Cryptosporidium parvum is a protozoan pathogen of humans and livestock worldwide. Its ability to infect a wide range of species raises questions as to the involvement of a specific host cell receptor for parasite-host recognition. To investigate the mechanism of parasite-host cell recognition, we have developed an in vitro cell suspension binding assay to investigate adhesion of C. parvum sporozoites to host cells. Morphologic features of binding events observed with this assay were identical to those described in natural infections. Glycoconjugates, Madin Darby bovine kidney (MDBK) cell fractions, and plasma membrane vesicles (PMVs) were screened for their ability to block binding of sporozoites to MDBK cells. Mucins, MDBK cell fractions, and PMVs exhibited dose-dependent inhibition of sporozoite binding. The major inhibitory fraction from MDBK cells was found to be insoluble in aqueous medium, nonsaponifiable, and lacking carbohydrate moieties, nitrogen, and phosphorus. Its inhibitory effect was resistant to heat, protease digestion, and glycosidase treatment, suggesting that the inhibitory activity is a lipid or a lipid-like component. The inhibitory activity was purified from MDBK cells, and in larger amounts from bovine small intestinal mucosa, by organic solvent extraction, semipreparative high-pressure liquid chromatography, and preparative high-performance thin-layer chromatography. Biochemical analyses, thin-layer chromatography staining techniques, mass spectrometry, and elemental analysis were used to partially characterize the purified lipid. These results indicate that a host intestinal lipid(s) or a lipidlike component(s) may play an important role in the early stages of host cell invasion by C. parvum.

Three dogs treated for systemic blastomycosis with intravenous amphotericin B (one case) or amphotericin B lipid complex (two cases) developed mild to severe calcinosis cutis two to six weeks after the initiation of treatment. Abnormalities in serum calcium and phosphorus during treatment for blastomycosis or at the time of diagnosis of calcinosis cutis were slight or absent. The calcification was not associated with lesions of cutaneous blastomycosis. Calcification was limited to the skin in two cases and may have also involved the kidneys in one. The calcinosis cutis resolved completely in all three dogs with no (two cases) or only palliative (one case) therapy.

Declines of amphibians have been attributed to many factors including habitat degradation. The introduction of grass carp (Ctenopharyngodon idella) as a biological agent for aquatic plant control in ponds and lakes managed narrowly for human recreation has likely contributed to amphibian declines through massive plant removal and associated habitat simplification and thus degradation. This research examined the interactions among grass carp and three Midwestern aquatic plants (Jussiaea repens, Ranunculus longirostris, and R. flabellaris) that may be of value in rehabilitation of habitats needed by amphibians. The feeding preference study found that C. idella avoided eating both J. repens and R. longirostris. Ranunculus species studied to date contain a vesicant toxin called ranunculin that is released upon mastication. The study that compared the effects of R. flabellaris, J. repens and a control food administered by tube feeding to C. idella found significant lesions only in the mucosal epithelium of the individuals exposed to R.flabellaris. The avoidance by C. idella of J. repens and R. longirostris in the feeding preference study, and the significant toxicity of R. flabellaris demonstrated by the dosing study, indicate these plants warrant further examination as to their potential effectiveness in aquatic amphibian habitat rehabilitation.

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