Coexistence of free-living amoebae and bacteria in selected South African hospital water distribution systems.

Pathogenic free-living amoebae (FLA), such as Naegleria fowleri, Balamuthia mandrillaris and Acanthamoeba species isolated from aquatic environments have been implicated in central nervous system, eye and skin human infections. They also allow the survival, growth and transmission of bacteria such as Legionella, Mycobacteria and Vibrio species in water systems. The purpose of this study was to investigate the co-occurrence of potentially pathogenic FLA and their associated bacteria in hospital water networks in Johannesburg, South Africa. A total of 178 water (n = 95) and swab (n = 83) samples were collected from two hospital water distribution systems. FLA were isolated using the amoebal enrichment technique and identified using PCR and 18S rDNA sequencing. Amoebae potentially containing intra-amoebal bacteria were lysed and cultured on blood agar plates. Bacterial isolates were characterized using the VITEK®2 compact System. Free-living amoebae were isolated from 77 (43.3 %) of the samples. Using microscopy, PCR and 18S rRNA sequencing, Acanthamoeba spp. (T3 and T20 genotypes), Vermamoeba vermiformis and Naegleria gruberi specie were identified. The Acanthamoeba T3 and T20 genotypes have been implicated in eye and central nervous system infections. The most commonly detected bacterial species were Serratia marcescens, Stenotrophomonas maltophilia, Delftia acidovorans, Sphingomonas paucimobilis and Comamonas testosteroni. These nosocomial pathogenic bacteria are associated with systematic blood, respiratory tract, the urinary tract, surgical wounds and soft tissues infections. The detection of FLA and their associated opportunistic bacteria in the hospital water systems point out to a potential health risk to immune-compromised individuals.

Detection of free-living amoebae using amoebal enrichment in a wastewater treatment plant of Gauteng Province, South Africa.

Free-living amoebae pose a potential health risk in water systems as they may be pathogenic and harbor potential pathogenic bacteria known as amoebae resistant bacteria. Free-living amoebae were observed in 150 (87.2%) of the environmental water samples. In particular, Acanthamoeba sp. was identified in 22 (12.8%) using amoebal enrichment and confirmed by molecular analysis. FLA were isolated in all 8 stages of the wastewater treatment plant using the amoebal enrichment technique. A total of 16 (9.3%) samples were positive for FLA from influent, 20 (11.6%) from bioreactor feed, 16 (9.3%) from anaerobic zone, 16 (9.3%) from anoxic zone, 32 (18.6%) from aerators, 16 (9.3%) from bioreactor effluent, 11 (6.4%) from bioreactor final effluent, and 45 (26.2%) from maturation pond. This study provides baseline information on the occurrence of amoebae in wastewater treatment plant. This has health implications on receiving water bodies as some FLA are pathogenic and are also involved in the transmission and dissemination of pathogenic bacteria.

Identification methods for Legionella from environmental samples.

Laboratories responsible for Legionella diagnostics around the world use a number of different culturing methods of non-equivalent sensitivities and specificities, to detect Legionella species in environmental samples. Specific countries usually standardize and use one approved method. For example, laboratories in Australia use the Australian Standard (AS) method and those in Europe, the International Standard method (ISO). However, no standard culturing methods have been established in South Africa to date. As a result, there is uncertainty about the true prevalence and most common species of Legionella present in the South African environment. In an attempt to provide guidelines for the development of a standard method specific for South Africa, the ISO, AS and a most probable number method were evaluated and compared. In addition, the effect of sample re-incubation with autochthonous amoebae on culture outcome was studied. Samples were collected from four environments, representing industrial water, mine water and biofilm. The samples were concentrated by membrane filtration and divided into three portions and cultured without pretreatment, after acid treatment and after heat treatment, on four culture media namely alphaBCYE, BMPA, MWY and GVPC agar. A selective approach, incorporating heat treatment, but not acid treatment, combined with culture on alphaBCYE and GVPC or MWY, was most appropriate for legionellae detection in the samples evaluated. Legionellae were cultured from 82% of the environmental samples we evaluated. In 54% of the samples tested, legionellae were present in numbers equal to or exceeding 10(2) colony-forming units per milliliter (cfu/ml). Legionella pneumophila serogroups (SGs) 1-14 were the most prevalent species and were present as single, or a combination of two or more SGs in a number of samples tested. Re-incubation of sample concentrates with autochthonous amoebae improved the culturability of legionellae in 50% of cultures on alphaBCYE and 25% on GVPC.

Exposures to Legionella pneumophila and Chlamydia pneumoniae in South African Mine Workers.

Exposures of South African mine workers to Legionella pneumophila and Chlamydia pneumoniae were studied over a period of six months. Seroprevalence data were compared with data from a group of factory workers. Antibodies to L. pneumophila serogroups 1-4 were demonstrated in 36% of the mine workers and 10% of the factory workers (p < 0.0001). Seroconversion occurred in 18% of the mine workers during the study period. Antibodies to C. pneumoniae were present in 66% of the mine workers and 22% of the factory workers (p < 0.0001). Seroconversion was demonstrated in 17% of the mine workers during the six months of follow-up. A significant association between the presence of respiratory symptoms and seroconversion was demonstrated (p < 0.025). The data suggest that mine workers are at increased risk for infections with L. pneumophila and C. pneumoniae. Legionella pneumophila; Chlamydia pneumoniae; mine workers; factory workers; risk factors; seroprevalence; seroconversion

'Atypical' bacteria are a common cause of community-acquired pneumonia in hospitalised adults.

OBJECTIVES: To assess the proportion of cases of community-acquired pneumonia caused by 'atypical' bacteria, including the recently discovered Chlamydia pneumoniae, and to compare the clinical, radiographic and laboratory features of patients with and without 'atypical' bacteria. METHODS: A prospective serological study was carried out on consecutive adult pneumonia patients from July 1987 to July 1988. Acute and convalescent sera were tested in batches for antibodies against Legionella pneumophila serogroup 1, C. pneumoniae, Chlamydia psittaci, Coxiella burnetii (phase-2 antigen) and Mycoplasma pneumoniae (IgG and IgM). Records and chest radiographs were examined retrospectively. RESULTS: Acute and convalescent sera were available from 113 patients. The records of 4 patients could not be traced and 17 patients did not fulfil the inclusion criteria. Thirty-two of these 92 patients (35.9%) were found to be infected with 'atypical' bacteria. The two most common organisms were C. pneumoniae (20.7%) and L. pneumophila (8.7%). There were no differences in the clinical and radiographic features of patients with and without 'atypical' bacteria. Clinicians prescribed erythromycin or tetracyclines with equal frequency in the two groups. CONCLUSIONS: 'Atypical' bacteria, especially C. pneumoniae, are a common cause of community-acquired pneumonia in adults in South Africa. This is the first demonstration of an aetiological role of C. pneumoniae in this country. We confirmed the finding of other studies that there are no clinical, radiographic or laboratory features characteristic of 'atypical' bacterial infection in hospitalised patients. This has major implications for therapy, as these organisms respond to erythromycin and tetracyclines, but not to beta-lactam antibiotics.