Fluorescence Spectroscopy for the Diagnosis of Endometritis in the Mare.
Authors: D'Agostino Andrea, Di Palma Tommaso, Cecchini Gualandi Stefano, Boni Raffaele
Journal: Animals : an open access journal from MDPI
Summary
# Fluorescence Spectroscopy for Diagnosing Equine Endometritis Endometritis remains a significant cause of subfertility in mares, yet reliable diagnostic methods remain limited in practice. Researchers exploited the innate ability of polymorphonuclear leucocytes to generate hydrogen peroxide as an immune response, developing a fluorescence-based assay to detect this oxidative marker in endometrial cells collected via uterine flushing. The H2DCF-DA staining protocol, normalised against cellular content using hydroethidine, demonstrated significantly elevated hydrogen peroxide levels in endometrial samples from mares with confirmed endometritis (mean 6.31 ± 1.92) compared to unaffected controls (3.12 ± 1.26, p = 0.001); at a cutoff threshold of 4.4, the assay achieved 88.5% sensitivity and 92.3% specificity. Beyond the fluorescence findings, positive samples also displayed macroscopic differences—greater fluid volume and cloudiness on uterine flushing—suggesting practitioners might identify endometritis through combined assessment of flush appearance alongside cellular analysis. Whilst false negatives (11.5%) warrant consideration, this approach offers an objective, standardised diagnostic complement to conventional cytology, potentially improving selection of mares requiring treatment before breeding and reducing subfertility-related losses.
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Practical Takeaways
- •Fluorescence spectroscopy of endometrial cells offers a quantitative diagnostic method for endometritis that could improve breeding soundness evaluations in mares before insemination
- •The assay's relatively low false negative rate (11.5%) makes it potentially useful as a screening tool, though interpretation should be combined with clinical findings given the 7.7% false positive rate
- •This technique measures immune cell activation rather than just pathogen presence, potentially identifying inflammation that may impair fertility even in culture-negative cases
Key Findings
- •Oxygen peroxide content of endometrial cells was significantly higher in endometritis-positive samples (6.31 ± 1.92) compared to negative samples (3.12 ± 1.26, p = 0.001)
- •Uterine fluid amount and turbidity were significantly higher in endometritis-positive mares (p < 0.01)
- •Using a fluorescence cutoff level of 4.4, the assay achieved 92.3% accuracy with 7.7% false positives and 11.5% false negatives
- •Fluorescence spectroscopy using H2DCF-DA staining can diagnose endometritis by measuring polymorphonuclear leukocyte-derived hydrogen peroxide in endometrial cells