did the other pathogenic spirochetes. S. innocens 22/7 was also tested for pathogenicity in hysterotomy-derived pigs that had previously been artificially colonized with a spirochete-free intestinal flora and shown to be susceptible to swine dysentery. Despite effective colonization, strain 22/7 did not produce any disease, nor was there any exacerbation of large intestinal pathology or clinical signs when pigs with an experimentally induced existing colitis caused by Yersinia pseudotuberculosis were superinfected with strain 22/7. Certain non-S. hyodysenteriae spirochetes are therefore capable of inducing disease in gnotobiotic pigs, but their role as primary or opportunistic pathogens in conventional pigs remains equivocal.     

Infection of pigs with the Cameroon isolate (Cam/82) of African swine fever virus

African swine fever (ASF) was produced in eight pigs by exposure to donors infected with the Cameroon/82 isolate of African swine fever virus. The primary clinical sign was pyrexia of more than 40 degrees C first observed 10 to 13 days post-exposure (dpe) in all pigs; other clinical signs were rarely observed. The most frequent post-mortem lesion was haemorrhage in the visceral lymph nodes. Other lesions included excess fluid in the abdominal cavity and petechial haemorrhages in the kidneys. Viraemia was first observed 1 to 2 days before the onset of pyrexia and maximal titres of more than 10(7.5) HAD50 per ml occurred 11 to 14 dpe. Virus excretion by the pharyngeal route was observed at 2 to 4 days before the onset of pyrexia and continued throughout the course of infection. Susceptible pigs, mixed directly with infected ones, contracted infection within 2 h; transmission time increased to 2 to 6 h when recipient pigs were separated by wire mesh from the infected pigs. The comparatively low mortality, ill-defined clinical signs and clinical recovery of many of the infected pigs show that the Cam/82 ASF virus is of relatively low virulence and thereby resembles recent European, South American and Caribbean isolates.     

Ultrastructural study of the renal tubular system in acute experimental African swine fever: virus replication in glomerular mesangial cells and in the collecting ducts

Summary Despite the considerable attention given to kidney lesions in African swine fever (ASF), a number of questions remain to be answered. Structural and ultrastructural examination showed that a highly virulent isolate of ASF virus (Malawi 83) replicated in glomerular mesangial cells and renal collecting duct epithelial cells, with hyperplasia of the latter in infected pigs. Replication in mesangial cells may be due to their contact with the bloodstream, as well as to their phagocytic capacity and high metabolism rate. Virus replication in macrophages and endothelial cells of interstitial capillaries, and the necrosis of these infected cells gave rise to a large number of free virus in interstitial tissue. This, together with the lesser thickness of the basal membrane of collecting ducts in comparison to the rest of the tubular system, probably facilitates ASFV infection of tubular epithelial cells. Virus replication in these cells may account for the presence of virus in the urine of pigs with acute ASF where haematuria is not observed.     

Use of a swine bioassay and a RT-PCR assay to assess the risk of transmission of swine hepatitis E virus in pigs

The objective of this study was to assess the risk of transmission of swine hepatitis E virus (swine HEV) to na?ve pigs by inoculation with tissues or feces collected from pigs infected experimentally with swine HEV. Seventy-five, 3-week-old pigs were assigned randomly to 24 groups of 3-4 pigs and inoculated with homogenates of tissues (liver, heart, pancreas, or skeletal muscle) or a suspension of feces from swine HEV-infected pigs collected at 3, 7, 14, 20, 27, or 55 days post inoculation (DPI). Each inoculum was prepared as a 10% suspension (w/v) in PBS buffer and tested by a semi-quantitative RT-PCR for swine HEV RNA and by the swine bioassay. The inoculation route was intravenous for liver, heart and pancreas, and via stomach tube for skeletal muscle and fecal suspension. The liver homogenate inocula and feces collected at 3-7 and 14-20 DPI were positive for swine HEV RNA by RT-PCR. The pigs inoculated with liver homogenates collected at 3-7 and 14-20 DPI developed anti-HEV antibodies and swine HEV RNA was detected in their sera. Pigs inoculated with heart, pancreas, skeletal muscle homogenates or fecal suspensions failed to develop anti-HEV antibodies. These findings suggest that there is a potential risk of transmission of swine HEV via liver tissue from infected pigs in the early stages (3-20 DPI) of infection and the in vitro RT-PCR assay correlates well with the swine bioassay.     

Experimental transmission of porcine circovirus type 2 (PCV2) in weaned pigs: a sequential study

Weaned specific pathogen-free pigs were inoculated intranasally with porcine circovirus type 2 (PCV2) and killed in groups of two or three animals at 6, 13, 20, 27 and 34 days post-inoculation (dpi), together with appropriate uninfected controls, for examination by histopathological, immunohistochemical (immunogold silver staining; IGSS), polymerase chain reaction (PCR) and viral isolation techniques. Serum samples were also collected for detection of antibodies. No major clinical signs were observed in infected pigs, and gross lesions were essentially limited to the lungs and lymph nodes of some of the animals. Histologically, no lesions were seen at 6 dpi, but bronchointerstitial pneumonia was invariably noted from 13 dpi onwards. Granulomatous inflammation, with or without intracytoplasmic inclusions, was present in lymphoid tissues (e.g. lymph nodes, thymus, spleen and tonsil) from day 20 onwards, being most severe at days 20 and 27 dpi. Liver inflammation was present at days 13, 20 and 27 dpi. Virus was demonstrated in the tissues by isolation and PCR methods throughout the experiment. PCV2 antigens were detected by IGSS in bronchial and bronchiolar epithelial cells, in mononuclear cells and multinucleated giant cells within inflammatory lesions, and in mononuclear cells of apparently normal tissues (e.glamina propria of the small intestine and the bronchus-associated lymphoid tissue). The lesions were consistent with those of postweaning multisystemic wasting syndrome (PMWS), although not all previously reported PMWS lesions were seen. PCV2 antibodies were detected in infected pigs from day 13 onwards. The results demonstrated widespread distribution of PCV2 after infection and persistence of the virus in vivo for at least 34 days. It would appear that PCV2 can induce PMWS lesions in weaned pigs in the absence of porcine parvovirus and other common swine pathogens.     

Enteropathogenicity of various isolates of Treponema hyodysenteriae.

Isolates of Treponema hyodysenteriae from 25 geographically separated outbreaks of swine dysentery were tested for their ability to produce the disease. Clinical signs and lesions typical of acute swine dysentery were produced in 52 of 68 (75%) susceptible specific pathogen-free pigs that had been orally inoculated with pure cultures of 23 of 25 beta-hemolytic isolates. In addition, 13 weakly beta-hemolytic isolates of nondysentery origin with morphology similar to T. hyodysenteriae did not produce disease when orally inoculated into susceptible specific pathogen-free pigs. Two of these latter isolates, Puppy and B296, and one pathogenic, beta-hemolytic isolate failed to produce disease when orally inoculated into puppies.     

Mycoplasma hyopneumoniae Potentiation of Porcine Reproductive and Respiratory Syndrome Virus-Induced Pneumonia

An experimental model that demonstrates a mycoplasma species acting to potentiate a viral pneumonia was developed. Mycoplasma hyopneumoniae, which produces a chronic, lymphohistiocytic bronchopneumonia in pigs, was found to potentiate the severity and the duration of a virus-induced pneumonia in pigs. Pigs were inoculated with M. hyopneumoniae 21 days prior to, simultaneously with, or 10 days after inoculation with porcine reproductive and respiratory syndrome virus (PRRSV), which induces an acute interstitial pneumonia in pigs. PRRSV-induced clinical respiratory disease and macroscopic and microscopic pneumonic lesions were more severe and persistent in M. hyopneumoniae-infected pigs. At 28 or 38 days after PRRSV inoculation, M. hyopneumoniae-infected pigs still exhibited lesions typical of PRRSV-induced pneumonia, whereas the lungs of pigs which had received only PRRSV were essentially normal. On the basis of macroscopic lung lesions, it appears that PRRSV infection did not influence the severity of M. hyopneumoniae infection, although microscopic lesions typical of M. hyopneumoniae were more severe in PRRSV-infected pigs. These results indicate that M. hyopneumoniae infection potentiates PRRSV-induced disease and lesions. Most importantly, M. hyopneumoniae-infected pigs with minimal to nondetectable mycoplasmal pneumonia lesions manifested significantly increased PRRSV-induced pneumonia lesions compared to pigs infected with PRRSV only. This discovery is important with respect to the control of respiratory disease in pigs and has implications in elucidating the potential contribution of mycoplasmas in the pathogenesis of viral infections of other species, including humans.     

Experimental infection of newly weaned pigs with human and porcine strains of Serpulina pilosicoli.

Cultures of Serpulina pilosicoli 95/1000, isolated from a pig with porcine intestinal spirochetosis (PIS), and S. pilosicoli WesB, isolated from an Aboriginal child with diarrhea, were used to infect 5-week-old newly weaned pigs. Four of 12 pigs infected with strain 95/1000 and 2 of 12 pigs infected with strain WesB became colonized and developed watery, mucoid diarrhea within 2 to 11 days postinfection. Affected pigs all had moderate subacute mucosal colitis, with gross and histological changes similar to those previously reported in both natural and experimentally induced cases of PIS. Silver-stained histological sections of the colon and cecum from affected pigs demonstrated spirochetes within dilated intestinal crypts, where they were associated with neutrophilic exocytosis and mucus secretion. Sections from one pig infected with strain 95/1000 showed large numbers of spirochetes attached by one end to the colonic epithelium, a feature consistent with PIS. This study confirms the role of S. pilosicoli in the etiology of PIS and provides evidence that S. pilosicoli strains of human origin have pathogenic potential in an animal model.     

Reovirus type I infection of small intestinal epithelium in suckling mice and its effect on M cells

In 10-day-old mice, reovirus serotype I (reo I) selectively adheres to the apical surface of M cells and penetrates the intestinal epithelium via M cells overlying Peyer's patches before causing disseminated infection. Recently, reo I enteritis has been described in adult mice. We wished to determine if reo I enteritis also occurs in suckling mice and, if so, to determine which major epithelial cell types become infected and where the virus enters epithelial cells other than M cells. Transmission electron microscopy revealed that after oral inoculation of 10-day-old mice, peak infection of M cells preceded that of absorptive and undifferentiated crypt cells. The percentage of M cells in the dome epithelial cell population was reduced more than 4-fold between 4 and 72 hours after reo I inoculation compared with saline-inoculated mice. By 6 days after inoculation, reo I replication was no longer observed and there was a more than 2-fold increase in M cells overlying Peyer's patches domes of reo I-inoculated mice compared with saline controls. By 13 days, control and infected mice had similar percentages of M cells. When incubated with isolated intestinal epithelial sheets, reo I adhered selectively to and was endocytosed via the basal plasma membrane of absorptive cells. Thus, reo I initially penetrates the intestinal epithelium via the apical surface of M cells which become infected. Virions subsequently enter absorptive and crypt cells via their basal surfaces. During the first 3 days of enteritis, the M cell population becomes markedly depleted which may affect the permeability of the mucosal barrier to microorganisms and other antigens as well as influence the host immune response.