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Applied and Environmental Microbiology, November 2003, p. 6758-6761, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6758-6761.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

New Method Using Sedimentation and Immunomagnetic Separation for Isolation and Enumeration of Cryptosporidium parvum Oocysts and Giardia lamblia cysts

Jaime Massanet-Nicolau*

Thames Water Utilities, Reading, Berkshire RG2 0JN, United Kingdom

Received 16 January 2003/ Accepted 17 August 2003

A new method for the isolation of Cryptosporidium parvum oocysts and Giardia lamblia cysts from biosolid samples has been developed that utilizes sedimentation and immunomagnetic separation. The method was used to recover stained cysts and oocysts (spike organisms) from primary settled sewage sludge, anaerobically digested sewage sludge, and bovine manure. Recovery efficiencies associated with this method were approximately 40 to 60% and were significantly greater than those associated with similar methods based on sucrose flotation (P < 0.001). The recovery efficiency of the sedimentation-based method showed no significant reduction as a result of sample storage for up to 21 days (P > 0.05). Recovery efficiencies were determined by spiking samples with prestained cysts and oocysts, allowing them to be differentiated from those naturally present in the biosolid samples. The prestained cysts and oocysts had been fixed in 5% formalin, and the recovery efficiencies associated with this method may be different from recovery efficiencies for fresh cysts or oocysts.

* Mailing address: Thames Water Utilities, Spencer House, Manor Farm Rd., Reading, Berkshire RG2 0JN, United Kingdom. Phone: 0118 923 623 1. Fax: 0118 923 631 1. E-mail: jaime.massanet-nicolau{at}thameswater.co.uk.

Applied and Environmental Microbiology, November 2003, p. 6758-6761, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6758-6761.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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