Evaluation of intestinal protozoan morphology in polyvinyl alcohol preservative: comparison of zinc sulfate- and mercuric chloride-based compounds for use in Schaudinn's fixative

Abstract

As a result of disposal problems related to the use of mercury compounds, many laboratories have considered switching from mercuric chloride-based Schaudinn's and polyvinyl alcohol (PVA) stool preservatives to other non-mercury-based preservatives. The primary use for PVA-preserved specimens is the permanent stained smear, the most important technique in the routine ova and parasite examination for the identification and confirmation of intestinal protozoa. A comparison of organism recovery and morphology of the intestinal protozoa was undertaken with PVA containing either a zinc sulfate base or the "gold standard" mercuric chloride base. Paired positive fecal specimens (106 from 64 patients) were collected and examined microscopically by the trichrome stain technique. There were 161 instances in which organism trophozoite and/or cyst stages were identified and 3 in which human cells were identified. Morphology, clarity of nuclear and cytoplasmic detail, overall color differences, and the ease or difficulty in detecting intestinal protozoa in fecal debris, as well as the number of patients with a missed diagnosis, were assessed from the permanent stained smear. Overall organism morphology of the intestinal protozoa preserved in zinc sulfate-PVA was not always equal in nuclear and cytoplasmic detail or range of color after permanent staining to that seen with mercuric chloride-PVA. However, the same organisms were usually identified in both specimens, with the exception of situations in which organism numbers were characterized as rare (no organisms per 10 oil immersion fields at x1,000 magnification but at least one organism in the smear) [9 of 161 (5.6%)] or the organism was missed because of poor morphologic detail [12 of 161 (7.5%)]. In only six of these cases [6 of 161 (3.7%)] did the results involve pathogens. The patient diagnosis was missed in four cases of amebiasis and two cases of giardiasis; in both situations the organism numbers were rare. There were no discrepant results with Dientamoeba fragilis. Overall agreement between the two PVA-based results was 87.0% (140 of 161); when the instances of rare organisms were disregarded, the overall agreement was 92.5% (149 of 161). On the basis of these findings, zinc-PVA is viable substitute for mercuric chloride-PVA used for trichrome permanent stained smears.