Abstract
BackgroundBovine mastitis is an important health and cost factor in the milk industry. To elucidate whether isolated perfused bovine udders can be used to study early inflammatory events of mastitis, 1 mg of lipopolysaccharide (LPS) was instilled into quarters of 10 isolated perfused bovine udders. Three hours and 6 h after LPS instillation, tissue samples were taken from the gland cistern and base of the udder, subsequently stored in RNAlater and processed for the determination of inflammation-dependent gene regulation by real-time RT-qPCR. Gene expression analysis was performed using delta-delta Ct method. To translate mRNA results to protein, IL-1ß and IL-6 were determined in tissue homogenate by ELISA.ResultsThe instillation of 1 mg LPS lead to an increased expression of pro-inflammatory cytokines and chemokines like TNF-α, CCL20, CXCL8 as well as of IL-1 ß, IL-6 and IL-10, lingual antimicrobial peptide (LAP) and S100A9. However, the degree of elevation differed slightly between gland cistern and udder base and markedly between 3 and 6 h after instillation, with a distinct increase in mediator expression after 6 h. IL-1β protein increased in a time-dependent manner, whereas IL-6 was unchanged within 6 h of LPS instillation.ConclusionCompared to in vivo studies with instillation of LPS into udders of living cows, a similar inflammation-dependent gene regulation profile can be mimicked in the isolated perfused bovine udder, indicating a supplementation of animal experiments.Graphical abstract
Highlights
Bovine mastitis is an important health and cost factor in the milk industry
The hypothesis of the current study was that investigations of bovine mastitis using the model of the isolated perfused bovine udder may bridge a gap between simple two-dimensional cell cultures and complex in vivo experiments
It was stressed in former studies that the response of mammalian epithelial cells (MECs) to LPS or inactivated bacteria do not mimic all responses observed in the infected udder in vivo
Summary
Bovine mastitis is an important health and cost factor in the milk industry. To elucidate whether isolated perfused bovine udders can be used to study early inflammatory events of mastitis, 1 mg of lipopolysaccharide (LPS) was instilled into quarters of 10 isolated perfused bovine udders. Mastitis, defined as an inflammatory process of udder tissue mainly due to bacterial infection, is a major health and cost burden in dairy cattle worldwide [1, 2]. Intramammary infections with Escherichia coli, Streptococcus uberis and Staphylococcus aureus in particular differ in their immunological response and can lead to severe and or long lasting damage of the tissue, recent gene expression profiles of tissue from mastitis cows have been performed, early events of infection and inflammation in particular are still not fully understood [4, 5]. We wanted to perform a proof of principle study to find out whether the isolated perfused bovine udder may be a suitable tool to study early inflammatory events in mammary tissue.
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