Isolation of parasites on fruits and vegetables.

ABSTRACT

The current FDA method to recover parasites from fruits and vegetables is derived from procedures used to isolate parasitic protozoa from water. A 1kg portion of fruit or vegetable is divided into 200 g subportions. The subportions are sequentially processed in a sonic cleaning bath with 1.5 liters of detergent solution (1% sodium dodecyl sulfate, 0.1% Tween 80) and sonicated for 10 minutes. As each subsample is removed, it is thoroughly drained. After this sonic treatment, the wash water is collected in a polypropylene beaker, transferred to 50 ml polypropylene centrifuge tubes and centrifuged for 15 min at 1500 x g. The sediment is consolidated into one tube along with two rinsings of each tube. The final sediment is fixed in 4% formaldehyde for 10 minutes before examination for parasites. Indirect fluorescent antibody is applied to stain the parasites (Giardia spp. and/or Cryptosporidium spp.) by using commercial kits when available. If a large quantity of extraneous matter is contained in the sediment it may be reduced by layering on Sheather's fluid and centrifuging at 1500 x g for 15 minutes. The supernatant is collected and washed twice in distilled water. This procedure is adequate for protozoa and nonoperculate helminth eggs; operculate helminth eggs may be cleaned by extraction with ethyl acetate. When cabbage and lettuce were seeded at 1 organism/g, the rate of recovery for Cryptosporidium parvum with the FDA method was 1%. When cabbage was seeded at 1 egg/g and 10 eggs/g, the average rate of recovery of decorticated eggs of Ascaris sp. or untreated Trichuris sp. was 10%.(ABSTRACT TRUNCATED AT 250 WORDS)