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The cotton-top tamarin (CTT; Saguinus oedipus) is an endangered New World primate that
develops a highly prevalent idiopathic colitis resembling human ulcerative colitis. This study
found that enteropathogenic Escherichia coli (EPEC) caused acute colitis in CTTs, which was
associated with ulcerative colitis. EPEC clinical isolates revealed localized adherence patterns
by HEp-2 assay and were devoid of Shiga-toxin production. Sequencing of the eae gene
(GenBank accession no. AF319597) revealed 99.2% identity to sequences of human isolates
(GenBank AF116899) and corresponded to the e intimin gene subtype. Detection of intimin
sequences by polymerase chain reaction on primary fecal cultures indicated widespread EPEC
infection in the CTT colony. Prospective analysis revealed that animals with fecal cultures
positive for intimin sequences had a higher frequency of active colitis (75.0% vs. 27.2%; P !
.005, x2 test) and higher histological scores of colonic inflammation (0.875 vs. 0.455, respec-
tively; P ! .05, Mann-Whitney rank sum test).
The cotton-top tamarin (CTT; Saguinus oedipus) is an en- disease [2]. Circumstantial evidence has implicated Campylo-
dangered primate native to the forests of northern Colombia. bacter jejuni, corona-like viruses, and Helicobacter species as
A highly prevalent idiopathic colitis resembling human ulcer- agents that may play a role in producing this disease syndrome
ative colitis has been recognized in CTTs housed at the New [2, 3].
England Regional Primate Research Center (NERPRC) and An infectious etiology has also been suggested as a cofactor
elsewhere since the 1970s. This idiopathic colitis shares similar of ulcerative colitis in humans, and enteroadherent Escherichia
clinical, endoscopic, and histopathological features with ulcer- coli has been implicated in the pathogenesis of this disease. The
ative colitis in humans and has been widely used as a nonhuman adhesive properties of E. coli isolates from patients with ulcer-
primate model to investigate immunopathogenesis, risk factors, ative colitis differ from those of isolates from healthy control
and novel therapeutic interventions for the human condition patients, and intimin-positive strains have been identified in
[1]. CTTs with chronic colitis have a high incidence of colonic some affected persons [4, 5]. Furthermore, treatment of patients
carcinoma. Although the etiology of CTT colitis and colonic with ulcerative colitis by means of probiotic nonpathogenic E.
carcinoma is unknown, several lines of evidence suggest that coli has an effect similar to that of mesalazine in maintaining
environmental factors play a role: A prospective study of the remission [6]. Here we describe enteropathogenic E. coli (EPEC)
epizootiology of colitis and adenocarcinoma showed that infection in CTTs and its association with ulcerative colitis in
chronic mucosal changes were modified by diet [2]. Further- this species.
more, the marked decreased incidence of both colitis and car-
cinoma of animals raised in isolation from the rest of the colony
Methods
suggested the existence of an infectious agent as a cofactor in
Animals and housing. CTTs were housed at the NERPRC. All
CTTs were housed in pairs or grouped into small extended families
Received 2 April 2001; revised 24 May 2001; electronically published 8 consisting of 1 adult breeding male CTT, 1 adult breeding female
August 2001.
Animals were housed at the New England Regional Primate Research
CTT, and 2–6 siblings !18 months old, as described elsewhere [7].
Center in accordance with standards of the American Association for Ac- Average colony census during the study period was ∼200 CTTs.
creditation of Laboratory Animal Care and of the Harvard Medical School’s Bacteriology. Rectal swabs of clinically affected animals were
Animal Care and Use Committee. cultured by standard techniques. Samples were plated on Mac-
Financial support: National Institutes of Health (RR-00168).
Conkey, Hektoen, and blood agar at 37⬚C in 5% CO2 and on
Reprints or correspondence: Dr. Keith G. Mansfield, Harvard Medical
School, New England Regional Primate Research Center, PO Box 9102, Campylobacter agar at 42⬚C with 5% O2, 85% N, and 10% CO2.
Southborough, MA 01772-9012 (Keith_Mansfield@HMS.Harvard.edu). Plates were evaluated at 24, 48, and 72 h, and individual bacterial
colonies were identified by standard biochemical techniques (API
The Journal of Infectious Diseases 2001; 184:803–7
䉷 2001 by the Infectious Diseases Society of America. All rights reserved. Rapid 20E; BioMerieux Vitek). A HEp-2 adherence assay was done
0022-1899/2001/18406-0021$02.00 as described elsewhere [8]. Bacterial isolates were serotyped at the
804 Mansfield et al. JID 2001;184 (15 September)
Pennsylvania State University E. coli Reference Laboratory (Uni- to the apical cytoplasmic membrane with pedestal formation
versity Park, PA). Shiga-toxin production was assessed by Vero cell and rearrangement of the underlying cytoskeleton. Morpho-
cytotoxicity assay, as described elsewhere [9]. logical alterations were limited to the large intestine, which was
Polymerase chain reaction (PCR) and sequencing. For char-
diffusely involved in all cases. In contrast, the small intestine
acterization of individual bacterial clones, DNA was isolated from
was spared and was normal in all animals.
single colonies and grown overnight in broth. PCR utilizing primers
Rectal swabs and fecal samples were obtained from all clin-
directed at the intimin (eae), Shiga-like toxin (stx1 and stx2), and
Figure 1. Enteropathogenic Escherichia coli (EPEC) infection of colonic epithelium in cotton-top tamarins. A, Irregular colonic surface epi-
thelium accompanied by marked epithelial crypt cell hyperplasia (hematoxylin-eosin stain; magnification, ⫻85). Inset, Bacilli intimately associated
with apical cytoplasmic membrane (toludene blue stain; magnification, ⫻428). B, Ultrastructural features of EPEC infection. Adherent bacilli
with effacement of normal microvillus border (magnification, ⫻5000). Inset, Pedestal formation with rearrangement of underlying cytoskeleton
(magnification, ⫻13,000).
this animal, the lesion was present multifocally in 1 of 3 colonic after the first survey. Of 35 animals tested, intimin sequences
sections examined. In the remaining intimin-positive animals, were detected by PCR on DNA isolated from primary fecal
inflammatory changes did not differ from those typically at- cultures from 12 (34.3%) animals. Despite the absence of severe
tributed to idiopathic ulcerative colitis in CTTs. acute colitis, the incidence of infection was not statistically dif-
The colony was retested for the presence of EPEC 7 months ferent from that 7 months earlier.
806 Mansfield et al. JID 2001;184 (15 September)
variation among type IV pilin (bfpA) genes from diverse enteropathogenic top tamarins (Saguinus oedipus oedipus) living wild in their natural hab-
Escherichia coli strains. Infect Immun 2000; 68:7028–38. itat. Dig Dis Sci 1998; 43:1443–53.
13. Mansfield KG, Lin KC, Newman J, et al. Identification of enteropathogenic 15. Barnett FD, Philpott D, Abul-Milh M, Huesca M, Sherman PM, Lingwood
Escherichia coli in simian immunodeficiency virus–infected infant and CA. Phosphatidylethanolamine recognition promotes enteropathogenic E.
adult Rhesus macaques. J Clin Microbiol 2001; 39:971–6. coli and enterohemorrhagic E. coli host cell attachment. Microb Pathog
14. Wood JD, Peck OC, Tefend KS, et al. Colitis and colon cancer in cotton- 1999; 27:289–301.