dIn this study, we introduce a multilocus sequence typing (MLST) scheme, comprised of seven single-copy housekeeping genes, to genetically characterize Trichomonas vaginalis. Sixty-eight historical and recent isolates of T. vaginalis were sampled from the American Type Culture Collection and female patients at area health care facilities, respectively, to assess the usefulness of this typing method. Forty-three polymorphic nucleotide sites, 51 different alleles, and 60 sequence types were distinguished among the 68 isolates, revealing a diverse T. vaginalis population. Moreover, this discriminatory MLST scheme retains the ability to identify epidemiologically linked isolates such as those collected from sexual partners. Population genetic and phylogenetic analyses determined that T. vaginalis population structure is strongly influenced by recombination and is composed of two separate populations that may be nonclonal. MLST is useful for investigating the epidemiology, genetic diversity, and population structure of T. vaginalis.
RDEC-1 is a piliated strain of Escherichia coli that was isolated from and produces diarrhea in rabbits without invading the mucosa or synthesizing one of the classical enterotoxins. Previous histological and fluorescent-antibody studies of RDEC-1 diarrhea revealed an acute inflammatory response and large numbers of RDEC-1 associated with (adhering to) the mucosal surface of the ileum, cecum, and colon. The purpose of the present investigation was to further elucidate the histopathology by scanning (SEM) and transmission (TEM) electron microscopy. SEM revealed aggregates of bacteria on the surface of the gut; their distribution was patchy in the ileum and diffuse in the cecum and colon. Bacteria were in contact with each other and appeared to be closely associated with the epithelial surface. TEM showed that the brush border region of the epithelial cells was found to be in varying stages of degeneration, and the bacteria could not be seen adhering to the mucosal cells unless the brush border was absent. Bacteria were in close contact only with epithelial cells that had lost their brush border. The space between the bacteria and the epithelial cells was 11 nm, and it appeared to be filled, in most cases, with densely stained material. This E. coli rarely penetrated epithelial cells, but when it did; it was found in the supranuclear region and never reached the lamina propria. From previous and present studies, it seems probable that RDEC-1 produces diarrhea in rabbits by a mechanism that may be cytotoxic and differs from the classic mechanisms by which E. coli produces diarrhea.Escherichia coli has been shown to produce diarrhea by invasion of the intestinal mucosa or by synthesis of heat-labile or heat-stable enter- otoxins (8, 22, 26, 27). Cantey and Blake have recently described a piliated E. coli (RDEC-1) that reliably produces diarrhea in rabbits with-
A heat-labile material with cytotoxic and enterotoxic activities was isolated from axenically cultivated Entamoeba histolytica. The cytotoxin-enterotoxin was partially purified from the particulate-free supernatant of sonicated trophozoites by ammonium sulfate precipitation and gel filtration. Cytotoxic activity, assayed on monolayers of HeLa or BHK-21 cells, was restricted to proteins that eluted in the molecular weight range of 25,000--35,000 daltons. Cytotoxicity was demonstrated at protein concentrations as low as 2--4 microgram/ml, was heat-labile (75 C, 30 min), and was inhibited by specific immune IgG and by an undetermined factor in nonimmune serum. Enterotoxic activity of the partially purified toxin was demonstrated by induction of fluid secretion in ligated rabbit ileal loops. The cytotoxin-enterotoxin of E. histolytica may play an important role in the production of diarrhea and mucosal injury in amoebic colitis.
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