Browsing by Author "Gergely, Maroti"
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Item Analysis of Inflammatory and Regulatory Cytokines in the Milk of Dairy Cows with Mastitis : A Comparative Study with Healthy Animals(2024) Lohova, Elizabete; Pilmane, Māra; Šerstņova, Ksenija; Melderis, Ivars; Gontar, Lukasz; Kochanski, Maksymilian; Drutowska, Andzelika; Gergely, Maroti; Prieto Simón, Beatriz; Institute of Anatomy and AnthropologyBovine mastitis remains a major problem in the global dairy cattle industry. The acute invasion of udder by pathogens induces innate immune response as the first defence mechanism in subclinical and clinical mastitis. The aim of the study was to determine inflammatory and regulatory cytokines IL-2, IL-4, TGF-β1, IL-17A, beta-defensin 3 and IL-10 and their potential changes in milk of dairy cows with subclinical and clinical mastitis, and to compare the findings with healthy animals. Milk samples from 15 holstein Friesian breed cows were used in the study. Cows were divided into three groups based on their health status (5 healthy, 5 subclinical and 5 clinical animals). All samples were tested using immunohistochemistry to evaluate IL-2, IL-4, IL-10, IL17A, TGF-β1 and β-Def 3 proteins. Expression of all proteins was detected in all milk samples. High expression of IL-2, IL-4, IL17A, TGF-β1 was detected in healthy cows’ milk and in milk of cows with subclinical and clinical mastitis. However, expression of IL-10 and β-Def 3 in milk samples of healthy cows was significantly higher compared to the milk of cows with subclinical and clinical mastitis (p <.001). IL-10 and β-Def 3 can be considered as informative biomarkers in diagnosis of subclinical and clinical mastitis.Item Composition of mastitis causing microorganisms and cytokines in healthy cow’s milk : a pilot study(2023) Junga, Anna; Pilmane, Māra; Šerstņova, Ksenija; Lohova, Elizabeta; Melderis, Ivars; Gontars, Lukašs; Kochanski, Maksymilian; Drutowska, Andzelika; Gergely, Maroti; Prieto-Simona, Beatrise; Institute of Anatomy and AnthropologyThe aim of this study was to examine clinically healthy cow’s udder milk microbiota and presence of cytokines in different seasons. Milk samples taken from the cows were checked for the presence of Gram-positive and Gram-negative bacteria, and the somatic cell count was detected. Immunohistochemistry methods were performed to detect interleukin (IL) -2, IL-4, IL-8, IL-10, IL-12, IL-17a, â-defensin-3, transforming growth factor (TGF)-â1, interferon-ã and nuclear factor (NF)-êB presence in the milk. S. agalactiae, S. uberis, S. aureus, E. coli, and Klebsiella, Enterobacter, Citrobacter spp. were found in healthy cow’s milk. In the first round, the highest prevalence was observed for S. aureus. In the second round, the highest mean levels were observed for S. uberis, then followed S. aureus. IL-4, IL-17a and TGF-â1 demonstrated the highest expression in the milk samples. NF-êB had the lowest expression among all factors. The presence of a rich bacterial microbiome (mostly S.aureus, S.uberis) in the milk of healthy animals, as well as changing bacterial species between in spring and autumn seasons occur as a result of both the immune state of the animal and many external factors, which consequently affects the amount of expressed cytokines.Item Expression of anti-inflammatory markers IL-2, IL-10, TGF-β1, βDEF-2, βDEF-3 and Cathelicidin LL37 in dairy cattle milk with different health status of the udder(2022-07-10) Šerstņova, Ksenija; Pilmane, Māra; Vitenberga-Verza, Zane; Melderis, Ivars; Gontars, Lukašs; Kochanski, Maksymilian; Drutowska, Andzelika; Gergely, Maroti; Prieto-Simona, Beatrise; Institute of Anatomy and AnthropologyGreat economic losses to the dairy industry are associated with bovine mastitis, which results in poor milk quality and high treatment costs. Anti-inflammatory proteins play an important role in the suppression of the immune response against invading pathogenic microorganisms and are therefore being studied for possible use in the early diagnosis of mastitis. In our study, we used milk samples from 15 cows of Holstein Friesian breed with different health status (5 healthy, 5 subclinical, and 5 clinical animals), and tested them using immunohistochemical (IHC) analysis to evaluate the presence of IL-2, IL-10, TGF-β1, βDEF-2, DEF-3, and Cathelicidin LL37 proteins. The calculation of positively and negatively stained cells for each biomarker was performed using the semiquantitative counting method. We found the presence of all factors with the exception of Cathelicidin LL37, which was almost absent in milk samples of all animal groups. The significant decrease of IL-10, β-def2, and β-def3 expression levels within the 3 days of sampling, found in the milk of animals with sub- and clinical mastitis, indicates the loss of antiinflammatory protection of the affected cow’s udder. In contrast, the stable increase of IL-2 and TGF-β1 positive cells observed in the milk of mastitis-affected cows, and the similar expression of these factors in the milk of healthy animals, indicate the possible lack of involvement of these cytokines at an early stage of udder inflammation.Item Identification of Inflammatory and Regulatory Cytokines IL-1α-, IL-4-, IL-6-, IL-12-, IL-13-, IL-17A-, TNF-α-, and IFN-γ-Producing Cells in the Milk of Dairy Cows with Subclinical and Clinical Mastitis(2022-03-17) Vitenberga-Verza, Zane; Pilmane, Māra; Šerstņova, Ksenija; Melderis, Ivars; Gontar, Łukasz; Kochanski, Maksymilian; Drutowska, Andzelika; Gergely, Maroti; Prieto-Simona, Beatrise; Institute of Anatomy and AnthropologyIn naturally occurring bovine mastitis, effects of infection depend on the host inflammatory response, including the effects of secreted cytokines. Knowledge about the inflammatory and regulatory cytokines in milk cells of free-stall barn dairy cows and in naturally occurring mastitis is lacking as most studies focus on induced mastitis. Hereby, the aim of the study was to determine inflammatory and regulatory cytokines in the milk of dairy cows with subclinical and clinical mastitis. The following examinations of milk samples were performed: differential counting of somatic cells (SCC), bacteriological examination, and immunocytochemical analysis. Mean SCC increased in subclinical and clinical mastitis cases. The number of pathogenic mastitis-causing bacteria on plates increased in subclinical mastitis cases but decreased in clinical mastitis. The inflammatory and regulatory markers in the milk cells of healthy cows showed the highest mean cell numbers (%). In mastitis cases, immunoreactivity was more pronounced for IL-4, IL-6, IL-12, IL-13, IL-17A, TNF-, and IFN-y. Data about subclinical and clinical mastitis demonstrate inflammatory responses to intramammary infection driven by IL-1, IL-4, and IL-17A. Moreover, the host defense response in mastitis is characterized by continuation or resolution of initial inflammation. IL-12 and INF-immunoreactivity was recognized to differ mastitis cases from the relative health status.