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<br /> 0099-27AQ/�i/SO8.00+0 DOI: .
<br /> Copyright O 20Q3� American Soaery for Micmbiolo�r. All Rig6ts Raemd.
<br /> Infectious Cryptos�noridium parvum Oocysts in Fina1
<br /> Reclaimed E�uent
<br /> Angela L Gennaccaro,' Molly R. Mclaughtin,t Wa�ter Quiatero-Betancourt,
<br /> Deb�ra E. Huffman. and Joan B. Rose$
<br /> ' lkp�tment of Mmvu Scienc� Uni►+a�iry of South Florido,
<br /> St Petasbwg Flo�ida 33701
<br /> Received 6 Deoember 2001JAoxpted 2 hme 2003
<br /> Water samples collected thronghoat aeverai reclamation fadLttes were anal�xd for t6e pt�sentx of in[ec-
<br /> tious Crypro�epoiidiu� pa,�n,a by the focas deta�ion method—most-prob�blannmber odl cnlpu+e kchniqu�,
<br /> ResWts revealed the presence of intectlona C. parvrrm ooc�ets in qpgy o[ the Snal disinieMed e6nent aamples.
<br /> Sampled e�uent contsined on averag�e aeven infections oocyats per 100 litera. Tlens, reclaimal water is not
<br /> pathogen tree but aonfains in[ectious C. parrum.
<br /> Recla�ed water (treated wastevvater) is being ntilized in the water aampks for viable C parwm by usinB vital atai� 3596
<br /> United States and t6roughout the world as an alternative nun_ of the u�9uent samples and 4b96 of the effiueni samples :con-
<br /> potable water saurce. Ia the`United States, 18 states currently tained viable ooc.ysb (4). In the lest few yeata, the fucva da
<br /> have standards and anot6er 18 have guidelines for reclaimed tearon method-most-probable-n�ber (FDM-MPl� cell cu!-
<br /> yvater (8). These standards, for the moat part, are based on ���}� �� to test the ooryst mfoctHitY
<br /> totai suspended solids and fecal ooliforms. A variety of micro- bxause the P���Y �Play�ed methoda did not acwrateiy
<br /> bial pat6ogens are present in waatewater and can be detccted re$ect the mfaxious nature of the ooc.ysta (�. The objective of
<br /> in reclaimed waur. Therefore, advanced tsestment, mctudin8 tha psesent atndy was to demoostrate the prasenoe of infeo-
<br /> filtration and disinfection, is required to produoe rec]aimed � G�� o�s in Snal reclaimed e�uent from aa
<br /> water that does not have a negative �pact on pnblic health. In �aon facilitia in the United States by using tho FDM-
<br /> regulatory langnage. this means that pathogeaS are to be less MPN all cniture technique.
<br /> than the limit of detection of tha assay (8). Samples were coUected 6bm influent, seoondary eIDuent,
<br /> �YP�oSPo�mPa^'�, a coccidian protozoan parasitq is a ��tion, aad final disinfected effiuent wate:s. Sfr.ra�a-
<br /> potential contaminant of recJaimed water. G parvum oo�ysts �� �� ��e United States, utiliziag a variery �
<br /> have been found to be .pe�isteat in the environment and re- �� y ��� ���,_ or decp�bed sand and antbiaciu
<br /> sistant to chlorination. Becanae of thia, physical ramovai by �ters or fabric disk filtecs) and disinfection methods (ehiorine
<br /> chemical pretreatment and filtration ia the primary meaas of
<br /> reducing the lcvel of oocysts in environmental water (�, q g� °r W radiation), w�ere monitored. Three facilitiea were
<br /> possble risk to human health e�rists if Sitration fails to fimction monitored five times wer a 1-year time period. Three addi-
<br /> e�ciently. This risk is greater still with redaimed watez,,as to aOO �� � mO��� �r a 5-month time period,
<br /> date no monitoring for C. pmvu►n ocx.ysts has been nqnired OII6 tO � �� °f �P� d��IIB O° the site, was 8ltered
<br /> and little information is awdilable on tho 8ltration effioienry in ��� �� �� �'� ��° ��
<br /> these facilities. Reantly, in the state of Florida, moaitoring for A°° Arbor, Mich.). After Sitration, elution, and antr3fugatIon
<br /> protozoan parasites, includiag Cryptosporidium. onoe avery 2 �D8 t6e guidelines provided by EPA method 1623 (9j, oocysts
<br /> years for larger facilities and once every 5 years for �aller wen concentrated into a pellet by immunoma�etic aeparation
<br /> facilities has been mandated Sampling is recammended at a ��� Anti-Crypto Rit; Dynat Biotech,. 7nG„ Iake SnF
<br /> single point followiag disinfection (2). ass. N.Y.)-
<br /> In oae studY. C Pa+vum oocysts were deteded in untreated Fialf of the concentrated pellet was anal�zed by the FDM-
<br /> wastewater (67'% of the samples were positive) and in re- MPN all ailtan technique (�. (The other half of the con-
<br /> daimed water (2546 of final e�nent samples were poaitive) (5). antrated pellet was processed by microscopic screening of t6e
<br /> However, only the presenoe of the oocysts was ova(uated using sample after staining witfi uwnodonal aadbodies [anti.Cyypto.
<br /> fluorescenx microscopy. Robertson et al. evaluated waato- �: Waterborne InG, New Orieaz� La.) tagged with flu•
<br /> on�cein isothiocyanate, nsing en indirecx fluoresceat-anb'body
<br /> assay [IFA] W determine total crnmb of oocyats.) The oorxxn-
<br /> ' Corresponding sut6or. Ma�7ing addrea� Departmeat of Mariae trate was ficst bleaGh treated at Toom tempetature for 8 min
<br /> Scienx, Universiry of South Florida, lI0 7th Ave. South, St petea- with a lOS9'o solution of 496 aodium
<br /> burg, FL 33701. Phone: (727) 553-3415. Fm� (727.) 553-1189. &mail: �tp.�� �� ���rite in phoi-
<br /> �►SW88(s�nadne.uaf.edu. P (PBS: PH 72) to elmtinate baeterial oon-
<br /> t Present addreas: United States Geob8ical SutveY. Ceaur for tamination as well as to trigger excystatioa of the oocysts. The
<br /> Coastal and Regional Muiae Sh�dies, SL Petcsbucg, FL 337p1. �ple was then washed once qrith PB$ aDd oenQrifyged, sYt�c
<br /> # Pre.�ent address: D�parement of Fis6eries and Wildlife, Michigan whicb t6t pellet was suspeaded 'm 1 ml of pn�varmed growth
<br /> State Univeisiry, Eaat Lanaing, MI 48824. medium (RPMI 1640 [F'isher ScientiSc, Pittabur�, Pa] plaa
<br /> 4983
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