title: Cryptosporidium oocysts and Giardia cysts-A practical and sensitive method for counting and manipulating small numbers.
abstract: A rapid and inexpensive method is described for accurate and reproducible counting and manipulating small numbers of Cryptosporidium oocysts and Giardia cysts. From a suspension of oocysts or cysts at concentration from 1000 to 5000/mL (1-5/μL), replicate 5μL droplets are micro pipetted onto the edge of a microscope slide. Unstained oocysts or cysts in each droplet can be counted in a few minutes and replicated for statistical strength. The concentration of the suspension can then be verified by pipetting desired volumes containing approximately desired numbers onto confined 13mm membranes for IFA staining and counting with replication as desired. Requiring only a micropipette, analytical balance, and a microscope, this provides a useful tool accessible to virtually any microbiology laboratory.
authors.affiliation: Civil, Mining, & Environmental Engineering, University of Wollongong, Wollongong, NSW 2522, Australia. Electronic address: jongerth@uow.edu.au.
authors.forename: Jerry E
authors.initials: JE
authors.lastname: Ongerth
authors.affiliation: Civil, Mining, & Environmental Engineering, University of Wollongong, Wollongong, NSW 2522, Australia.
authors.forename: Frhat M A
authors.initials: FMA
authors.lastname: Saaed
mesh_terms: D000818:Animals; D003458:Cryptosporidium; D005872:Giardia; D033661:Oocysts / Q000382:microbiology*
keywords: Counting; Cryptosporidium oocysts; Giardia cysts; Manipulating; Small numbers