350
|
ABDELNABY ET AL.
FIGURE 2 Ultrasonogram showing uterine body- coloured area with red and blue colours refereed to blood flow away and toward the uterus in order to get spectral graph in from of peak velocity point (PV) and end point (EV) before (a) and after treatment (b) note all Doppler parameters do not significantly differ before and after treatment.
and histopathological samples were collected from all infected mares on day 0 (third ovulation).
Inseminations of affected mares
The only mares determined to have chronic endometritis (treated mares) were inseminated at the third ovulation using a stallion ' s fro- zen/thawed semen, which had a 35%– 40% pregnancy rate. Every 6 h while in mares in oestrus, the dominant follicle was assessed by ultrasound, and within 6 h after ovulation, mares were inseminated intrauterine using frozen/thawed semen (in one straw [0.5 mL; Go- vaere et al., 2014 ]), with a check of pregnancy on days 9, 14 and 20.
Blood sampling and hormonal evaluation
Routine jugular venipuncture was performed to collect serum for hormonal analysis. Following that, the blood was coagulated and cen- trifuged at 1900 g . For hormonal analysis, serum was kept at −20°C. A commercially available kit (DRG International, Inc.) Estradiol 17β (E2) and progesterone (P4) were analysed using a commercially avail- able assay kit by enzyme- bound immune adsorption as determined earlier in the current study ' s laboratory (Abdelnaby, Abo El- Maaty, & El- Badry, 2020 ; Abdelnaby, Abo El- Maaty, Ragab, & Seida, 2020 ).
Histopathological evaluations
All mares with endometritis had a similar endometrial histopathol- ogy. At least 2 blinded independent pathologists read the biopsies
and provide agreement. Uterine biopsies were collected days before treatment (day −1), and after treatment (day 0 = 3rd ovulation) sam- ples were fixed in 10% neutral- buffered formalin. The fixed tissues were then dehydrated, cleared, and embedded in paraffin. Section- ing of paraffin blocks at a thickness of 3– 4 μm was done using a ro- tatory microtome (Leica, made in Germany). Tissue sections were stained using haematoxylin and eosin and Masson ' s trichrome to de- tect the presence of fibrosis under a light microscope Olympus BX43 with a camera (Suvarna et al., 2012 ).
Statistical analysis
All data were first assessed for normality. The data were expressed as mean ± standard error of the mean. Coloured areas of the uterus and uterine thickness and blood flow estimated by pulse wave Dop- pler are assessed using repeated measures of ANOVA using SPSS software 20. The effect of the days was calculated. In addition, Dun- can ' s multiple range test was determined to differentiate p < 0.05 between significant means.
RESULTS
Histopathological evaluation before and after treatment
Uterine biopsy samples before treatment showed a marked degree of degenerative changes as seen in glandular atrophy and fibrosis with newly formed blood vessels between the organised fibrous tissue bundles (Figure 3 ), on the contrary to the biopsy after the
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80 |
Page 81 |
Page 82 |
Page 83 |
Page 84 |
Page 85 |
Page 86 |
Page 87 |
Page 88 |
Page 89 |
Page 90 |
Page 91 |
Page 92
