Phenology of five tick species in the central Great Plains.

The states of Kansas and Oklahoma, in the central Great Plains, lie at the western periphery of the geographic distributions of several tick species. As the focus of most research on ticks and tick-borne diseases has been on Lyme disease which commonly occurs in areas to the north and east, the ticks of this region have seen little research attention. Here, we report on the phenology and activity patterns shown by tick species observed at 10 sites across the two states and explore factors associated with abundance of all and life specific individuals of the dominant species. Ticks were collected in 2020-2022 using dragging, flagging and carbon-dioxide trapping techniques, designed to detect questing ticks. The dominant species was A. americanum (24098, 97%) followed by Dermacentor variabilis (370, 2%), D. albipictus (271, 1%), Ixodes scapularis (91, <1%) and A. maculatum (38, <1%). Amblyomma americanum, A. maculatum and D. variabilis were active in Spring and Summer, while D. albipictus and I. scapularis were active in Fall and Winter. Factors associated with numbers of individuals of A. americanum included day of year, habitat, and latitude. Similar associations were observed when abundance was examined by life-stage. Overall, the picture is one of broadly distributed tick species that shows seasonal limitations in the timing of their questing activity.

New hard tick (Acari: Ixodidae) reports and detection of Rickettsia in ticks from Sierra Nevada de Santa Marta, Colombia.

The Sierra Nevada de Santa Marta (SNSM), located in northern Colombia, is considered a geographical island with high levels of biodiversity and endemism. However, little is known about tick species and their associated microorganisms at the SNSM. In this study we sampled host-seeking ticks in areas of the town of Minca within the SNSM. We collected 47 ticks identified as Amblyomma pacae, Amblyomma longirostre, Amblyomma ovale, Amblyomma mixtum, Haemaphysalis juxtakochi, Ixodes sp. cf. Ixodes affinis and Ixodes sp. Of these ticks, we tested for Rickettsia spp. by amplifying the gltA, SCA1, and 16S rRNA genes via PCR. Rickettsia amblyommatis was detected in one pool of 3 larvae and in a female of A. pacae. Additonally, we isolated Rickettsia sp. belonging to the group of spotted fevers in larvae of A. longirostre. This study reports new findings of six species of ticks and two species of Rickettsia within the SNSM.

Survey of Rickettsia spp. in ticks (Acari: Ixodidae) infesting opossums (Didelphis albiventris) and capybaras (Hydrochoerus hydrochaeris) from an urban park in southeastern Brazil.

Ticks are hematophagous arthropods and, during feeding, may transmit pathogens to vertebrate hosts, including humans. This study aimed to investigate the presence of Rickettsia spp. in ticks collected between 2010 and 2013 from free-ranging capybaras (Hydrochoerus hydrochaeris) and opossums (Didelphis albiventris) that inhabit Sabiá Park in Uberlândia, Brazil. Overall, 1,860 ticks were collected: 1,272 (68.4%) from capybaras (487 of the species Amblyomma sculptum, 475 adults and 12 nymphs; 778 Amblyomma dubitatum, 727 adults and 51 nymphs; and seven larva clusters of the genus Amblyomma); and 588 (31.6%) from opossums (21 A. sculptum, one adult and 20 nymphs; 79 A. dubitatum, all nymphs; 15 Ixodes loricatus, 12 adults and three nymphs; 457 Amblyomma sp. larva clusters; 15 Ixodes sp. larva clusters; and one Argasidae larva cluster). Out of 201 DNA samples tested for the presence of Rickettsia spp. DNA using polymerase chain reaction (PCR) 12 showed amplification of a gtlA gene segment that was specific to Rickettsia bellii, a bacterium non-pathogenic to humans. As there has been a report showing serological evidence of infections caused by Rickettsia species of the spotted fever group (SFG) in capybaras and opossums in the park, including Rickettsia rickettsii, the etiological agent of Brazilian spotted fever, and considering the presence of A. sculptum ticks, which are aggressive to humans, as well as these vertebrate hosts, which are amplifiers of R. rickettsii, it is important to monitor the presence of SFG rickettsiae in the Sabiá Park, which is visited daily by thousands of people.

High-resolution prediction models for Rhipicephalus microplus and Amblyomma cajennense s.l. ticks affecting cattle and their spatial distribution in continental Ecuador using bioclimatic factors.

In Ecuador, the main tick species affecting cattle are Rhipicephalus microplus and Amblyomma cajennense sensu lato. Understanding their spatial distribution is crucial. To assess their distribution, data from 2895 farms visited between 2012 and 2017 were utilized. Ticks were collected during animal inspections, with each farm's location georeferenced. Bioclimatic variables and vapor pressure deficit data were obtained from Climatologies at High resolution for the Earth´s Land Surface Areas (CHELSA) dataset. They were overlaid to develop predictive maps for each species using Random Forest (RF) models. The cross-validation results for RF prediction models showed high accuracy for both R. microplus and A. cajennense s.l. presence with values of accuracy = 0.97 and 0.98, sensitivity = 0.96 and 0.99, and specificity = 0.96 and 0.93, respectively. A carefully selected subset of bioclimatic variables was used to describe the presence of each tick species. Higher levels of precipitation had positive effect on the presence of R. microplus but a negative effect on A. cajennense s.l. In contrast, isothermality (BIO3) was more important for the presence of A. cajennense s.l. compared to R. microplus. As a result, R. microplus had a broader distribution across the country, while A. cajennense s.l. was mainly found in coastal areas with evident seasonality. The coexistence of both species in some regions could be attributed to transitional zones, whereas high altitudes limited tick presence. This information can aid in developing appropriate tick management plans, particularly considering A. cajennense s.l.'s broad host range species and R. microplus's specificity for cattle. Moreover, the predictive models can identify areas at risk of associated challenging hemoparasite, requiring special attention and mitigation measures.

Effective control of the motile stages of Amblyomma americanum and reduced Ehrlichia spp. prevalence in adults via permethrin treatment of white-tailed deer in coastal Connecticut, USA.

The lone star tick, Amblyomma americanum, is a common human-biting species whose range has been largely restricted to the southeastern United States, until recent detections of established populations on Long Island, New York and throughout coastal southern New England. We evaluated the effectiveness of topical treatment of 10 % permethrin delivered via 4-poster devices to white-tailed deer, Odocoileus virginianus, in the management of a newly discovered A. americanum population in Norwalk, Connecticut. Using a high-density deployment of one 4-poster device/12.7 ha, we were successful in significantly reducing densities of host-seeking adults (93 % reduction), nymphs (92 %), and larvae (96 %) from 2018 to 2020. We also documented a significant reduction (87 %) in parasitizing adults and nymphs on white-tailed deer from 2018 to 2019. The prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii combined in host-seeking adults declined significantly from 47 % at the time the A. americanum population was discovered in 2017 to 7% in 2020. However, the prevalence in nymphs remained static (∼9%) throughout the study period. These data demonstrate that, when properly deployed in a density-dependent manner in terms of deer abundance, 4-poster devices can effectively manage parasitizing and host-seeking A. americanum populations and reduce the prevalence of two ehrlichial species of public health importance.

Validating Species Distribution Models With Standardized Surveys for Ixodid Ticks in Mainland Florida.

Tick-borne pathogens are of growing concern. The U.S. Centers for Disease Control and Prevention (CDC) developed guidelines standardizing surveys of tick vectors to better monitor the changes in their occurrences. Unbiased surveillance data, from standardized surveys, are presumed critical to generate valid species distribution models (SDMs). We tested previously generated SDMs from standardized protocols for three medically important ticks [Amblyomma americanum (Linnaeus, Ixodida, Ixodidae), Ixodes scapularis (Say, Ixodida, Ixodidae), and Dermacentor variabilis (Say, Ixodida, Ixodidae)]. These previous models ruled out a quarter to half of the state as having these species, with consensus occurrence in about a quarter of the state. New surveys performed throughout 2019 on 250 transects at 43 sites indicated the rule-out functions were 100% accurate for I. scapularis and D. variabilis and 91.9% for A. americanum. As SDM concordance increased, the proportion of transects yielding ticks increased. Independent surveys of SDMs provide external validation-an aspect missing from many SDM studies.

Established Population of the Gulf Coast Tick, Amblyomma maculatum (Acari: Ixodidae), Infected with Rickettsia parkeri (Rickettsiales: Rickettsiaceae), in Connecticut.

We identified an established population of the Gulf Coast tick (Amblyomma maculatum Koch) infected with Rickettsia parkeri in Connecticut, representing the northernmost range limit of this medically relevant tick species. Our finding highlights the importance of tick surveillance and public health challenges posed by geographic expansion of tick vectors and their pathogens.

Transcriptome analysis of Ehrlichia ruminantium in the ruminant host at the tick bite site and in the tick vector salivary glands.

Heartwater is a non-contagious tick-borne disease of domestic and wild ruminants. Data regarding the complex processes involved during pathogen-vector-host interaction during Ehrlichia ruminantium infection is lacking and could be improved with knowledge associated with gene expression changes in both the pathogen and the host. Thus, in the current study, we aimed to identify E. ruminantium genes that are up-regulated when the pathogen enters the host and before the disease is established. Identification of such genes/proteins may aid in future vaccine development strategies against heartwater. RNA-sequencing was used to identify E. ruminantium genes that were exclusively expressed at the tick bite site in sheep skin biopsies (SB) and in adult tick salivary glands (SG). RNA was extracted from pooled samples of the SB or SG collected at different time points during tick attachment and prior to disease manifestation. Ribosomal RNA (rRNA) was removed and the samples were sequenced. Several E. ruminantium genes were highly expressed in all the samples while others were exclusively expressed in each. It was concluded that E. ruminantium genes that were exclusively expressed in the SB or both SB and SG when compared to the transcriptome datasets from bovine elementary bodies (BovEBs) from cell culture may be considered as early antigenic targets of host immunity. In silico immunogenic epitope prediction analysis and preliminary characterization of selected genes in vitro using ELIspot assay showed that they could possibly be ideal targets for future vaccine development against heartwater, however, further epitope characterization is still required.

Low Abundance of Three Tick Species in the Piedmont of North Carolina.

Multiple species of ticks, including Ixodes scapularis (Say, Ixodida:Ixodidae), Amblyomma americanum (L., Ixodida:Ixodidae), and Dermacentor variabilis (Say, Ixodida:Ixodidae), occur in high and increasing abundance in both the northeast and southeast United States. North Carolina is at the nexus of spread of these species, with high occurrence and abundance of I. scapularis to the north and A. americanum to the south. Despite this, there are few records of these species in the Piedmont of North Carolina, including the greater Charlotte metropolitan area. Here, we update the known occurrence and abundance of these species in the North Carolina Piedmont. We surveyed for ticks using cloth drags, CO2 traps, and leaf litter samples at a total of 79 sites within five locations: Mecklenburg County, South Mountains State Park, Stone Mountain State Park, Duke Forest, and Morrow Mountain State Park, all in North Carolina, during the late spring, summer, and fall seasons of 2019. From these surveys, we had only 20 tick captures, illuminating the surprisingly low abundance of ticks in this region of North Carolina. Our results indicate the possibility of underlying habitat and host factors limiting tick distribution and abundance in the North Carolina Piedmont.

Rickettsia amblyommatis isolated from Amblyomma mixtum (Acari: Ixodida) from two sites in Panama.

Rickettsia amblyommatis is widely distributed in the Americas, and has been reported to infect different species of ticks within its distribution. In Panama, R. amblyommatis is the most common Rickettsia and its presence was molecularly detected in nine species of ticks and one flea species. This work described the isolation of R. amblyommatis in Vero cells by shell vial technique, from Amblyomma mixtum ticks collected from a captive tapir from Gamboa (Colon province), and a horse from El Valle de Antón (Cocle province). These represent the first isolations of R. amblyommatis in Panama.

Locomotion activity and its effects on the survival of Amblyomma sculptum (Acari: Ixodidae) nymphs under laboratory conditions.

The active locomotion of ticks is directly associated with the epidemiology of tick-borne diseases, as it has important implications for the interaction of ticks with their hosts and their dispersion in the environment. In an attempt to elucidate the factors involved in the dispersion of Amblyomma sculptum, the present work aimed to characterize different aspects of the active locomotion of A. sculptum nymphs under laboratory conditions. To this end, nymphs were placed on a string at a 70° inclination and their walking activity was recorded daily along with their survival period. During their lifetime, ticks walked an average of 110 m. Their locomotion was not in a straight line and nymphs changed direction 142 times throughout their lifetimes. The mean distance walked per experimental day was 1.8 m, while the average walking distance before changing direction was 52 cm. The distance walked per experimental day reduced over time. The survival of ticks was affected by walking; resting nymphs survived for over 6 months, while the survival of those that walked daily was reduced to approximately 62 days. The results showed that A. sculptum nymphs were able to cover distances of over 100 m throughout their lifetimes, but they walked short distances at a time and constantly changed direction. This behavior indicates that ticks are not able to disperse over long distances by means of active locomotion.

Rickettsia spp. and Ehrlichia spp. in Amblyomma ticks parasitizing wild amphibious sea kraits and yellow-margined box turtles in Okinawa, Japan.

Recently, several tick-borne pathogens were detected in reptile-associated ticks. However, studies on the microorganisms in reptile-associated ticks in Japan are limited. This molecular survey thus aimed to identify and characterize tick-borne pathogens (Rickettsiaceae and Anaplasmataceae) in reptile-associated ticks in Japan. In total, 77 Amblyomma nitidum and 104 Amblyomma geoemydae were collected from wild amphibious sea kraits (Laticauda semifasciata, Laticauda colubrina, and Laticauda laticaudata) and from yellow-margined box turtles (Cuora flavomarginata evelynae), respectively. Conventional polymerase chain reaction was performed using the DNA extracted from the ticks to detect the selected pathogens. Sequencing analysis of four Rickettsia genes (gltA, ompA, ompB, and sca4) led to the identification of a putative novel Rickettsia sp. and Rickettsia aeschlimannii-like rickettsia in A. nitidum and A. geoemydae, respectively. Sequencing analysis of gltA and groEL of Anaplasmataceae revealed that the Ehrlichia spp. in these ticks were novel and related to Candidatus Ehrlichia occidentalis. This is the first study on the microorganisms in A. nitidium and the first record of Rickettsia and Ehrlichia in A. geoemydae. Further studies are required to understand their pathogenicity to humans and animals and their life cycle in the wild.

Morphological and molecular characterization supporting Amblyomma mixtum presence in Cuba.

Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) is a widely distributed tick taxon. Recent studies have reassessed this taxon as a complex of six species. Amblyomma mixtum Koch, 1844 has been suggested by some authors as the only species of this complex that is present in Cuba. Other authors have pointed a niche overlapping for A. mixtum and A. cajennense s.s. in the country. Detailed taxonomic studies on the Cuban species belonging to this complex are needed in order to evaluate their current distribution according to the recent classification. This study aimed to characterize Cuban populations from the A. cajennense complex by using tick samples obtained from 3 occidental provinces and 1 central province of the country. Morphological identification and measurements of the main relevant taxonomic structures were conducted by using Scanning Electron Microscopy. Phylogenetic analyzes were carried out with 16S ribosomal RNA, internal transcribed spacer 2 and the subunit I of mitochondrial cytochrome c oxidase gene sequences. The results of these studies demonstrated that all samples belonged to the species A. mixtum (Koch, 1844). This study constitutes the first molecular characterization of this Amblyomma species in Cuba. Further studies will be necessary in order to corroborate if A. cajennense s.s. is also present in the island.

Molecular phylogeny of Amblyomma exornatum and Amblyomma transversale, with reinstatement of the genus Africaniella (Acari: Ixodidae) for the latter.

The phylogeny of the hard tick genus Amblyomma Koch, 1844 deserves special attention, because several poorly studied tick species associated with reptiles still bear the name of this genus, although they may not belong to it. This study focuses on the phylogeny of two such species with uncertain taxonomic status, i.e., Amblyomma transversale (Lucas, 1845) and Amblyomma exornatum Koch, 1844, analyzing two mitochondrial (cytochrome c oxidase subunit 1 and 16S rRNA) and two nuclear (18S and 28S rRNA) genes. In the cox1 phylogenetic analysis, both Am. transversale and Am. exornatum were part of a sister group to all other Metastriata, whereas in the 16S rRNA gene analysis, Am. transversale belonged to a sister group to three subfamilies (Amblyomminae Neumann, 1911; Haemaphysalinae Hoogstraal and Aeschlimann, 1982; Bothriocrotoninae Klompen, Dobson and Baker, 2002), and Am. exornatum formed a sister group to other Amblyomminae. However, based on the 18S and 28S rRNA genes, Am. transversale belonged to a sister group of either Bothriocrotoninae alone or of both Bothriocrotoninae and Haemaphysalinae, respectively. In the latter two phylogenetic analyses Am. exornatum always clustered within Amblyomminae. Morphological comparisons revealed that Am. transversale has at least four unique characters and shares a high number of traits with the genera Robertsicus Barker and Burger, 2018 and Archaeocroton Barker and Burger, 2018, as well as with the subgenus Alloceraea Schulze, 1918 (represented by Haemaphysalis inermis Birula, 1895). These results justify that the genus Africaniella Travassos Dias, 1974 should be reinstated, and the species name of Am. transversale should be used as Africaniella transversale (Lucas, 1845).

Molecular detection of a Borrelia sp. in nymphs of Amblyomma brasiliense ticks (Acari: Ixodidae) from Iguaçu National Park, Brazil, genetically related to Borrelia from Ethiopia and Côte d'Ivoire.

Gram-negative spirochetes of the genus Borrelia are transmitted to vertebrate hosts through the tick bite during blood intake. Pathogenic Borrelia species may cause relapsing fever or Lyme borreliosis in humans. Our study aimed to molecularly detect and characterize bacteria of this genus in ticks collected in the Iguaçu National Park, located in southern Brazil. Ticks were collected from the environment (free living) and from hosts from May 2015 to July 2017. In total, 829 ticks were tested, being 741 from the environment and 88 from hosts; 128 ticks were larvae, 523 were nymphs, and 178 were adults (80 males and 98 females). The species identified were: Amblyomma brasiliense (42.9 %), Amblyomma coelebs (16.8 %), Amblyomma sp. (15.0 %), Amblyomma incisum (10.3 %), Amblyomma ovale (8.7 %), Haemaphysalis juxtakochi (5.5 %), Rhipicephalus microplus (0.5 %), Amblyomma longirostre (0.2 %), and Rhipicephalus sanguineus sensu lato (0.1 %). DNA extraction was performed with pools of larvae or nymphs, or individually with adults, for a total of 394 samples. The PCR technique used to detect Borrelia DNA was performed with two rounds of amplification reactions targeting a segment of the flagellin B gene (flaB). Amplification occurred in only one DNA sample, which was obtained from nymphs of A. brasiliense collected from the vegetation of a trail. The nucleotide sequence analysis revealed 90.8 % identity to a sequence of Borrelia sp. from Côte d'Ivoire, and 89.1 % identity to a sequence from Ethiopia. Phylogenetic analysis revealed that the Borrelia sequence from A. brasiliense was distinct from the Borrelia species of the groups belonging to Lyme borreliosis, relapsing fever and the one associated with reptile and echidna as hosts. The sequence is likely from a putative new species of Borrelia and was detected for the first time in A. brasiliense ticks.

Capybaras (Hydrochoerus hydrochaeris) as amplifying hosts of Rickettsia rickettsii to Amblyomma sculptum ticks: Evaluation during primary and subsequent exposures to R. rickettsii infection.

Rickettsia rickettsii is the agent of Brazilian spotted fever (BSF), the most lethal tick-borne disease in the western hemisphere. Amblyomma sculptum is the main vector of R. rickettsii in southeastern Brazil. Capybaras act as primary hosts for this tick species, and as amplifying hosts for R. rickettsii, creating new infected lineages of A. sculptum in BSF-endemic areas. In a recent study, we demonstrated that capybaras successively exposed to R. rickettsii-infected A. sculptum ticks developed a mean rickettsemic period of 9.2 days during primary infection, but no rickettsemia during subsequent expositions, when capybaras were immune to R. rickettsii. During the primary and subsequent infections, capybaras were also infested with uninfected A. sculptum ticks. These infestations compose the present study, which aimed to evaluate: (i) if either rickettsemic or non-rickettsemic capybaras could serve as infection sources of R. rickettsii for A. sculptum larvae and nymphs; (ii) the vector competence of the subsequent nymphs and adults (molted from the engorged ticks collected from capybaras); and (iii) if there were R. rickettsii-acquisition by uninfected ticks co-feeding with infected ticks on immune capybaras (without rickettsemia). Through experimental infection of capybaras with R. rickettsii via tick feeding, simulating a natural condition, we demonstrated that primarily infected capybaras developed rickettsemia that resulted in successful acquisition feeding of larvae and nymphs of A. sculptum, since part of these ticks maintained R. rickettsii transstadially, and the resultant molted ticks (either nymphs or adults) successfully transmitted the bacterium by feeding on susceptible rabbits. Contrastingly, all rabbit infestations with ticks derived from acquisition feeding on R. rickettsii-immune capybaras (including when acquisition ticks fed in direct contact with donor ticks) evidenced absence of R. rickettsii transmission due to lack of clinical signs and antibody response in those rabbits. Our results indicate that capybaras could serve as R. rickettsii-amplifying hosts for A. sculptum ticks only during the capybara's primary infection, but not during subsequent infections on immune capybaras. Finally, the probable co-feeding nonsystemic transmission of R. rickettsii seems to be irrelevant in the context of BSF epidemiology, in areas where capybaras are incriminated as main amplifying hosts of R. rickettsii for A. sculptum ticks.

Free-living ticks (Acari: Ixodidae) in the Iguaçu National Park, Brazil: Temporal dynamics and questing behavior on vegetation.

Basic knowledge of species, ecological preferences and behavior of ticks in preserved rainforests are scanty. We herein describe species richness, seasonal activity, altitude influence and questing height of ticks along 3375 m of animal trails within the largest remain of inland Atlantic rainforest in Brazil, the Iguaçu National Park (INP). Altogether, 2954 ticks from nine tick species were collected; Amblyomma brasiliense Aragão (49.09% of the specimens), Amblyomma coelebs Neumann (29.08%), Amblyomma incisum Neumann (12.49%), Haemaphysalis juxtakochi Cooley (5.21%), Amblyomma ovale Koch (2.64%), Amblyomma longirostre Koch (0.20%), Ixodes fuscipes Fonseca (0.17%), Ixodes loricatus Neumann (0.07%) and Amblyomma dubitatum Neumann (0.03%) (Ixodida: Ixodidae). There was an evident seasonal influence on tick host-questing activity, with each tick stage prevailing in a season before the following one, and a general trend of one tick generation per year. Adult tick questing height mode on vegetation showed a correspondence between tick species and preferred host size and height. Adult ticks quested higher than immatures, but several larval clusters were found high enough to seek for large-sized hosts as well. Nymphal and adult tick questing height increased significantly at higher altitudes within the park. Amblyomma incisum and A. coelebs were more abundant at higher and lower altitudes, respectively. The knowledge on tick species richness, ecological preferences and behavior herein obtained provides baseline information to understand tick-borne disease epidemiology occurring at landscapes under a strong anthropogenic impact.

Evaluation of oral fluralaner (Bravecto®) for efficacy against nymphs of Amblyomma americanum and Rhipicephalus sanguineus (sensu lato).

BACKGROUND: Amblyomma americanum and Rhipicephalus sanguineus (sensu lato) nymphs commonly feed on and transmit pathogens to dogs (Canis familiaris). Control of immature and adult tick life stages is necessary to fully protect animals. We evaluated efficacy of oral fluralaner (Bravecto®) against induced infestations with A. americanum and R. sanguineus (s.l.) nymphs on dogs in two experiments. METHODS: In each experiment, 10 dogs were administered oral fluralaner chewable tablets one time on Day 0 at a targeted minimum dose of 25 mg/kg body weight and 10 dogs remained non-treated controls. Dogs were infested with two groups of 50 A. americanum nymphs and two groups of 50 R. sanguineus (s.l.) nymphs on Days -1, 6, 28, 56 and 84. At 48 h and 72 h post-infestation, nymphs were collected from dogs, assessed as live or dead, and enumerated into categories defining attachment and engorgement status. Fluralaner efficacy was determined in separate analyses against all live nymphs and against live-fed nymphs, i.e. live nymphs that were attached to dogs at the time of collection and/or were engorged. Fluralaner was considered effective when mean numbers of live ticks were reduced in fluralaner-treated dogs by ≥ 90%. RESULTS: Fluralaner efficacy against all live and live-fed A. americanum nymphs in the first experiment was > 94% on all collection days. Efficacy against all live R. sanguineus (s.l.) nymphs in the first experiment was > 96% on all collection days  excluding the 48 h counts for infestations on Days 28 (83.7%), 56 (82.9%) and 84 (86.7%); efficacy against live-fed R. sanguineus (s.l.) nymphs was > 95% on all 48 h/72 h count days. Fluralaner efficacy against all live A. americanum nymphs in the second experiment was > 93% on all collection days for 8 weeks excluding the 48 h count for infestation on Day 56 (87.8%); efficacy against live-fed A. americanum nymphs was > 91% on all count days for 8 weeks. Efficacy against all live R. sanguineus (s.l.) nymphs in the  second experiment was > 91% on all 72 h collection days  except for infestations on Days 28 (76.8%) and 56 (86.3%); efficacy against live-fed R. sanguineus (s.l.) nymphs was 100% on all 72 h count days. CONCLUSIONS: A single administration of oral fluralaner to dogs is effective against A. americanum and R. sanguineus (s.l.) nymphs for up to 12 weeks.

Lethal Effects of a Silica Gel + Pyrethrins (Drione) on Amblyomma americanum (Ixodida: Ixodidae) Larvae and Nymphs.

Ixodids (hard ticks) ingest blood from host animals, and they can transmit pathogenic organisms that induce medical and veterinary diseases. As resistance to synthetic conventional acaricides becomes more common, alternative tactics are coming under heightened scrutiny. Laboratory bioassays were used to assess the efficacy of CimeXa, a commercially available silica gel desiccant dust product, and Drione, a commercial product containing silica gel + pyrethrins and a synergist, piperonyl butoxide, against lone star tick, Amblyomma americanum (L.) (Ixodida: Ixodidae), larvae and nymphs. Both life stages were completely killed by CimeXa by 24 h, and Drione caused total larval mortality within 1 h when they were briefly immersed in the dusts and when they crawled across dust-treated substrate; nymphs were completely killed by 4 h after the same kinds of exposure. Mortality of A. americanum larvae and nymphs occurred after the pests crawled across dried aqueous suspensions of the products, but this was not as efficient and fast-acting as when the immature life stages were exposed to dry dusts. Further, dried aqueous suspensions of Drione were not substantially more lethal than dried aqueous suspensions of CimeXa. CimeXa and Drione will provide prophylactic control on vegetation and animals for as long as the silica gel remains without being physically removed. Both of the dust-based products will likely also be effective against other problematic ixodid species. Advantages and disadvantages, and potential uses, of desiccant dust-based acaricides are discussed.

Novel PCR exclusion assay to detect spotted fever group rickettsiae in the lone star tick (Amblyomma americanum).

The lone star tick (Amblyomma americanum) is the most common and abundant human-biting tick in the southeastern United States where spotted fever rickettsioses frequently occur. However, the role of this tick in transmitting and maintaining pathogenic and non-pathogenic spotted fever group rickettsiae (SFGR) remains poorly defined. This is partially due to the high prevalence and abundance of Rickettsia amblyommatis in most populations of A. americanum. Many molecular assays commonly employed to detect rickettsiae use PCR primers that target highly conserved regions in the SFGR so low abundance rickettsia may not be detected when R. amblyommatis is present. It is costly and inefficient to test for low abundance rickettsial agents with multiple individual specific assays even when they are multiplexed, as most samples will be negative. Real time PCR assays may also be hampered by inadequate limits of detection (LODs) for low abundance agents. We exploited the absence of an otherwise relatively SFGR-conserved genome region in R. amblyommatis to design a hemi-nested PCR-assay which has a sensitivity of 10 copies in detecting the presence of most SFGR, but not R. amblyommatis in DNA of infected lone star ticks. This deletion is conserved in 21 isolates of R. amblyommatis obtained from multiple states. We demonstrated the assay's utility by detecting a pathogenic SFGR, Rickettsia parkeri, in 15/50 (30 %) of field collected A. americanum ticks that were previously screened with conventional assays and found to be positive for R. amblyommatis. These co-infected ticks included 1 questing female, 6 questing nymphs, and 8 attached males. The high prevalence of R. parkeri among host-attached ticks may be due to several variables and does not necessarily reflect the risk of disease transmission from attached ticks to vertebrate hosts. This novel assay can provide accurate estimates of the prevalence of less common SFGR in A. americanum and thus improve our understanding of the role of this tick in the maintenance and transmission of the SFGR commonly responsible for human rickettsioses.