Duration of tick attachment necessary for transmission of Anaplasma phagocytophilum by Ixodes scapularis (Acari: Ixodidae) nymphs.

This study assessed the duration of tick attachment necessary for a successful transmission of Anaplasma phagocytophilum by an infected I. scapularis nymph. Individual nymphs were placed upon BALB/c mice and allowed to feed for predetermined time intervals of 4 to 72 h. Ticks removed from mice at predetermined intervals were tested by PCR for verification of infection and evaluation of the bacterial load. The success of pathogen transmission to mice was assessed by blood-PCR at 7, 14 and 21 days postinfestation, and IFA at 21 days postinfestation. Anaplasma phagocytophilum infection was documented in 10-30 % of mice, from which ticks were removed within the first 20 h of feeding. However, transmission success was ≥70% if ticks remained attached for 36 h or longer. Notably, none of the PCR-positive mice that were exposed to infected ticks for 4 to 8 h and only half of PCR-positive mice exposed for 24 h developed antibodies within 3 weeks postinfestation. On the other hand, all mice with detectable bacteremia after being infested for 36 h seroconverted. This suggests that although some of the ticks removed prior to 24 h of attachment succeed in injecting a small amount of A. phagocytophilum, this amount is insufficient for stimulating humoral immunity and perhaps for establishing disseminated infection in BALB/c mice. Although A. phagocytophilum may be present in salivary glands of unfed I. scapularis nymphs, the amount of A. phagocytophilum initially contained in saliva appears insufficient to cause sustainable infection in a host. Replication and, maybe, reactivation of the agent for 12-24 h in a feeding tick is required before a mouse can be consistently infected.

Ecology of Bats, Their Ectoparasites, and Associated Pathogens on Saint Kitts Island.

Ectoparasites of bats and bat-associated pathogens are poorly studied in the Lesser Antilles Islands. We report on an 11-mo field study on Saint Kitts Island of bat populations, their associated ectoparasites, and pathogens. We report on five ectoparasite species, including four Streblidae (Diptera) and a Spinturnicidae (Acari). Several genotypes of unnamed Bartonella were isolated from bats and ectoparasites. Microfilaria of an undetermined Litomosoides spp. were detected in blood from Artibeus jamaicensis Leach (Chiroptera: Phyllostomidae) (and associated ectoparasites: Trichobius intermedius Peterson and Hurka (Diptera: Streblidae) and Periglischrus iheringi Oudemans (Acari: Spinturnicidae)). In addition, an Ehrlichia sp. and Rickettsia africae were detected in the blood of several bat species. Our study is one of the first surveys of ectoparasite-borne pathogens in wild mammals from St. Kitts.

Rickettsia parkeri Rickettsiosis, Arizona, USA.

In the United States, all previously reported cases of Rickettsia parkeri rickettsiosis have been linked to transmission by the Gulf Coast tick (Amblyomma maculatum). Here we describe 1 confirmed and 1 probable case of R. parkeri rickettsiosis acquired in a mountainous region of southern Arizona, well beyond the recognized geographic range of A. maculatum ticks. The likely vector for these 2 infections was identified as the Amblyomma triste tick, a Neotropical species only recently recognized in the United States. Identification of R. parkeri rickettsiosis in southern Arizona demonstrates a need for local ecologic and epidemiologic assessments to better understand geographic distribution and define public health risk. Education and outreach aimed at persons recreating or working in this region of southern Arizona would improve awareness and promote prevention of tickborne rickettsioses.

Prevalence of Tick-Borne Pathogens in Host-Seeking Amblyomma americanum (Acari: Ixodidae) and Odocoileus virginianus (Artiodactyla: Cervidae) in Florida.

Amblyomma americanum (L.), the lone star tick, is an aggressive tick that is expanding its geographic range within the United States. This tick is the vector for the human and veterinary pathogens Ehrlichia chaffeensis and Ehrlichia ewingii and is associated with other microbes of unspecified pathogenicity including Rickettsia amblyommii, Panola Mountain Ehrlichia, and Borrelia lonestari In Florida, there has been sparse contemporary data on the prevalence of these organisms in host-seeking lone star ticks. To determine the prevalence of this tick and associated microbes in North Central Florida state parks, ∼1,500 lone star tick specimens were collected between 2010 and 2012 analyzed by polymerase chain reaction (PCR) sequencing. Additionally, 393 white-tailed deer, Odocoileus virginianus (Zimmerman), samples were analyzed for pathogen prevalence using molecular methods and serology. In lone star ticks, 14.6, 15.6, and 57.1% were positive for E. chaffeensis, E. ewingii, and Rickettsia spp. DNA, respectively. Panola Mountain Ehrlichia or B. lonestari DNA were each detected in nearly 2% of tick specimens. In white-tailed deer, 7.3% were PCR positive for E. chaffeensis, 6.0% for E. ewingii, and 3.2% for rickettsial species. Approximately 45% of white-tailed deer specimens had antibodies to Ehrlichia spp., and <1% had antibodies to Borrelia burgdorferi In summary, E. chaffeensis, E. ewingii, and spotted fever group rickettsia are highly prevalent in host-seeking lone star ticks and in white-tailed deer in Florida. The molecular and serological evidence of these microbes underscore their zoonotic potential in this region.

Panola Mountain Ehrlichia in Amblyomma maculatum From the United States and Amblyomma variegatum (Acari: Ixodidae) From the Caribbean and Africa.

Panola Mountain Ehrlichia (PME) has been suggested as an emerging pathogen of humans and dogs. Domestic goats and white-tailed deer (Odocoileus virginianus) are also susceptible and likely serve as reservoirs. Experimentally, both the lone star tick (Amblyomma americanum (L.)) and the Gulf Coast tick (Amblyomma maculatum Koch) can transmit PME among deer and goats. In the current study, we detected PME in adult wild-caught A. maculatum from the United States and Amblyomma variegatum (F.) from the Caribbean and Africa. This significantly expands the range, potential tick vectors, and risk for exposure to PME.

Tick-borne pathogens and disease in dogs on St. Kitts, West Indies.

Between 2009 and 2011, we conducted a case-control study of ticks and tick-associated pathogens affecting dogs on the island of St. Kitts, eastern Caribbean, including 55 cases of clinically suspected tick-borne disease (TBD) and 110 presumably healthy animals presented for elective surgeries. Rhipicephalus sanguineus caused year-round infestations of dogs, and 36% of the dogs in the study were infested at the time of examination. Overall, 62% of suspected TBD cases and 24% of presumably healthy dogs tested positive by PCR for infections with: Anaplasma platys (0% and 4%), Babesia canis vogeli (20% and 6%), Babesia gibsoni (18% and 5%), Ehrlichia canis (35% and 7%), and Hepatozoon canis (5% and 2%). Co-infections were documented in 15% of these PCR-positive dogs. Antibodies against A. platys or E. canis were noted in 36% of the dogs. Thrombocytopenia was the most common sign of infection, followed by anemia. This is the first detection of A. platys, B. canis vogeli, or H. canis on St. Kitts and the first detection of B. gibsoni in the Caribbean. We conclude that tick-borne pathogens of dogs are highly prevalent in this region and may present in dogs that appear healthy, in spite of hematologic abnormalities that may increase surgical risk.

Coxiella burnetii, the agent of Q fever, in domestic sheep flocks from Wyoming, United States.

Coxiella burnetii, the agent of Q fever, is an intracellular bacterial pathogen. It has a nearly cosmopolitan distribution. We conducted a serological survey of domestic sheep herds for infections with C. burnetii in Wyoming following reports of abortion and open ewes. Based on the serologic evidence, there was no link between reproductive problems and exposure to C. burnetii. However, the overall prevalence of C. burnetii in WY sheep was 7%, which indicates that the agent is present in the environment and could pose a threat to public health.

Detection of Coxiella burnetii in commercially available raw milk from the United States.

Unpasteurized (raw) milk can be purchased in 39 U.S. states, with direct consumer purchase for human consumption permitted in 29 of those 39 states. Raw milk (n=21; cow, 14; goat, 7) was purchased in 12 states, and Coxiella burnetii, the agent of Q fever, was detected in 9 of 21 (42.9%) samples tested by polymerase chain reaction. Viability of the pathogen was demonstrated by isolation of the agent in tissue culture. The demonstration of viable C. burnetii in commercially available raw milk poses a potential public health risk.

Presence of Coxiella burnetii DNA in the environment of the United States, 2006 to 2008.

Coxiella burnetii is an obligate intracellular bacterium that causes the zoonotic disease Q fever. Because C. burnetii is highly infectious, can survive under a variety of environmental conditions, and has been weaponized in the past, it is classified as a select agent and is considered a potential bioweapon. The agent is known to be present in domestic livestock and in wild animal populations, but the background levels of C. burnetii in the environment have not been reported. To better understand the amount of C. burnetii present in the environment of the United States, more than 1,600 environmental samples were collected from six geographically diverse parts of the United States in the years 2006 to 2008. DNA was purified from these samples, and the presence of C. burnetii DNA was evaluated by quantitative PCR of the IS1111 repetitive element. Overall, 23.8% of the samples were positive for C. burnetii DNA. The prevalence in the different states ranged from 6 to 44%. C. burnetii DNA was detected in locations with livestock and also in locations with primarily human activity (post offices, stores, schools, etc.). This study demonstrates that C. burnetii is fairly common in the environment in the United States, and any analysis of C. burnetii after a suspected intentional release should be interpreted in light of these background levels. It also suggests that human exposure to C. burnetii may be more common than what is suggested by the number of reported cases of Q fever.

Isolation of Rickettsia parkeri and identification of a novel spotted fever group Rickettsia sp. from Gulf Coast ticks (Amblyomma maculatum) in the United States.

Until recently, Amblyomma maculatum (the Gulf Coast tick) had garnered little attention compared to other species of human-biting ticks in the United States. A. maculatum is now recognized as the principal vector of Rickettsia parkeri, a pathogenic spotted fever group rickettsia (SFGR) that causes an eschar-associated illness in humans that resembles Rocky Mountain spotted fever. A novel SFGR, distinct from other recognized Rickettsia spp., has also been detected recently in A. maculatum specimens collected in several regions of the southeastern United States. In this study, 198 questing adult Gulf Coast ticks were collected at 4 locations in Florida and Mississippi; 28% of these ticks were infected with R. parkeri, and 2% of these were infected with a novel SFGR. Seventeen isolates of R. parkeri from individual specimens of A. maculatum were cultivated in Vero E6 cells; however, all attempts to isolate the novel SFGR were unsuccessful. Partial genetic characterization of the novel SFGR revealed identity with several recently described, incompletely characterized, and noncultivated SFGR, including "Candidatus Rickettsia andeanae" and Rickettsia sp. Argentina detected in several species of Neotropical ticks from Argentina and Peru. These findings suggest that each of these "novel" rickettsiae represent the same species. This study considerably expanded the number of low-passage, A. maculatum-derived isolates of R. parkeri and characterized a second, sympatric Rickettsia sp. found in Gulf Coast ticks.

Seroprevalence of Q fever in the United States, 2003-2004.

We performed serum testing for IgG antibodies against Coxiella burnetii (phase I and phase II) and analyzed questionnaire data from 4,437 adults > or = 20 years of age who participated in the National Health and Nutrition Examination Survey 2003-2004 survey cycle. National Q fever seroprevalence was determined by enzyme-linked immunosorbent assay and confirmed by using immunofluorescent antibody testing. Overall seroprevalence for Coxiella burnetii was 3.1% (95% confidence interval [CI] = 2.1-4.3%) among 4,437 adults > or = 20 years of age. Coxiella burnetii age-adjusted antibody prevalence was higher for men than for women (3.8%, 95% CI = 2.7-5.2% versus 2.5%, 95% CI = 1.5-3.7%, respectively, P < 0.05). Mexican Americans had a significantly higher antibody prevalence (7.4%, 95% CI = 6.6-8.3%) than either non-Hispanic whites (2.8%, 95% CI = 1.7-4.3%) or non-Hispanic blacks (1.3%, 95% CI = 0.6-2.5%) (P < 0.001). Multivariate analysis showed that the risk for Q fever antibody positivity increased with age and was higher among persons who were foreign-born, male, and living in poverty. These findings indicate that the national seroprevalence of Q fever in the United States is higher than expected on the basis of case numbers reported to the Centers for Disease Control and Prevention from state health departments. Potential differences in risk for exposure by race/ethnicity warrant further study.

Serological evidence of typhus group rickettsia in a homeless population in Houston, Texas.

We tested sera from 176 homeless people in Houston for antibodies against typhus group rickettsiae (TGR). Sera from 19 homeless people were reactive to TGR antigens by ELISA and IFA. Two people had antibodies against Rickettsia prowazekii (epidemic typhus) and the remaining 17 had antibodies against Rickettsia typhi (murine typhus).

The first report of human illness associated with the Panola Mountain Ehrlichia species: a case report.

INTRODUCTION: Two species of Ehrlichia are known to cause human illness. Several other species have been discovered in ticks and animals, and recent reports suggest that some of these Ehrlichia species might be human pathogens. We report here the first association of a recently discovered pathogen, the Panola Mountain Ehrlichia species, with a case of human illness. CASE PRESENTATION: A 31-year-old man from Atlanta, Georgia (GA) in the United States of America (USA) presented with a persistent sore neck of 3 weeks duration following a tick bite. DNA from the Panola Mountain Ehrlichia species, which was recently discovered in a goat in Georgia, was detected in an acute blood sample. Serologic testing was inconclusive. Polymerase chain reaction tests for other tick-borne diseases found in this region were negative. The patient rapidly improved in response to doxycycline therapy. CONCLUSION: Detection of Ehrlichia DNA in an acute blood sample meets the Centers for Disease Control and Prevention laboratory confirmation criteria for ehrlichiosis, and response to doxycycline provides supporting clinical evidence. The Panola Mountain Ehrlichia species, an emerging pathogen transmitted by ticks in the eastern USA, should be considered as a possible cause of tick-borne illness in this region.

Natural and experimental infection of white-tailed deer (Odocoileus virginianus) from the United States with an Ehrlichia sp. closely related to Ehrlichia ruminantium.

An Ehrlichia sp. (Panola Mountain [PM] Ehrlichia sp.) closely related to Ehrlichia ruminantium was recently detected in a domestic goat experimentally infested with lone star ticks (LSTs, Amblyomma americanum) collected from Georgia, USA. The infected goat exhibited pyrexia and mild clinical pathologic abnormalities consistent with ehrlichiosis. At least two other Ehrlichia species (Ehrlichia chaffeensis and Ehrlichia ewingii) are maintained in nature by a cycle involving LSTs as the primary vector and white-tailed deer (Odocoileus virginanus) as a known or suspected reservoir. To investigate the possibility that white-tailed deer are potential hosts of the PM Ehrlichia sp., whole blood samples collected from 87 wild deer from 2000 to 2002 were screened with a species-specific nested PCR assay targeting the citrate synthase gene. In addition, two laboratory-raised white-tailed deer fawns were each infested with 120 wild-caught LST adults from Missouri, USA, and blood samples were periodically collected and tested for the PM Ehrlichia sp. Of 87 deer tested from 20 locations in the southeastern United States, three (3%) deer from Arkansas, North Carolina, and Virginia were positive for the PM Ehrlichia sp. Wild-caught ticks transmitted the PM Ehrlichia sp. to one of two deer fawns, and colony-reared nymphal LSTs acquired the organism from the deer, maintained it transstadially as they molted to adults, and transmitted the PM Ehrlichia sp. to two naïve fawns. These findings indicate that white-tailed deer are naturally and experimentally susceptible to infection with an Ehrlichia sp. closely related to E. ruminantium and are able to serve as a source of infection to LSTs.

Geographic distribution and genetic diversity of the Ehrlichia sp. from Panola Mountain in Amblyomma americanum.

BACKGROUND: A novel Ehrlichia, closely related to Ehrlichia ruminantium, was recently discovered from Panola Mountain State Park, GA, USA. We conducted a study to determine if this agent was recently introduced into the United States. METHODS: We developed a sensitive PCR assay based on the conserved gltA (citrate synthase) gene and tested DNA samples extracted from 1964 field-collected and 1835 human-biting Amblyomma americanum from 23 eastern states of the USA. RESULTS: The novel agent was detected in 36 ticks collected from 10 states between 1998 and 2006. Infected ticks were collected both from vegetation (n = 14, 0.7%) and from humans (n = 22, 1.2%). Fragments of the conserved gltA gene and the variable map1 gene were sequenced from positive samples. Two distinct clades, with 10.5% nucleic acid divergence over the 730 bp map1 sequence, were identified. CONCLUSION: These data suggest that the Panola Mountain Ehrlichia was not recently introduced to the United States; this agent has an extensive distribution throughout the range of its tick vector, has been present in some locations for several years, and displays genetic variability. Furthermore, people in several states were exposed to this agent through the bite of infected ticks, underscoring the potential public health risk of this emerging ehrlichiosis.

Two USA Ehrlichia spp. cause febrile illness in goats.

Ehrlichia spp. are not currently recognized as a cause of illness in goats in the USA, but three Ehrlichia are enzootic in lone star ticks (Amblyomma americanum) in the eastern USA, and related bacteria in other countries cause illness in goats. We exposed naïve goats to Ehrlichia-infected Amblyomma and demonstrated that infection and clinical illness can be caused by two USA species, E. ewingii and the recently discovered Panola Mountain Ehrlichia sp. Clinical features in all five goats are described; ehrlichioses were associated with pyrexia, serous nasal discharge, inappetance, lethargy, decreased alkaline phosphatase, and, in most cases, neutropenia. Goats remained chronically infected for several months following exposure to ehrlichiae and transmitted the pathogens to uninfected ticks. In the eastern USA, undifferentiated febrile illness in goats might be caused by previously unrecognized ehrlichial infections, and pastures housing-infected goats could become infested with a large number of infected ticks.

Rickettsial pathogens in the tropical rat mite Ornithonyssus bacoti (Acari: Macronyssidae) from Egyptian rats (Rattus spp.).

We collected and tested 616 tropical rat mites (Ornithonyssus bacoti (Hirst)) from rats (Rattus norvegicus (Berkenhout) and R. rattus (Linnaeus)) throughout 14 governorates in Egypt and tested DNA extracts from pools of these mites for Bartonella spp., Coxiella burnetii, and Rickettsia spp. by PCR amplification and sequencing. Three different mite-associated bacterial agents, including one Bartonella and two Rickettsia spp., were detected in eight pools of mites. Further research could demonstrate the vector potential of mites and pathogenicity of these agents to humans or animals.

Population survey of Egyptian arthropods for rickettsial agents.

Between June 2002 and July 2003, 987 fleas, representing four species, and 1019 ticks, representing one argasid and eight ixodid species, were collected from Egyptian animals. These arthropods were tested for rickettsial agents using polymerase chain reaction. DNAs from Anaplasma and Ehrlichia spp. were detected in 13 ticks. Previously undescribed Bartonella spp. were detected in 21 fleas. Coxiella burnetii was detected in two fleas and 20 ticks. Rickettsia typhi was detected in 27 fleas from 10 cities. Spotted fever group rickettsiae were detected in both fleas and ticks and included Rickettsia aeschlimanii and an unnamed Rickettsia sp.