Related Experiment Video
Updated: May 17, 2026

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification
Published on: September 15, 2012
Periovulatory changes in the endoscopic appearance of the reproductive tract and teasing behavior in the bitch
1Guide Dogs National Breeding Centre, Leamington Spa, Warwickshire, UK. rachel.moxon@guidedogs.org.uk
Abstract:
A variety of diagnostic techniques have been applied to address the problem of determining optimal mating time in the bitch, although some have not been objectively investigated. Nine healthy bitches were examined daily using routine clinical methods as part of their normal breeding management with the aim of establishing which method was most useful. Accordingly, measurement of the degree of vulval swelling, endoscopic measurement of the vaginal mucosal color and appearance, and objective evaluation of behavior of the bitch were related to the time of ovulation (calculated when progesterone reached 5 ng/mL; Day 0) and the duration of the fertilization period (Day 2 to Day 5) determined by measurement of plasma progesterone concentration and assessment of vaginal cytology. There was a surprisingly large individual variation in measurements of vulval swelling, such that although trends were apparent this technique was not precise enough to establish the optimal mating time. However, measuring color saturation and vaginal mucosal fold width was successful. A consistent change in the mucosal color saturation was observed that related to the beginning (Day -5) and duration of the fertile period although there was no difference in measurement of color saturation between Day -5 and Day 2 (P = 0.31), and Day 2 and Day 5 (P = 0.43). The vaginal mucosal folds were smaller in width at the beginning of the fertilization period (Day 2) than at the beginning of the fertile period (Day -5) (P = 0.008), and all bitches had a consistent appearance of mucosal contours on Day 3. Assessment of sexual behavior was not considered precise enough to enable determination of the optimal time for mating because of the large amount of individual variation in behavioral scores. There was no difference in female behavior scores between Day -5 and Day 2 (P = 0.29), or between Day 2 and Day 5 (P = 0.06). Using objective techniques this study has quantified changes in the color and fold width of the vaginal mucosa that have previously only been subjectively reported. These changes can be used to identify the fertilization period and it is possible, with future refining of the technique, that endoscopic examination could replace vaginal cytology to be used in conjunction with plasma progesterone concentration when examining bitches to determine the optimal time to breed.
Related Concept Videos
Oogenesis
Ovarian Cycle
Hormonal Regulation of the Menstrual Cycle
At puberty, GnRH begins a pulsatile release pattern, which triggers the anterior pituitary gland to secrete follicle-stimulating hormone (FSH) and luteinizing hormone (LH). The frequency and amplitude of GnRH pulses vary across the menstrual cycle, with faster pulses favoring LH release and slower pulses favoring FSH release.
Hormonal Control of the Ovarian Cycle
Before puberty, the hypothalamus releases GnRH in a low frequency, low amplitude pulsatile manner. This along with the immature hypothalamic-pituitary-gonadal axis activity, results in low estrogen levels and the absence of a fully functional ovarian cycle. At puberty, GnRH secretion increases in both frequency and...
Secretory Phase
Following ovulation, the corpus luteum, a temporary endocrine structure, produces progesterone and estrogens. These hormones stimulate the growth and coiling of endometrial...

