HISTOPATHOLOGY OF DONKEYS WITH ENDOMETRITIS
characteristic histopathological lesions of each Kenney- Doig cate- gory as possible was selected as the representative sample of the corresponding grade. Case information of the selected jennies is listed (Table 2 ). The
endometrial biopsy of jenny A, B, C and D were rated as I, IIA, IIB, and III respectively. Few abnormalities were observed in the endo- metrium of jenny A (Figure 1 ). The endometrial epithelium of jenny A was intact and lined with columnar epithelial cells and the stratum spongiosum was loose inferring the presence of mucosal oedema. The endometrial glands were evenly distributed in the stroma and looked normal. There was no obvious abnormality of lymphatics and blood vessels. In jenny B, the endometrial biopsy was rated as IIA, which exhibited mild stromal oedema, mild infiltration of lympho- cytes and eosinophils in the stratum compactum and spongiosum as well as minimal fibrosis of the glands (Figure 2 ). In jenny C, the endometrial epithelium was lined up with cuboidal epithelial cells. The endometrial biopsy illustrated mild stromal oedema and mod- erate infiltration of lymphocytes in both the stratum compactum and spongiosum. Mild periglandular fibrosis was identified in jenny C, which was graded as IIB (Figure 3 ). Jenny D was graded as III, and the endometrial biopsy showed a diffuse, moderate to severe infiltration of lymphocytes and eosinophils within the stratum com- pactum and spongiosum. Glandular density decreased with moder- ate periglandular fibrosis and frequent areas of glandular nesting. No obvious abnormalities of blood vessels and lymphatics were ob- served (Figure 4 ). Among all jennies, different degrees of endometrial inflamma-
tion were microscopically observed with 20 (41.7%) jennies hav- ing mild inflammation, 20 (41.7%) having moderate inflammation, and 8 (16.7%) having severe inflammation. To further describe the cytology results, jenny A showed mild inflammation (5– 15% PMN) with small amount of segmented non- degenerative neutro- phils and accumulated debris (Figure 5a ). The cytology of jenny B (Figure 5b ) and jenny C (Figure 5c ) had moderate inflammation (15– 30% PMN)
illustrating some segmented non- degenerative
to degenerative neutrophils. There were large and moderate amounts of debris in the cytology background of jenny B and jenny C respectively. Severe inflammation (>30% PMN) was ob- served in the cytology of jenny D (Figure 5d ) with minimal de- bris in the background. Low columnar endometrial epithelial cells were shown in cytology of jenny A and B, while in jenny C and D, endometrial epithelial cells were cuboidal and cell clusters were observed. Additionally, a small amount of red blood cells was ob- served in the cytology of jenny A and C.
DISCUSSION
Overall, 2 of 48 cases were graded as category I indicating a healthy endometrium according to the Kenny- Doig scoring system, while they were both repeat breeders with cloudy LVL appearance and their cytological findings showed mild inflammation. They were finally diagnosed with endometritis due to a history of infertility,
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cytologic results related to inflammation as well as a positive bacte- rial culture result in one of two jennies. Similar results have also been reported that some mares with a biopsy score I have >5% PMN in cytology indicating the presence of mild inflammation of endome- trium (Buczkowska et al., 2016 ). Endometrial cytology is usually used for the evaluation of inflammation by the presence of PMNs. Mild peracute inflammation may only exhibit changes in cytology instead of obvious histopathological alterations. As a result, we applied the criteria described by de Amorim et al. ( 2016 ) to diagnose endometri- tis in jennies, which combined clinical finding and laboratory data to improve the accuracy of diagnosis. Historically there was a dilemma when using cytology as a diag-
nostic tool in endometritis because of the lack of a uniform standard. Existing criteria include PMN counts per hpf or per slide, neutrophil to epithelial cell ratio, or the proportion of neutrophils (Card, 2005 ). A recent study pointed out that a ratio of 1% or more of neutrophils to epithelial cells was the most sensitive criterion in mares, but it does not establish degrees of severity associated with the number of neutrophils >1% (de Amorim et al., 2016 ). We applied the criteria described by Card ( 2005 ) along with endometrial biopsy to evaluate endometrial inflammation. Because of different stages of the oestrous cycle, endometrial
morphology of the healthy jennies varies during endometrial biopsy evaluation. Hence, it is critical to determine the jennies ' stage of the oestrous cycle before biopsy sampling. In this study, irrespective of the pathological findings, evidence of the oestrus stage in cytologi- cal and histopathological evaluation was observed. In the endome- trial cytology of the jennies, the impact of oestrus was mainly on the changes in cell morphology. The endometrial epithelial cells tend to be cuboidal in dioestrus, while columnar in oestrus (Quartuccio et al., 2020 ). In the histopathologic findings of this study, the oestrus cycle mainly affected the morphology of the endometrial stroma and glands. Varying degrees of oedema within the stratum spon- giosum associated with elongated nuclei in the columnar epithelial cells were observed in jennies A and B, which were consistent with oestrus- associated changes detected via transrectal ultrasonog- raphy. The stromal oedema of jenny C was mainly localised, and in combination with its stage of oestrus, this tissue oedema may have been artefactually caused by improper handling during tissue prepa- ration and slide production. The endometrium of jenny D showed characteristic dioestrus biopsy findings, including the absence of significant oedema in the endometrium, cuboidal epithelial cells with round nuclei and a wider glandular lumen (Snider et al., 2011 ). When analysing the endometrial cytology and biopsy samples,
it was found that donkeys with more severe clinical signs such as the presence of vaginal discharge and poor reproductive perfor- mance were often associated with more severe inflammatory cy- tological findings and higher endometrial biopsy grades. In those endometrial biopsies with infiltration of inflammatory cells, vari- ous types of inflammatory cells were observed including eosino- phils, macrophages, neutrophils, and lymphocytes. Inflammation cell
infiltration is more common in the endometrium of jennies than of mares, which is the characteristic of higher grades in the
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