No abstract

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.

Groundwater in the Mooi River catchment is prone to mining, agricultural, municipal and septic tank pollution. In this study physico-chemical and microbiological parameters were determined using appropriate methods. Bacterial isolates were identified by 16S rRNA sequencing (heterotrophic plate count (HPC) bacteria and amoeba-resistant bacteria (ARB)) and multiplex polymerase chain reaction (Escherichia coli). Antibiotic resistance tests were also performed. Physico-chemical parameters were generally within target water quality ranges for drinking water. HPC bacteria ranged between 10(5) and 10(7) colony-forming units (cfu)/ml. E. coli were enumerated from Trimpark, School and Cemetery. The Blaauwbank borehole was negative for faecal streptococci. Pseudomonas spp. were most abundant in the bulk water. Opportunistic pathogens isolated included Pseudomonas aeruginosa, Acinetobacter, Aeromonas, Alcaligenes, Flavobacterium, Bacillus cereus and Mycobacterium spp. Varying patterns of antibiotic resistance were observed. Most HPC bacterial isolates were resistant to cephalothin and/or amoxicillin and a few were resistant to erythromycin and streptomycin. Pseudomonas spp. was also the most abundant ARB. Other ARBs included Alcaligenes faecalis, Ochrobactrum sp. and Achromobacter sp. ARBs were resistant to streptomycin, chloramphenicol, cephalothin, and/or amoxicillin compared to HPCs. The presence of E. coli and ARB in these groundwater sources indicates potential human health risks. These risks should be further investigated and quantified, and groundwater should be treated before use.

The purpose of this study was to investigate the occurrence of free-living amoebae in the water system of a teaching hospital in Johannesburg (South Africa). Water and biofilm samples were collected from the theatres, theatre sterilisation service unit, central sterilisation service unit and endoscopy/bronchoscopy unit. The samples were filtered and seeded on non-nutrient agar spread with heat-killed Escherichia coli. Of the 71 samples collected, 63 (88.7%) were positive for free-living amoeba. Acanthamoeba spp., Balamuthia spp. and Hartmanella spp. were identified by morphology. The presence of free-living amoeba in the hospital water network may be a potential health risk.

This study investigated the occurrence of free-living amoebae (FLA) in a public hospital in South Africa. A total of 97 water and biofilm samples from the municipal water inlet of the hospital, theatres, theatre sterilization service unit, central sterilization service unit, endoscopy/gastroscopy unit, intensive care unit and the renal unit were collected and examined for the presence of FLA using an amoebal co-culture and molecular techniques. Of the 97 samples, 77 (79.4%), 40 (52%) water and 37 (48.1%) biofilm, contained FLA. The genera Acanthamoeba, Vermamoeba (formerly Hartmanella) and Naegleria were detected by morphology, 18S rRNA PCR (polymerase chain reaction) and sequence analyses. Further sequence analysis of the five Acanthamoeba-positive isolates revealed a close resemblance with the potentially pathogenic T20 genotype. These results show a potential health risk to immuno-compromised patients and health care workers as some of the species detected are pathogenic and may harbor potential intracellular bacteria responsible for nosocomial infections. To date, this is the first report on the detection of potentially pathogenic amoebae from South African hospital water systems.