Pyometra Group Research Articles

Expression patterns and distribution of aquaporin water channels in cervix as a possible mechanism for cervical patency in bitches affected by pyometra

Pyometra is a life-threatening disease, the severity of which depends on cervical patency status. This study investigated cervical inflammation status as well as the expression patterns and localization of aquaporin (AQP1, AQP2, AQP3, AQP5, and AQP9), and hormone receptors in cervical tissue that influences canine pyometra. Of the 36 animals enrolled in the study, 24 were diagnosed with pyometra and separated into two groups: open cervix pyometra and close cervix pyometra, while 12 healthy animals presented for elective ovariohysterectomies were allocated into the control group. Surgical treatment was performed for treatment of pyometra. After each operation, cervix samples were collected and analyzed for AQP and hormone receptor expression patterns determined by qPCR and protein expression by means of immunohistochemistry. Blood samples were also collected to determine serum progesterone concentrations. AQP9 expression was downregulated approximately 3-fold while and PGR expression was downregulated more than 2 fold in both pyometra groups compared to the control group. AQP3 and AQP5 gene expression levels were upregulated more than 3 fold in the open-cervix pyometra group than the closed-cervix pyometra group (P < 0.05). This is the first study to describe the expression patterns and immunolocalization of AQPs in canine cervical tissue based on pyometra patency status and to report AQP3 and AQP5 expression in cervical tissue linked to cervical patency.

Evaluation of calcium, phosphorus and some biochemical parameters in dogs with open and closed cervix pyometra

The aim of this study was to evaluate the changes in calcium, phosphorus and some biochemical parameters in dogs with open and closed cervix pyometra, which was then compared with a control group. A total of 62 bitches of age group 5-10 years old irrespective of breed were enrolled into the study. Control group consisted of 22 bitches which were clinically healthy and in luteal phase of the estrus cycle. On clinical examination, pyometra was diagnosed in 40 bitches while 23 out of 40 bitches had open-cervix pyometra and 17 of 40 bitches had closed-cervix pyometra. Evaluation of haematological changes revealed that there was increased white blood cell (WBC), total protein (TP), globulin, alkaline phosphatase (ALKP) while decreased glucose (Glu), albumin (Alb) and albümin globulin ratio in pyometra cases (p&amp;lt;0.001). The highest WBC and ALKP levels were detected in closed-cervix pyometra group (p&amp;lt;0.001). Calcium (Ca+2) levels had significant rise in open-cervix pyometra cases (p&amp;lt;0.05) while phosphorus (Phos) levels had tend to increase in closed-cervix pyometra cases (p=0.08). Also, Ca was directly correlated with Glu, Alb and TP while inversely correlated with WBC and blood urea nitrogen/creatinine (BUN/Crea). However, Phos was significantly associated with BUN and Crea (p&amp;lt;0.001; R=0.915 and R=0.860, respectively). As a result of this study, it was observed that the increased Ca levels in open-cervix cases and the elevated Phos levels in closed-cervix cases, were associated with the biochemical parameters related to hepatic and renal dysfunction in dogs with pyometra.

Exploring oxidative stress, immunological and metabolic biomarkers in dairy cows with postpartum pyometra.

Pyometra is a prevalent and severe infectious disease that affects the reproductive systems of cattle worldwide. This study's main goal was to investigate the biomarkers for oxidative stress (OS), adiponectin, leptin and neopterin (NPT) in cows suffering from postpartum pyometra. The study also aimed to determine which bacteria were most commonly implicated in the development of the disease. A total of 74 cows with pyometra were examined and compared to a control group of healthy cows (n = 20). In comparison to the healthy control and post-treatment groups, the pyometra group showed higher mean values of leptin, adiponectin and malondialdehyde (MDA). In contrast, the glutathione (GSH) and superoxide dismutase (SOD) mean values were lower in the pyometra group as compared to the post-treatment and control groups. NPT levels in the post-treatment groups were lower than those in cows with pyometra but comparable to the healthy control group (p > .05). When compared to the other biomarkers, NPT, leptin and adiponectin showed higher sensitivity and specificity in identifying pyometra cases (AUC ≥0.99). The predominant bacterial isolates from the ptomtra-affected cows consisted of Escherichia coli (N = 29; 39.2%), Arcanobacterium pyogenes (N = 27; 36.5%) and Fusobacterium necrophorum (N = 13; 17.6%). Mixed infection was determined in nine samples (12.2%). Conclusively, OS, adiponectin, leptin and NPT play crucial roles in comprehending the development of postpartum pyometra in cows and have the potential to serve as biomarkers for the disease.

Effect of pyometra on vascularity alterations, oxidative stress, histopathology and inflammatory molecules in feline

This study aimed to investigate blood flow, hemodynamical features by Doppler ultrasound, the oxidative stress biomarkers from serum samples, and histopathology from uterine tissue, in healthy queens and queens with pyometra. Twenty queens were categorized into two groups, according to signs, history, and ultrasound findings, as pyometra and control healthy queens. Doppler ultrasonography, total antioxidant capacity (TAC), malondialdehyde (MDA), albumin, bacteriological isolation, histopathology, and immunohistochemistry of tumor necrosis factor-alpha (TNF-α) and nuclear factor kappa B (NF-κB) P65 were performed. Uterine diameter and thickness increased significantly in the pyometra group compared to control. Uterine peak velocity and flow rate were significantly higher in the control group. The pyometra group showed a significant decrease in albumin, TAC, and a significant increase in MDA. Fibrosis and mononuclear inflammatory cell infiltration were seen in the pyometra samples. The mean area percentage of TNF-α expression in the uteri of the pyometra group was higher. The expression of NF-κB P65 in the uteri in the pyometra group was significantly higher. Doppler ultrasonography can provide valuable information for diagnosing pyometra in queens by elevating the uterine thickness with reducing blood flow rate. Oxidative stress, TNF-α, and NF-κB expression alterations varied between pyometra and control groups.

Proteínas de fase aguda, perfis hematológicos e bioquímicos séricos de cadelas em diestro, mucometra e piometra

ABSTRACT Blood samples from 200 female dogs divided into 6 groups (diestrus, mucometra, pyometra) were evaluated, with the pyometra group categorized according to the ASA classification (American Society of Anesthesiologists), that is, from ASA II to V, totaling six groups. Aiming to analyze the acute phase proteins (APP), hematological and serum biochemical profiles of the female dogs in the study, establishing the differential diagnosis and prognosis according to the group. The SDS-PAGE method was used for protein fractionation, complete blood count using an automated hematological analyzer and histogram, biochemical tests performed using a semi-automatic spectrophotometer and measurement of serum concentrations of sodium and ionic calcium using the selective ion method. The results showed that female dogs with pyometra categorized in the ASA II to V classification revealed biochemical alterations between the study groups, as well as the acute phase proteins, presented variations according to the systemic involvement, degree of inflammatory response and ASA classification of the female dogs with pyometra, in which ceruloplasmin (p=0.07) and transferrin (p=0.07) did not show statistical significance, but showed an increase in their concentration according to the inflammatory evolution, albumin (p&lt;0.0001), IgGCP (p &lt;0.0001), haptoglobin (p&lt;0.0001), alpha acid glycoprotein (p&lt;0.0001), IgGCL (p&lt;0.0001) and the one identified by its atomic weight 23,000 Da (p=0.0031), enabling the assessment of the acute phase response and aiding in the early detection of the systemic inflammatory response and better therapeutic guidance for the patient.

Cystic endometrial hyperplasia-pyometra syndrome impairs the preantral follicle reserve in domestic bitches (Canis familiaris)

Cystic endometrial hyperplasia (CEH)-pyometra syndrome is the most common uterine pathological condition reported in breeding bitches, however, their described effects on fertility are limited to uterine disorders and conception rates. As the preantral follicle population represents the available reserve of gametes recruited during the lifespan, the aim of this study was to evaluate the effects of CEH-pyometra syndrome on the: (i) preantral follicle morphology, (ii) developing follicle rates, and (iii) preantral follicle and stromal cell densities. Ovarian fragments from bitches subjected to elective or therapeutic ovariohysterectomy were allocated according to uterine diagnosis as follows: control (n = 7, clinically healthy), CEH-mucometra (n = 8, uterine lumen filled with a sterile mucus), and pyometra (n = 17, presence of a purulent mucus) groups. Overall, the control group had 3.4 and 4.1-fold higher probability (P < 0.0001) of the presence of normal preantral follicles compared with CEH-mucometra and pyometra groups, respectively. Moreover, ovarian fragments from the pyometra group showed an increase in the percentage of developing follicles (P < 0.05) compared to the control. Both CEH-mucometra and pyometra groups showed lower (P < 0.05) preantral follicle and stromal cell densities (P < 0.05) compared to the control. In summary, the CEH-pyometra syndrome decreased the percentage of morphologically normal follicles and enhanced the developing follicle rates. Additionally, a reduction of preantral follicle and stromal cell densities suggests that the inappropriate uterine environment induced by CEH-pyometra syndrome can lead to premature depletion of ovarian reserve.

The assessment of haematologic and serum chemistry parameters in canine pyometra: a systematic review and meta-analysis.

Pyometra is common in non-spayed adult female dogs requiring early diagnosis and treatment to increase the survival rate. The diagnosis of pyometra is mainly based on clinical examination and anamnesis. Radiography, ultrasonography and laboratory analyses are recommended to support a diagnosis. The aim of this study was to assess blood parameters associated with pyometra by performing a systematic review and meta-analysis. The review was conducted in accordance with PRISMA guidelines. A search of three databases (PubMed, Google Scholar and CAB abstracts) was performed in July 2022. Studies providing information about laboratory parameters for both the pyometra group and healthy control group dogs were eligible for inclusion. Data extraction included the first author's name, publication year, country, number of participants in both groups, mean values of selected studies, standard deviation values, and blood parameters. The risk of bias for each study was evaluated, and a random-effects meta-analysis was performed. We included 44 studies which investigated 12 blood parameters. High heterogeneity was detected in all parameters in these meta-analyses. The following blood parameters were increased in dogs with pyometra: white blood cell (mean=27.75×109 L-1 , mean difference (MD)=17.16, 95% confidence interval (CI) 14.85 to 19.47), monocytes (mean=2.06×1012 /L, MD=1.37, 95% CI 0.99 to 1.74), blood urea nitrogen (mean=41.42 mg/dL MD=18.06, 95% CI 12.26 to 23.85), alkaline phosphatase (mean=212.78 IU/L, MD=137.51, 95% CI 81.81 to 88.62), and aspartate aminotransferase (mean=48.31 IU/L, MD=16.96, 95% CI 10.61 to 23.30). The following parameters were reduced: red blood cell (mean=5.42 1012 /L, MD=-1.37, 95% CI -1.68 to -1.05), haemoglobin (mean=121.20 g/L, MD=-30.57, 95% CI -39.70 to 21.45), albumin (mean=23.71 g/L, MD = -8.16, 95% CI -11.46 to -4.86). Lymphocyte, creatinine, urea, and alanine transaminase parameters were increased in some studies and decreased in others. In canine pyometra, some blood parameters are consistently increased, some consistently decreased, and some increased or decreased depending on the study.

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.

Could homocysteine represent a negative acute phase reactant in canine infections-a pilot study?

Homocysteine (Hcy) was investigated as the biomarker of cardiac, renal, and gastrointestinal disorders in dogs. Data about low Hcy concentrations in the systemic inflammatory response syndrome raised a hypothesis that Hcy in dogs is a negative acute-phase reactant. This survey compared Hcy concentrations, serum amyloid A (SAA), and the routine laboratory parameters between healthy (HD, N=6) and dogs with inflammation of different extent: mild ( dirofilariosis (DIR), N=31), moderate (babesiosis (BAB), N=12), and severe (pyometra (PYO), N=8). The BAB and PYO groups had lower Hcy er than HD. Also, the levels in the PYO group were below those in the DIRO group. SAA had the inverse pattern. Across the groups, Hcy and SAA levels correlated negatively (ρ = -0.502, P&lt;0.001). Hcy and SAA correlated with the erythrocyte count, hematocrit, hemoglobin and mean cellular hemoglobin concentrations, and neutrophil count, with correlations being positive for Hcy and negative for SAA. Among all dogs, hemoglobin was the only independent predictor of Hcy concentration. Hcy levels in canine infections, decreased as acute-phase reaction (APR) intensified. Also, they were related with the hematology changes accompanying the APR. Further studies will establish the clinical potential of these alterations.

Evaluation of infrared thermography findings in postoperative follow-up in dogs with pyometra

In this study, the usability of infrared thermography (IRT) applications in the postoperative follow-up after the sterilization operation was investigated in dogs with pyometra. The study was carried out on healthy animals (Group 2, n=7) diagnosed as pyometra (Group 1, n=7) and undergone elective ovariohysterectomy, consisting of 14 female dogs. Serum CRP levels were measured from the patients before the operation and on the 1st, 4, and 7th days postoperatively. Thermographic images from the eye and incision area were recorded, and the results were analyzed statistically. There were no intraoperative and postoperative complications in both study groups, and all dogs in Group 1 recovered uneventfully. Serum CRP levels were higher at 0, 4th, and 7th days in Group 1 (P0.05) in both groups, a decrease was observed on the 4th and 7th days (P0.05). While serum CRP values reached the maximum level on the 1st day after surgery in the healthy and patient groups, it was seen as a milder increase due to the inflammatory reaction in the pyometra group. Due to the uncomplicated completion of the study, CRP values tended to decrease on the 4thday, and no significant difference was observed in the thermographic images. However, it is not reflected in the statistical data; in the pyometra group, the eye on the 1st and 4th days. The abdominal temperature remained higher than the control group on all other days. It is thought that the use of thermography, which is a non-invasive and practical application, in addition to serum CRP measurement after ovariohysterectomy, in similar studies to be applied in larger populations in the future, will help determine the prognosis.

Doppler Velocimetry of Uterine Artery in Bitches with Cystic Endometrial Hyperplasia-Pyometra Complex

Doppler ultrasound is a useful diagnostic tool to evaluate uterine blood flow in different canine reproductive states. The aim of this study was to describe and compare uterine blood flow in bitches suffering from pyometra according to the ultrasonographic appearance of uterus. Seventy-five bitches with pyometra confirmed clinically, haematologically and ultrasonographically were included in the study. Ten healthy bitches with no haematological and ultrasonographic abnormality of uterus were also included in the study as control group. The bitches with pyometra were divided into four groups based on the sonographic characterization of uterus and its luminal contents, viz., B (few small cysts, normal endometrial surface, anechoic uterine content, n=11), C (many large cysts, irregular surface and hypertrophic endometrium, n=24), DH (many large cysts, irregular surface and hypertrophic endometrium, hyperechoic content, n=31) and DA (many large cysts, irregular surface and atrophic endometrium, hyperechoic content, n=9). The uterine horn diameter and uterine wall thickness was measured with two-dimensional ultrasonography. With the use of pulsed-wave Doppler, flow velocity waveforms of uterine artery were obtained. Parameters of blood flow velocity such as peak systolic velocity (PSV), end diastolic velocity (EDV), as well as haemodynamic parameters such as resistance index (RI), pulsatility index (PI) and systolic-diastolic ratio (S/D) were calculated electronically. Peak systolic velocity and end diastolic velocity were significantly higher in pyometra group than control with highest velocities recorded in DA group (P&lt;0.05). The RI, PI and S/D were significantly lower in pyometra group than in control (P&lt;0.05). The RI was significantly lower in group C and DA compared to groups B and DH (P&lt;0.05). However, no significant difference was reported in PI and S/D among the groups of pyometra (P&gt;0.05). It was concluded that the uterine artery blood flow velocity of bitches suffering from pyometra differs, not only from normal bitches, but also varies with the type of pyometra with varying pathological nature. Furthermore, haemodynamic parameters appear as useful markers to differentiate uterine pathologic conditions and to understand the pathological type of pyometra.

Activity of the glycosidases β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase, and sialidase in uterine tissues from female dogs in diestrus with and without pyometra

This study aimed to compare the activity of selected glycosidases (β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase, and sialidase) in homogenates of uterine tissues obtained from female dogs with and without pyometra. In addition, it examined the availability of substrates for these glycosidases in the homogenates. The study was carried out on female dogs undergoing ovariohysterectomy for pyometra (n = 10) and clinically healthy dogs (n = 10) undergoing elective spaying. The activity of β-galactosidase, α-l-fucosidase, and β-N-acetyl-hexosaminidase was analyzed using a spectrofluorometer and that of sialidase using a colorimetric method. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis with Alcian Blue (AB) and Periodic Acid–Schiff (PAS) staining was performed to determine the presence of substrates for these glycosidases in the homogenates of uterine tissues. The results revealed that the activity of all the examined glycosidases was significantly higher (P < 0.05) in the uterine tissues isolated from dogs with pyometra in comparison to healthy dogs. The electrophoretic patterns of the selected samples showed several proteins, which contained different sugar moieties stained by AB and PAS and the profiles differed significantly between the pyometra group and the healthy group. Densitometric analysis of AB staining showed patterns between 233 and 148, 86 and 55, and 43 and 20 kDa, which differed markedly in sugar content between the examined groups of animals. Similarly, PAS staining analysis revealed patterns of different molecular weights, between 233 and 117 and between 55 and 32 kDa, which also differed in sugar content. These findings suggest that canine pyometra is accompanied by the increase in the activity of selected glycosidases in the uterus. This could potentially modify the glycan structures of uterine glycoproteins and in result their biological functions. Further studies are needed to elucidate the potential role of the increased activity of glycosidases in the pathogenesis of this disease.

Uterine artery hemodynamics in female dogs with open- and closed-cervix pyometra

Although pyometra is a common disease, the mechanisms that determine cervical opening remain unknown. Knowing that the vascular structures are crucial in pathophysiology, it was observed need for hemodynamic studies assessing uterine artery of female dogs with pyometra and its relation to the neck opening. Thirty-five female dogs were selected and separate into three groups: control group (CG) (n = 12), open-cervix pyometra group (OCG) (n = 11) and closed-cervix pyometra group (CCG) (n = 12), with the objective of evaluating and comparing the hemodynamic changes of the uterine artery [peak systolic velocity (PSV), end diastolic velocity (EDV), and resistance index (RI)] in female dogs with open- and closed-cervix pyometra and correlate them with measurements of uterine diameter (UD) and endometrial thickness (ET). The correlation analysis showed that, with the exception of PSV, the hemodynamics indices were associated with UD and ET, presenting a moderate and positive correlation between UD and EDV (r = 0.62; P&lt;0.01), a moderate and negative correlation between UD and RI (r =-0.68; P&lt;0.01) and also moderate and negative correlation between ET and RI (r = -0.62; P&lt;0.01). These results suggest that alterations of uterine artery hemodynamics are similar in dog females with open- or closed-cervix pyometra, although the UD and the ET can influence in the uterine perfusion.

Pyometra-associated insulin resistance assessment by insulin binding assay and tyrosine kinase activity evaluation in canine muscle tissue

Diestrus is associated with insulin resistance in bitches and pyometra can further impair insulin sensitivity. This study aimed to compare insulin sensitivity, insulin binding, and tyrosine kinase activity in bitches in anestrus, diestrus, or with pyometra. Patients submitted to elective ovariohysterectomy were divided into anestrus (n = 11) or diestrus (n = 13) according to reproductive history, vaginal cytology, and uterine histology. The group pyometra (n = 8) included bitches diagnosed with the disease based on clinical presentation and abdominal ultrasound findings and further confirmed by uterine histopathology. All patients were submitted to an intravenous glucose tolerance test (IVGTT) before ovariohysterectomy, and rectus abdominis muscle samples were collected during surgery for plasmatic membrane suspension preparation. Muscle-membranes were submitted to cold saturation insulin binding assay for dissociation constant (Kd) and maximum binding capacity (Bmax) determination, as well as exogenous substrate Poly (Glu: Tyr 4:1) phosphorylation assay for basal tyrosine kinase evaluation. Bitches with pyometra showed higher basal insulin (P < 0.001) and higher area under the curve (AUC) for insulin (P = 0.01) and glucose (P < 0.001) response during the IVGTT in comparison with bitches in anestrus or diestrus. Diestrus (P < 0.0001) and pyometra (P = 0.001) were associated with reduced tyrosine kinase activity in comparison with anestrus. No differences were documented in Kd and Bmax results for the low-affinity/high-capacity insulin receptors; however, high-affinity/low-capacity insulin receptors showed higher Kd and Bmax results in bitches in diestrus or with pyometra (P < 0.05) in comparison with anestrus. Despite the pyometra group showed the highest Kd values (P < 0.01), its Bmax results did not differ from the diestrus group (P > 0.05). Diestrus’ higher Kd values and reduced tyrosine kinase activity in muscle tissue were compensated by increased total insulin binding capacity. Absent differences in IVGTT results between diestrus and anestrus bitches corroborate this finding. However, in bitches with pyometra, the highest Kd values were not compensated by increased total insulin binding capacity. This finding was associated with insulin resistance and glucose intolerance in IVGTT results. Moreover, pyometra resolution restored insulin sensitivity and glucose tolerance. These features can play a key role in pyometra-associated CDM, as well as in diabetic remission after pyometra resolution.

The deleterious effect of postpartum pyometra on the reproductive indices, the metabolic profile, and oxidant/antioxidant parameters of dairy cows.

Background and Aim:Postpartum uterine infectious diseases, such as pyometra, have discrepancy with both health and, subsequently, productivity of dairy cows due to its high prevalence and the high cost of treatment. Therefore, this study investigates the influence of pyometra on the reproductive indices, the metabolic profile, and oxidant/antioxidant parameters of the pyometric animal compared to those of healthy ones.Materials and Methods:The study included 30 cows. The animals were differentiated into two groups of 15 cows each: A group of pyometra and a control group. All pyometric cows were subjected to breeding soundness examination after the end of pyometra and were compared to the control group. Blood samples were obtained to assess the levels of glucose, non-esterified fatty acids (NEFA), triglycerides (TGs), cholesterol, albumin, total protein, alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase (ALP), blood urea nitrogen (BUN), creatinine, calcium (Ca), phosphorus, sodium, potassium, progesterone hormone (P4), malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), and superoxide dismutase.Results:Results revealed significant prolonged duration of first estrus, the days open, and the required number of services due to pyometra. The pyometra group yielded increased levels of NEFA, TGs, ALP, BUN, creatinine, MDA, and progesterone hormone. In addition, significant decrease in the levels of glucose, cholesterol, albumin, Ca, phosphorus, sodium, TAC, GPx, and superoxide dismutase was observed in the pyometra group. Finally, no difference in the concentrations of total protein, ALT, AST, and potassium was observed in the pyometra group.Conclusion:The reproductive indices was adversely influenced in cows with postpartum pyometra, and metabolic profile, involving energy balance signals and liver function indicators, revealed differences between the two groups. Increased levels of oxidative stress parameters and decrease levels of antioxidant levels were also found, suggesting that pyometra is an incentive for oxidative stress. Overall, checking the energy balance, metabolic imbalances, and oxidant/antioxidant profile, accompanied with pre-emptive procedures during the postpartum period, is essential and can reduce the chances of such diseases and possible noxious results in highly productive cows.

Acute phase proteins and antioxidant responses in queens with pyometra

Acute phase proteins (APP) and biomarkers of oxidative status have proved to be clinically useful biomarkers of pyometra in different species. Despite pyometra is considered one of the most important feline reproductive diseases, information about the APP response and the oxidative status in queens with pyometra is lacking. This study aimed to evaluate the APP and the antioxidant responses at diagnosis and in the post-operative period in feline pyometra. Serum concentrations of serum amyloid A (SAA), haptoglobin (Hp), albumin, total serum thiols (Thiol) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity (TEAC) assessed by two different methodologies (TEAC1 and TEAC2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in 23 queens with pyometra at diagnosis and in 13 healthy control queens submitted to elective ovariohysterectomy. The APP and antioxidants were also evaluated in 11 queens of the pyometra group at days two and 10 after surgery. At diagnosis, queens with pyometra had serum concentrations of SAA, Hp, and FRAP significantly higher (P < 0.001, P < 0.001 and P < 0.05, respectively), and of albumin, Thiol, CUPRAC and TEAC2 significantly lower (P < 0.001, P < 0.001, P < 0.001 and P < 0.01, respectively) than controls. Moreover, concentrations of APP and antioxidants were significantly different (with a tendency to return to physiologic levels) at day 10 after surgery than before surgery. Significant associations were found between APP and antioxidants. According to these results, an APP response and the development of oxidative stress were detected in queens with pyometra. In addition, APP and antioxidants tended to return to physiologic values after surgery in the queens that recovered from the disease. Therefore, our results suggest that APP and selected antioxidants, such as Thiol and CUPRAC, could be potentially useful biomarkers in diagnosis and assessment of the post-operative period in feline pyometra.

Cystic endometrial hyperplasia-pyometra syndrome in bitches: identification of hemodynamic, inflammatory, and cell proliferation changes.

Cystic endometrial hyperplasia (CEH)-pyometra syndrome is one of the most common diseases of noncastrated female dogs. However, determination of etiological mechanisms and differential diagnosis of CEH-pyometra syndrome are undefined. The aim of this study is to compare immunohistochemically the expression of cyclooxygenase-2 (COX-2) inflammatory mediator, Ki-67 antigen proliferation marker, vascular endothelial growth factor (VEGF-A) angiogenesis mediator and its FLT-1 and KDR receptors, and correlate with Doppler velocimetry of uterine artery and endometrial vascularization in bitches with CEH-pyometra syndrome. Bitches were allocated into CEH-mucometra Group (n = 13), Pyometra Group (n = 11), and Control Group (n = 8). Pyometra Group presented cytoplasmatic staining intensity for COX-2, VEGF-A, and FLT-1 and KDR receptors in luminal epithelium cells significantly higher compared to CEH-mucometra and Control groups. For the glandular epithelium, Pyometra Group had higher immunostaining score for VEGF-A and its receptors (FLT-1 and KDR). Hemodynamic indexes showed negative correlation with VEGF-A and its receptors as well as with COX-2. On the other hand, uterine vascularization score showed positive correlation in relation to immunostaining of COX-2, VEGF-A, and receptors in the endometrium luminal epithelium. In conclusion, uterus of bitches with CEH-pyometra syndrome show inflammatory process characterized by COX-2 expression, resulting in greater expression of proliferative Ki-67 marker as tissue response against the infectious agent. Furthermore, the increased VEGF-A expression and its receptors in CEH-pyometra reflect the increased blood flow and lower vascular resistance. Therefore, canine pyometra is characterized by an inflammatory, proliferative, and vascular disorder.

Multiplex cytokine analyses in dogs with pyometra suggest involvement of KC-like chemokine in canine bacterial sepsis

Clinical diagnostic criteria for sepsis (systemic inflammatory response syndrome caused by infection) are unspecific and, therefore, biomarkers for sepsis diagnosis are needed for appropriate treatment and patient survival. Pyometra, a common disease caused by bacterial infection of the uterus, results in sepsis in nearly 60% of cases in dogs. We used dogs with pyometra as a natural model for sepsis and collected serum samples from 39 dogs, of which 22 with pyometra and 17 healthy controls. Dogs with pyometra were further grouped into dogs with sepsis (n=18) and without sepsis (n=4). Serum concentrations of a panel of cytokines, including keratinocyte-derived chemokine (KC)-like, granulocyte-macrophages colony stimulating factor (GM-CSF), interleukin (IL)-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, chemokine C-X-C motif ligand (CXCL)10 and tumor necrosis factor (TNF)-α, were measured using multiplex analyses. Serum C-reactive protein (CRP) levels were determined using an automated immunoturbidimetric assay. In addition to physical examination hematological and serum biochemical analyses were performed to evaluate the overall status of the dogs. Significantly higher concentrations of KC-like (757 vs 304pg/ml) were detected in dogs with pyometra as compared to healthy dogs. Within the pyometra group, dogs with sepsis compared to dogs without sepsis had a higher KC-like concentration (873 vs 300pg/ml). Hemoglobin levels were significantly lower in dogs with pyometra compared to healthy dogs, regardless of the presence or absence of sepsis, and correlated negatively with KC-like. KC-like concentrations correlated positively with CRP, number of hospitalization days, number of monocytes, concentrations of IL-8, and percentage band neutrophils. Our data suggest that bacterial infection triggers the expression of KC-like and further studies are warranted of KC-like as a possible biomarker for diagnosing sepsis and uterine bacterial infection in dogs.

Klinische symptomen als merker van bacteriële isolatie uit de uterus van honden verdacht van pyometra

In deze studie werden vrouwelijke honden met klinische klachten verenigbaar met pyometra prospectief geëvalueerd. Het signalement, de klinische symptomen, laboratoriumresultaten en chirurgische bevindingen werden vergeleken tussen honden met en zonder bacteriële isolatie op basis van aërobe technieken. Patiënten met een positieve bacteriële isolatie werden in de pyometragroep geklasseerd, terwijl patiënten met een negatieve bacteriële isolatie als honden met mucometra werden gegroepeerd. Honderdveertig honden (118 met pyometra en 22 met mucometra) voldeden aan de inclusiecriteria. De toediening van een antibioticum vooraleer door te verwijzen was geassocieerd met verlengde klinische symptomen bij de patiënten van de pyometragroep (12 ± 2 dagen versus 7 ± 1 dagen; P = 0,006). Bij de pyometrapatiënten werden klinische symptomen, zoals koorts, anorexia en ongemak bij abdominale palpatie vaker waargenomen dan bij de mucometrapatiënten. Het totale aantal leukocyten was de enige parameter die significant verschilde (P=0,01). Hoewel er tussen de twee groepen geen verschil in kleur en consistentie van de baarmoedervloeistof werd opgemerkt, was de baarmoeder van de honden met pyometra zwaarder dan bij de honden met mucometra (851,80 ± 800,30 g versus 263,50 ± 297,10 g). E. coli was de vaakst geïsoleerde bacterie (92/123).

Cytometric analysis of surface molecules of leucocytes and phagocytic activity of granulocytes and monocytes/macrophages in cows with pyometra.

Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p<0.001). Significantly (p≤0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p<0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.