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Following intravaginal inoculation, a<jats:named-content content-type=\"genus-species\">C. muridarum<\/jats:named-content>strain deficient in plasmid-encoded pGP3 or pGP4 but not pGP5, pGP7, or pGP8 failed to spread to the mouse gastrointestinal tract, although mice infected with these strains developed productive genital tract infections. pGP3- or pGP4-deficient strains also failed to colonize the gastrointestinal tract when delivered intragastrically. pGP4 regulates pGP3, while pGP3 does not affect pGP4 expression, indicating that pGP3 is critical for<jats:named-content content-type=\"genus-species\">C. muridarum<\/jats:named-content>colonization of the gastrointestinal tract. Mutants deficient in GlgA, a chromosome-encoded protein regulated by pGP4, also consistently colonized the mouse gastrointestinal tract. Interestingly,<jats:named-content content-type=\"genus-species\">C. muridarum<\/jats:named-content>colonization of the gastrointestinal tract positively correlated with pathogenicity in the upper genital tract. pGP3-deficient<jats:named-content content-type=\"genus-species\">C. muridarum<\/jats:named-content>strains did not induce hydrosalpinx or spread to the GI tract even when delivered to the oviduct by intrabursal inoculation. 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