Background -Acid fast cell wall deficient forms (CWDF) of bacteria have been grown from blood, bronchial washings, and ocular anterior chamber fluid from patients with sarcoidosis. A monoclonal antibody raised against Mycobacterium tuberculosis whole cell antigen (H37RV) was used to characterise further CWDF grown from the blood of patients with sarcoidosis. Methods -Blood from 20 patients with active sarcoidosis and from 20 controls was cultured using methods favourable for the growth of CWDF. Isolates were further characterised by indirect fluorescent antibody analysis using a monoclonal antibody highly reactive with M tuberculosis. Results -CWDF were grown from the blood of 19 of 20 subjects with sarcoidosis. All isolates stained positively with the monoclonal antibody and with a modified Kinyoun stain. No organisms were grown from the blood of controls.Conclusions -These data demonstrate that CWDF can be grown from the blood of nearly all patients with active sarcoidosis. The results confirm that the organisms are mycobacterial in origin and are similar, if not identical, to M tuberculosis. Their role in the pathogenesis of sarcoidosis is unknown. (Thorax 1996;51:530-533)

Serratia marcescens is recognized as an important and potentially hazardous nosocomial pathogen. The organism has been implicated here as the first reported case of S. marcescens meningitis associated with skin disinfection. A quaternary ammonium compound (QAC—Benzalkonium Chloride), was used to sterilize the skin prior to injection in a physician's office. Epidemiological studies were initiated. Six spray bottles containing disinfectant, the opened stock bottle of QAC, and an unopened bottle of disinfectant were all cultured. S. marcescens was noted growing in the spray bottles as well as in the opened stock bottle. Anti-biograms of the patient and epidemiological isolates are essentially the same. It is our contention as well as that of the Centers for Disease Control that an appropriate skin disinfectant such as Tincture of Chlorhexidine, Iodophors, or Tincture of Iodine should be used, and that physicians performing surgical techniques in the office be aware of the potential hazard of contamination. The consequences of nosocomial infection with resistant organisms warrant every precaution by health care professionals.

Since culture of Borrelia burgdorferi from patients with chronic Lyme disease has been an extraordinarily rare event, clarification of the nature of the illness and proving its etiology as infectious have been difficult. A method for reliably and reproducibly culturing B. burgdorferi from the blood of patients with chronic Lyme disease was therefore sought by making a controlled blood culture trial studying 47 patients with chronic Lyme disease. All had relapsed after long-term oral and intravenous antibiotics. 23 patients with other chronic illness formed the control group. Positive cultures were confirmed by fluorescent antibody immuno-electron microscopy using monoclonal antibody directed against Osp A, and Osp A PCR. 43/47 patients (91%) cultured positive. 23/23 controls (100%) cultured negative. Although persistent infection has been, to date, strongly suggested in chronic Lyme disease by positive PCR and antigen capture, there are major problems with these tests. This new method for culturing B. burgdorferi from patients with chronic Lyme disease certainly defines the nature of the illness and establishes that it is of chronic infectious etiology. This discovery should help to reestablish the gold standard in laboratory diagnosis of Lyme disease.

Cell wall deficient (CWD)-forms or L-forms of bacteria are characterized by a complete or partial loss of cell wall components and by the change of cellular shape. CWD-forms (spheroplasts) of bacteria are commonly prepared in vitro and are used for various practical applications. However, very little is known about the conditions required for the formation of CWD-forms of bacteria and mycobacteria in vivo and their significance in aetiology of various chronic diseases of humans (tuberculosis, sarcoidosis, and Crohn's disease) and animals (paratuberculosis). It is quite difficult to detect CWD-forms of mycobacteria in biological material and the possibilities of their detection by microscopy, culture, DNA hybridization techniques and other methods are limited. This obviously leads to a relatively small amount of published data on the detection and the isolation of CWD-forms of mycobacteria both from human or veterinary biological material, which can lead to the conclusion of a non-infectious origin of the diseases. The present review also includes studies performed by authors from the former Soviet Union and likely represents the first complete summarization of their knowledge in this sphere. However, certain results may be viewed as somewhat controversial. Accordingly, more attention should be paid to the research of cell wall deficient forms, not only in association with chronic and latent mycobacterial infections. List of abbreviations: AFR = acid-fast rods; AG = arabinogalactan; AT = antituberculotics; BCG = Bacillus Cal-mette-Guerin; CFU = colony forming units; CWD-forms = cell wall deficient forms; DNA = deoxyribonucleic acid; E = ethambutol; ELISA = enzyme-linked immunosorbent assay; EM = electron microscopy; GPL = glyc-opeptidolipids; H = isoniazid; HEYM = Herrold's egg yolk medium; HPC = hexadecylpyridinium chloride (N-cetylpyridinium chloride monohydrate); LAM = lipoarabinomannan; LOS = extractable glycolipids; M. = Myco-bacterium; MIC = minimum inhibition concentration; M7H9 = Middlebrook 7H9; PAS = para-amino-salycilic acid; PCR = polymerase chain reaction; PGL = phenolic glycolipids; pv. = pseudovar; R = rifampicin; RFLP = restriction fragment lenght polymorphism; SDS PAGE = sodium dodecyl sulphate polyacrylamide gel electrophoresis; S = streptomycin; TSB = tryptic soy broth; Z = pyrazinamide; ZN = Ziehl-Neelsen

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