Abstract
Caseous lymphadenitis (CLA) is a chronic disease that affects small ruminants and causes economic losses in the associated breeding system. The causative agent of CLA is Corynebacterium pseudotuberculosis, a Gram-positive bacterium that exhibits tropism for external and internal lymph nodes and induces abscess formation in the host. Bacterial communities often produce a biofilm matrix that serves various functions, including protection against hostile environmental conditions, antibiotics, and the host immune response. Although biofilm formation has been reported for C. pseudotuberculosis, not all strains demonstrate this property in culture. In this work, we report the first comparative proteomic analysis of one biofilm-forming (CAPJ4) and one biofilm-non-forming strain (CAP3W) of C. pseudotuberculosis isolated from goats. Bacterial whole cell protein extracts were obtained for mass spectrometry analyses. Using LC-MS/MS, our studies reveal three and four proteins exclusively found in the CAPJ4 and CAP3W proteome, respectively. In addition, label-free quantitative analysis identified 40 proteins showing at-least 2-fold higher values in CAPJ4 compared CAP3W proteome Notably, CAPJ4 differentially synthesized the penicillin-binding protein, which participates in the formation of peptidoglycans. CAPJ4 also exhibited upregulation of N-acetylmuramoyl-L-alanine amidase and galactose-1-phosphate uridylyltransferase, which are involved in biofilm formation and exopolysaccharide biosynthesis. Here, we demonstrate that biofilm formation in C. pseudotuberculosis is likely associated with specific proteins, some of which were previously shown to be associated with virulence and biofilm formation in other organisms. Our findings may drive studies related to the bacterial mechanisms involved in the biofilm formation, in addition to providing targets for the treatment of CLA.
Highlights
Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA), a disease that primarily affects goats and sheep [1]
Differences in the functional genomes of CAP3W and CAPJ4 isolates were assessed at the protein level via proteomic analysis which detected 1,032 and 1,031 proteins in CAP3W and CAPJ4, respectively (Figure 2A)
Using label-free quantification, we evaluated the relative abundance of differentially expressed proteins in the proteomes of the CAPJ4 strain in comparison to that of the CAP3W strain (Figure 2B)
Summary
Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA), a disease that primarily affects goats and sheep [1]. Infection in humans by this pathogen, has been reported [2, 3]. This disease can be found worldwide; it is concentrated in regions that rely heavily on the commercialization of products derived from small ruminants [4]. CLA presents with symptoms characterized by the development of abscesses in visceral and superficial lymph nodes of animals [5]. Bacterial biofilms are composed of microbial communities embedded in an extracellular polymeric matrix and can attach to various surfaces [6]. Pathogens may form biofilms in animals, which can lead to chronic diseases characterized by the presence of abscesses [7]
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