Proliferative enteropathy involving Lawsonia intracellularis infection in rabbits (Oryctlagus cuniculus)

Five rabbits suffering from diarrhea were diagnosed with proliferative enteropathy (PE). Histopathology revealed a thickened mucosa consisting of hyperplastic intestinal epithelium and infiltration of inflammatory cells mainly consisted of macrophages. In the affected epithelial cytoplasm, numerous curved bacillus-like organisms were observed in the Warthin-Starry silver stain and electron microscopy observation. In polymerase chain reactions, Lawsonia intracellularis-specific DNA fragment were amplified from affected ileal tissue extracted DNA in each case and present 5 cases were confirmed to be L. intracellularis infection. Serum collected from the affected rabbit was immunohistochemically reactive with L. intracellularis in tissue sections from pigs with porcine proliferative enteropathy, as well as with tissue sections from the five affected rabbits. Thus, serum obtained from the affected rabbit may be applicable to immunohistochemical detection for L. intracellularis infection in other species.     

Comparison of Histopathologic Findings in Duodenal and Ileal Endoscopic Biopsies in Dogs with Chronic Small Intestinal Enteropathies

Background

The current tendency when investigating dogs with chronic upper gastrointestinal signs is to perform endoscopy and biopsy only the duodenum. This approach could lead to overlooking important ileal lesions and affect the clinical management.

Objectives

To compare concurrent duodenal and ileal endoscopic biopsies in dogs with chronic enteropathies and evaluate their correlation with clinicopathologic findings.

Animals

Thirty‐eight dogs with chronic enteropathies.

Methods

Duodenal and ileal biopsies were retrospectively reviewed. Nine histologic variables, 5 structural (villous stunting, epithelial injury, crypt distension, lacteal dilatation, and mucosal fibrosis) and 4 inflammatory (intraepithelial lymphocytes, lamina propria lymphocytes and plasma cells, eosinophils, and neutrophils) were scored. Clinical severity scores and relevant clinicopathologic variables were evaluated.

Results

There was only slight agreement between duodenal and ileal histologic scores (κ = 0.003). There was slight agreement between the presence of any of the morphological and inflammatory variables, with the exception of mucosal fibrosis (κ = 0.44). Statistically significant correlation was found between clinical severity and duodenal crypt distension (P = .031), ileal lacteal dilatation (P = .038), and ileal mucosal lymphoplasmacytic inflammation (P = .035). A significant correlation was found between hypoalbuminemia and ileal lacteal dilatation (P = .033) and number of ileal intraepithelial lymphocytes (P = .019). A statistically significant correlation was found between hypocobalaminemia and number of ileal intraepithelial lymphocytes (P = .012).

Conclusions and Clinical Importance

When investigating dogs with chronic upper gastrointestinal signs, the collection of concurrent duodenal and ileal endoscopic biopsies is recommended.     

Evaluation of porcine ileum models of enterocyte infection by Lawsonia intracellularis

The early interaction of Lawsonia intracellularis with host cells was examined with the use of porcine ileum models. Two conventional swine were anesthetized, and ligated ileum loops were prepared during abdominal surgery. The loops were inoculated with 108 L. intracellularis or saline. After 60 min, samples of each loop were processed for routine histologic and electron microscopic study. Histologic and ultrathin sections of all the loops appeared normal, with no apposition of bacteria and host cells or bacterial entry events in any loop. Portions of ileum from a single gnotobiotic piglet were introduced as xenografts into the subcutis of each flank of 5 weaned mice with severe combined immunodeficiency disease. After 4 wk, 108 L. intracellularis were inoculated into each of 4 viable xenografts with a sterile needle; the other 3 viable xenografts received saline. Histologic and ultrathin sections of all the xenografts 3 wk after inoculation showed relatively normal porcine intestinal architecture, with normal crypts, crypt cell differentiation, and low villous structures; the xenografts treated with the bacteria also showed intracytoplasmic L. intracellularis within crypt and villous epithelial cells. Thus, entry of L. intracellularis into target epithelial cells and multiplication may not be sufficient alone to directly cause cell proliferation. A proliferative response may require active division of crypt cells and differentiation in conjunction with L. intracellularis growth.     

Porcine proliferative enteritis: serological, microbiological and pathological studies from three field epizootics.

Three outbreaks of porcine proliferative enteritis were evaluated clinically, pathologically, microbiologically and serologically. The disease was characterized by a chronic intermittent diarrhea. Pathological lesions included a thickened, turbid ileum with the microscopic appearance of proliferating ileal crypt epithelial cells. Comma shaped intracytoplasmic organisms were observed in the apical portions of the proliferating crypt epithelial cells with a Warthin-Starry silver stain. Microbiologically, both Campylobacter sputorum subspecies mucosalis and Campylobacter hyointestinalis, were cultured from ileal specimens of seven pigs with lesions of porcine proliferative enteritis. Microagglutination antibody titers were determined on sera from 12 of 14 pigs with porcine proliferative enteritis and on sera from 91 clinically normal swine. Pigs with porcine proliferative enteritis had a low antibody titer to subspecies mucosalis that ranged from 1-3 with a mean of 2.17. A varied C. hyointestinalis titer from 3-7 with mean of 4.83 was determined. Titers to either subspecies mucosalis and C. hyointestinalis were higher in non-porcine proliferative enteritis pigs. The results indicate that the presence of a positive titer to either C. hyointestinalis or subspecies mucosalis in swine is not indicative of clinical disease. The isolation of C. hyointestinalis from diseased ileal specimens (porcine proliferative enteritis) confirms previous reports implicating this agent in the disease.     

Immunohistochemistry and polymerase chain reaction for the detection of Lawsonia intracellularis in porcine intestinal tissues with proliferative enteropathy

Detection method of Lawsonia intracellularis was studied in formalin-fixed paraffin-embedded intestinal tissues from 5 naturally infected pigs by immunohistochemistry with a monoclonal antibody against outer membrane protein of L. intracellularis. Warthin-Starry silver stain revealed clusters of argyrophilic, slightly curved rod-shaped organisms in the apical cytoplasm of enterocytes. Immunohistochemical staining with a L. intracellularis-specific monoclonal antibody confirmed the presence of the organism in the apical cytoplasm of hyperplastic enterocytes. The presence of L. intracellularis in the ileum of pig with proliferative enteropathy was confirmed by polymerase chain reaction (PCR) further on the basis of amplification of 319 base pair products specific for porcine L. intracellularis chromosomal DNA. Immunohistochemistry and PCR may be a complementary method to confirm the diagnosis of L. intracellularis infection in pigs.     

Tonsillar changes in pigs given pseudorabies (Aujeszky's disease) virus

All of the eight 5-day-old pigs orally given pseudorabies (Aujeszky's disease) virus developed tonsillitis. The initial changes occurred in the subepithelial area between the lymphoid nodule and the crypt epithelium, showing a characteristic pattern of necrosis. The necrosis became more severe and gained access into the lymphoid nodule and crypt epithelium. Coincident with the histopathologic changes, numerous specific immunofluorescences were detected, first in the nucleus and in some parts of the cytoplasm of cells distributed in the subepithelial area. The fluorescence subsequently spread into adjacent lymphoid nodules and crypt epithelial cells. Ultrastructurally, many enveloped virus particles were detected in the center of the necrosis. Thereafter, the crypt epithelial cells also underwent degeneration, and a small number of virus particles were detected in the nucleus of the degenerating epithelial cells. In the more advanced stage, the enveloped virus particles were discharged into the crypt lumen.     

Pathogenesis of digestive tract lesions in duck plague.

White Pekin ducklings were inoculated orally with duck plague virus. Tissues from the digestive tract were collected daily after inoculation and examined by light, electron and fluorescent microscopy. There were necrosis and degeneration of stratified squamous epithelium of the esophagus and cloaca, epithelium of intestinal crypt and esophageal submucosal glands, macrophages in the lamina propria, and submucosal fibrocytes and lymphocytes. Submucosal hemorrhages occurred after degeneration and necrosis of lymphocytes, macrophages, fibrocytes and epithelial cells. Viral antigens were detected in all these cells by use of fluorescein-labeled antibodies. With the electron microscope, nucleocapsids were seen in the nuclei, budding through the inner nuclear membrane; enveloped virions were present in cytoplasmic vacuoles of macrophages, epithelial cells and fibrocytes. In lymphocytes, nucleocapsids were also in the nuclei, but karyorrhexis and cytolysis occurred before viral maturation was completed.     

Equine proliferative enteropathy: a cause of weight loss, colic, diarrhoea and hypoproteinaemia in foals on three breeding farms in Canada

Proliferative enteropathy (PE) is a transmissible enteric disease caused by Lawsonia intracellularis. An outbreak of equine PE was diagnosed in foals from 3 breeding farms. Most foals had been weaned prior to the appearance of clinical signs, which included depression, rapid and marked weight loss, subcutaneous oedema, diarrhoea and colic. Poor body condition with a rough haircoat and a potbellied appearance were common findings in affected foals. Respiratory tract infection, dermatitis and intestinal parasitism were also found in some foals. Haematological and plasma biochemical abnormalities included hypoproteinaemia, transient leucocytosis, anaemia and increased serum creatinine kinase concentration. Postmortem diagnosis of PE was confirmed on 4 foals based on the presence of characteristic intracellular bacteria within the apical cytoplasm of proliferating crypt epithelial cells of the intestinal mucosa, using silver stains, and by results of PCR analysis and immunohistochemistry. Antemortem diagnosis of equine PE was based on the clinical signs, hypoproteinaemia and the exclusion of common enteric infections. Faecal PCR analysis was positive for the presence of L. intracellularis in 6 of 18 foals tested while the serum of all 7 foals with PE serologically evaluated had antibodies against L. intracellularis. Most foals were treated with erythromycin estolate alone or combined with rifampin for a minimum of 21 days. Additional symptomatic treatments were administered when indicated. All but one foal treated with erythromycin survived the infection. This study indicates that equine PE should be included in the differential diagnosis of outbreaks of rapid weight loss, diarrhoea, colic and hypoproteinaemia in weanling foals.     

Pathology of infectious diarrhea of infant rats (IDIR) induced by an antigenically distinct rotavirus

Suckling rats were inoculated with a group B rotavirus to determine the progression of the morphologic changes induced in the intestine by this virus. Several changes were observed by light microscopy 1 day after viral inoculation: shortening of small intestinal villi, villous epithelial necrosis, and villous epithelial syncytia. The lesions were most often present in the distal small intestine, although other small intestinal segments were affected to a lesser degree. By day 3 post-inoculation, epithelial necrosis, and syncytia were no longer present; however, the villous epithelium was disorganized and irregularly vacuolated, and intestinal crypt epithelium was hyperplastic. Alterations in villous height to crypt depth ratios were present in portions of the small intestine for the remainder of the 12-day study period. Epithelial syncytia appeared to form by the breakdown of the lateral interdigitating membranes of the absorptive villous epithelium. Viral particles, abundant in the syncytia, appeared to form from amorphous or reticular arrays of viral precursor material. Group B rotaviral antigens, as detected by indirect immunofluorescence, were present in large amounts in the small intestinal villous epithelium only on the first day after viral inoculation. These studies show that two important diagnostic features of group B rotaviral infections of rats, epithelial syncytia and viral antigen as determined by immunofluorescence, are present only on the first day of disease. These findings should be taken into consideration when attempting to diagnose disease induced by this agent.     

Regional Ileitis in Pigs: Morphological and Pathogenetical Aspects

Twenty-seven weaned pigs with a wasting appearance were investigated. From a morphological point of view the pigs were divided into three groups. In the group of pigs with macro-scopical signs of regional ileitis, affected tissue showed substantial epithelial proliferation, and electron microscopic studies revealed the presence of microorganisms within the cytoplasm of the epithelial cells. There was a loss of, or only faint, enzymatic activity of alkaline phosphatase in the mucosal epithelium. In another group of five pigs there was no, or only slight, light microscopical signs of regional ileitis but presence of intracellular microorganisms. The enzymatic activity of the ileal epithelium was low. Low enzymatic activity of the ileal epithelium was observed in a third group of wasting pigs, which had no histological or electron microscopical signs of regional ileitis.     

Experimentally induced porcine proliferative enteritis in specific-pathogen-free pigs

Thirty-three, 10-week-old, specific-pathogen-free pigs were randomly allotted to 3 treatment groups: group 1--intragastrically given homogenized intestinal mucosa (crude inoculum) from pigs with naturally occurring proliferative enteritis; group 2--given cultures of Campylobacter sputorum subsp mucosalis; and group 3--controls. One pig from each group was killed 4, 7, 10, 14, 18, 21, 24, 28, 31, 36, and 38 days after inoculation. The earliest intestinal lesion observed in groups 1 and 2 was leukocytic exudate within crypt lumina and focal inflammation of the surrounding lamina propria. The lesions occurred primarily over ileal aggregated lymphoid nodules (Peyer's patches). These changes were followed by focal proliferation of immature crypt epithelial cells and infiltration of increasing numbers of macrophages into the lamina propria. Campylobacter sp-like organisms were observed within the cytoplasm of affected epithelial cells by light and electron microscopies. Lesions progressed to diffuse crypt cell proliferation, elongation of crypts, and loss of villi. Mucosal necrosis was not a prominent feature.     

Cryptosporidiosis in guinea pigs: a retrospective study

Cryptosporidiosis was diagnosed in 81 guinea pigs (Cavia porcellus) from 1979 through 1985 at a research animal diagnostic laboratory. Most of the guinea pigs were juveniles of Hartley stock and originated from 6 commercial laboratory animal suppliers or from one pet store supplier. Common clinical signs reported were failure to gain weight, weight loss, diarrhea, and death. At necropsy, macroscopic findings included emaciation, hyperemia of the small intestine, serosal edema of the cecal wall, and increased fluidity of ingesta throughout the intestines. Oval to round cryptosporidia (1 to 4 microns) were seen microscopically within or on the brush border of mucosal epithelial cells from the duodenum through the cecum. Acute histologic lesions consisted of necrosis and sloughing of enterocytes at the villus tips, inflammation, hyperemia and edema of the lamina propria, and hyperplasia of crypt epithelium. More chronic lesions consisted of marked villus bridging or villus fusion and blunting, metaplasia of the mucosal epithelium, and lymphocytic infiltration of the lamina propria.     

What is your diagnosis? Perifemoral mass in a cow

Abstract: A 15‐year‐old female Simmental cross‐breed cow was presented to the Purdue University Veterinary Teaching Hospital for evaluation of a perifemoral soft tissue mass. Impression smears made from an excisional biopsy contained a population of pleomorphic mesenchymal cells with abundant, periodic acid–Schiff‐positive (PAS), intracytoplasmic granular material, and rare elongated multinucleated cells consistent with strap‐like cells. A second population of small round cells suggestive of lymphocytes or progenitor cells was also noted. A cytologic diagnosis of sarcoma was made, with rhabdomyosarcoma considered most likely based on the large amount of PAS‐positive material (presumed to be glycogen) and the rare strap‐like cells. Histopathologic sections contained an unencapsulated, densely cellular neoplasm composed of haphazardly arranged highly pleomorphic mesenchymal cells and a few small round cells. The mesenchymal cells were positive for vimentin, non‐specific muscle actin, and myoglobin, and negative for phosphotungstic acid‐hematoxylin, smooth muscle actin, and desmin. Glycogen granules were confirmed by transmission electron microscopy. A diagnosis of pleomorphic rhabdomyosarcoma was made. While cytologic findings may suggest rhabdomyosarcoma, cytologic features can be highly variable, and a definitive diagnosis usually requires cytochemical and immunohistochemical staining.     

Proliferative enteritis associated with Lawsonia intracellularis in a Japanese macaque (Macaca fuscata).

A 2.5-yr-old, intact male Japanese macaque (Macaca fuscata) was observed to have a thickened ileum during exploratory laparotomy. Lawsonia intracellularis-associated proliferative enteritis was diagnosed using histopathology (Warthin-Starry stain), immunohistochemistry, and polymerase chain reaction analysis of the ileal biopsy. The animal developed transient diarrhea and severe hypoproteinemia 16 days after surgery but recovered with intensive treatment using azithromycin. Given the fact that very specific tests are required for identifying this organism, L. intracellularis may be underdiagnosed in nonhuman primates.     

Experimental hamster enteritis: an electron microscopic study.

Hamster enteritis (HE) was experimentally produced in weanling hamsters by orally inoculating healthy hamsters with suspensions of ilea obtained from hamsters with HE. Control groups of hamsters were inoculated orally with suspensions of ilea from healthy hamsters. Electron microscopy was done on ilea from 6 control hamsters, 31 hamsters with experimentally produced HE, and 4 hamsters with naturally occurring HE. Ultrastructural changes were not observed in the absorptive epithelium of control animals. Two different intracytoplasmic bacterial organisms were observed in epithelial cells of hamsters with experimentally produced HE. Organisms that were observed early in the disease process were identified as Escherichia coli. Organisms ultrastructurally similar to Campylobacter spp were observed later in the disease and were only within hyperplastic epithelial cells. The hyperplastic ileal epithelium of hamsters with naturally occurring HE contained campylobacter-like organisms.     

Cell differentiation in intestinal adenomatosis of pigs studied by histochemistry of laminin and enzymes of epithelial and subepithelial tissue

The distribution of enzymes and laminin was examined in ileal tissue from pigs suffering from intestinal adenomatosis to reveal the nature of the lesion. A disruption of the normal and specific pattern of distribution was found. Thus, the normal ileal epithelium was characterised by brush border enzymes: alkaline phosphatase, magnesium-dependent adenosine triphosphatase (Mg-ATPase), fluoride resistant acid phosphatase and 5'-nucleotidase; enzymes of the basolateral border: Mg-ATPase; and cytoplasmic enzymes: beta-glucuronidase, non-specific esterase and acid phosphatase. Subepithelial fibroblasts seemed to be characterised by 5'-nucleotidase. Laminin was present as a continuous band under the surface and crypt epithelium, somewhat thicker in the former. In contrast, the branching proliferating crypts of intestinal adenomatosis largely lacked enzymes characteristic of both villus and crypt cells. Reactions for the subepithelial components, laminin and fibroblasts were also reduced. The deficient differentiation of the epithelial as well as subepithelial components in porcine intestinal adenomatosis distinguish the condition from crypt hyperplasia and indicate an adenoma-like character.     

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.     

Apoptosis of villous epithelial cells and follicle-associated epithelial cells in chicken cecum

The process of the disappearance of epithelial cells was examined in chicken cecal villi and follicle-associated epithelium (FAE). The apoptotic epithelial cells with intense DNA-fragmentation and their exfoliation were found in the villous tips. The epithelial cells with weak DNA-fragmentation were seen in the upper portion of the villi and their sparse exfoliations were also found there. Numerous epithelial cells in the intestinal lumen expressed the apoptotic features. A row of apoptotic epithelial cells with DNA-fragmentation was also found in the apical FAE, whereas no M cells exhibited any apoptotic signs. In all cecal regions, CD3+, CD8+, and TCR2+ lymphocytes were predominant in the epithelium at the upper portion of the villi and the FAE. CD4+ lymphocytes were mainly seen in the lamina propria. TCR1+ lymphocytes were not abundant in comparison with TCR2+ lymphocytes in the epithelium. TCR3+ T lymphocytes were rarely detected. These results suggest that the chicken cecal epithelial cells exfoliated into the lumen after the induction of the apoptosis, and that the induction may be involved with CD3+, CD8+, and TCR2+ lymphocytes. No death in M cells suggests that M cells may transform into microvillous epithelial cells.