Limits...
New molecular microbiology approaches in the study of Campylobacter fetus.

Kienesberger S, Gorkiewicz G, Wolinski H, Zechner EL - Microb Biotechnol (2011)

Bottom Line: Application of comparative and functional genomics approaches will be essential to understand the molecular basis of this pathogen's interactions with various hosts.Here we report recent progress in genome analyses of C. fetus ssp. fetus and C. fetus ssp. venerealis, and the development of molecular tools to determine the genetic basis of niche-specific adaptations.To move forward in understanding the mechanisms underlying C. fetus virulence, current efforts focus on developing suitable in vitro models to reflect host- and tissue-specific aspects of infection.

View Article: PubMed Central - PubMed

Affiliation: 1Institute of Molecular Biosciences, University of Graz, Humboldtstrasse 50/1, A-8010 Graz, Austria.

ABSTRACT
Campylobacter fetus infection is a substantial problem in herds of domestic cattle worldwide and a rising threat in human disease. Application of comparative and functional genomics approaches will be essential to understand the molecular basis of this pathogen's interactions with various hosts. Here we report recent progress in genome analyses of C. fetus ssp. fetus and C. fetus ssp. venerealis, and the development of molecular tools to determine the genetic basis of niche-specific adaptations. Campylobacter research has been strengthened by the rapid advancements in imaging technology occurring throughout microbiology. To move forward in understanding the mechanisms underlying C. fetus virulence, current efforts focus on developing suitable in vitro models to reflect host- and tissue-specific aspects of infection.

Show MeSH

Related in: MedlinePlus

© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3815791&req=5

Mentions: Intracellular C. fetus can be visualized using differential immunofluorescence staining (Fig. 3) as previously established for other pathogens (Elsinghorst, 1994; Dehio et al., 1997; Pentecost et al., 2006). A standard staining procedure for labelling intracellular bacteria (Srivastava and Isberg, 2002) was adapted for C. fetus using monoclonal anti‐C. fetus serotype A antibodies (Brooks et al., 2002) and Alexa Dye‐conjugated secondary antibodies (Invitrogen). Campylobacter fetus‐infected Caco‐2 cells were treated sequentially with gentamicin, then with primary and secondary antibodies. To distinguish intracellular from extracellular bacteria the host cells were then permeabilized with methanol followed by restaining with the same primary but a different secondary antibody. Microscopy readily localizes intracellular bacteria within the infected mammalian cells when clinical or field isolates of either C. fetus subspecies are investigated. By contrast the type strains C. fetus ssp. venerealis ATCC 19438 and C. fetus ssp. fetus ATCC 27374 are non‐invasive and serve as negative controls for intracellular visualisation of bacteria. An example of Caco‐2 cells infected with the clinical isolate C. fetus ssp. fetus 82‐40 is shown in Fig. 3. Intracellular bacteria are stained with red only. Solely green or yellow (red plus green) dually labelled bacteria (circle) identify cells in extracellular locations. This immunofluorescence approach combined with confocal microscopy is at an early stage yet it is proving informative for visualizing C. fetus–host cell interactions. The development of techniques including differential staining of cell compartments or structures will offer clearer, more detailed localization data for bacteria within infected cells (Watson and Galan, 2008). Moreover, the capacity to differentiate intracellular and extracellular bacteria is important to defining the phenotypes of mutants functionally deficient for host cell entry versus those defective in intracellular viability.


New molecular microbiology approaches in the study of Campylobacter fetus.

Kienesberger S, Gorkiewicz G, Wolinski H, Zechner EL - Microb Biotechnol (2011)

© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815791&req=5

Mentions: Intracellular C. fetus can be visualized using differential immunofluorescence staining (Fig. 3) as previously established for other pathogens (Elsinghorst, 1994; Dehio et al., 1997; Pentecost et al., 2006). A standard staining procedure for labelling intracellular bacteria (Srivastava and Isberg, 2002) was adapted for C. fetus using monoclonal anti‐C. fetus serotype A antibodies (Brooks et al., 2002) and Alexa Dye‐conjugated secondary antibodies (Invitrogen). Campylobacter fetus‐infected Caco‐2 cells were treated sequentially with gentamicin, then with primary and secondary antibodies. To distinguish intracellular from extracellular bacteria the host cells were then permeabilized with methanol followed by restaining with the same primary but a different secondary antibody. Microscopy readily localizes intracellular bacteria within the infected mammalian cells when clinical or field isolates of either C. fetus subspecies are investigated. By contrast the type strains C. fetus ssp. venerealis ATCC 19438 and C. fetus ssp. fetus ATCC 27374 are non‐invasive and serve as negative controls for intracellular visualisation of bacteria. An example of Caco‐2 cells infected with the clinical isolate C. fetus ssp. fetus 82‐40 is shown in Fig. 3. Intracellular bacteria are stained with red only. Solely green or yellow (red plus green) dually labelled bacteria (circle) identify cells in extracellular locations. This immunofluorescence approach combined with confocal microscopy is at an early stage yet it is proving informative for visualizing C. fetus–host cell interactions. The development of techniques including differential staining of cell compartments or structures will offer clearer, more detailed localization data for bacteria within infected cells (Watson and Galan, 2008). Moreover, the capacity to differentiate intracellular and extracellular bacteria is important to defining the phenotypes of mutants functionally deficient for host cell entry versus those defective in intracellular viability.

Bottom Line: Application of comparative and functional genomics approaches will be essential to understand the molecular basis of this pathogen's interactions with various hosts.Here we report recent progress in genome analyses of C. fetus ssp. fetus and C. fetus ssp. venerealis, and the development of molecular tools to determine the genetic basis of niche-specific adaptations.To move forward in understanding the mechanisms underlying C. fetus virulence, current efforts focus on developing suitable in vitro models to reflect host- and tissue-specific aspects of infection.

View Article: PubMed Central - PubMed

Affiliation: 1Institute of Molecular Biosciences, University of Graz, Humboldtstrasse 50/1, A-8010 Graz, Austria.

ABSTRACT
Campylobacter fetus infection is a substantial problem in herds of domestic cattle worldwide and a rising threat in human disease. Application of comparative and functional genomics approaches will be essential to understand the molecular basis of this pathogen's interactions with various hosts. Here we report recent progress in genome analyses of C. fetus ssp. fetus and C. fetus ssp. venerealis, and the development of molecular tools to determine the genetic basis of niche-specific adaptations. Campylobacter research has been strengthened by the rapid advancements in imaging technology occurring throughout microbiology. To move forward in understanding the mechanisms underlying C. fetus virulence, current efforts focus on developing suitable in vitro models to reflect host- and tissue-specific aspects of infection.

Show MeSH
Related in: MedlinePlus