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Chapter 21 : Advances in Vancomycin Resistance:

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Abstract:

This chapter concentrates on the mechanisms and biological features of vancomycin resistance in methicillin- and cephem-resistant (MRSA). Vancomycin-resistant (VRSA) strains isolated from three American patients contained plasmids carrying the gene complex. The gene complex is carried on a transposon which seems to have been transferred by conjugation from vancomycin-resistant enterococcus (VRE) that coexisted in the patients’ bodies. The enzyme encoded by the gene, together with those encoded by the adjacent genes , , and , replaces D-alanyl-D-alanine residues of peptidoglycan by D-alanyl-D-lactate. In the cell wall of Mu50, peptidoglycan crosslinking is significantly decreased as compared to hetero-vancomycin-intermediate (VISA) strain Mu3 or vancomycin susceptible (VSSA) strains. That is, there are more D-alanyl-D-alanine false targets in the cell-wall peptidoglycan layers of Mu50 than in those of control strains. The vancomycin resistance of VISA is not due to the acquisition of a resistance gene from another bacterial species. It is generated spontaneously from VSSA strains in vitro, though the development of the VISA phenotype does not occur through a single-step selection process. A remarkable feature of hetero-VISA is that it can be obtained from the VSSA strain by selection with beta-lactam antibiotics. A high-thoroughput sequencing strategy for regulator genes in combination with microarray transcription profiling in many isogenic VISA and hetero-VISA combinations might reveal several alternative series of regulator mutations.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21

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beta-Lactam Antibiotics
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Staphylococcus aureus
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Glycopeptide Antibiotics
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Figures

Image of Figure 1
Figure 1

Thick cell wall prevents the saturation of peptidoglycan layers with vancomycin. The Mu50 cells with thin (A) and thick cell wall (B) were prepared by incubating the cells in resting media (RMs) with different nutrient compositions (11). RM does not support the cell growth. The numbers under the panels are mean and SD of cell-wall thickness in nm. (C) The cells were incubated in RM containing 30 mg/liter of vancomycin, and the time course of vancomycin consumption was monitored by HPLC. Symbols: squares, the cell densities of thin (open) and thick (closed) cell wall. Circles, vancomycin concentration in the culture supernatant of the cells with thin (open) and thick (closed) cell wall. Note that the cells with thick cell walls consume twice as much vancomycin as those with thin cell walls. Consumption of vancomycin by the cells with thick cell walls does not reach maximum before 60 min, whereas that by the latter cells reaches maximum before 5 min.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21
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Image of Figure 2
Figure 2

Analysis of vancomycin-resistant subpopulations of strains. Plating efficiency test is performed with maximum inoculum size of about 10 CFU onto BHI agar plates of 8.5 cm in diameter containing various concentrations of vancomycin. Strains analyzed are Mu50 (open circle), PC27-14 (closed circle), PC27 (open triangle), Mu3 (closed triangle), H1 (open square), FDA209P (closed square). Note that PC27 and PC27-14 were obtained from Mu3 cell population based on their colony sizes alone without recourse to drug selection.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21
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Image of Figure 3
Figure 3

Heterogeneous colony sizes in Mu3 cell population. Mu3 cells were spread on brain heart infusion agar plate and incubated for 24 h at 37°C before the photo was taken. Inset: magnification of the area in the square. RC, regular sized colony; SC, small colony; PC, pin-point colony.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21
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Figure 4

Novel detection method of VISA strains. The chemiluminescence method to detect metabolism of viable cells after 3 h exposure of the cells to vancomycin. Three representative VISA strains, Mu50, MI, and AMC11094, are clearly distinguished from VSSA strains FDA209P (thin line) and NCTC8325 (dotted line). Hetero- VISA strain Mu3 has a marginal pattern and is not well differentiated from VSSA with this protocol.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21
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Tables

Generic image for table
Table 1

Isolation of hetero-VISA, VISA, and VRSA strains from various countries

UK, United Kingdom; MI, Michigan; NJ, New Jersey; NY, New York; IL, Illinois; PA, Pennsylvania; and MN, Minnesota.

with reduced vancomycin susceptibility.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21
Generic image for table
Table 2

Hetero-VISA strain Mu3 is composed of cells of various colony size classes

CSC, colony size class: RC, regular colony formed after 24 h; SC, small colony after 24 h; PC, pinpoint colony formed after 24 h; C2, colony formed after 48 h; C3, colony formed after 72h.

C2 and C3 are mostly unstable. One stable strain each was analyzed in this study.

Citation: Hiramatsu K, Kapi M, Tajima Y, Cui L, Ito T, Trakulsomboon S. 2005. Advances in Vancomycin Resistance: , p 289-298. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch21

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