Infection of Immunosuppressed C57BL/6N Adult Mice with a Single Oocyst of Cryptosporidium parvum

The present study was designed to determine the minimum number of Cryptosporidium parvum oocysts capable of producing patent infections in immunosuppressed C57BL/6N adult mice. Sixty-four female mice were divided into 6 groups of 8 mice each, except group 1 that contained 24 mice. Mice in groups 1–3 were immunosuppressed with dexamethasone and inoculated with 1, 5, and 10 oocysts per mouse, respectively. The accuracy of the inoculum size was microscopically confirmed. Mice in groups 4–6 served as controls: they received either only oocyst inoculation (group 4), or immunosuppression (group 5), or no treatments (group 6). Fecal oocyst shedding was monitored daily for each mouse using an indirect immunofluorescent assay. Parasite colonization in the terminal ileum of each mouse was evaluated histologically. Four of 24 mice in group 1 developed patent infections, with a prepatent period of approximately 6 days. All mice in groups 2 and 3 developed patent infections, with prepatent periods ranging from 4 to 7 days. Mice in groups 4–6 remained uninfected. Parasite colonization was observed in the terminal ilea of all mice in groups 1–3 that shed fecal oocysts. The present study experimentally demonstrates that a single viable oocyst can induce patent C. parvum infections in immunosuppressed C57BL/6N adult mice and indicates that this mouse model could be used for the parasite genotype or isolate cloning.

[1]  M. Healey,et al.  Experimental Cryptosporidium parvum infections in immunosuppressed adult mice , 1992, Infection and immunity.

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

[3]  K. Shianna,et al.  Randomly Amplified Polymorphic DNA PCR Analysis of Bovine Cryptosporidium parvum Strains Isolated from the Watershed of the Red River of the North , 1998, Applied and Environmental Microbiology.

[4]  Y. Horii,et al.  Persistent infection of Strongyloides venezuelensis and normal expulsion of Nippostrongylus brasiliensis in Mongolian gerbils, Meriones unguiculatus, with reference to the cellular responses in the intestinal mucosa , 1993, Parasite immunology.

[5]  J. Court,et al.  The efficacy of benzimidazole anthelmintics against late fourth stage larvae of Trichostrongylus colubriformis in gerbils and Nippostrongylus brasiliensis in rats. , 1985, Veterinary parasitology.

[6]  R. Tarazona,et al.  Cryptosporidium parvum infection in experimentally infected mice: infection dynamics and effect of immunosuppression. , 1998, Folia parasitologica.

[7]  N. A. Sinclair,et al.  Effects of ozone, chlorine dioxide, chlorine, and monochloramine on Cryptosporidium parvum oocyst viability , 1990, Applied and environmental microbiology.

[8]  T. Nolan,et al.  Strongyloides stercoralis: the first rodent model for uncomplicated and hyperinfective strongyloidiasis, the Mongolian gerbil (Meriones unguiculatus). , 1993, The Journal of infectious diseases.

[9]  K. Friedhoff Manual of veterinary parasitological laboratory techniques: Ministry of Agriculture, Fisheries and Food, Agricultural Development and Advisory Service, Technical Bulletin No. 18, 2nd edition, 1977, iii + 129 pp., 16 figs., XI plates, 9 tables, Price £3.50 , 1978 .

[10]  V. Pandey,et al.  Biological parameters of Trichostrongylus colubriformis in Meriones unguiculatus. , 1999, Veterinary parasitology.

[11]  D. Lindsay,et al.  Infection dynamics of Cryptosporidium parvum (Apicomplexa: Cryptosporiidae) in neonatal mice (Mus musculus). , 1986, The Journal of parasitology.

[12]  S. Yang,et al.  Infectivity of preserved Cryptosporidium parvum oocysts for immunosuppressed adult mice. , 1996, FEMS immunology and medical microbiology.

[13]  L. Sekla,et al.  Purification of Cryptosporidium oocysts and sporozoites by cesium chloride and Percoll gradients. , 1987, The American journal of tropical medicine and hygiene.

[14]  H. Robinson,et al.  Murine infection model for maintenance and amplification of Cryptosporidium parvum oocysts , 1995, Journal of clinical microbiology.

[15]  S. Yang,et al.  Therapeutic efficacy of paromomycin in immunosuppressed adult mice infected with Cryptosporidium parvum. , 1995, The Journal of parasitology.

[16]  S. Cifelli,et al.  Efficacy of thiabendazole, levamisole hydrochloride and the major natural avermectins against Trichostrongylus colubriformis in the gerbil (Meriones unguiculatus). , 1981, Research in veterinary science.

[17]  H. Wędrychowicz,et al.  Trichostrongylus colubriformis in rabbits: some quantitative aspects and pathogenesis of single and multiple infections , 1988 .

[18]  M. Shirley,et al.  Eimeria tenella: infection with a single sporocyst gives a clonal population , 1996, Parasitology.

[19]  J. P. Davis,et al.  A massive outbreak in Milwaukee of cryptosporidium infection transmitted through the public water supply. , 1994, The New England journal of medicine.

[20]  Takahiko Kobayashi,et al.  Sensitive enzyme-linked immunosorbent assay (ELISA) method to measure parasite-specific antibodies of Mongolian gerbils , 1994 .

[21]  S. Yang,et al.  The immunosuppressive effects of dexamethasone administered in drinking water to C57BL/6N mice infected with Cryptosporidium parvum. , 1993, The Journal of parasitology.

[22]  S. Wright,et al.  Infective Dose Size Studies on Cryptosporidium parvum Using Gnotobiotic Lambs , 1993 .

[23]  J. Cullen,et al.  Intrahepatic postsinusoidal venous obstruction in a dog. , 1991, Journal of veterinary internal medicine.

[24]  S. Tzipori,et al.  A study of Cryptosporidium parvum genotypes and population structure. , 1998, Memorias do Instituto Oswaldo Cruz.

[25]  L. Cohn,et al.  The influence of corticosteroids on host defense mechanisms. , 1991, Journal of veterinary internal medicine.

[26]  N. Tsuji,et al.  Long-lasting parasitism of Strongyloides venezuelensis in Mongolian gerbils (Meriones unguiculatus). , 1993, The Journal of parasitology.

[27]  L. Perryman,et al.  Susceptibility of major histocompatibility complex (MHC) class I- and MHC class II-deficient mice to Cryptosporidium parvum infection , 1994, Infection and immunity.

[28]  丸山 治彦 Sensitive Enzyme-linked Immunosorbent Assay(ELISA) Method to Measure Parasite-specific Antibodies of Mongolian Gerbils , 1994 .

[29]  J. Beesley,et al.  An attempt to produce Ostertagia circumcincta infections in Mongolian gerbils. , 1988, Veterinary parasitology.

[30]  L. Perryman,et al.  Infectivity and neutralization of Cryptosporidium parvum sporozoites , 1987, Infection and immunity.

[31]  H. Wędrychowicz,et al.  Local and systemic antibody responses in gerbils following vaccination with irradiated or non-irradiated Trichostrongylus colubriformis larvae , 1986, Journal of Helminthology.

[32]  H. Robinson,et al.  Differentiation between human and animal isolates of Cryptosporidium parvum using molecular and biological markers , 1998, Parasitology Research.

[33]  D. E. Thompson,et al.  Activity of three anthelmintics against mixed infections of two Trichostrongylus species in gerbils, sheep, and goats. , 1967 .

[34]  R. Connan Experiments with Trichostrongylus colubriformis (Giles, 1892) in the guinea-pig. I. The effect of the host response on the distribution of the parasites in the gut. , 1966, Parasitology.

[35]  Is cryptosporidium clonal? , 1997, Parasitology today.

[36]  H. Wędrychowicz,et al.  Influence of adjuvants on immunity in rabbits vaccinated with infective larval somatic proteins of Trichostrongylus colubriformis. , 1990, Veterinary parasitology.

[37]  C W Daniels,et al.  Dose response of Cryptosporidium parvum in outbred neonatal CD-1 mice , 1993, Applied and environmental microbiology.

[38]  D. Emery,et al.  A serial study of rejection of Trichostrongylus colubriformis by immune sheep. , 1992, International journal for parasitology.