Comparison of the cytobrush, cottonswab, and low-volume uterine flush techniques to evaluate endometrial cytology for diagnosing endometritis in chronically infertile mares.
Abstract: Endometritis is the most important cause of infertility in barren mares. The quick method of endometrial cytology (EC) has a relatively high reliability in diagnosing endometrial inflammation in the mare. For reliable cytological results, a collection technique that yields many well-preserved cells representative of a large uterine surface area without causing harm to the reproductive tract is required. The aim of the study was to compare three usually employed techniques for collection of endometrial and inflammatory cells (guarded cotton swab, uterine lavage, and cytobrush) in chronically infertile mares. Twenty Standardbred mares were used. In each mare, samples for EC were collected, first by a cotton swab (DGS), then by a cytobrush (CB), and finally by low volume flush (LVF). The slides were stained using the Diff Quick stain. The following parameters were assessed for each tested technique: background content of the slides; quality of the cells harvested; total cellularity; neutrophils; ratio PMN/uterine epithelial cells; inflammatory cells; vaginal epithelium cells. Categorical variables were compared using contingency tables and Pearson Chi-square tests, whereas continuous variables were compared using one-way analysis of variance (ANOVA); P<0.05 was considered significant. Samplings by DGS and CB resulted easy and quick to perform via a single operator in all cases. LVF was performed easily, but required the presence of 2-3 players and took more time. The background content of the slides prepared by DGS appeared proteinaceous, slides prepared by LVF appeared contaminated by red blood cells or debris, whereas slides prepared by CB appeared clear. All smears showed a good total cellularity. The CB yielded significantly more cells (P<0.0001) than DGS and LVF. The DGS produced significant more cells than LVF (P<0.0001). The DGS produced significantly more (P=0.003) intact cells than CB and LVF. Distorted cells were significantly (P=0.001) more frequent in smears by LVF. The CB harvested significantly (P=0.009) more fragmented cells. CB and LVF produced significantly (P<0.0001; P=0.02) more PMNs/HPF than DGS. In smears collected by LVF the proportion of PMNs/uterine epithelial cells was significantly (P=0.0062; P=0.0023) higher than in smears by CB and DGS. CB collected a significantly higher (P=0.0011) proportion of PMNs than DGS. Acute endometritis was diagnosed in 50% (10/20) of the mares by DGS cytological samples, 25% (5/20) by CB, and 75% (15/20) by LVF. Inflammatory cells other than PMN (lymphocytes, macrophages, eosinophils) were collected exclusively by CB method. Epithelial cells from the vagina were only detected in LVF slides. The agreement of the diagnosis of endometritis between the three techniques of collection and between the different criteria adopted to evaluate smears obtained with the same technique was poor (k≤0.3). In conclusion, results show that cytobrush and flush specimens were superior in all parameters to cotton swab smears. Even though the cytobrush technique requires specialized equipment, sample collection by this method was easier, more consistent, and quicker than the lavage method, indicating that the brush would be the preferred collection method for use on field in the mare. More studies are needed to establish criteria for interpretation of inflammation in the mare on cytobrush samples.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2011-08-23 PubMed ID: 21855980DOI: 10.1016/j.theriogenology.2011.07.020Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article evaluates the effectiveness of three different methods for diagnosing endometritis in mares that have chronic fertility issues. The techniques that were compared are the cytobrush, cotton swab, and low-volume uterine flush methods, with the findings suggesting that the cytobrush and flush specimens were superior compared to cotton swab smears.
Methodology
- The research was conducted on 20 Standardbred mares, each of which had samples collected for endometrial cytology using a cotton swab, a cytobrush, and a low volume flush method, with the cotton swab method being used first, followed by the cytobrush, and then the flush method last.
- The slides from the tests were stained using the Diff Quick stain technique, and several parameters assessed for each sampling technique, including the background content of the slides, the quality of the harvested cells, total cellularity, neutrophils, the ratio of PMN/uterine epithelial cells, inflammatory cells, and vaginal epithelium cells.
- The categorical variables were compared with contingency tables and Pearson Chi-square tests, with continuous variables being compared using a one-way analysis of variance. A result was considered significant if P<0.05.
Findings
- The samplings done by cotton swab and cytobrush could be performed quickly and easily by a single operator, while the lavage method required assistance from 2-3 additional individuals and took more time.
- Cotton swab sampling resulted in a proteinaceous slide background, lavage prepared slides had contamination from red blood cells or debris, and cytobrush prepared slides were clear.
- All smear tests showed good total cellularity, but the cytobrush method yielded significantly more cells than the other two methods, and the lavage method produced the least cells.
- The cotton swab technique produced more intact cells than the other two methods, while the lavage method provided more distorted cells, and the cytobrush method harvested the most fragmented cells.
- Diagnostics of acute endometritis were made via 50% of cotton swab samples, 25% of cytobrush samples, and 75% of lavage samples.
- Only the cytobrush method was able to collect inflammatory cells other than PMN, and vaginal epithelial cells were only detected in slides from the lavage method.
- The diagnostic agreement for endometritis between the three collection techniques was poor (k≤0.3).
Conclusion
- Based on the results, the cytobrush and flush specimens were superior in all parameters compared to the cotton swab smears. Even though the cytobrush technique requires specialized equipment, sample collection by this method was easier, more consistent, and quicker than the lavage method, indicating that the brush would be the preferred collection method for use in the field with mares.
- The researchers concluded by suggesting that further studies are needed to establish criteria for interpreting inflammation in mare samples obtained by using the cytobrush method.
Cite This Article
APA
Cocchia N, Paciello O, Auletta L, Uccello V, Silvestro L, Mallardo K, Paraggio G, Pasolini MP.
(2011).
Comparison of the cytobrush, cottonswab, and low-volume uterine flush techniques to evaluate endometrial cytology for diagnosing endometritis in chronically infertile mares.
Theriogenology, 77(1), 89-98.
https://doi.org/10.1016/j.theriogenology.2011.07.020 Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, Unit of Obstetrics, University of Napoli Federico II, Via F. Delpino 1, 80137 Napoli, Italy.
MeSH Terms
- Animals
- Diagnostic Techniques, Obstetrical and Gynecological / instrumentation
- Diagnostic Techniques, Obstetrical and Gynecological / veterinary
- Endometritis / diagnosis
- Endometritis / pathology
- Endometritis / veterinary
- Endometrium / pathology
- Female
- Horse Diseases / pathology
- Horses
- Infertility, Female / etiology
- Infertility, Female / pathology
- Infertility, Female / veterinary
Citations
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- Sikora M, Król J, Nowak M, Stefaniak T, Aubertsson G, Kozdrowski R. The usefulness of uterine lavage and acute phase protein levels as a diagnostic tool for subclinical endometritis in Icelandic mares. Acta Vet Scand 2016 Sep 7;58(1):50.
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- Del Prete C, Attolini E, Merlo B, Iacono E, Nocera FP, De Martino L, Longobardi C, Damiano S, Longobardi V, Cocchia N, Pasolini MP. Post-Insemination Infusion of Wharton's Jelly Mesenchymal Stromal/Stem Cells-Derived Conditioned Medium: A Novel Approach for Improving Pregnancy Outcomes in Problem Mares. Vet Sci 2025 May 16;12(5).
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