Salmonella L-forms: formation in human bile in vitro and isolation culture from patients' gallbladder samples by a non-high osmotic isolation technique  D.N. Wang, W.J. Wu, T. Wang, Y.Z. Pan, K.L. Tang, X.L. She, W.J. Ding, H. Wang  Clinical Microbiology and Infection  Volume 21, Issue 5, Pages 470.e9-470.e16 (May 2015) DOI: 10.1016/j.cmi.2014.12.016 Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions

Fig. 1 Stable L-forms isolated from the bile-inducer cultures of Salmonella spp. The obvious growth phenomenon of stable L-form cells derived from (a) Salmonella Typhi and (b) Salmonella Paratyphi A in PG liquid medium (light microscopy, 200 ×). (c) An L-form cell derived from Salmonella Typhi, its rough surface can be seen and is approx 5.2 μm in diameter (scanning electron microscopy (SEM), 12 000 ×). (D) Two L-form cells derived from Salmonella Paratyphi A, they have a rough surface and are arranged in pairs and are respectively approximately 8.5 μm and 11.0 μm in diameter (SEM, 5000 ×). Clinical Microbiology and Infection 2015 21, 470.e9-470.e16DOI: (10.1016/j.cmi.2014.12.016) Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions

Fig. 2 The results of agarose gel electrophoresis of the multi-PCR assay for the genes of the stable L-forms isolated from the bile-inducer cultures of Salmonella Typhi and Salmonella Paratyphi A. The multi-PCR assay for the genes of Salmonella L-forms. Lane M: DNA ladder maker. Lane 1: invA and flic-d genes of bacterial form of Salmonella Typhi. Lane 2: invA and flic-d genes of the stable L-forms isolated from the bile-inducer cultures of the Salmonella Typhi. Lane 3: invA and flic-a genes of bacterial form of Salmonella Paratyphi A. Lane 4: invA and flic-a genes of the stable L-forms isolated from the bile cultures of Salmonella Paratyphi A. Lane 5: Negative control. Clinical Microbiology and Infection 2015 21, 470.e9-470.e16DOI: (10.1016/j.cmi.2014.12.016) Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions

Fig. 3 Stable L-form cells isolated from human gallbladder. (a) The stable L-form cells with obvious growth phenomena in PG liquid medium (light microscopy, 200 ×). (b) The L-form cells have rough surfaces and are approximately 2.2–5.0 μm in diameter. They are arranged in short chains or irregularly, interconnected by their surface structures (scanning electron microscopy (SEM), 8000 ×). (c) The two L-form cells are approximately 3.2–4.1 μm in diameter and some bud-like structures can also be seen on their surfaces (SEM, 20 000 ×). (d) The large L-form is approx 5.2 μm in diameter and many small L-form cells (elementary bodies) are spherical, ovoid or short rods (indicated by the arrow) and are approx 0.12–0.38 μm in diameter, most of them are arranged in irregular clusters (SEM, 10 000 ×). Clinical Microbiology and Infection 2015 21, 470.e9-470.e16DOI: (10.1016/j.cmi.2014.12.016) Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions