Analysis of Inflammatory and Regulatory Cytokines in the Milk of Dairy Cows with Mastitis: A Comparative Study with Healthy Animals.

Bovine 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-ß1, 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-ß1 and ß-Def 3 proteins. Expression of all proteins was detected in all milk samples. High expression of IL-2, IL-4, IL17A, TGF-ß1 was detected in healthy cows' milk and in milk of cows with subclinical and clinical mastitis. However, expression of IL-10 and ß-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 ß-Def 3 can be considered as informative biomarkers in diagnosis of subclinical and clinical mastitis.

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.

In 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-α, and IFN-γ. Data about subclinical and clinical mastitis demonstrate inflammatory responses to intramammary infection driven by IL-1α, 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-γ immunoreactivity was recognized to differ mastitis cases from the relative health status.

Dynamics and Diversity of Microbial Contamination in Poultry Bedding Materials Containing Parts of Medicinal Plants.

Microorganisms thriving in poultry bedding materials during their exploitation are involved in the development of several diseases and disfunctions of animals. They can also contaminate food products and pose risks to the environment and human health. This study provides an analysis of dynamics and diversity in microbiological contamination observed during the exploitation of poultry bedding materials containing parts of medicinal plants: Satureja hortensis, Origanum vulgare, Melissa officinalis, Salvia officinalis, and Thymus vulgaris, compared with standard types of beddings: straw chaff and straw pellets. The research was carried out in two 42-day experimental cycles involving in total 2400 broiler chickens. Each week, the total count of mesophilic bacteria, fungi and yeasts, the presumptive presence and count of Staphylococcus sp., Escherichia sp., Listeria sp., Salmonella sp., and Candida sp. were determined by culturing on selective media, along with pH and moisture measurements. After 35 days of the experiment, a reduction of the total count of mesophilic bacteria above 1 log compared to the control (11.86 vs. 13.02 log CFU/g) was observed. As the count of yeasts decreased after 21 days, an increase in the total count of bacteria was reported, which indicates a strong competition between microorganisms. The results improve our understanding of the temporal effects of using materials containing parts of medicinal plants on the microbial contamination in poultry litter.