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Transcriptomic analysis of circulating leukocytes obtained during the recovery from clinical mastitis caused by Escherichia coli in Holstein dairy cows

Cheng, Zhangrui; Palma-Vera, Sergio; Buggiotti, Laura; Salavati, Mazdak; Becker, Frank; Werling, Dirk; Wathes, D Claire; consortium, GplusE


Zhangrui Cheng

Sergio Palma-Vera

Laura Buggiotti

Mazdak Salavati

Frank Becker

Dirk Werling

D Claire Wathes

GplusE consortium


The risk and severity of clinical infection with Escherichia coli as a causative pathogen for bovine mastitis is influenced by the hosts’ phenotypic and genotypic variables. We used RNA-Seq analysis of circulating leukocytes to investigate global transcriptomic profiles and genetic variants from Holstein cows with naturally occurring cases of clinical mastitis, diagnosed using clinical symptoms and milk microbiology. Healthy lactation-matched cows served as controls (CONT, n = 6). Blood samples were collected at two time periods during the recovery phase post diagnosis: EARLY (10.3 ± 1.8 days, n = 6) and LATE (46.7 ± 11 days, n = 3). Differentially expressed genes (DEGs) between the groups were identified using CLC Genomics Workbench V21 and subjected to enrichment analysis. Variant calling was performed following GATKv3.8 best practice. The comparison of E. coli(+) EARLY and CONT cows found the up-regulation of 1090 DEGs, mainly with immune and inflammatory functions. The key signalling pathways involved NOD-like and interleukin-1 receptors and chemokines. Many up-regulated DEGs encoded antimicrobial pep-tides including cathelicidins, beta-defensins, S100 calcium binding proteins, haptoglobin and lactoferrin. Inflammation had largely resolved in the E. coli(+) LATE group, with only 29 up-regulated DEGs. Both EARLY and LATE cows had up-regulated DEGs encoding ATP binding cassette (ABC) transporters and haemoglobin subunits were also up-regulated in LATE cows. Twelve candidate genetic variants were identified in DEGs between the infected and CONT cows. Three were in contiguous genes WIPI1, ARSG and SLC16A6 on BTA19. Two others (RAC2 and ARHGAP26) encode a Rho-family GTPase and Rho GTPase-activating protein 26. These results show that the initial inflammatory response to E. coli continued for at least 10 days despite prompt treatment and provide preliminary evidence for genetic differences between cows that may pre-dispose them to infection.


Cheng, Z., Palma-Vera, S., Buggiotti, L., Salavati, M., Becker, F., Werling, D., …consortium, G. (in press). Transcriptomic analysis of circulating leukocytes obtained during the recovery from clinical mastitis caused by Escherichia coli in Holstein dairy cows. Animals, 12(16), 2146.

Journal Article Type Article
Acceptance Date Aug 18, 2022
Online Publication Date Aug 21, 2022
Deposit Date Aug 22, 2022
Publicly Available Date Aug 22, 2022
Print ISSN 2076-2615
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 12
Issue 16
Pages 2146
Keywords E. coli mastitis; cow; mammary gland; antimicrobial peptides; ABC transporters; MHC system
Publisher URL


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