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Ex vivo tumor necrosis factor-α response of blood leukocytes in Danish Holstein Friesian cows stimulated by Gram-positive and Gram-negative bacteria isolated from mastitic milk

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  • Momena Khatun
  • ,
  • Birthe Marie Damgaard
  • ,
  • Jens Bech Andersen
  • Christine Maria Røntved, CMR On-Site, Denmark

A whole blood stimulation assay was used to investigate the effects of parity, number of weeks after calving and Gram-positive and Gram-negative bacteria on the ex vivo TNF-α responsiveness of Danish Holstein-Friesian cows of first to third lactation (n = 28). Blood samples were collected in weeks 2, 3, 5 and 8 after parturition and stimulated with Escherichia coli LPS (10 μg/mL), Staphylococcus aureus peptidoglycan (PGN, 10 μg/mL) and dead Escherichia coli, Streptococcus uberis, Staphylococcus aureus, and Streptococcus dysgalactiae at a concentration of 2.5 × 10 6/mL. The antibiotic polymyxin-B (100 μg/mL) was added to the Gram-positive bacteria to avoid the influence of environmental endotoxin by ELISA test. Overall, parity had no effect, whereas number of weeks after calving altered the TNF-α responsiveness of the majority of the stimulants. Ex vivo, Gram-positive bacteria always resulted in a higher TNF-α response than Gram-negative bacteria with large differences within the individual cows. High correlations were found within the Gram-negative stimulants panel (r = 0.83) and within the Gram-positive (r = 0.81 to 0.86) stimulants panel except PGN. The higher TNF-α responsiveness by Gram-positive bacteria is in agreement with in vitro studies in human but in contrast to the in vivo TNF-α responsiveness in bovine udder.

Original languageEnglish
Article number110204
JournalVeterinary Immunology and Immunopathology
Number of pages6
Publication statusPublished - Apr 2021

    Research areas

  • TNF-α, Ex vivo whole blood assay, Mastitis bacteria, Parity, Early lactation, TNF-alpha, WHOLE-BLOOD, DAIRY-COWS, CYTOKINE PRODUCTION, BOVINE MASTITIS, DIFFERENT PATTERNS, INTRAMAMMARY INFECTION, STAPHYLOCOCCUS-AUREUS, TNF-ALPHA, STREPTOCOCCUS-DYSGALACTIAE, INFLAMMATORY MEDIATORS, Cattle Diseases/microbiology, Milk/microbiology, Lactation, Gram-Positive Bacteria/classification, Cattle, Female, Mastitis, Bovine/microbiology, Tumor Necrosis Factor-alpha/analysis, Leukocytes/immunology, Animals, Denmark, Gram-Negative Bacteria/classification

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