Łukasz Kurek

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.

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.

Biophysical parameters of rats’ skin after the administration of methimazole

Abstract The paper describes the influence of oral administration of methimazole on biophysical skin parameters. Wistar rats of different sex (220-260 g) were used in the experiment. Biophysical skin parameters, such as transepidermal water loss (TEWL), corneometry, and pH were examined at seven-day intervals. Significant changes in the parameters were observed on the 7th d of methimazole administration. The changes were observed in both sex but males appeared to be less sensitive in that respect. Changes in the parameters in the females showed rapid mechanisms, which normalised transepidermal water loss and skin hydration, as well as restored skin barrier functions. TEWL, skin hydration, and skin pH measurements allow an early assessment of skin barrier dysfunction after administration of this drug.

Content of selected amino acids in the gastrocnemius muscle during experimental hypothyroidism in rats

Abstract Introduction: Thyroid hormones affect protein turnover, and in the case of hypothyroidism a decrease in protein synthesis and reduced release of certain amino acids from skeletal muscles are observed. Changes in the amino acid system of skeletal muscles may be responsible for the occurrence of muscle disorders. Material and Methods: The study measured the content of selected amino acids in the gastrocnemius muscle of Wistar rats during experimental hypothyroidism induced by oral administration of methimazole at a concentration of 0.05% in drinking water for 90 d. The rats were divided into four groups: E1 (n = 6) - experimental males, E2 (n = 6) - experimental females, C1 (n = 6) - control males, and C2 (n = 6) control females. Results: A statistically significant reduction occurred in leucine, isoleucine, and 1-methylhistidine levels in males, and 1-methylhistidine in females, in comparison to the control groups. Conclusion: The hypothyroidism-induced changes in amino acid content may be responsible for the occurrence of skeletal muscle function disorders.

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.

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.

Relationship between Total Homocysteine, Folic Acid, and Thyroid Hormones in Hypothyroid Dogs

Background Both elevated homocysteine and decreased folic acid concentrations are observed in human patients with hypothyroidism and can influence the development of numerous secondary disorders. Objectives The aim of the study was to assess total homocysteine concentration in serum and to examine its relationship with the concentration of folic acid and thyroid hormones (tT4 and fT4). Animals Ten healthy and 19 hypothyroid client‐owned dogs. Methods Dogs with clinical signs of hypothyroidism had the diagnosis confirmed by additional tests. Total homocysteine, folic acid, total thyroxine, and free thyroxine concentrations in serum were evaluated. Results Hypothyroid dogs were diagnosed with increased homocysteine (median 22.20 μmol/L; range, 16.50–37.75) and decreased folic acid (median 20.62 nmol/L; range, 10.54–26.35) concentrations, as compared to healthy dogs (11.52 μmol/L; range, 10.00–16.65 and 30.68 nmol/L; range, 22.84–38.52, respectively). In sick dogs, total homocysteine was inversely correlated with folic acid (ρ = −0.47, P < 0.001), total thyroxine (ρ = −0.69, P = 0.0092), and free thyroxine (ρ = −0.56, P = 0.0302). Conclusions Hypothyroidism in dogs causes hyperhomocysteinemia. Concomitant mild folic acid decrease in hypothyroid dogs might be as a result of hyperhomocysteinemia.

Reproductive system condition in dairy cows with left-sided displacement of the abomasums

Abstract The aim of the study was to evaluate selected parameters representing reproductive system conditions in cows suffering from left-sided displacement of the abomasum (LDA). Eighty Holstein- Friesian cows were divided into control (n=40) and LDA (n=40) groups. Haematological and biochemical evaluations were performed in the control group and in cows with LDA before and after surgical treatment. Cytological and ultrasound examinations of the uterus were performed at 21 (baseline), 35 and 63 days after parturition. In the LDA group, significantly decreased concentrations of calcium, magnesium, phosphorus, copper, potassium, glucose and total cholesterol were stated, while aspartate aminotransferase activity, white blood cell count, erythrocyte count, haematocrit, haemoglobin content and concentration of β-hydroxybutyrate, free fatty acids and bilirubin were increased (all P<0.05). Percentage of neutrophils in the uterus and the uterus diameter were increased in the LDA group at the baseline and at 35 days after parturition (P<0.001). The neutrophil percentage was also increased at 63 days after parturition in the LDA group (P<0.001). Number of days to first oestrus, number of services per conception and calving-to-conception interval were higher in the LDA group (P<0.001). This study has shown reproductive system changes and impaired fertility in dairy cows as the consequence of LDA occurrence. Thus, LDA treatment in cows should be combined with diagnostic evaluation of the uterus and reproductive system to improve reproductive performance. As indicated in the cytological examination, a subclinical inflammatory process may last even 60 days after parturition, leading to substantial impairment of reproductive function in dairy cows.

Cytological image of the endometrium in cows in follicular and luteal phases of the ovarian cycle and in cows with follicular and luteal ovarian cysts

Abstract The experiment was conducted on 30 Holstein-Friesian cows: 10 cows in the follicular phase of the cycle and in the luteal phase 10 d later, 10 cows with follicular cysts, and 10 with luteal cysts. The presence of the ovarian structures was confirmed by ultrasonography. Serum levels of progesterone and 17β-oestradiol were tested with ELISA. Samples for cytological examination were collected from the uterus of all cows using a cytological brush. Following staining, the smears were evaluated in terms of quality and percentages of endometrial cells. In the follicular phase of the oestrous cycle, cells of type A - superficial cells (64.6 ± 4.48) were proportionally the largest group of cells. Cells of type C - basal cells (19.8 ± 2.75) were also present. In the luteal phase, the highest percentage of cells was of type B - intermediate cells (76.9 ± 4.26). When follicular cysts were present on the ovaries, the cytology resembled the follicular phase of the cycle, but with many younger type C cells (33.1 ± 4.11). In the case of luteal cysts on the ovaries, the cytology was similar to that of the luteal phase of the cycle, however with a lower percentage of type B cells (58.1 ± 5.71), and a slightly higher percentage of the other types. The differences in the cytological image of the uterus when different ovarian structures are present, depend on the hormonal activity of those structures. Due to the lack of literature data, the results of the study are important as a model, and may substantially facilitate identification of phases of the oestrus cycle, or the pathologies described, as well as indicate the current status of the endometrium