Adam 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.

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.

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.

Mitochondrial D-loop mutations can be detected in sporadic malignant tumours in dogs

Abstract The aim of this study was to identify mutations in the D-loop region of mtDNA in head, neck, and limb tumours in dogs, and determination of their relationship with the process of neoplastic transformation. Blood and tumour tissue samples from 19 dogs with diagnosed sporadic malignant tumours were analysed. DNA extraction, amplification, and sequencing of the mtDNA D-loop, and bioinformatic analyses were performed. Five mutations and 19 polymorphisms were observed in 68.42% of all tumours. Polymorphic variants were noted in 42.86% of the head and neck tumours and in 58.33% of the limb tumours. Mutations were observed in 21.05% of dogs. The mutations were found in 28.57% of the head and neck tumours and in 16.66% of the limb tumours. The mutations were identified in 50% of the studied epithelial cancers. In the mesenchymal tumours, no mutations in the D-loop region were observed. Mitochondrial haplotype A17 was found in over 40% cases of limb tumours. No association between the age, breed, sex, type of tumour, and detected polymorphic variants were observed. Different mutational changes in the D-loop sequences of mtDNA identified in the blood and tumour tissues may indicate a relationship between the type of tumour and individual changes in the D-loop nucleotide sequences of mtDNA.

Morphological, densitometric and mechanical properties of pelvic limb bones in 14-month-old female ostriches (Struthio camelus)

Limited information on physiological characteristic of bones in ostrich skeleton are available, even though bone weaknesses in ostriches are commonplace. Thus, the aim of this study was to evaluate morphological, densitometric, and mechanical properties of pelvic limb long bones (femur, tibia, and tarsometatarsus) in 14-month-old female ostriches (Struthio camelus var. domesticus). After the slaughter procedure, all bones were isolated, cleaned of soft tissues, and bone weight and length were determined. Using dual energy X-ray absorptiometry (DEXA) bone mineral density (BMD) and bone mineral content (BMC) were determined. Volumetric bone mineral density (vBMD) and geometrical parameters of the bones were determined using the quantitative computed tomography (QCT) method. Maximum elastic strength and ultimate strength of the bones were determined using three-point bending test. Significant differences were revealed between morphological, densitometric, and mechanical properties of femur, tibia, and tarsometatarsus in female ostriches at the slaughter age of 14 months. Elaborated experimental model and determination of morphological, densitometric, and mechanical properties of femur, tibia, and tarsometatarsus in female ostriches may serve for further studies on metabolic regulation of skeletal system properties with environmental, physiological, dietary, pharmacological, and toxicological factors.

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.

Serum concentration of magnesium in dogs suffering from tumors of the perianal glands

The aim of this study was to determine serum magnesium concentrations in dogs with benign and malignant tumors of the perianal glands. The magnesium concentration was determined in samples taken from 45 tumor-affected and 17 healthy males. Twenty one dogs suffered from malignant tumors (Malignant group), while benign changes were diagnosed in 24 cases (Non-malignant group). All dogs with neoplastic disease were treated with antiandrogens and antiestrogens. Determination of magnesium concentrations in serum obtained from dogs before the start of treatment (baseline), and one and six months later was performed using atomic absorption spectrophotometry. One-way ANOVA and the post-hoc Tukeys HSD test were used to compare differences between the groups investigated and time-related changes. Significantly higher serum magnesium concentrations were found in the group with malignant tumors when compared to the control and non-malignant groups (P<0.001). Serum magnesium concentrations in dogs with non-malignant neoplastic changes had increased gradually by 17% and 41% at one and six months when compared to the baseline values (P<0.05). In conclusion, the malignant neoplastic process in dogs was associated with a higher baseline serum magnesium concentration. Thus, determination of the serum magnesium concentration might be helpful for diagnostic differentiation between malignant and benign perianal tumors in dogs.