Piotr Brodzki

Inflammatory cytokines and acute-phase proteins concentrations in the peripheral blood and uterus of cows that developed endometritis during early postpartum.

The aim of the study was to evaluate the level of proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin-6 [IL-6]), anti-inflammatory cytokine (interleukin-10 [IL-10]), and acute-phase proteins (haptoglobin [Hp] and serum amyloid A [SAA]) in serum and uterine washings in cows that developed endometritis during the early postpartum period. The study was carried out on 40 cows. The experimental group consisted of 20 cows with subclinical endometritis and the control group of 20 cows without endometritis. Analyses in both groups of cows were carried out at 5, 22, and 40 days postpartum (DPP). Experimental material consisted of the blood serum and uterine washings. The levels of the following cytokines: TNF-α, IL-6, IL-10 and acute-phase proteins: Hp and SAA were determined using ELISA. Our study reported that the levels of TNF-α, IL-6, IL-10, Hp, and SAA at 22 DPP were higher in cows with subclinical endometritis (P < 0.001). The levels of TNF-α (P = 0.01), IL-6 and IL-10 (P = 0.001), and Hp (P < 0.001) at 40 DPP were higher in cows with subclinical endometritis compared to healthy cows. The level of IL-10 in uterine washings at 5 DPP was higher (P = 0.001), whereas of SAA was lower (P = 0.01) in cows with subclinical endometritis. At 22 DPP, the levels of IL-6, IL-10, and Hp were higher (P < 0.001) in cows with endometritis. At 40 DPP, the level of TNF-α was lower, whereas these of IL-10 and Hp were elevated (P < 0.001) in cows with endometritis compared to healthy cows. The results indicate that the evaluation of the levels of cytokines and Hp in serum, but primarily in uterine washings, can be an important diagnostic indicator in cows that developed subclinical endometritis. High levels of IL-10 in cows with subclinical endometritis may contribute to the weakening of local resistance mechanisms of the uterus and lead to the persistence of the inflammation in the postpartum period. The present study also shows that the simultaneous examination of selected parameters of antagonistic interactions allows for better assessment of the current state of local immunity in the uterus.

Inflammatory cytokine and acute phase protein concentrations in the peripheral blood and uterine washings of cows with subclinical endometritis in the late postpartum period

The aim of the study was to evaluate the concentrations of proinflammatory cytokines: tumor necrosis factor (TNF-α) and interleukin-6 (IL-6), anti-inflammatory cytokine interleukin-10 (IL-10), and acute phase proteins (APPs)—haptoglobin (Hp) and serum amyloid A (SAA) in serum and uterine washings of cows with subclinical endometritis, and compare them to healthy animals. The study was performed on 24 cows on day 60 after delivery. The cows were divided into two groups based on the results of cytological tests: 12 cows with subclinical endometritis and 12 healthy cows. Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines in the study material were determined with ELISA: TNF-α, IL-6, IL-10 and APPs - Hp and SAA. The results show that the levels of TNF-α (p < 0.01), IL-6, IL-10 as well as SAA and Hp were significantly higher in the serum of cows with subclinical endometritis compared to the controls (p < 0.001). Uterine washings had significantly higher levels of IL-6, IL-10, and Hp in the experimental cows compared to the controls (p < 0.001). The demonstrated differences in the concentration of cytokines and APP between cows with subclinical endometritis and healthy cows, in both the serum and uterine washings, may suggest the usefulness of these parameters in the diagnosis of subclinical endometritis in cows in the late postpartum period.

Determination of selected parameters for non-specific and specific immunity in cows with subclinical endometritis

Endometritis in dairy cow herds is a serious economic problem all over the world due to the large economic losses. The aim of the study was a comparative evaluation of selected indicators of non-specific and specific immunity in cows with subclinical endometritis and in cows without inflammation of the uterus. The study was performed on 40 cows on day 65 after delivery. Based on the results of cytological tests, the cows were divided into two groups: experimental (subclinical endometritis) and control (20 cows in each group). A flow cytometric analysis was performed for the leukocyte surface molecules CD4, CD8, CD14, CD21, CD25. Moreover the phagocytic activity of granulocytes and monocytes/macrophages in peripheral blood and uterine washings was determined. It has been demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower for cows with subclinical endometritis when compared to cows undergoing a normal puerperal period (p<0.001). A significant (p≤0.001) decrease in the percentage of CD4+, CD14+, CD25+ and CD4+CD25+ leukocytes was also observed in peripheral blood of the cows from the experimental group. In uterine washings a significant decrease (p<0.001) in CD21+ and increase in CD8+ lymphocytes was detected. The results indicate that dysfunction of cell immunity coexisting with subclinical endometritis may be the main factor causing advanced inflammation of the uterus. Knowledge of immunological mechanisms observed in cows with subclinical endometritis could aid in choosing the right adjuvant therapy using immunomodulating agents.

Sperm quality and selected biochemical parameters of seminal fluid in dogs with benign prostatic hyperplasia

Benign prostatic hyperplasia (BPH) in dogs is most commonly associated with age and increasing concentrations of dihydrotesterone, a hormone that stimulates growth and secretion of the prostatic epithelial cells. During this process, the biochemical composition of prostatic secretion changes, which can affect the quality of semen and limit the ability of the sperm to contribute to fertilization. Therefore, the present study was conducted to examine possible correlation between BPH and biological quality of semen. The study was performed in 11 sexually mature dogs of various breeds. Animals were divided into two groups: healthy dogs (Group I; n = 5; mean age 4.32; SEM = 1.28) and dogs with BPH (Group II n = 6; mean age 6.16; SEM = 0.65). Semen and prostate secretions were collected and evaluated in this study. Standard semen examinations were conducted in the ejaculates collected; moreover, the extent of apoptosis and DNA defragmentation was determined. The selected biochemical parameters were determined in the prostate secretion. According to the examination results, there were no significant differences in standard semen parameters between the two groups of dogs. Nevertheless, morphological tests of semen in dogs with BPH demonstrated elevated percentages of primary defects in spermatozoa. A significant increase (P = 0.01) in DNA defragmentation of sperm was found in dogs with BPH. Moreover, changes in the biochemical composition of prostate secretion were demonstrated. In dogs with BPH, pH of prostate secretions was greater (P = 0.03), concentrations of cholesterol increased while concentrations of Zn and Cu decreased. The study findings reveal that BPH does not change semen quality in dogs.

Microbiological quality of milk sold directly from producers to consumers

The aim of this study was to determine the microbiological quality of raw cow milk from direct sale points. Raw cow milk samples were collected from 5 randomly selected direct sale points for microbiological evaluation. The samples were analyzed to determine total aerobic bacterial count, somatic cell count (SCC), counts of Enterobacteriaceae, Enterococcus, Escherichia coli, and Staphylococcus, and presence of Salmonella, Listeria monocytogenes, and inhibitory substances. The mean counts of total aerobic bacterial in samples from all direct sale points were between 9.2×10(4) and 3.6×10(7) cfu/mL. Milk samples collected from 5 direct sale points revealed counts Enterobacteriaceae ranging from 6.4×10(1) to 1.7×10(6) cfu/mL. Escherichia coli were detected in 12 milk samples with counts ranging from 5.0×10(0) to 1.1×10(2) cfu/mL. Staphylococcus spp. bacteria were found in all milk samples, at counts ranging from 1.6×10(3) to 5.1×10(4) cfu/mL. Listeria monocytogenes bacteria were detected in 1 sample, and SCC in all samples ranged from 78,000 to 1,730,000/mL. The examined samples did not contain Salmonella rods or inhibitory substances. In the samples examined in this study, international hygiene standards were exceeded for total aerobic bacterial count (n=48) as well as for SCC (n=19). Two milk samples contained pathogenic bacteria (Listeria monocytogenes and Staphylococcus aureus) that pose a potential hazard for consumer health.

Phenotyping of leukocytes and granulocyte and monocyte phagocytic activity in the peripheral blood and uterus of cows with endometritis

This study was a comparative evaluation of selected immunological parameters in peripheral blood and uterine wash samples from cows with a normal postpartum period compared with cows with endometritis. We aimed to determine the usefulness of these parameters in monitoring the puerperium. In total, 40 cows were included in the study: 20 had endometritis (experimental group), and 20 did not have uterine inflammation (control group). Animals were chosen on the basis of cytological and bacteriological test results. The tests were conducted 5, 22, and 40 days postpartum. In both groups, flow cytometric analysis of the surface molecules CD4, CD8, CD21, CD25, and CD14 in the peripheral blood and uterine washings was performed. Granulocyte and monocyte phagocytic activity was determined using a commercial Phagotest kit that was adapted for flow cytometry. The percentage of phagocytic granulocytes and monocytes in both the peripheral blood and the uterine washings was significantly lower for cows in the experimental group compared with the control group (P < 0.01). A significant decrease (P < 0.01) in the percentage of CD4+, CD25+, CD14+, and CD4 + CD25(high) leukocyte subpopulations was also observed in the peripheral blood of cows with endometritis. A significant decrease (P < 0.01) in CD21+ lymphocytes and an increase in CD8+ lymphocytes was detected in uterine washings. The results of this work indicate that cell immunity dysfunction may be the main factor causing advanced inflammation of the uterus in endometritis. Knowledge of the immunological mechanisms observed in cows with endometritis might aid in choosing the correct immunomodulating agent-based adjuvant therapy.

Effect of Uterine Inflammatory Status as well as Calcium and Magnesium Concentrations on the Uterine Involution Process in Dairy Cows

Abstract The aim of the study was a comparative analysis of uterine horn diameter, calcium and magnesium concentrations in blood serum and uterine washings in cows both with and without endometritis. The study was performed on 28 cows at 5, 22 and 42 DP (day post partum), of which 14 cows had been diagnosed with endometritis and the other 14 had no endometritis. The animals were selected based on clinical symptoms and cytological examination. Uterine horn diameter was determined in both examined groups of cows using an ultrasonographic device. The level of calcium and magnesium in the blood serum and uterine washings, and protein in uterine washings was determined using a BS-160 Mindray analyzer. Uterus diameter was similar in both examined groups of cows only at 5 DP, and on subsequent days of examination this parameter was higher in the endometritis group compared to the control (P<0.001). Calcium levels in the blood serum of cows with endometritis were lower than in cows without endometritis, both at 22 DP (P<0.001) and at 42 DP (P=0.004). The levels of calcium, magnesium and total proteins in uterine washings were higher in cows with endometritis compared to the group without endometritis, both at 22 and at 42 DPP (P<0.001). The results of this study confirm that the process of uterine involution is slower in cows with endometritis compared to cows without endometritis, and the myometrium of inflamed uterus does not utilize sufficient amounts of calcium and magnesium, which means that the concentrations of these elements in uterine fluid are higher than in cows without endometritis.

Total antioxidative capacity and zinc concentration in dogs suffering from perianal tumours

Abstract The aim of the study was to determine total antioxidative capacity (TAC) and zinc concentration in serum of dogs suffering from perianal tumours just before the start of the antihormonal treatment (AHT) and one and six months later. The study was performed on 45 dogs divided into two groups: control group suffering from non-malignant tumours (N = 24) and a group with malignant neoplastic changes (N = 21). Serum TAC and zinc concentrations were measured using photometric and atomic absorption spectrophotometric methods. Six months after the start of the AHT, TAC was significantly lower by 10.6% in dogs with malignant tumours when compared to controls (P = 0.03). In the non-malignant group, serum zinc concentration was higher before the treatment than in the malignant group, while the opposite results were observed six months later (P < 0.001). In the non-malignant group, gradually decreasing values of serum zinc concentration at each stage of the investigation were observed, while the opposite results were obtained in the malignant group (P < 0.05). The obtained results indicate that malignant neoplastic process is associated with significantly reduced TAC. Determination of serum zinc concentration in dogs with non-malignant and malignant perianal tumours may have practical diagnostic and prognostic values and may serve towards increasing the effectiveness of AHT monitoring.

Endometrial Cytology at Luteal and Follicular Phases of the Ovarian Cycle in Cows

Abstract The aim of the study was to examine cytological changes in the uterus in cows during the follicular and luteal phases of the ovarian cycle, as well as to compare two different methods (brush and flushing) used for cytological material collection and to evaluate their usefulness for monitoring of the endometrium. Ovarian cycle phases were confirmed by ultrasound and by the level of sex hormones (17-β-estradiol and progesterone). The following types of cells were identified in the cytological smears: type I - surface cells; type II - intermediate cells; type III - basal cells; polymorphonuclear leukocytes (PMNs); L - lymphocytes. The number of type I and III cells was statistically significantly higher in the follicular phase than in the luteal phase, both in smears prepared using a brush (P<0.001) and by uterine flush (P=0.003). The number of type II cells was statistically significantly higher in the luteal phase than in the follicular phase in both methods (P<0.001). The results of the study show that phases of the ovarian cycle in cows can be identified based on changes in the quality and percentage of different types of endometrial cells in a cytological examination.

Cytological evaluation of inflammation of the uterus and influence of endometritis on selected reproductive parameters in dairy cows

Abstract There were two aims of this study. One was to evaluate the postpartum state of the reproductive system in cows based on ultrasonography, bacterial culture, and cytological examination of the uterus. The other was to determine whether postpartum endometritis affects the subsequent state of the endometrium, and, in consequence, selected reproductive parameters in cows. The study was conducted on 60 cows: the experimental group of 30 cows with endometritis, and 30 cows free of uterine inflammation (control). The percentage of leukocytes in both groups was similar only on day 5 of postpartum. In all subsequent tests (26, 40, 61 d postpartum), the percentage of leukocytes in the experimental group was statistically significantly higher than in the control (P < 0.001), both in samples collected with a brush and in lavage samples. Involution of the uterus in the experimental group was also slower (P < 0.001). The analysed reproductive parameters were markedly less favourable in the experimental group than in the control. The study showed that postpartum inflammation of the uterus can persist for a long time in the form of endometritis, causing substantial deterioration of reproductive parameters in cows. The authors suggest that cytological evaluation of the uterus, preferably using a brush, should be performed as soon as possible after parturition, even before day 21, up to which time puerperal metritis may still persist. Evaluation of the inflammatory process based on the number of leukocytes and the quality of endometrial cells is important.