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

Momena Khatun*, Birthe Marie Damgaard, Jens Bech Andersen, Christine Maria Røntved

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

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
Volume234
Number of pages6
ISSN0165-2427
DOIs
Publication statusPublished - Apr 2021

Keywords

  • 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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