Legionella gresilensis sp. nov. and Legionella beliardensis sp. nov., isolated from water in France.

Novel Legionella-like isolates, strains Montbéliard A1T and Gréoux 11 D13T, isolated from two different French water sources, were studied taxonomically and phylogenetically. Morphological and biochemical characterization revealed that they were Gram-negative, aerobic, non-spore-forming bacilli with a cut-glass appearance that grew only on L-cysteine-supplemented buffered charcoal yeast extract agar. Phenotypic characterization using fatty acid and ubiquinone profiles and SDS-PAGE analysis confirmed that they were closely related, but distinct from, other species of the genus Legionella, since serotyping could not relate them to any existing serogroup. Genotypic profiles generated by randomly amplified polymorphic DNA and 16S-23S rDNA spacer region PCR analyses were unique for each of these isolates. DNA-DNA relatedness values of strains Montbéliard A1T and Gréoux 11 D13T to each other and to other Legionella type strains were less than 25%. Phylogenetic affiliation of these organisms obtained by 16S rDNA sequence comparisons confirmed that they were distinct from any other known Legionella species. All the above results confirm that these strains constitute two novel species for which the names Legionella gresilensis sp. nov. (type strain Gréoux 11 D13T = ATCC 700509T = CIP 106631T) and Legionella beliardensis sp. nov. (type strain Montbéliard A1T = ATCC 700512T = CIP 106632T) are proposed.

Isolation of Legionella oakridgensis from two patients with pleural effusion living in the same geographical area.

Two cases of Legionnaire's disease caused by Legionella oakridgensis were diagnosed at the university hospital in Nantes, France. The two patients' isolates were identified by means of phenotyping and genotyping methods. Epidemiological investigations concluded that the first case was hospital acquired while the second case was considered community acquired.

Legionella taurinensis sp. nov., a new species antigenically similar to Legionella spiritensis.

A group of 42 Legionella-like organisms reacting specifically with Legionella spiritensis serogroup 1 antisera were collected throughout Europe by the Centre National de Référence (French National Reference Centre) for Legionella. This group of isolates differed somewhat from L. spiritensis in terms of biochemical reactions, ubiquinone content and protein profile. The latter two analyses revealed that one of these L. spiritensis-like isolates, Turin I no. 1T, was highly related, but not identical to any of the red autofluorescent species of Legionella. In fact, this strain was the first of these particular isolates recognized to emit a red autofluorescence when exposed to UV light. Profile analysis of randomly amplified polymorphic DNA established that the red autofluorescent L. spiritensis-like isolates constituted a homogeneous group distinct from Legionella rubrilucens and Legionella erythra. DNA-DNA hybridization studies involving the use of S1 nuclease confirmed that the indicated group of isolates are a new species of Legionella, for which the name Legionella taurinensis is proposed with strain Turin I no. 1T (deposited as ATCC 700508T) as the type strain.

Identification of Legionella species by random amplified polymorphic DNA profiles.

Random amplified polymorphic DNA (RAPD) was used for the identification of Legionella species. Primer SK2 (5'-CGGCGGCGGCGG-3') and standardized RAPD conditions gave the technique a reproducibility of 93 to 100%, depending on the species tested. Species-specific patterns corresponding to the 42 Legionella species were consequently defined by this method; the patterns were dependent on the recognition of a core of common bands for each species. This specificity was demonstrated by testing 65 type strains and 265 environmental and clinical isolates. No serogroup-specific profiles were obtained. A number of unidentified Legionella isolates potentially corresponding to new species were clustered in four groups. RAPD analysis appears to be a rapid and reproducible technique for identification of Legionella isolates to the species level without further restriction or hybridization.

Species identification of Legionella via intergenic 16S-23S ribosomal spacer PCR analysis.

Species identification of Legionella in routine laboratory testing is hampered by the lack of highly discriminatory phenotypic tests. Amplification polymorphism of the intergenic 16S-23S spacer regions (ISR) has been previously developed for identification of species within the Legionellaceae [Hookey, J.V., Birtles, R.J. & Saunders, N.A. (1995). J Clin Microbiol 33, 2377-2381], but it did not provide enough resolution to distinguish all members of the bluish-white autofluorescent species and the red autofluorescent group of the Legionellaceae. By choosing new primers that target regions 4 (positions 1521-1541 of Escherichia coli 16S rRNA gene) and 6 (positions 114-132 of E.coli 23S rRNA gene) within the rDNA operon close to the 16S-23S intergenic spacer, 34 profiles were determined among the 79 type and reference strains representing 42 species that were tested. Analysis of the RFLP generated after Hinfl restriction digestion of the PCR products further improved the method, allowing complete discrimination among the species and subspecies of Legionella tested. Twenty-three well-identified strains from unrelated origins belonging to seven species gave amplification patterns identical to that of their type strain. The technique was also tested on 80 field isolates that could not be unequivocally assigned to groups by phenotypic methods. Seventy-two per cent (58/80) of these isolates had a profile identical to that of a type strain, while 27% (22/80) may correspond to new taxa since their ISR-PCR profiles did not match any of the known profiles.

Comparative analysis of infrequent-restriction-site PCR and pulsed-field gel electrophoresis for epidemiological typing of Legionella pneumophila serogroup 1 strains.

Two methods were compared for the analysis of 48 unrelated and epidemiologically related Legionella pneumophila serogroup 1 isolates. These are the infrequent-restriction-site PCR (IRS-PCR) assay with adapters designed for XbaI and PstI restriction sites and the pulsed-field gel electrophoresis (PFGE) analysis determined after DNA restriction with SfiI. Both methods demonstrated a high level of discrimination with a similar capacity for differentiating 23 of the 24 unrelated isolates. PFGE analysis and IRS-PCR assay were both able to identify epidemiologically related isolates of L. pneumophila from three outbreaks. Hence, IRS-PCR assay appears to be a reproducible (intergel reproducibility, 100%) and discriminative (discriminatory index, > or = 0.996) tool for typing of Legionella. Compared to PFGE, however, IRS-PCR presented an advantage through ease of performance and with attributes of rapidity and sensitivity of target DNA.

[Legionnaires' disease in travellers]. / Légionelloses du voyageur.

The outbreak of pneumonia involving delegates to the 1976 American Legion convention at a Philadelphia hotel was the first example of travel-associated legionnaires' disease. Travel is now well known as a common risk factor for legionnaires' disease. This travel-associated disease is a preoccupation among European countries because of morbidity among citizens of the European Union. The definition of the case of legionellosis is a patient who presents an acute lower respiratory tract infection with focal signs of pneumonia and/or radiological features, and microbiological evidence of Legionella infection. A case is considered to be travel associated if the patient has spent one or more nights away from home during the ten days before becoming ill. An European Surveillance Scheme for Travel-Associated Legionnaires' Disease was established in 1987 to identify clusters and outbreaks of cases of the disease. This group centralizes the case reports of twenty-nine collaborating centres in twenty-five countries. Outbreaks of legionnaires' disease were described in hotels, camps or cruise ships. In 1996, the number of travel-associated cases of legionnaires' disease represented 16% of the total number cases. The increase of the number of reported cases may reflect improved surveillance and increased ascertainment. In Europe in 1996, the diagnosis of legionellosis was confirmed by detection of Legionella pneumophila sero-group 1 antigen in urine (36%), seroconversion (fourfold rise in antibody titre, 33%) and culture of the organism (16%). Fifteen per cent of legionellosis was diagnosed by the identification of a single high antibody titre. In France a coordination between Public Health Institutions (Réseau National de Santé Public and DDASS), clinicians, laboratories and National Reference Center was established to improve prevention and control of legionnaires' disease outbreaks. Legislation obliges to report each case. When more two cases in the same area are notified an epidemiological investigation must be done. The knowing of the source of the contamination and its eradication allows to prevent new cases and outbreaks. Outbreaks of Legionnaires' disease are even now mediatic and this fact leads to maintain attention for the quality of diagnosis and epidemiology investigation due to touristic and economic consequences for the implicated countries.

The first clinical isolate of Legionella parisiensis, from a liver transplant patient with pneumonia.

A bluish white autofluorescent strain of Legionella was isolated from the tracheal aspirate of a female liver transplant patient who developed hospital-acquired pneumonia. This strain had biochemical characteristics compatible with those of L. cherrii, L. anisa, and L. parisiensis and could not be differentiated from L. bozemanii and L. parisiensis by the direct fluorescent-antibody assay. Phylogenetic analysis of partial 16S rRNA gene sequences of this strain (ATCC 700174) revealed the closest homology to the species L. parisiensis (99.5%). An L. parisiensis species-specific profile was also identified by a random amplified polymorphic DNA technique. This is the first report of L. parisiensis isolation from humans.

Randomised, double-blind, placebo-controlled study of pivagabine in neurasthenia.

One hundred and eighteen patients with neurasthenia, as defined by ICD 10 (International Classification of Diseases), participated in a randomised, double-blind, placebo-controlled trial of pivagabine (4-[(2,2-dimethyl-1-oxopropyl)amino]butanoic acid, CAS 69542-93-4, Tonerg). Pivagabine 1800 mg/d was administered orally for four weeks. At the end of the trial, active medication was significantly superior to placebo on the Clinical Global Impression (CGI) improvement of illness scale. In addition, pivagabine treatment reduced the physical and mental fatigability of patients, and increased their sense of well-being.

Reporting, filing, and retrieval of coronary angiographic findings: a new personal computer-based system.

A software system for semiautomatic reporting and automatic filing of coronary angiographies has been developed and is suitable for an IBM Personal Computer (PC) with graphic tablet and printer. Coronary angiographic findings referring to the native anatomy, pathology, and post-surgical status can be easily recorded. The data is automatically coded and filed according to category of information. This system can analyze previously filed data and perform searches under one or more angiographic findings. This allows for a comparison of different series of patients.

Effects of steroid hormones on muscle reinnervation after nerve crush in rabbit.

The ability of an association of three steroid hormones to influence the reinnervation process and the trophism of rabbit muscles denervated by crush of the sciatic nerve was investigated. The beginning of reinnervation was established with electromyographic recordings from the tibialis anterior muscle. The distance from the site of crushing to the point where the motor nerve enters the tibialis anterior muscle was then measured in each animal, and the nerve regeneration velocity (mm/day) was calculated: a slightly but significantly higher (P less than 0.001) mean value was found in treated animals compared with untreated ones. When soleus and extensor digitorum longus (EDL) muscles were histochemically examined 50 days after lesion, a larger mean diameter of type 2c fibers was found in treated than in untreated animals, pointing out a possible useful effect of the treatment. On the contrary, the size reduction of EDL type 2b fibers was more pronounced in treated rabbits, indicating a catabolic influence of the drugs on this fiber type.