Vaginal candidiasis can be induced in normal rats by estrogenization. Within 48 hr of estrogen administration, the epithelial cells of the vagina are fully keratinized and thus facilitate C. albicans adherence and infection. Oophorectomy is used to increase the animal's susceptibility to infection. This protocol describes vaginal candidiasis in oo-phorectomized Wistar rats upon estrogenization. An advantage of this model of infection is that a large amount of vaginal fluids can be recovered.
Additional Materials (see Basic Protocol 2)
Female oophorectomized Wistar rats, 80 to 100 g (Charles River) Estradiol benzoate (Sigma) dissolved in sterile saline immediately before use Eppendorf Multipipet 4780 (or equivalent) with 2.5-ml calibrated syringe (Combitip plus) and sterile tips
1. Inject female oophorectomized Wistar rats subcutaneously (unit 1.6) with 0.5 mg estradiol benzoate in 0.1 ml sterile saline 6 days prior to vaginal inoculation, and repeat every other day until completion of the experiment.
2. Six days after initiation of estradiol treatment, prepare a suspension of 0.1 ml sterile PBS containing 107 C. albicans blastoconidia, 3153A (see Support Protocol 1).
3. Inoculate 0.1 ml yeast cell suspension into the vagina using a 2.5-ml calibrated syringe. Insert the tip to the end of the vaginal lumen, close to the cervix.
To favor intravaginal contact and adsorption of fungal cells, hold the animal head down for 1 min after inoculation.
4. Proceed with lavage, cfu enumeration, and histological examination (see Basic Protocol 3, steps 4 to 7), but wash the vagina with 500 |l PBS.
For vaginal scrapings with the use of a calibrated plastic loop, see De Bernardis et al. (1999).
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