Improved immunofluorescence assay for detection of Giardia and Cryptosporidium from asymptomatic adult cervine animals

Abstract

We tested an improved immunofluorescence assay (IFA) in detecting Giardia and Cryptosporidium from feces of asymptomatic adult cervine animals. Samples were concentrated by sucrose flotation before being stained by fluorescent monoclonal antibody and examined microscopically. The detection limit was determined as 500 G. intestinalis cysts or 200 C. parvum oocysts/g of sample. Among the 82 samples collected from adult fallow deer, Columbian black-tailed deer, and Tule elk in northern California, 3 (3.7%) contained G. intestinalis cysts, which were confirmed by a species-specific polymerase chain reaction (PCR) following immunomagnetic capture (IC) of cysts. C. parvum oocysts were detected in a total of 13 (15.9%) samples, and oocysts from 2 such samples were smaller than oocysts from the other 11 samples. C. parvum identification was also confirmed by specific IC-PCR and sequencing of the PCR product. In addition, a C. muris-like organism was detected in 2 (2.4%) samples. Findings obtained with the improved IFA confirmed that cysts/oocysts may pass unnoticed in adult cervine animals and that subclinically infected individuals could serve as potential carriers of infection for humans and other animals via contaminated feces or water.