title: Surveillance of parasitic Legionella in surface waters by using immunomagnetic separation and amoebae enrichment.
abstract: Free-living amoebae (FLA) are potential reservoirs of Legionella in aquatic environments. However, the parasitic relationship between various Legionella and amoebae remains unclear. In this study, surface water samples were gathered from two rivers for evaluating parasitic Legionella. Warmer water temperature is critical to the existence of Legionella. This result suggests that amoebae may be helpful in maintaining Legionella in natural environments because warmer temperatures could enhance parasitisation of Legionella in amoebae. We next used immunomagnetic separation (IMS) to identify extracellular Legionella and remove most free Legionella before detecting the parasitic ones in selectively enriched amoebae. Legionella pneumophila was detected in all the approaches, confirming that the pathogen is a facultative amoebae parasite. By contrast, two obligate amoebae parasites, Legionella-like amoebal pathogens (LLAPs) 8 and 9, were detected only in enriched amoebae. However, several uncultured Legionella were detected only in the extracellular samples. Because the presence of potential hosts, namely Vermamoeba vermiformis, Acanthamoeba spp. and Naegleria gruberi, was confirmed in the samples that contained intracellular Legionella, uncultured Legionella may survive independently of amoebae. Immunomagnetic separation and amoebae enrichment may have referential value for detecting parasitic Legionella in surface waters.
authors.affiliation: Department of Ophthalmology, Cheng Hsin General Hospital , Taipei, Taiwan, Republic of China.
authors.forename: Tsui-Kang
authors.initials: TK
authors.lastname: Hsu
authors.forename: Shu-Fen
authors.initials: SF
authors.lastname: Wu
authors.forename: Bing-Mu
authors.initials: BM
authors.lastname: Hsu
authors.forename: Po-Min
authors.initials: PM
authors.lastname: Kao
authors.forename: Chi-Wei
authors.initials: CW
authors.lastname: Tao
authors.forename: Shu-Min
authors.initials: SM
authors.lastname: Shen
authors.forename: Wen-Tsai
authors.initials: WT
authors.lastname: Ji
authors.forename: Wen-Chien
authors.initials: WC
authors.lastname: Huang
authors.forename: Cheng-Wei
authors.initials: CW
authors.lastname: Fan
mesh_terms: D000656:Amoeba / Q000254:growth & development / Q000382:microbiology*; D000818:Animals; D015373:Bacterial Typing Techniques; D003470:Culture Media; D004784:Environmental Monitoring / Q000379:methods*; D005618:Fresh Water / Q000469:parasitology; D018189:Immunomagnetic Separation / Q000379:methods; D007875:Legionella / Q000145:classification / Q000302:isolation & purification* / Q000502:physiology; D013696:Temperature; D014871:Water Microbiology; D060753:Water Quality
chemical_list: D003470:Culture Media
keywords: Extracellular Legionella; Free-living amoebae; Immunomagnetic separation (IMS); Intracellular Legionella