Temporal occurrence of Cryptosporidium in the Manila clam Ruditapes philippinarum in northern Adriatic Italian lagoons.

In order to evaluate the temporal occurrence of Cryptosporidium oocysts in Ruditapes philippinarum clams bred along the northeastern Italian Adriatic coast and molecularly characterize the isolates, 2,160 specimens (180 clams per month) were collected from three clam farms from January to December 2004. Two farms (sites A and B) were located in Venice (Chioggia, Veneto region) and one (site C) in the Marano Lagoons (Friuli Venezia Giulia region). Clams from 36 pools (i.e., one pool of 60 clams per month per site) were subjected to a high-sensitivity seminested PCR assay specific for a 360-bp diagnostic region internal to the Cryptosporidium spp. outer wall protein gene. Positive amplicons were sequenced and analyzed. Cryptosporidium DNA was found in clams from seven pools (sites A and B) during 1 month of sampling at site A and 6 months of sampling at site B, with Cryptosporidium hominis and Cryptosporidium parvum being detected. The expected infection rate of the clams was 0.36%. Site B showed a significantly higher expected infection rate (1.15%) than did the other sites (A = 0.14% and C = 0%). Given its high sensitivity and specificity, this seminested PCR assay can be considered a reliable tool for detecting and distinguishing species within the Cryptosporidium genus. The seasonal pattern of contamination and the related public health risks are of particular concern.

[1]  G. Prioli. La molluschicoltura in Italia , 2008 .

[2]  J. McLauchlin,et al.  Characterisation of a Cryptosporidium isolate from water buffalo (Bubalus bubalis) by sequencing of a fragment of the Cryptosporidium oocyst wall protein gene (COWP). , 2005, Veterinary parasitology.

[3]  D. Traversa,et al.  Cryptosporidium parvum oocysts in seawater clams (Chamelea gallina) in Italy , 2005 .

[4]  S. Enomoto,et al.  Cryptosporidium and cryptosporidiosis. , 2005, Advances in parasitology.

[5]  Adriano Sfriso,et al.  The changing state of contamination in the Lagoon of Venice. Part 1: organic pollutants. , 2005, Chemosphere.

[6]  R. Fayer,et al.  Cryptosporidium: a water-borne zoonotic parasite. , 2004, Veterinary parasitology.

[7]  R. Fayer,et al.  Zoonotic protozoa: from land to sea. , 2004, Trends in parasitology.

[8]  D. Otranto,et al.  Genotyping of Cryptosporidium Isolates from Chamelea gallina Clams in Italy , 2004, Applied and Environmental Microbiology.

[9]  J. McLauchlin,et al.  Detection of Cryptosporidium and Giardia in molluscan shellfish by multiplexed nested-PCR. , 2004, International journal of food microbiology.

[10]  Una Ryan,et al.  Cryptosporidium Taxonomy: Recent Advances and Implications for Public Health , 2004, Clinical Microbiology Reviews.

[11]  M. Panaro,et al.  Prevalence of Cryptosporidium in children with enteritis in Southern Italy , 1996, European Journal of Epidemiology.

[12]  Paul Monis,et al.  Cryptosporidium and Giardia-zoonoses: fact or fiction? , 2003, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[13]  J. Martinez-Urtaza,et al.  Contamination of bivalve molluscs by Cryptosporidium oocysts: the need for new quality control standards. , 2003, International journal of food microbiology.

[14]  R. Guerrant,et al.  Cryptosporidiosis: epidemiology and impact. , 2002, Microbes and infection.

[15]  Ó. García-Martín,et al.  Survival of Cryptosporidium parvum oocysts recovered from experimentally contaminated oysters (Ostrea edulis) and clams (Tapes decussatus) , 2002, Parasitology Research.

[16]  N. Beeching,et al.  Cryptosporidium muris Infection in an HIV-Infected Adult, Kenya , 2002, Emerging infectious diseases.

[17]  J. Marshall,et al.  Experimental infection of a lamb with Cryptosporidium parvum genotype 1 , 2001, Veterinary Record.

[18]  I. Sulaiman,et al.  Sequence Differences in the Diagnostic Target Region of the Oocyst Wall Protein Gene ofCryptosporidium Parasites , 2000, Applied and Environmental Microbiology.

[19]  S J Upton,et al.  Epidemiology of Cryptosporidium: transmission, detection and identification. , 2000, International journal for parasitology.

[20]  E. Costas,et al.  Detection of Infectious Cryptosporidium parvum Oocysts in Mussels (Mytilus galloprovincialis) and Cockles (Cerastoderma edule) , 2000, Applied and Environmental Microbiology.

[21]  Paul Monis,et al.  Cryptosporidium spp. in Domestic Dogs: the “Dog” Genotype , 2000, Applied and Environmental Microbiology.

[22]  G A Buck,et al.  Animal propagation and genomic survey of a genotype 1 isolate of Cryptosporidium parvum. , 2000, Molecular and biochemical parasitology.

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

[24]  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.

[25]  T. Graczyk,et al.  Detection of Cryptosporidium oocysts and Giardia cysts in the tissues of eastern oysters (Crassostrea virginica) carrying principal oyster infectious diseases. , 1998, The Journal of parasitology.

[26]  T. Graczyk,et al.  Survival of Infectious Cryptosporidium parvum Oocysts in Seawater and Eastern Oysters (Crassostrea virginica) in the Chesapeake Bay , 1998, Applied and Environmental Microbiology.

[27]  M. Cranfield,et al.  Recovery of Waterborne Cryptosporidium parvum Oocysts by Freshwater Benthic Clams (Corbicula fluminea) , 1998, Applied and Environmental Microbiology.

[28]  A. Crisanti,et al.  PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin. , 2006, FEMS microbiology letters.

[29]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[30]  P. Pezzotti,et al.  Clinical cryptosporidiosis and human immunodeficiency virus (HIV)-induced immunosuppression: findings from a longitudinal study of HIV-positive and HIV-negative former injection drug users. , 1997, The Journal of infectious diseases.

[31]  M. Cranfield,et al.  In Vitro Interactions of Asian Freshwater Clam (Corbicula fluminea) Hemocytes and Cryptosporidium parvum Oocysts , 1997, Applied and environmental microbiology.

[32]  A. Crisanti,et al.  PCR-RFLP analysis of the oocyst wall protein (COWP) gene discriminates between and , and between isolates of human and animal origin , 1997 .

[33]  T. Graczyk,et al.  Potential Role of the Eastern Oyster, Crassostrea virginica, in the Epidemiology of Cryptosporidium parvum , 1997, Applied and environmental microbiology.

[34]  L. Robertson,et al.  Survival of Cryptosporidium parvum oocysts under various environmental pressures , 1992, Applied and environmental microbiology.