Tick Dermacentor Research Articles

Responses of Amblyomma americanum and Dermacentor variabilis to odorants that attract haematophagous insects

Carbon dioxide (CO(2) ), 1-octen-3-ol, acetone, ammonium hydroxide, L-lactic-acid, dimethyl trisulphide and isobutyric acid were tested as attractants for two tick species, Amblyomma americanum and Dermacentor variabilis (Acari: Ixodidae), in dose-response bioassays using Y-tube olfactometers. Only CO(2) , acetone, 1-octen-3-ol and ammonium hydroxide elicited significant preferences from adult A. americanum, and only CO(2) was attractive to adult D. variabilis. Acetone, 1-octen-3-ol and ammonium hydroxide were separately evaluated at three doses against CO(2) (from dry ice) at a field site supporting a natural population of A. americanum nymphs and adults. Carbon dioxide consistently attracted the highest number of host-seeking ticks. However, for the first time, acetone, 1-octen-3-ol and ammonium hydroxide were shown to attract high numbers of A. americanum. Further research is needed to determine the utility of these semiochemicals as attractants in tick surveillance and area-wide management programmes.

Efficacy of an imidacloprid/flumethrin collar against fleas, ticks, mites and lice on dogs

BackgroundThe studies reported here were conducted to ascertain the efficacy of imidacloprid/flumethrin incorporated in a slow-release matrix collar, against infestations of dogs by fleas, ticks, mites and lice. Efficacy was evaluated against the flea Ctenocephalides felis felis, the ticks Rhipicephalus sanguineus, Ixodes ricinus, Ixodes scapularis, Dermacentor reticulatus and Dermacentor variabilis, the mite Sarcoptes scabiei and the biting louse Trichodectes canis.MethodsGroups of collar-treated dogs (n = 7–10) were infested with fleas and/or ticks at monthly intervals at least, over a period of up to 8 months. Efficacy against fleas was evaluated 24 to 48 h after treatment and 24 h after each re-infestation. Efficacy against ticks was evaluated at 48 h (acaricidal), 6 h (repellent) and 48 h (sustained) after infestation. The effect of regular shampooing or immersion in water on the efficacy of the collars was also tested. Efficacy against flea larvae was assessed by incubating blanket samples after dog contact with viable flea eggs. Effectiveness against lice and mites was evaluated after treatment of naturally infested animals. With the exception of the mites, efficacy was calculated by comparison with untreated negative control groups.ResultsEfficacy against fleas (24 h) generally exceeded 95%, and against flea larvae it exceeded 99% for 8 months. Sustained acaricidal (48 h) efficacy, covering a period of 8 months was 100% against I. ricinus, starting 2 days after treatment (in vivo), and 100% against I. scapularis (in vitro), above 97% against R. sanguineus, generally above 97% against D. reticulatus and above 90% for D. variabilis.Repellent (6 h) efficacy 2 days after treatment and continuing for 8 months was consistently 100% against I. ricinus, and above 90% against R. sanguineus.Regular shampooing affected efficacy against fleas and ticks to a lesser extent than regular immersion in water.The collars eliminated Trichodectes canis within 2 days and Sarcoptes scabiei within 3 months.ConclusionThe rapid insecticidal and acaricidal properties of the medicated collars against newly-acquired infestations of fleas and ticks and their sustained high levels of preventive efficacy have been clearly shown. Consequently they have the potential to prevent the transmission of vector-borne diseases and other conditions directly associated with infestation throughout an entire season of parasite abundance.

Acetylcholinesterase inhibition by nootkatone and carvacrol in arthropods

The essential oils from many botanicals have been screened for insecticidal activity. Two constituents of the Alaskan yellow cedar tree, the monoterpenoid carvacrol and the sesquiterpenoid nootkatone, both are toxic against several arthropods. The mode of action through which nootkatone and carvacrol exert their insecticidal activity remains uncertain. It has been hypothesized that they may inhibit acetylcholinesterase enzyme activity. The degree of acetylcholinesterase inhibition of carvacrol and nootkatone was compared to that of carbaryl, a known acetylcholinesterase inhibitor, in the house fly (Musca domestica), yellow fever mosquito (Aedes aegypti), American dog tick (Dermacentor variabilis) and American cockroach (Periplaneta americana). The concentration of carbaryl, at which 50% of the acetylcholinesterase activity was inhibited (IC50), was less than 2μM in all four arthropod models. Carvacrol was observed to cause slight inhibition of the acetylcholinesterase enzyme in house flies, ticks and cockroaches, but it did not inhibit the mosquito acetylcholinesterase enzyme. Nootkatone did not inhibit the acetylcholinesterase enzyme in any of the four arthropod models tested. From this study, we conclude that the acetylcholinesterase inhibition is not likely the primary mode of action for insecticidal activity by nootkatone or carvacrol.

Intraurban Near-Water Areas and Their Role in Distribution of Ticks (Acari, Ixodidae) in Megapolis the Example of Kyiv

Intraurban Near-Water Areas and Their Role in Distribution of Ticks (Acari, Ixodidae) in Megapolis the Example of Kyiv In 2002-2009, we studied 12 islands within the city of Kyiv, and found four indigenous species of ticks (Dermacentor reticulatus Fabricius, 1794; Ixodes ricinus Linnaeus, 1758; I. apronophorus Schulze, 1924; Rhipicephalus rossicus Jakimov et Kohl-Jakimova, 1911) and one invasive species (Hyalomma marginatum Koch, 1844). For A-E-gradient, established by man's impact degree, we revealed atypical distribution of species: their majority is noted in two pole areas - where the man impact is maximum and minimum. The investigated near-water areas are a kind of survival areas and sources of ixodides diversity in megalopolis.

Association of different genetic types of Francisella-like organisms with the rocky mountain wood tick (Dermacentor andersoni) and the American dog tick (Dermacentor variabilis) in localities near their northern distributional limits.

Dermacentor andersoni and Dermacentor variabilis from allopatric and sympatric populations near their northern distributional limits were examined for the presence of Francisella species using molecular techniques that targeted 373 bp of the 16S rRNA gene. Although there was no evidence for the presence of Francisella tularensis in any tick, Francisella-like endosymbionts (FLEs) were common in D. andersoni and D. variabilis adults and immatures. A significantly greater proportion of female ticks contained FLEs compared to male ticks. In addition, significantly more D. variabilis adult individuals contained multiple FLE sequence types than did D. andersoni adults. Ten different types of FLEs were identified based on the sequence data, which has implications for diagnostic tests and epidemiological studies of F. tularensis in tick populations in Canada. The three most prevalent types of FLEs have been detected previously in D. andersoni or D. variabilis from other parts of their distributional ranges, whereas the other seven FLE types have not been reported previously. A comparison of the FLEs from both allopatric and sympatric populations of these two tick species provided insight into the relative host-specificity and the modes of transmission of these tick-borne bacteria. In general, each FLE type was specific for one tick species, suggesting vertical transmission of each bacterium. However, there were a few instances of potential cross-transfer of two FLE types to the other tick species at locations where D. andersoni and D. variabilis occurred in sympatry, suggesting that there may be occasional horizontal transmission of some FLEs.

Infection Prevalences of Common Tick-borne Pathogens in Adult Lone Star Ticks (Amblyomma americanum) and American Dog Ticks (Dermacentor variabilis) in Kentucky

Rocky Mountain spotted fever, Lyme disease, and ehrlichiosis are tick-borne diseases that are reported annually in Kentucky. We conducted a survey to describe infection prevalence of tick-borne pathogens in Amblyomma americanum and Dermacentor variabilis ticks collected in Kentucky. During 2007-2008, we collected 287 ticks (179 D. variabilis and 108 A. americanum) from canine, feral hog, horse, raccoon, white-tailed deer, and human hosts in six counties in Kentucky. Ticks were screened for Rickettsia spp., Borrelia spp., and Ehrlichia spp. by using polymerase chain reaction. Forty-one (14.3%) ticks (31 A. americanum and 10 D. variabilis) were polymerase chain reaction-positive for a Rickettsia spp. Fourteen (4.9%) ticks (6 A. americanum and 8 D. variabilis) were positive for E. chaffeensis, and 4 A. americanum (1.4%) were positive for E. ewingii. One (0.4%) A. americanum was positive for Borrelia lonestari. Although Rocky Mountain spotted fever is diagnosed in Kentucky, no R. rickettsii was found in ticks in this study.

Dermacentor tick attached to tympanic membrane

In June, 2008, a 47-year-old woman presented to us with a 1-day history of crackling sensations in her left ear. On otoscopic examination, a tick was seen on the left tympanic membrane (figure). An otorhinolaryngologist removed the tick with endoscopic-guided forceps. There was no bleeding, perforation, hearing loss, or other sequelae. The crackling sensations resolved immediately. The tick was identified as Dermacentor variabilis, commonly known as the American dog tick. Our patient lived in a rural area of Minnesota, USA where the tick-borne diseases lyme and human anaplasmosis are endemic.

Tick-borne lymphadenopathy (TIBOLA) acquired in Southwestern Germany

BackgroundTick-borne lymphadenopathy (TIBOLA) was first described in 1997 in a patient in France. The causative agent, Rickettsia slovaca, is transmitted by Dermacentor ticks.Case presentationIn southwestern Germany we encountered a patient with a tick bite at the dorsal scalp that resulted in an eschar and nuchal lymphadenopathy. Additionally, fever, malaise as well as elevated inflammatory markers and transaminases occurred. The characteristic clinical picture along with positive antibody testing for rickettsiae of the tick-borne spotted fever group strongly suggest the diagnosis TIBOLA.ConclusionHuman rickettsioses are emerging infections. Clinicians should be aware of TIBOLA as a newly described rickettsial disease. As in our case, TIBOLA may be encountered in regions/countries where R. slovaca and Dermacentor ticks are prevalent but autochthonous acquisition was not described before.

A hidden beneficial: biology of the tick-wasp Ixodiphagus hookeri in Germany

The parasitic wasp Ixodiphagus hookeri parasitizes hard ticks and is therefore considered as a potential candidate for biological control of ticks. However, there are still considerable gaps of knowledge about the biology of I. hookeri, especially for European populations. Thus, the present study was performed to assess important life-history parameters of the parasitoid in Germany. Field studies accomplished in three successive years revealed that unfed Ixodes ricinus nymphs are infested by the parasitoid at a low but constant rate of 1.9–3.8% and that adult wasps are present only during a short period in late summer. The mean developmental time of wasps in I. ricinus nymphs ranged from 28 to 70 days under constant laboratory conditions and was prolonged in the second half of the year. Bioassays on parasitization and host preference behaviour showed that unfed nymphs of the host species I. ricinus are significantly preferred in experiments, in which unfed and engorged larvae as well as fully engorged nymphs were offered as alternatives. The marsh tick Dermacentor reticulatus was not accepted as an alternative host. Our data show that the investigated I. hookeri populations differ markedly from populations in other regions of the world in many aspects. The adaptation of different strains to local conditions explains the limited success of imported strains in earlier biological control attempts and highlights the importance of doing research to enhance the control potential of native strains.

Dermohypodermite de la face révélatrice d’un syndrome TIBOLA

Tick-borne lymphadenopathy (TIBOLA) is an emerging rickettsiosis in Europe, transmitted by the Dermacentor tick. This syndrome is defined as the association of an inoculation eschar on the scalp that may be surrounded by an erythema scalp, fever, and painful cervical lymphadenopathy in colder months. Children and women are at higher risk for TIBOLA. We report the case of a 9 year-old French child with an acute hemifacial edema and erythema revealing TIBOLA. Early empirical antibiotic therapy should be prescribed in any suspected TIBOLA, before confirmation of the diagnosis. The recommended treatment is doxycycline or macrolide.

Hemelipoglycoprotein from the ornate sheep tick, Dermacentor marginatus: structural and functional characterization

BackgroundTick carrier proteins are able to bind, transport, and store host-blood heme, and thus they function also as antioxidants. Nevertheless, the role of carrier proteins in ticks is not fully understood. Some of them are found also in tick males which do not feed on hosts to such an extent such as females (there are differences in male feeding in different tick species) and thus they are not dealing with such an excess of heme; some of the carrier proteins were found in salivary glands where the processing of blood and thus release of heme does not occur. Besides, the carrier proteins bind relatively low amounts of heme (in one case only two molecules of heme per protein) compared to their sizes (above 200 kDa).The main aim of this study is the biochemical characterization of a carrier protein from the ornate sheep tick Dermacentor marginatus, hemelipoglycoprotein, with emphasis on its size in native conditions, its glycosylation and identification of its modifying glycans, and examining its carbohydrate-binding specificity.ResultsHemelipoglycoprotein from D. marginatus plasma was purified in native state by immunoprecipitation and denatured using electroelution from SDS-PAGE separated plasma. The protein (290 kDa) contains two subunits with molecular weights 100 and 95 kDa. It is glycosylated by high-mannose and complex N-glycans HexNAc2Hex9, HexNAc2Hex10, HexNAc4Hex7, and HexNAc4Hex8. The purified protein is able to agglutinate red blood cells and has galactose- and mannose-binding specificity. The protein is recognized by antibodies directed against plasma proteins with hemagglutination activity and against fibrinogen-related lectin Dorin M from the tick Ornithodoros moubata.It forms high-molecular weight complexes with putative fibrinogen-related proteins and other unknown proteins under native conditions in tick plasma. Feeding does not increase its amounts in male plasma. The hemelipoglycoprotein was detected also in hemocytes, salivary glands, and gut. In salivary glands, the protein was present in both glycosylated and nonglycosylated forms.ConclusionA 290 kDa hemelipoglycoprotein from the tick Dermacentor marginatus, was characterized. The protein has two subunits with 95 and 100 kDa, and bears high-mannose and complex N-linked glycans. In hemolymph, it is present in complexes with putative fibrinogen-related proteins. This, together with its carbohydrate-binding activity, suggests its possible involvement in tick innate immunity. In fed female salivary glands, it was found also in a form corresponding to the deglycosylated protein.

Species Diversity of Ixodid Ticks Feeding on Humans in Amasya, Turkey: Seasonal Abundance and Presence of Crimean-Congo Hemorrhagic Fever Virus

Ticks (Acari:Ixodidae) are important pests transmitting tick-borne diseases such as Crimean-Congo hemorrhagic fever (CCHF) to humans. Between 2002 and 2009, numerous CCHF cases were reported in Turkey, including Amasya province. In the current study, species diversity, seasonal abundance of ticks, and presence of CCHF virus (CCHFV) in ticks infesting humans in several districts of Amasya province were determined. In the survey, a total of 2,528 ixodid ticks were collected from humans with tick bite from April to November 2008 and identified to species. Hyalomma marginatum (18.6%), Rhipicephalus bursa (10.3%), Rhipicephalus sanguineus (5.7%), Rhipicephalus (Boophilus) annulatus (2.2%), Dermacentor marginatus (2.5%), Haemaphysalis parva (3.6%), and Ixodes ricinus (1.6%) were the most prevalent species among 26 ixodid tick species infesting humans in Amasya province. Hyalomma franchinii Tonelli & Rondelli, 1932, was a new record for the tick fauna of Turkey. The most abundant species were the members of Hyalomma and Rhipicephalus through summer and declined in fall, whereas relative abundances of Ixodes and Dermacentor ticks were always low on humans in the province. Of 25 Hyalomma tick pools tested, seven pools were CCHFV positive by reverse transcription-polymerase chain reaction. Results indicated diversity of ixodid tick species infesting humans was very high, abundance of ticks changed by season, and ticks infesting humans had potential for transmitting CCHFV.

Full-length sequence, regulation and developmental studies of a second vitellogenin gene from the American dog tick, Dermacentor variabilis

Vitellogenin (Vg) is the precursor of vitellin (Vn) which is the major yolk protein in eggs. In a previous report, we isolated and characterized the first Vg message from the American dog tick Dermacentor variabilis. In the current study, we describe a second Vg gene from the same tick. The Vg2 cDNA is 5956 nucleotides with a 5775 nt open reading frame coding for 1925 amino acids. The conceptual amino acid translation contains a 16-residues putative signal peptide, N-terminal lipid binding domain and C-terminal von Willebrand factor type D domain present in all known Vgs. Moreover, the amino acid sequence shows a typical GLCG domain and several RXXR cleavage sites present in most isolated Vgs. Tryptic digest-mass fingerprinting of Vg and Vn recognized 11 fragments that exist in the amino acid translation of DvVg2 cDNA. Injection of virgin females with 20 hydroxyecdysone induced DvVg2 expression, vitellogenesis and oviposition. Using RT-PCR, DvVg2 expression was detected only in tick females after mating and feeding to repletion. Northern blot analysis showed that DvVg2 is expressed in fat body and gut cells of vitellogenic females but not in the ovary. DvVg2 expression was not detected in adult fed or unfed males. The characteristics that distinguish Vg from other similar tick storage proteins like the carrier protein, CP (another hemelipoglycoprotein) are discussed.

Rabbit Tularemia and Hepatic Coccidiosis in Wild Rabbit

Rabbit Tularemia and Hepatic Coccidiosis in Wild Rabbit

Fatal bovine anaplasmosis in a herd with new genotypes of Anaplasma marginale, Anaplasma ovis and concurrent haemoplasmosis

Haematological and molecular analysis of blood samples was carried out during an outbreak of bovine anaplasmosis in Hungary. Acute disease was observed in five animals, two of which died. Anaplasma-carrier state was diagnosed in 69 (92%) of cattle. Further evaluation of 24 blood samples revealed concurrent infections with Mycoplasma wenyonii and ‘ Candidatus M. haemobos’ in 22 and 21 animals, respectively. In addition, two cows were identified with rickettsaemia. Regarding molecular investigation of potential hard tick vectors, Haemaphysalis inermis and Dermacentor marginatus males collected from the animals were PCR-negative. However, in one pool (out of 18) of Ixodes ricinus males, and in six pools (out of 18) of D. reticulatus males the msp4 gene of Anaplasma marginale was detected. In the same I. ricinus pool Anaplasma ovis was also identified. All ticks were negative for haemoplasmas. Anaplasma sequences yielded 97–99% homology to sequences deposited in the Genbank. This is the first report of fatal bovine anaplasmosis associated with divergent A. marginale genotypes and concurrent ‘ Candidatus M. haemobos’ infection, as well as of an A. ovis strain in ticks collected from cattle.

Differences in prevalence of Borrelia burgdorferi and Anaplasma spp. infection among host-seeking Dermacentor occidentalis, Ixodes pacificus, and Ornithodoros coriaceus ticks in northwestern California

Previous studies revealed that the Pacific Coast tick ( Dermacentor occidentalis) is infected occasionally with the agents of Lyme disease ( Borrelia burgdorferi) or human granulocytic anaplasmosis ( Anaplasma phagocytophilum) and that it is an inefficient experimental vector of B. burgdorferi. The relationship of the pajahuello tick ( Ornithodoros coriaceus) to each of these bacterial zoonotic agents has not been reported. The primary bridging vector of both bacterial zoonotic agents to humans is the western black-legged tick ( Ixodes pacificus). Because of the spatial and temporal overlap of D. occidentalis and O. coriaceus populations with those of I. pacificus in natural foci of B. burgdorferi and A. phagocytophilum in northwestern California, we conducted field and laboratory studies to determine if the Pacific Coast tick or the pajahuello tick potentially may serve as secondary vectors of either bacterium. Our findings reconfirmed that wild-caught D. occidentalis ticks are infected infrequently with B. burgdorferi or A. phagocytophilum, but some adult ticks from dense woodlands or chaparral were found to contain two important veterinary pathogens for the first time ( Anaplasma bovis, A. ovis). The high prevalence of A. bovis infection (4.3%, n = 185 ticks) within chaparral-derived ticks suggests that D. occidentalis could be an efficient vector of this rickettsia. Experimental attempts to transmit borreliae or Anaplasma spp. that may have been present in >100 wild-caught D. occidentalis adults to naïve rabbits were unsuccessful. Anaplasma spp. were not detected in O. coriaceus, but one (4.3%) of 23 nymphs was infected with B. bissettii. This finding and an antecedent report of a B. burgdorferi-like spirochete from the same tick species demonstrate that O. coriaceus sometimes acquires and transstadially passes Lyme disease group spirochetes. I. pacificus nymphs inhabiting a woodland nidus of B. burgdorferi and A. phagocytophilum had a 5-fold higher prevalence of borreliae than adult ticks from the same generational cohort. In contrast to the results of preceding studies carried out at the same site, none of the nymphal or adult ticks was PCR-positive for A. phagocytophilum. This suggests that the distribution of this rickettsia is highly focal or variable from year-to-year within this particular woodland.

Detection of Theileria and Babesia in brown brocket deer (Mazama gouazoubira) and marsh deer (Blastocerus dichotomus) in the State of Minas Gerais, Brazil

Intraerythrocytic protozoan species of the genera Theileria and Babesia are known to infect both wild and domestic animals, and both are transmitted by hard-ticks of the family Ixodidae. The prevalences of hemoprotozoa and ectoparasites in 15 free-living Mazama gouazoubira, two captive M. gouazoubira and four captive Blastocerus dichotomus from the State of Minas Gerais, Brazil, have been determined through the examination of blood smears and the use of nested polymerase chain reaction (nPCR). The cervid population was inspected for the presence of ticks and any specimens encountered were identified alive under the stereomicroscope. Blood samples were collected from all 21 animals, following which blood smears were prepared, subjected to quick Romanowsky staining and examined under the optical microscope. DNA was extracted with the aid of commercial kits from cervid blood samples and from tick salivary glands. The nPCR assay comprised two amplification reactions: the first was conducted using primers specific for a 1700bp segment of the 18S rRNA gene of Babesia and Theileria species, whilst the second employed primers designed to amplify a common 420bp Babesia 18S rRNA fragment identified by aligning sequences from Babesia spp. available at GenBank. The ticks Amblyomma cajennense, Rhipicephalus microplus and Dermacentor nitens were identified in various of the cervids examined. Of the animals investigated, 71.4% (15/21) were infected with hemoprotozoa, including Theileria cervi (47.6%), Theileria sp. (14.3%), Babesia bovis (4.8%) and Babesia bigemina (4.8%). However, only one of the infected wild cervids exhibited accentuated anaemia (PCV=17%). This is first report concerning the occurrence of Theileria spp. in Brazilian cervids.

Seasonal abundance and activity of the tick Dermacentor silvarum in northern China

Studies on the seasonal abundance and activity of larvae, nymphs and adults of Dermacentor silvarum (Ixodida: Ixodidae), both on and off the host, were carried out over a 2-year period from February 2008 to January 2010 in northern China. During the study period, feeding ticks were removed weekly from domestic sheep and free-living ticks were collected weekly by flag-dragging in four different habitat types. Rodents were captured and inspected for immature ticks from June to September 2008. The results of this study suggest that the ticks mainly reside in shrubs and complete only one generation per year with sequential seasonal distribution and little overlap between the activity of the larvae and nymphs. Adults were most prevalent from late February to late May and peaked in number in mid-April. Larvae were found from early June to early September and peaked in number in mid-July. Nymphs were mainly distributed from late June to late September and peaked in mid-August. Adult and nymphal D. silvarum were found primarily on the ears of sheep, but no larvae were found on sheep. Additionally, an overwintering male adult population was detected on sheep after October, but no free-living adults were found by dragging.

A Kunitz Protease Inhibitor from Dermacentor variabilis , a Vector for Spotted Fever Group Rickettsiae, Limits Rickettsia montanensis Invasion

A defining facet of tick-Rickettsia symbioses is the molecular strategy employed by each partner to ensure its own survival. Ticks must control rickettsial colonization to avoid immediate death. In the current study, we show that rickettsial abundance in the tick midgut increases once the expression of a Kunitz-type serine protease inhibitor from the American dog tick (Dermacentor variabilis) (DvKPI) is suppressed by small interfering RNA (siRNA). A series of in vitro invasion assays suggested that DvKPI limits rickettsial colonization during host cell entry. Interestingly, we observed that DvKPI associates with rickettsiae in vitro as well as in the tick midgut. Collectively, our data demonstrate that DvKPI limits host cell invasion by Rickettsia montanensis, possibly through an association with the bacterium.

Distribution and prevalence of Cytauxzoon felis in bobcats (Lynx rufus), the natural reservoir, and other wild felids in thirteen states

Cytauxzoon felis, a protozoan parasite of wild and domestic felids, is the causative agent of cytauxzoonosis in domestic and some exotic felids in the United States. The bobcat (Lynx rufus) is the natural reservoir for this parasite, but other felids such as Florida panthers (Puma concolor coryii) and domestic cats may maintain long-term parasitemias and serve as reservoirs. Experimentally, two tick species, Dermacentor variabilis and Amblyomma americanum, have demonstrated the ability to transmit C. felis. These two tick species have overlapping distributions throughout much of the southeastern United States. The objective of the current study was to determine the distribution and prevalence of C. felis in free-ranging bobcat populations from 13 states including California, Colorado, Florida, Georgia, Kansas, Kentucky, Missouri, North Carolina, North Dakota, Ohio, Oklahoma, South Carolina, and West Virginia. These states were selected because of differential vector presence; D. variabilis is present in each of these states except for the region of Colorado sampled and A. americanum is currently known to be present only in a subset of these states. Blood or spleen samples from 696 bobcats were tested for C. felis infection by a polymerase chain reaction (PCR) assay which targeted the first ribosomal internal transcribed spacer region (ITS-1). Significantly higher prevalences of C. felis were detected from Missouri (79%, n=39), North Carolina (63%, n=8), Oklahoma (60%, n=20), South Carolina (57%, n=7), Kentucky (55%, n=74), Florida (44%, n=45), and Kansas (27%, n=41) compared with Georgia (9%, n=159), North Dakota (2.4%, n=124), Ohio (0%, n=19), West Virginia (0%, n=37), California (0%, n=26), and Colorado (0%, n=67). In addition to bobcats, seven cougars (Puma concolor) from Georgia, Louisiana, and North Dakota and one serval (Leptailurus serval) from Louisiana were tested for C. felis. Only one cougar from Louisiana was PCR positive, which represents the first report of an infected cougar outside of the Florida panther population. These data also indicate that C. felis is present in North Dakota where infection has not been reported in domestic cats. Based on a nonparametric analysis, prevalence rates were significantly higher in states where there are established populations of A. americanum, which supports recent data on the experimental transmission of C. felis by A. americanum and the fact that domestic cat clinical cases are temporally associated with A. americanum activity. Collectively, these data confirm that bobcats are a common reservoir for C. felis and that A. americanum is likely an epidemiologically important vector.