Dataset Information

Actinobaculum massiliense proteome profiled in polymicrobial urethral catheter biofilms

ABSTRACT: Actinotignum massiliense, a Gram-positive, facultatively anaerobic coccoid rod, is a rare human pathogen able to infect the urinary tract and belongs to the order of Actinomycetales. We identified A. massiliense as a resident of microbial biofilms growing on indwelling urethral catheter surfaces that were isolated from two patients with neurogenic bladders. These catheter biofilms (CBs) also harbored common uropathogens such as Proteus mirabilis and Aerococcus urinae, supporting the notion that A. massiliense depends on other co-colonizing microbes for survival. We isolated the bacterium from an anaerobically grown culture of a clinical sample, identified the species by 16S rRNA gene sequencing and verified this result via shotgun proteomics. Bacterial proteomes were profiled from the in vitro grown strain and four clinical ‘in vivo’ samples. The quantified proteomes allowed us to infer metabolic pathways and virulence/survival factors of importance in the CB milieu. Two putative subtilisin-like proteases, two Rib/Esp surface antigen repeat-containing proteins, a papain-like cysteine protease and a metal/heme/oligopeptide uptake system were highly expressed in vivo, but less so in vitro. We predict these proteins to be critical for adhesion and growth in CBs attacked by the host’s innate immune system or to improve bacterial fitness. Mixed acid fermentation following uptake and metabolism of xylose and glucuronate, sugars highly represented in proteoglycans and glycoglycerolipids of the urothelial mucosa and, in the case of glucuronate, shed into urine via renal xenobiotic conjugates, is inferred to be a major pathway for A. massiliense to generate energy under microaerobic conditions in CBs. The bacteria also appear to have active pathways for storage and utilization of glycogen as a carbon resource. Finally, we identified a putative polyketide synthase which may generate a secondary metabolite that interacts with either the host or co-colonizing organisms to enable A. massiliense survival in CBs.

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Urine

SUBMITTER: Yanbao Yu

LAB HEAD: Yanbao Yu

PROVIDER: PXD011327 | Pride | 2019-11-12

REPOSITORIES: Pride

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Actinobaculum massiliense Proteome Profiled in Polymicrobial Urethral Catheter Biofilms.

Yu Yanbao Y Tsitrin Tamara T Singh Harinder H Doerfert Sebastian N SN Sizova Maria V MV Epstein Slava S SS Pieper Rembert R

Proteomes 20181209 4

Actinobaculum massiliense, a Gram-positive anaerobic coccoid rod colonizing the human urinary tract, belongs to the taxonomic class of Actinobacteria. We identified A. massiliense as a cohabitant of urethral catheter biofilms (CB). The CBs also harbored more common uropathogens, such as Proteus mirabilis and Aerococcus urinae, supporting the notion that A. massiliense is adapted to a life style in polymicrobial biofilms. We isolated a clinical strain from a blo ...[more]

PMID: 30544882

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Project description:Biofilms are a communal of one or several kinds of microorganisms that growing on of both non-living and biotic surfaces by the production of multi-layers high-abundance extracellular matrix (ECM) to survive in the harsh environments (1-4). Biofilms consist of 85% (by volume) of matrix materials and 15% microbial cells (5). Biofilm ECM, consists of proteins, polysaccharides and/or extracellular DNAs, is important for biofilm integrity that increases environmental adaptability and induces antimicrobial resistance (6-7). Surface-adherent (sessile) bacteria in biofilms are more difficult to eradicate as minimum inhibitory concentrations (MICs) of antibiotics against bacterial biofilms is 100-1000 folds higher than the free living (planktonic) form that resulting in recurrent infections (8). Biofilms also possibly form nidus at the surface for the attachment of other pathogens lead to biofilms of multiple bacteria or multi-organisms (9ref). The communication, among organisms within biofilm, controls density of cell population refers to as “quorum sensing” (10) and different combinations of organisms in biofilms with either multiple bacteria or multi-species might induce different biofilm properties. While catheter-related colonization of Gram-positive bacteria from skin microbiota such as Streptococcus spp. and Staphylococcus spp. is common, biofilms in the inner lumen of catheter consist of both Gram-positive and Gram-negative bacteria. Because i) translocation of gut microbiota (eg. Enterococcus spp., Gram-negative bacteria and Candida albicans) into blood circulation during sepsis is one of the common causes of severe sepsis , ii) mixed systemic infection between bacteria and Candida spp. is even more severe than the infection by each organism in separation and iii) biofilms could be formed during bacteremia and fungemia, the biofilms from mixed species between bacteria and Candida spp. during sepsis is possible. In addition, central venous catheter-related candidiasis is common in intensive care units (ICU) patients refer to as “Candida catheter-related bloodstream infection (CRCBSI)”. Moreover, synergistic interaction between Candida albicans and several Gram-negative bacteria such as Escherichia coli (in peritonitis), Pseudomonas aeruginosa (in cystic fibrosis and ventilation associated pneumonia) and Acinetobacter baumannii (in ventilation associated pneumonia) has been mentioned. Hence, the collaboration between bacteria and Candida spp. might affect biofilm production as C. albicans in coexistence with the sessile microbes possibly enhance biofilms production that is detectable by crystal violet color (16,7). It is interesting to note that C. albicans are normal microbiota in human intestine and gut-translocation from intestine into blood circulation during severe sepsis (gut leakage) is demonstrated. Furthermore, both Gram-negative bacteria and C. albicans are the most and the second most predominant intestinal human microbiota, respectively, in which the natural interactions between these organisms is possible. Accordingly, catheter-related bacteremia is common among patients in ICU. Gut translocation of Candida spp. during sepsis, due to gut leakage, might induce the collaboration between bacteria and fungi results in persistent infection (20-21Chen L, 2011, 66//Wu H, 2015, 1, IJOS). Although the understanding in the interaction between organisms in biofilms should be beneficial for eradication strategies, the data of biofilms from the combination between gut-derived bacteria and fungi is still limited. As such, production of exo-polymers for biofilm-forming is a pathogenic virulent factor because biofilms is one of the important defend-mechanisms against host immune responses and antibiotics. Because i) antibiotic resistance caused by biofilm is a current serious medical problem , ii) the eradication of both bacterial and fungal biofilms is difficult and iii) antimicrobial treatment without biofilms-removal resulting in recurrent or persistent infection (23ref), biofilm prevention agent is needed (2410). Here, we explored i) the interaction between Gram-negative bacteria and C. albicans, in vitro, ii) macrophage responses against biofilm components, iii) biofilms in catheter-insertion mouse model and an evaluation on an interesting anti-biofilm.

Dataset Information

Actinobaculum massiliense proteome profiled in polymicrobial urethral catheter biofilms

Publications

<i>Actinobaculum massiliense</i> Proteome Profiled in Polymicrobial Urethral Catheter Biofilms.

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