Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, remains prevalent in North American elk, white-tailed deer and mule deer. A natural case of CWD in reindeer (Rangifer tarandus tarandus) has not been reported despite potential habitat overlap with CWD-infected deer or elk herds. This study investigates the experimental transmission of CWD from elk or white-tailed deer to reindeer by the oral route of inoculation. Ante-mortem testing of the three reindeer exposed to CWD from white-tailed deer identified the accumulation of pathological PrP (PrPCWD) in the recto-anal mucosa associated lymphoid tissue (RAMALT) of two reindeer at 13.4 months post-inoculation. Terminal CWD occurred in the two RAMALT-positive reindeer at 18.5 and 20 months post-inoculation while one other reindeer in the white-tailed deer CWD inoculum group and none of the 3 reindeer exposed to elk CWD developed disease. Tissue distribution analysis of PrPCWD in CWD-affected reindeer revealed widespread deposition in central and peripheral nervous systems, lymphoreticular tissues, the gastrointestinal tract, neuroendocrine tissues and cardiac muscle. Analysis of prion protein gene (PRNP) sequences in the 6 reindeer identified polymorphisms at residues 2 (V/M), 129 (G/S), 138 (S/N) and 169 (V/M). These findings demonstrate that (i) a sub-population of reindeer are susceptible to CWD by oral inoculation implicating the potential for transmission to other Rangifer species, and (ii) certain reindeer PRNP polymorphisms may be protective against CWD infection.
A rapid lateral flow assay for detection of bovine antibody to Anaplasma marginale was developed. The assay used a recombinant peptide of major surface protein 5 as the antigen and a monoclonal antibody specific for bovine IgG(1) conjugated with colloidal gold beads for detection. Serum and anticoagulated blood samples were obtained from cattle in an area where anaplasmosis was endemic. The samples were selected based on positive identification of the organism in blood smears. The unclotted blood samples were used for PCR determination of the presence of A. marginale while the sera were tested by a commercial competitive enzyme immunoassay (CELISA) and by the lateral flow assay (LFA). Similar samples, collected at a Canadian sales barn, were tested by the CELISA and LFA and 10% were tested by PCR for the presence of A. marginale nucleic acid. In addition, stored serum samples from a second endemic area were tested by CELISA and LFA. Of the 114 smear positive samples, all were positive by CELISA and LFA. All samples were also positive by PCR. Samples from Canadian sources (n=524) were negative in the CELISA but 11 sera gave false positive reactions in the LFA. All samples tested were PCR negative. Of 113 samples from herds with anaplasmosis, 53 were positive in the CELISA and 50 were LFA positive.
The lateral flow assay (LFA) is a rapid diagnostic test which may be performed under most conditions and is especially useful for field applications. This type of assay was applied to the detection of antibody to bovine Anaplasma marginale using sera from endemic areas and from areas which have been free from infection for more than 25 years. Briefly, the test uses recombinant A. marginale major surface protein 5 peptide (Msp5), immobilized on a cellulose acetate membrane. A serum sample is added to a pad containing a monoclonal antibody specific for bovine IgG(1), conjugated with colloidal gold, located at one end of the strip. The sample and gold conjugate are wicked along the membrane and if antibody is present in the serum, a visible line will form between the Msp5-antibody-conjugate immune complex in minutes. An additional band of recombinant protein A/G was added to the membrane as a positive control reaction of the monoclonal antibody conjugate. For comparison, direct examination of blood smears and a nested polymerase chain reaction (PCR) were performed on some of the samples. Using samples from herds in one endemic area, the PCR gave a sensitivity value of 9.2% while a commercial competitive enzyme immunoassay (CELISA) gave a sensitivity value of 17.2% and the LFA values of 20.5%. In a second endemic area, selected samples, all positive by direct examination gave a 71.7% sensitivity values with the PCR, 94.5% with the CELISA and 95.5% with the LFA. Using sera from a disease-free area, the specificity values were 100% for the PCR (testing a proportion of randomly selected samples), 99.5% for the CELISA and 98.0% for the LFA. It is envisaged that the validated LFA will be a useful tool for screening cattle moving from an area with infection to a disease-free area.
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