Characterization of Mannheimia haemolytica biofilm formation in vitro

Boukahil, Ismail; Czuprynski, Charles J.

doi:10.1016/j.vetmic.2014.11.012

Cited by 24 publications

(37 citation statements)

References 29 publications

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“…In this study, we demonstrate that M. haemolytica can form biofilms on bovine respiratory epithelial cells. These findings are consistent with a previous report that M. haemolytica cells adhere and form a biofilm-like structure in vivo (Morck et al, 1989), and on plastic in vitro (Boukahil and Czuprynski, 2015). Other members of the Pasteurellaceae family, including A. pleuropneumoniae and H. somni, have been shown to form biofilms in vitro and in vivo (Sandal et al, 2011;Tremblay et al, 2013).…”

Section: Discussionsupporting

confidence: 92%

“…We previously reported that M. haemolytica produces the greatest amount of biofilm on polystyrene after a 48 h incubation in vitro (Boukahil and Czuprynski, 2015). The extended incubation period posed a challenge to our goal to form biofilm on epithelial cells because unfixed epithelial cells detached from the well when incubated with M. haemolytica for this amount of time (unpublished).…”

Section: Haemolytica Forms Biofilm On Bovine Bronchial Epithelial mentioning

confidence: 99%

“…The EPS in H. somni biofilms was found to consist largely of polysaccharides, which contribute to persistence of the bacteria in its biofilm state during systemic infection (Elswaifi et al, 2012). In a previous study, we showed that M. haemolytica will form biofilm on plastic in vitro, and that protein is the major constituent in the EPS (Boukahil and Czuprynski, 2015).…”

Section: Introductionmentioning

confidence: 95%

“…Minimum biofilm eradication concentration (MBEC) assays were performed as described previously (Boukahil and Czuprynski, 2015), with some modifications. BBEC were washed once with 1 ml of sterile water, fixed in wells of a 24 well plate, rinsed three times with 1 ml water, and M. haemolytica biofilms allowed to form as described above in the biofilm assay.…”

Section: Antibiotic Susceptibility Assaysmentioning

confidence: 99%

“…To effectively study biofilm formation in the context of the host, it is necessary to examine their interactions with underlying epithelial cells (Tran et al, 2014). Some bacterial species require 48 h to form a biofilm (Boukahil and Czuprynski, 2015), which presents a technical challenge because the underlying host cells will detach as a result of bacterial overgrowth in a closed system. By fixing epithelial cells in tissue culture wells, we have devised a system to study biofilm formation directly on bovine respiratory epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

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Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells

Boukahil

Czuprynski

2016

Veterinary Microbiology

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Mannheimia haemolytica is the most important bacterial agent associated with the bovine respiratory disease complex (BRDC), which causes worldwide economic losses to the cattle industry. M. haemolytica cells initially colonize the tonsillar crypts in the upper respiratory tract of cattle, from where they can subsequently descend into the lungs to cause disease. Many bacteria exist as biofilms inside their hosts. We hypothesize that M. haemolytica colonization of cattle during its commensal state may include biofilm formation. To begin to assess this possibility, we developed an in vitro system to study biofilm formation directly on bovine respiratory epithelial cells. Using fixed primary bovine bronchial epithelial cells, we observed M. haemolytica biofilm formation after a 48h incubation period at 37°C. Addition of mucin, the main component of mucus present in the upper respiratory tract, decreased M. haemolytica biofilm formation on bovine epithelial cells. We investigated the effects of prior viral infection of the epithelial cells on subsequent biofilm formation by M. haemolytica and found negligible effects. Utilization of this model system will provide new insights into the potential role of biofilm formation by M. haemolytica in the pathogenesis of BRDC.

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Section: Discussionsupporting

confidence: 92%

Section: Haemolytica Forms Biofilm On Bovine Bronchial Epithelial mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Section: Antibiotic Susceptibility Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells

Boukahil

Czuprynski

2016

Veterinary Microbiology

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Exopolysaccharide Production and Biofilm Formation by Histophilus somni

Petruzzi

Inzana

2015

Current Topics in Microbiology and Immunology

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The biofilm matrix of Histophilus somni is a complex architecture that differs substantially in structure between a pathogenic and commensal isolate examined. Overall, most pathogenic isolates produce more biofilm than commensal isolates. A major component of the biofilm is exopolysaccharide (EPS), which is also produced in greater quantity in the pathogenic isolate than in the commensal isolate studied. The EPS is composed of a D-mannan polymer, with occasional galactose residues present on side chains, similar in composition to that of yeast mannan. When grown in the presence of sialic acid, the biofilm EPS becomes sialylated and the amino sugars N-acetylglucosamine and N-acetylgalactosamine can be detected. In vitro biofilm formation follows a typical 4-stage growth curve, characterized by attachment, growth, maturation, and detachment. Following experimental challenge, formation of an H. somni biofilm has been demonstrated in cardiopulmonary tissue, often with Pasteurella multocida cohabitating the biofilm. A recently developed diagnostic test can detect antibodies to the EPS only in animals with systemic disease due to H. somni and is therefore capable of distinguishing between healthy animals colonized with H. somni and animals with systemic disease.

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Identification of a Hemagglutinin from Gallibacterium anatis

et al. 2016

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Gallibacterium anatis has the ability to hemagglutinate rabbit erythrocytes; however, no bacterial component has yet been associated with this function. In the present work, a protein of approximately 65 kDa with hemagglutinating activity for glutaraldehyde-fixed chicken erythrocytes was purified by ion interchange chromatography from G. anatis F149(T) secreted proteins. The protein was recognized by a rabbit polyclonal serum against a hemagglutinin from Avibacterium paragallinarum. The 65 kDa purified protein presented identity with a G. anatis filamentous hemagglutinin by mass spectrometric analysis. As well, the bacterial surface of G. anatis was labeled by immune gold assays using a polyclonal serum against the 65-kDa protein. A similar protein was recognized in four other G. anatis strains by immunoblots using the same antiserum. The protein binds sheep or pig biotinylated fibrinogen, suggesting an interaction with basement membrane eukaryotic cells components, and the protein is present in G. anatis biofilms. Overall, the results suggest that the 65 kDa hemagglutinin is a common antigen and a potential virulence factor in G. anatis.

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Characterization of Mannheimia haemolytica biofilm formation in vitro

Cited by 24 publications

References 29 publications

Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells

Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells

Exopolysaccharide Production and Biofilm Formation by Histophilus somni

Identification of a Hemagglutinin from Gallibacterium anatis

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