Occurrence of Cryptosporidium and Giardia in sewage sludge and solid waste landfill leachate and quantitative comparative analysis of sanitization treatments on pathogen inactivation.

Circulation of Cryptosporidum and Giardia in the environment can be facilitated by spreading of sewage sludge on agricultural or livestock grazing lands or depositing in landfills. Solid waste landfill leachate and sewage sludge samples were quantitatively tested for C. parvum and C. hominis oocysts, and G. lamblia cysts by the combined multiplexed fluorescence in situ hybridization (FISH) and immunofluorescent antibody (IFA) method. Subsequently, the effects of four sanitization treatments (i.e., ultrasound and microwave energy disintegrations, and quicklime and top-soil stabilization) on inactivation of these pathogens were determined. The landfill leachate samples were positive for Giardia, and sewage sludge samples for both Cryptosporididium and Giardia. The overall concentration of G. lamblia cysts (mean; 24.2/g) was significantly higher (P<0.01) than the concentration of C. parvum and C. hominis oocysts (mean; 14.0/g). Sonication reduced the load of G. lamblia cysts to non-detectable levels in 12 of 21 samples (57.1%), and in 5 of 6 samples (83.3%) for C. parvum and C. hominis. Quicklime stabilization treatment was 100% effective in inactivation of Cryptosporidium and Giardia, and microwave energy disintegration lacked the efficacy. Top-soil stabilization treatment reduced gradually the load of both pathogens which was consistent with the serial dilution of sewage sludge with the soil substrate. This study demonstrated that sewage sludge and landfill leachate contained high numbers of potentially viable, human-virulent species of Cryptosporidium and Giardia, and that sonication and quicklime stabilization were the most effective treatments for sanitization of sewage sludge and solid waste landfill leachates.

[1]  Halshka Graczyk,et al.  Human-virulent microsporidian spores in solid waste landfill leachate and sewage sludge, and effects of sanitization treatments on their inactivation , 2007, Parasitology Research.

[2]  E. Pozio,et al.  Long-term survival of Cryptosporidium parvum oocysts in seawater and in experimentally infected mussels (Mytilus galloprovincialis). , 1999, International journal for parasitology.

[3]  C. Shiff,et al.  Environmental and geographical factors contributing to watershed contamination with Cryptosporidium parvum oocysts. , 2000, Environmental research.

[4]  M. Cranfield,et al.  Zoonotic transmission of Cryptosporidium parvum: Implications for water-borne cryptosporidiosis. , 1997, Parasitology today.

[5]  J. Rose,et al.  The infectivity of Cryptosporidium parvum in healthy volunteers. , 1995, The New England journal of medicine.

[6]  L Cole,et al.  Viability of Giardia cysts suspended in lake, river, and tap water , 1989, Applied and environmental microbiology.

[7]  R. Fayer,et al.  Cryptosporidium hominis n. sp. (Apicomplexa: Cryptosporidiidae) from Homo sapiens , 2002, The Journal of eukaryotic microbiology.

[8]  C Schönning,et al.  QMRA (quantitative microbial risk assessment) and HACCP (hazard analysis and critical control points) for management of pathogens in wastewater and sewage sludge treatment and reuse. , 2004, Water science and technology : a journal of the International Association on Water Pollution Research.

[9]  R. Cantúsio Neto,et al.  Occurrence of Giardia cysts and Cryptosporidium oocysts in activated sludge samples in Campinas, SP, Brazil. , 2004, Revista do Instituto de Medicina Tropical de Sao Paulo.

[10]  J. Schijven,et al.  Modelling the sewage discharge and dispersion of Cryptosporidium and Giardia in surface water. , 2001, Water research.

[11]  D. Veal,et al.  Determination of Cryptosporidium parvum oocyst viability by fluorescence in situ hybridization using a ribosomal RNA‐directed probe , 2004, Journal of applied microbiology.

[12]  T. Graczyk,et al.  Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife , 2006, Parasitology Research.

[13]  R C RENDTORFF,et al.  The experimental transmission of human intestinal protozoan parasites. II. Giardia lamblia cysts given in capsules. , 1954, American journal of hygiene.

[14]  J. Rose,et al.  Occurrence of Cryptosporidium oocysts in sewage effluents and selected surface waters. , 1987, The Journal of parasitology.

[15]  K. Schwab,et al.  Recovery, Bioaccumulation, and Inactivation of Human Waterborne Pathogens by the Chesapeake Bay Nonnative Oyster, Crassostrea ariakensis , 2006, Applied and Environmental Microbiology.

[16]  C. Gerba,et al.  Hazards from pathogenic microorganisms in land-disposed sewage sludge. , 1993, Reviews of environmental contamination and toxicology.

[17]  R. Fayer,et al.  Estimating viability of Cryptosporidium parvum oocysts using reverse transcriptase-polymerase chain reaction (RT-PCR) directed at mRNA encoding amyloglucosidase. , 2000, Journal of microbiological methods.

[18]  M. Hutchison,et al.  Fate of Pathogens Present in Livestock Wastes Spread onto Fescue Plots , 2005, Applied and Environmental Microbiology.

[19]  William D. Gollnitz,et al.  COMMERCIAL LABS: HOW ACCURATE ARE THEY? , 1994 .

[20]  P. Gale,et al.  Land application of treated sewage sludge: quantifying pathogen risks from consumption of crops , 2005, Journal of applied microbiology.

[21]  T. Graczyk,et al.  Human waterborne parasites in zebra mussels (Dreissena polymorpha) from the Shannon River drainage area, Ireland , 2004, Parasitology Research.

[22]  A. Thompson,et al.  Giardia and Cryptosporidium join the 'Neglected Diseases Initiative'. , 2006, Trends in parasitology.

[23]  M. Hutchison,et al.  Declines of zoonotic agents in liquid livestock wastes stored in batches on‐farm , 2005, Journal of applied microbiology.

[24]  B. Grimes,et al.  Combined FISH and mAb detection of cryptosporidium parvum and giardia lamblia carried by synanthropic flies , 2002 .

[25]  J. Rose,et al.  Assessment of Methods for Detection of Infectious Cryptosporidium Oocysts and Giardia Cysts in Reclaimed Effluents , 2003, Applied and Environmental Microbiology.

[26]  R. Fayer,et al.  The Effects of E‐beam Irradiation and Microwave Energy on Eastern Oysters (Crassostrea virginica) Experimentally Infected with Cryptosporidium parvum , 2005, The Journal of eukaryotic microbiology.

[27]  M. Hänninen,et al.  Comparative analysis of Cryptosporidium, Giardia and indicator bacteria during sewage sludge hygienization in various composting processes , 2004, Letters in applied microbiology.

[28]  Huw Smith,et al.  Occurrence of Cryptosporidium spp oocysts and Giardia spp cysts in sewage influents and effluents from treatment plants in England , 1997 .

[29]  H. Dupont,et al.  Virulence of three distinct Cryptosporidium parvum isolates for healthy adults. , 1999, The Journal of infectious diseases.

[30]  D. Veal,et al.  Oligonucleotide probes for specific detection of Giardia lamblia cysts by fluorescent in situ hybridization , 2001, Journal of applied microbiology.

[31]  Allen P. Miraflor,et al.  Human Enteropathogen Load in Activated Sewage Sludge and Corresponding Sewage Sludge End Products , 2007, Applied and Environmental Microbiology.

[32]  N. Ashbolt,et al.  Evaluation of fluorochromes for flow cytometric detection of Cryptosporidium parvum oocysts labelled by fluorescent in situ hybridization , 1998, Letters in applied microbiology.

[33]  Y. Boulard,et al.  Ultrastructural changes following treatment with a microwave pulse in the oocyst ofEimeria magna Perard, 1925 , 2004, Parasitology Research.

[34]  Thiriat,et al.  Determination of Giardia cyst viability in environmental and faecal samples by immunofluorescence, fluorogenic dye staining and differential interference contrast microscopy , 1998, Letters in applied microbiology.

[35]  J. Jofre,et al.  Occurrence of Cryptosporidium spp. oocysts in raw and treated sewage and river water in north‐eastern Spain , 2005, Journal of applied microbiology.

[36]  L. Robertson,et al.  Occurrence of Cryptosporidium Oocysts and Giardia Cysts in Sewage in Norway , 2006, Applied and Environmental Microbiology.

[37]  T. Graczyk,et al.  Cryptosporidium parvum and Giardia lamblia Recovered from Flies on a Cattle Farm and in a Landfill , 2004, Applied and Environmental Microbiology.

[38]  M. Hänninen,et al.  Simultaneous detection of Cryptosporidium parvum and Giardia in sewage sludge by IC‐PCR , 2001, Journal of applied microbiology.

[39]  B. Grimes,et al.  Detection of Cryptosporidium parvum and Giardia lamblia carried by synanthropic flies by combined fluorescent in situ hybridization and a monoclonal antibody. , 2003, The American journal of tropical medicine and hygiene.

[40]  M. Nadji,et al.  Methodology for Preservation of High Molecular-Weight RNA in Paraffin-Embedded Tissue: Application for Laser-Capture Microdissection , 2005, Diagnostic molecular pathology : the American journal of surgical pathology, part B.

[41]  M. Cranfield,et al.  Evaluation of commercial enzyme immunoassay (EIA) and immunofluorescent antibody (FA) test kits for detection of Cryptosporidium oocysts of species other than Cryptosporidium parvum. , 1996, The American journal of tropical medicine and hygiene.

[42]  M. Iturriza-Gómara,et al.  Detection of Viral, Bacterial, and Parasitological RNA or DNA of Nine Intestinal Pathogens in Fecal Samples Archived as Part of the English Infectious Intestinal Disease Study: Assessment of the Stability of Target Nucleic Acid , 2005, Diagnostic molecular pathology : the American journal of surgical pathology, part B.

[43]  I Ozturk,et al.  A new process for the combined treatment of municipal wastewaters and landfill leachates in coastal areas. , 2002, Water science and technology : a journal of the International Association on Water Pollution Research.

[44]  Gordon A. McFeters,et al.  Drinking Water Microbiology , 1990, Brock/Springer Series in Contemporary Bioscience.

[45]  M. Cranfield,et al.  Cryptosporidium parvum oocysts recovered from water by the membrane filter dissolution method retain their infectivity. , 1997, The Journal of parasitology.

[46]  M. Sobsey Field survey of enteric viruses in solid waste landfill leachates. , 1978, American journal of public health.

[47]  G. Glass,et al.  Quantitative assessment of viable Cryptosporidium parvum load in commercial oysters (Crassostrea virginica) in the Chesapeake Bay , 2006, Parasitology Research.

[48]  L. Ash,et al.  Parasites: A Guide to Laboratory Procedures and Identification , 1989 .

[49]  S. Sattar,et al.  Fate of Cryptosporidium oocysts, Giardia cysts, and microbial indicators during wastewater treatment and anaerobic sludge digestion. , 1999, Canadian journal of microbiology.

[50]  F. Antunes,et al.  Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies , 2005, Parasitology Research.