Characterization of a novel relapsing fever spirochete in the midgut, coxal fluid, and salivary glands of the bat tick Carios kelleyi.

Bat ticks, Carios kelleyi, from Iowa were examined for the presence of relapsing fever group borreliae. A novel spirochete was characterized by DNA sequence analysis of polymerase chain reaction amplicons for the 16S rRNA, flaB, and glpQ genes in either triturated tick pools or single ticks. All loci and the concatenated DNA sequence of 3,289 bases identified the Carios bacterium as a relapsing fever spirochete most closely related to, but distinct from, Borrelia turicatae. Spirochetes reactive with a Borrelia-specific monoclonal antibody were observed microscopically in the coxal fluid and salivary glands from one tick. These data confirm the presence of a novel species of relapsing fever spirochete in bat ticks and the potential for new enzootic foci for endemic relapsing fever that warrants further investigation. The name Borrelia johnsonii is proposed for this novel spirochete in honor of Dr. Russell C. Johnson.

Testing human sera for antibodies against avian influenza viruses: horse RBC hemagglutination inhibition vs. microneutralization assays.

BACKGROUND: The hemagglutination inhibition (HI) assay is a frequently used method to screen human sera for antibodies against influenza A viruses. Because HI has relatively poor sensitivity in detecting antibodies against avian influenza A strains, a more complicated microneutralization (MN) assay is often preferred. Recent research suggests that the sensitivity of the HI assay can be improved by switching from the traditionally used turkey, guinea pig, human, or chicken RBCs to horse RBCs. OBJECTIVE: To evaluate the performance of the horse RBC HI when screening for human antibodies against avian influenza types H3, H4, H5, H6, H7, H9, H11, and H12. STUDY DESIGN: We evaluated the reproducibility of horse RBC HI and its agreement with MN results using sera from people exposed or not exposed to wild and domestic birds. RESULTS: The horse RBC HI assay had high reliability (90%-100%) and good agreement with MN assay results (52%-100%). CONCLUSION: The horse RBC HI assay is reliable, less expensive, less complex, and faster than the MN assay. While MN will likely remain the gold standard serologic assay for avian viruses, the horse RBC HI assay may be very useful as a screening assay in large-scale epidemiologic studies.

Prevalence of Ehrlichia, Borrelia, and Rickettsial agents in Amblyomma americanum (Acari: Ixodidae) collected from nine states.

Ambyomma americanum (L.) (Acari: Ixodidae) is an aggressive tick that feeds on humans during all postembryonic life stages. In many regions of the United States, it is the tick most commonly found attached to humans. Public health interest has grown recently, due to the recognition of new human pathogens transmitted by A. americanum and the expanding distribution of the tick. A. americanum is a vector of several bacteria pathogenic to humans. Ehrlichia chaffeensis and Ehrlichia ewingii cause moderate-to-severe febrile illness. "Rickettsia amblyommii," a member of the spotted fever group Rickettsia, also has recently been implicated as a possible human pathogen based on serologic evidence from persons recovering from illness after a tick bite. We have determined the prevalence of infection of Ehrlichia chaffeensis, E. ewingii, "Borrelia lonestari", and R. amblyommii within A. americanum ticks from 29 sites in nine states. Overall infection prevalences were 4.7% for E. chaffeensis (range, 0-27%), 3.5% for E. ewingii (range, 0-18.6%), 2.5% for B. lonestari (range, 0-12.2%), and 41.2% for R. amblyommii (range, 0-84.0%). In addition, 87 ticks (4.3%) were infected with two or more bacteria. This report documents new distribution records for E. ewingii, B. lonestari, and R. amblyommii and underscores the nonhomogeneous distribution of pathogen foci of infection. Additional surveillance throughout the range ofA. americanum is warranted to increase physician and public awareness of the risk of disease to humans from exposure to the agents transmitted by this tick.

Avian influenza among waterfowl hunters and wildlife professionals.

We report serologic evidence of avian influenza infection in 1 duck hunter and 2 wildlife professionals with extensive histories of wild waterfowl and game bird exposure. Two laboratory methods showed evidence of past infection with influenza A/H11N9, a less common virus strain in wild ducks, in these 3 persons.

Serosurveillance for Anaplasma phagocytophilum antibodies in white-tailed deer (Odocoileus virginianus) in Iowa, USA.

Cases of human granulocytic anaplasmosis have increased in number and are being identified in new geographic areas since its discovery in 1994. White-tailed deer (WTD) become infected with the causative agent, Anaplasma phagocytophilum, and serve as natural sentinels for this organism. In order to determine if A. phagocytophilum is present in the state of Iowa, sera collected from 628 WTD in 2004 from 13 sites and from 282 WTD in 1999 from a single, common site were tested by enzyme-linked immunosorbent assay and Western immunoblotting. A seroprevalence of 9.1% was found among the 2004 samples, and there was no change in seropositivity rates from 1999 to 2004 at the single, common site. As A. phagocytophilum is another tick-borne human pathogen to be identified in the state of Iowa, this study has important implications for health care providers.

West Nile virus serosurveillance in Iowa white-tailed deer (1999-2003).

Sera from white-tailed deer (Odocoileus virginianus) were collected in Iowa during the winter months (1999-2003), 2 years before and after West Nile virus (WNV) was first reported in Iowa (2001), and were analyzed for antibodies to WNV. Samples from 1999 to 2001 were antibody negative by blocking enzyme-linked immunosorbent assay (bELISA) and plaque reduction neutralization test (PRNT(90)). Prevalence derived from bELISA (2002, 12.7%; 2003, 11.2%) and WNV PRNT(90) (2002, 7.9%; 2003, 8.5%) assays were similar. All samples were negative for antibodies against St. Louis encephalitis virus as determined by PRNT(90). Antibodies to flaviviruses were detected by indirect enzyme-linked immunosorbent assay (iELISA) prior to the first WNV cases reported in Iowa (1999-2001) with prevalence ranging from 2.2% to 3.2%, suggesting the circulation of an additional undescribed flavivirus prior to the introduction of WNV into the area. Flavivirus prevalence as determined by iELISA increased in 2002 and 2003 (23.3% and 31.9%, respectively). The increase in prevalence exceeded estimates of WNV prevalence, suggesting that conditions favored general flavivirus transmission (including WNV) during the 2002-2003 epizootic. These data indicate that serologic analysis of deer sera collected from hunter harvests may prove useful for surveillance and evidence of local transmission of WNV and other pathogens and identify white-tailed deer as a species for further studies for host competency.

Detection of Rickettsia, Borrelia, and Bartonella in Carios kelleyi (Acari: Argasidae).

Carios kelleyi (Colley & Kohls 1941), a tick associated with bats and bat habitats, has been reported to feed on humans, but there is little published data regarding the presence of vector-borne pathogens in these ticks. C. kelleyi nymphs and adults were collected from residential and community buildings in Jackson County, Iowa, and tested by polymerase chain reaction for Rickettsia, Borrelia, Bartonella, Coxiella, and Anaplasma. Rickettsia DNA was detected in 28 of 31 live ticks. Sequences of the 17-kDa and rOmpA genes suggest that this agent is a novel spotted fever group Rickettsia. Transstadial and transovarial transmission of this Rickettsia were demonstrated. The flagellin gene of a Borrelia, closely related to B. turicatae, was detected in one of 31 live ticks. The 16S-23S intergenic spacer region of Bartonella henselae also was detected in one of 31 live ticks. Coxiella or A. phagocytophilum DNA were not detected in these ticks.