The presence of virulence factor genes downregulates uterine AQP3 and alters glutathione peroxidase activity and uterine histopathology in canine pyometra.

Present study was designed to evaluate the role of virulence factor genes (papG, cnf1 and hylA) in the pathogenesis of canine pyometra. Antimicrobial susceptibility test and detection of virulence genes were performed Escherichia coli (E. coli) detected in uterine swab samples. Animals were divided into two groups based on the presence (VF+, n:14) or absence (VF-, n:7) of the virulence factor genes papG, cnf1 and hylA. Blood and tissue glutathione peroxidase activity, uterine histopathologic analysis and AQP3, ESR1, PGR, OXTR gene expressions were determined in both groups. Statistical analyses were performed using Stata version 15.1. All E. coli isolates were susceptible to amikacin, whereas resistant to ampicillin, amoxicillin/clavulanic acid and lincomycin. None of the isolates were susceptible to cefotaxime. E. coli isolates had at least one virulence gene. The most prevalent gene was fimH (100%), followed by fyuA (95.8%), usp (83.3%), sfa (75%), cnf1 and hlyA (70.8%) genes. Blood GPx activity was greater in VF+ animals. On the other hand, uterine tissue GPx activity was lower in VF+ group compared to the control group. Expression levels of AQP3 were upregulated more than fivefold in VF-dogs compared to the control group. In addition, AQP3 expression levels were found approximately threefold higher in VF (-) than VF (+) group (p < .05). Varying degree of inflammation noted for all animals with pyometra, but the presence of bacteria noted only in VF+ animals. In conclusion, the presence of virulence factor genes does not play a role in the histopathological degree of inflammation, the presence of bacteria was found to vary. Serum GPx activity increased in VF+ animals. While the hormone receptor expressions were similar, AQP expression was upregulated in the absence of virulence factor genes.

The role of nutritional-immunological indices in estimating serum LPS and antioxidant enzyme activity and sepsis status in female dogs with pyometra caused by E. coli.

The aim of this study was to diagnose pyometra and related sepsis status using cost-effective nutritional-immunological indices, antioxidants, and toxin levels in dogs and to investigate the utility of the indices in predicting toxin and antioxidant status. A total of 29 dogs were enrolled into the present study. Among these, 9 female dogs in their diestrus stages, were allocated for elective ovariohysterectomy. The pyometra group was also separated into two subgroups as Sepsis (+) and Sepsis (-). Blood samples were collected into two tubes containing EDTA for hematological analysis; without anticoagulant for serum progesterone, LPS concentration, and antioxidant levels at the time of diagnosis. Bacteriological and tissue samples of the uterus were collected after the ovariohysterectomy. Antioxidant activity, progesterone, and toxin concentration were determined by using commercial ELISA kits. Statistical analyses were performed using Stata version 16.1 and MedCalc 16 statistical software. Receiver operating characteristics curves were used for the threshold for evaluating pyometra and sepsis status. Pairwise comparisons were carried out of the area under the curve (AUC) for thresholds of nutritional immunologic indices (hemoglobin, albumin, lymphocyte, platelet (HALP) score; prognostic nutritional index (PNI); Albumin hemoglobin index (AHI)), serum LPS and antioxidant activity. Linear regression model was used for the estimation of serum LPS and antioxidant activity by using indices. Mean serum progesterone, LPS concentrations, and Nitric Oxide (NO) production were greater, while serum superoxide dismutase (SOD), tissue SOD, and glutathione peroxidase (GPx) activities were lower in dogs with pyometra. All nutritional-immunologic indices were lower in pyometra cases. Nutritional-immunologic indices (AUC of HALP:0.759; PNI:0.981; AHI 0.994), NO (AUC: 0.787) and SOD (AUC: 0.784) levels were useful for pyometra diagnosis. AHI and LPS were useful for the determination of sepsis status with the AUC values of 0.850 and 0.740, respectively. While AHI was useful for the estimation of serum LPS and NO concentration (p < 0.001), PNI was useful for serum SOD concentration (p = 0.003). In conclusion, PNI, HALP and AHI can be used in the diagnosis of pyometra, however, only AHI and LPS levels can be used in the diagnosis of sepsis. SOD and NO can be used to determine pyometra but have no effect on determining sepsis status. Additionally, the estimation of the levels of serum LPS, NO, and SOD activities can be done using the AHI and PNI values.

The evaluation of SIRS status with hemato-biochemical indices in bitches affected from pyometra and the Usefulness of these indices as a potential diagnostic tool.

The aims of this study were to establish a cut-off point by evaluating the usability of the hemato-biochemical indices to determine the severity of inflammation caused by pyometra, and to determine the practical usage of the indices as a diagnostic biomarker in cases of pyometra. Animals that were admitted to Ankara University Veterinary Faculty Obstetrics and Gynecology Clinic were included in the present study. Dogs suffering from pyometra were allocated to two groups according to their SIRS status (SIRS, n = 12; Non-SIRS (NSIRS, n = 16)). Healthy dogs presented for routine ovariohysterectomy were used as the control (n = 13). Blood samples were collected into two tubes containing EDTA for hematological analysis and without anticoagulant for biochemical tests before the beginning of treatment. Hemato-biochemical indices, which are Neutrophil-Lymphocyte Ratio (NLR), Monocyte-Lymphocyte Ratio (MLR), Neutrophil Percentage-Albumin Ratio (NPAR), Platelet-Lymphocyte Ratio and Systemic Immune-Inflammation Index, were calculated. All data were analyzed using the MIXED procedure of the SPSS. Receiver operating characteristics curves were used to determine the dog level threshold for evaluating the SIRS status and used to determine a predicted threshold with hemato-biochemical indices for differentiation between dogs with pyometra (regardless of SIRS status) and the control. Pairwise comparisons were carried out of the area under the curve (AUC) for thresholds of SII, NLR, PLR, MLR, NPAR. White blood cells, lymphocytes, monocyte, neutrophil, ALP, NLR, MLR, and NPAR levels were greater in dogs that suffer from SIRS (P < 0.05). Red blood cells and hematocrit levels were significantly higher in the control group (P < 0.05). Albumin levels were higher in the control group (P < 0.05). Moreover, NLR, MLR and NPAR levels were found to be significantly higher in the SIRS group (P < 0.05). Besides, NLR and MLR were the best predictors for diagnosis of the SIRS with the AUC values of 0.936, and 0.966 respectively. Furthermore, NLR, MLR and SII could be a useful diagnostic tool for pyometra with the AUC values of 0.955, 0.931, 0.716. The blood indices could be used as fast, easily accessible, and inexpensive markers for the evaluation of the SIRS status in dogs with pyometra together with diagnosis.