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Three diagnostic assays for detecting West Nile virus (WNV) in avian oral swabs were evaluated in California in 2004 and 2005: two commercial antigen-capture assays, VecTest and Rapid Analyte Measurement Platform (RAMP), and reverse transcriptase-polymerase chain reaction (RT-PCR) of oral swabs in a specialized viral transport medium (VTM). Results from this study demonstrated that VTM was excellent for transportation and maintenance of WNV in avian oral swab samples and allowed for detection by RT-PCR and subsequent confirmation by virus isolation. Oral swabs and kidney tissue in VTM tested by RT-PCR were found to have similar accuracy in detecting WNV in corvids. The two antigen-capture assays, VecTest and RAMP, provided few false positives for corvids, with over 95% specificity. When performed by multiple local agencies throughout the state, VecTest and RAMP were similarly sensitive for oral swabs of American Crows (Corvus brachyrhynchos) (70% and 64%, respectively). Data from known WNV positive corvid oral swabs in VTM tested by antigen-capture assays at a diagnostic laboratory suggested that RAMP was more sensitive than VecTest. Due to high probability of false negatives, neither test is recommended for use on non-corvids. While WNV antigen-capture assays were effective screening tools for corvids, they were markedly less sensitive for Western Scrub Jays (Aphelocoma californica).

West Nile virus (WNV) transmission generally involves a mosquito vector and an avian reservoir host, with mammals as incidental hosts. Although most mammalian WNV infections cause low or no morbidity or mortality, tree squirrels are susceptible to WNV-associated neurologic disease with infection prevalence comparable to that in dead birds. Positive species included fox squirrel (Sciurus niger), western gray squirrel (S. griseus), and eastern gray squirrel (S. carolinensis). Kidney tissue (dissected and swabbed), and oropharyngeal (oral) swab samples from tree squirrels submitted by California vector control and rehabilitation agencies were tested by reverse transcription-polymerase chain reaction; cycle threshold values were similar for all three samples, ranging from 21.9 to 26.5. Kidney tissue was more sensitive than oral swabs for detecting WNV in squirrels. Three of 36 live neurologic tree squirrels had viremia approximately 5 log(10) plaque-forming units/mL or greater, similar to WNV-infected birds. Tree squirrels are useful in WNV surveillance and provide localized evidence of WNV transmission to mammals.

ABSTRACT:Between 2 August and 22 September 2000, 37 hunter-killed tule elk (Cervus elaphus nannodes) were evaluated at the Grizzly Island Wildlife Area, California, USA, for evidence of paratuberculosis. Elk were examined post-mortem, and tissue and fecal samples were submitted for radiometric mycobacterial culture. Acid-fast isolates were identified by a multiplex polymerase chain reaction (PCR) that discriminates among members of the Mycobacterium avium complex (MAC). Histopathologic evaluations were completed, and animals were tested for antibodies using a Johne's enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion. In addition, 104 fecal samples from tule elk remaining in the herd were collected from the ground and submitted for radiometric mycobacterial culture. No gross lesions were detected in any of the hunter-killed animals. Mycobacterium avium subsp. paratuberculosis (MAP) was cultured once from ileocecal tissue of one adult elk and was determined to be a strain (A18) found commonly in infected cattle. One or more isolates of Mycobacterium avium subsp. avium (MAA) were isolated from tissues of five additional adult elk. Gastrointestinal tract and lymph node tissues from 17 of the 37 elk (46%) examined had histopathologic lesions commonly seen with mycobacterial infection; however, acid-fast bacteria were not observed. All MAC infections were detected from adult elk (P 5 0.023). In adult elk, a statistically significant association was found between MAA infection and ELISA sample-to-positive ratio (S/P)$0.25 (P50.021); four of five MAA culturepositive elk tested positive by ELISA. Sensitivity and specificity of ELISA S/P$0.25 for detection of MAA in adult elk were 50% and 93%, respectively. No significant associations were found between MAC infection and sex or histopathologic lesions. Bacteriologic culture confirmed infection with MAP and MAA in this asymptomatic tule elk herd. The Johne's ELISA was useful in signaling mycobacterial infection on a population basis but could not discriminate between MAA and MAP antibodies. The multiplex PCR was useful in discriminating among the closely related species belonging to MAC.