Extracellular excystation and development of Cryptosporidium: tracing the fate of oocysts within Pseudomonas aquatic biofilm systems

BMC Microbiology

Peta L Clode¹  Paul Monis²  Hanna Edwards⁴  Andrew Thompson⁴  Wan Koh³ 

[1] Centre for Microscopy, Characterisation and Analysis, The University of Western Australia, 35 Stirling Hwy, Crawley 6009, WA, Australia;South Australian Water Corporation, 250 Victoria Square, Adelaide 5000, SA, Australia;School of Occupational and Public Health, Ryerson University, 350 Victoria Street, Toronto M5B2K3, Ontario, Canada;School of Veterinary and Life Sciences, Murdoch University, South Street, Murdoch 6150, WA, Australia

关键词: Flow cytometry;    Scanning electron microscope;    Confocal microscope;    Excystation;    Extracellular multiplication;    Biofilms;    Cryptosporidium;   

Others  :  1137731
DOI  :  10.1186/s12866-014-0281-8

 received in 2014-07-30, accepted in 2014-10-30,  发布年份 2014

【 摘 要 】

Background

Aquatic biofilms often serve as environmental reservoirs for microorganisms and provide them with a nutrient-rich growth environment under harsh conditions. With regard to Cryptosporidium, biofilms can serve as environmental reservoirs for oocysts, but may also support the growth of additional Cryptosporidium stages.

Results

Here we used confocal laser scanning microscopy, scanning electron microscopy (SEM), and flow cytometry to identify and describe various Cryptosporidium developmental stages present within aquatic biofilm systems, and to directly compare these to stages produced in cell culture. We also show that Cryptosporidium has the ability to form a parasitophorous vacuole independently, in a host-free biofilm environment, potentially allowing them to complete an extracellular life cycle. Correlative data from confocal and SEM imaging of the same cells confirmed that the observed developmental stages (including trophozoites, meronts, and merozoites) were Cryptosporidium. These microscopy observations were further supported by flow cytometric analyses, where excysted oocyst populations were detected in 1, 3 and 6 day-old Cryptosporidium-exposed biofilms, but not in biofilm-free controls.

Conclusions

These observations not only highlight the risk that aquatic biofilms pose in regards to Cryptosporidium outbreaks from water distribution systems, but further indicate that even simple biofilms are able to stimulate oocyst excystation and support the extracellular multiplication and development of Cryptosporidium within aquatic environments.

【 授权许可】

   
2014 Koh et al.; licensee BioMed Central Ltd.

【 图 表 】

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