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The purpose of the study was to compare flow cytometric and haematologic variables in dogs with spontaneous endometritis pyometra complex (EPC) treated with aglepristone to healthy controls. Peripheral blood mononuclear cells: CD3, CD4, CD8, and B lymphocytes (CD21) were analysed by flow cytometry and white blood cell count. Significant differences were observed (P≤0.01) between control (C) and study (S) group in the total number of leukocytes, monocytes and granulocyte populations (without lymphocytes) on beginning, after 7 d, and return to reference value after 14 d of the treatment. The percentage of the T-cell (CD3+) at the beginning was 47.22 ±9.64% of total lymphocytes, in contrast to B lymphocytes (CD21+) that represented the smallest percentage of 14.24 ±7.74% (P≤0.01). The percentage of the lymphocytes CD4+ was 27.42 ±5.53% and CD8+ was 25.18 ±4.36%. The percentage of CD3+ lymphocytes was increasing throughout the experiment in group C and gradually decreased in group S from 14th to 28th d of dioestrus. No differences in the number of CD3+, CD4+, CD8+, and CD21+ lymphocytes between group C and S on the 14th and 28th d of dioestrus (P≥0.05) were observed. The number of CD8+ cells in group S decreased gradually from day 14 to 28 but no statistical differences were noted. Treatment of pyometra with aglepristone decreased the number of leukocytes, monocytes, and granulocytes to referential value but statistically significant influence on the level of subpopulations of T and B lymphocytes was not observed. The results enabled to estimate for the first time the number of lymphocyte subpopulations in dioestrus in both healthy bitches as well as in those suffering from pyometra.
In this study, inflammation in the blood of bitches with pyometra (PG) was compared before and 15 days after ovariohysterectomy (OVH). The results were compared with those for control dioestrus bitches (CG) to reveal the evidence of inflammation in blood after a routine surgery. Inflammation was tracked by tumour necrosis factor-alpha (TNF-α), interleukine-6 (IL-6) and C-reactive protein (CRP), using immunocytochemistry (ICC) and immunofluorescence capture of blood cells in cell blocks. ICC is performed mainly during routine cytological examinations, whereas the use of cell blocks in blood examination is uncommon. Insofar as we know, this is the first study using cell block techniques on canine blood samples. Three commercially available antibodies against TNF-α, IL-6 and CRP, forming two panels, were evaluated. A standard streptoavidin-biotin complex technique was used for ICC. TNF-α and IL-6 labelling was scored for colour and intensity, and CRP for immunofluorescence capture. TNF-α and IL-6 colour and intensity scores differed significantly between the PG and CG groups, and were higher in PG before OVH (P < 0.01, P < 0.01, P < 0.001, P < 0.01, respectively). IL-6 intensity was significantly greater in PG 15 days after OVH (P < 0.05). CRP capture in PG was strong before OVH and high in both groups 15 days later. Low-dose anti-inflammatory agents or an anti-cytokine therapy may be useful in pyometra treatment in the future because these treatments may offer protection from systemic inflammatory response syndrome before and after OVH.
This study aimed to compare the effectiveness of ovariohysterectomy operations with aglepristone and aglepristone+PGF₂ₐ treatment protocols, preceded by clinical, gynaecological, and ultrasonographical examinations and total blood and hormone analyses in bitches. Furthermore, the influence of initial progesterone (P4) concentrations and uteral diameters on the efficacy of pyometra treatment was determined. Thirty bitches with pyometra were divided into three equal groups: the surgically treated group (OP) and groups treated pharmacologically aglepristone (AL) and with aglepristone+PGF₂ₐ (AP). A dose of 10 mg/kg of aglepristone was administered subcutaneously on days 1, 2, and 7 and if needed on day 14 (groups AL and AP). The bitches of the AP group received additionally 0.25 mg/kg of PGF₂ₐ, dinoprost trometamin, once every 24 h between days 3 to 7. Eight bitches in the OP group (80%), five bitches (50%) in the AL group, and five bitches (50%) in the AP group recovered. The success ratios we established, ignoring the criteria for treatment acceptance, increased to 60% in the AL group and 83.3% in the AP group when the bitches with P4>2 ng/mL and without ovarian cysts were evaluated and the difference between success ratios of the two groups was found to be insignificant. It has been found that carrying out frequently repeated examinations, pharmacological treatments using aglepristone or aglepristone+PGF₂ₐ constitutes a safe alternative to ovariohysterectomy in bitches in the dioestrus phase with P4>2 ng/mL and without ovarian cysts.
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