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Understanding the relationships between landscape connectivity and abundance of Ixodes ricinus ticks.

The density of Ixodes ricinus ticks in a heterogeneous landscape of northern Spain was determined and associated with some aspects of habitat topology. The habitat mosaic was used to quantify connectivity between patches of different tick density. The analysis revealed that patches with high tick abundance are 'stepping-stone' territories that, when removed from the landscape, cause large changes in connectivity. Sites with medium tick abundance do not cause such a critical transition in connectivity. Patches with low tick abundance, but optimal abiotic conditions for survival are located within the minimum cost corridors network joining the patches, while those sites where the tick has been intermittently collected are located at variable distances from this network. These results suggest that tick distribution in a zone is highly affected not only by abiotic variables (vegetation and weather) but also by host movements. Whether these high-density 'stepping-stone' patches occur in other tick species needs to be evaluated because of the potential implications of these foci for human health.

Animals↗

The fossil record and the origin of ticks (Acari: Parasitiformes: Ixodida).

Ticks are obligate hematophagous ectoparasites of terrestrial vertebrates. Hypotheses on the origin of ticks have been proposed based on tick-host associations and the total-evidence approach analysis of morphological and molecular characters. Nevertheless, the origin of ticks remains a controversial issue. Here, I revised the tick fossil record including reports from the literature and the description of 7 new specimens. The analysis of fossil ticks provides few clues to tick evolution but does not contradict recent hypotheses based on total-evidence approach analysis that place the origin of ticks in the Cretaceous (65-146 mya) with most of the evolution and dispersal occurring during the Tertiary (5-65 mya).

Amber↗

Species of ticks on camels and their seasonal population dynamics in Eastern Ethiopia.

A study was conducted to identify the species of ticks found on camels (Camelus dromedarius) and their seasonal population dynamics in Eastern Ethiopia. Collection and identification of the ticks were undertaken at 2-month intervals from December 1997 to August 1999. On each occasion, all the visible adult ticks were collected from one side of the body of each of the same 17 camels. The most abundant species of ticks on the camels were Rhipiephalus pulchellus (85.2%), Hyalomma dromedarii (5.9%), Amblyomma gemma (4.0%) and Amblyomma variegatum (1.8%). The average tick load per camel was higher during rainy months than during dry months. The smallest number of ticks per camel was observed during the driest month (December), whereas the highest was recorded in the wettest month (August). Any strategy intended to mitigate problems of tick infestation of camels in this area should take into account the identified tick species and their season of abundance.

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Evidence of Bartonella sp. in questing adult and nymphal Ixodes ricinus ticks from France and co-infection with Borrelia burgdorferi sensu lato and Babesia sp.

Ticks are known vectors for a wide range of pathogenic microorganisms. Their role in the transmission of some others is so far only suspected. Ticks can transmit multiple pathogens, however, little is known about the co-existence of these pathogens within questing ticks. We looked for the presence of DNA from three micro-organisms, Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. which are known or suspected tick-borne pathogens, using a cohort of 92 questing Ixodes ricinus ticks collected from pastures in northern France. DNA was extracted from each individual tick and the presence of the three pathogens was investigated using Polymerase Chain Reaction (PCR) amplification. Nine among 92 samples (9.8%) demonstrated PCR products using Bartonella specific primers, 3 among 92 (3.3%) using Borrelia burgdorferi sensu lato specific primers and 19 among 92 (20.6%) using Babesia specific primers. Seven among 92 samples (7.6%) were PCR positive for at least two of the pathogens and one sample was positive for all three. Adult ticks (12/18; 67%) showed significantly higher infection rates compared to nymphs (11/74; 15%) for all three pathogens (P < 0.001). This study is the demonstration of the simultaneous presence of Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. in questing Ixodes ricinus ticks.

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Spatiotemporal variation in a Lyme disease host and vector: black-legged ticks on white-footed mice.

We monitored population density of white-footed mice (Peromyscus leucopus), burdens of immature black-legged ticks (Ixodes scapularis) on mice, and infection prevalence of host-seeking ticks on six forest plots in southeastern New York State from 1995 through 1999. Despite densities of mice that fluctuated two orders of magnitude, average larval and nymphal tick burdens per mouse remained remarkably constant. Spatial variability in mouse density and tick burdens was modest. The total number of larval and nymphal ticks that fed on the mouse population each year depended strongly on population density of mice; a steady increase was observed in both mouse density and total tick meals on mice from 1996 through 1999. The result was a steady increase in the infection prevalence of nymphal and adult ticks with the etiological agent of Lyme disease, Borrelia burgdorferi, over this time. We suggest that fluctuations in population density of mice, combined with possible regulation of tick burdens on mice, may influence risk of human exposure to Lyme disease.

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Rectal infusion and aspiration of material through the guts of ixodid ticks (Acari: Ixodidae).

A technique for inoculating and removing substances via the anus of vector ticks was devised to define features of vector competence precisely. Calibrated inocula (greater than 5 nanoliter) containing aqueous dye and polystyrene beads as well as infectious agents were infused into the rectal sacs of ticks using glass microcapillary pipettes placed within the expanded anal orifice. The guts of preadult and adult ticks, Ixodes dammini Spielman, Clifford, Piesman & Corwin, Dermacentor variabilis (Say), Hyalomma impeltatum Schulze & Schlottke, and Amblyomma americanum (L.), were thereby infused with these inocula. Distribution of inocula was determined by examining hemolymph and sectioned ticks and confirmed that material placed in the rectal sac spread throughout the midgut diverticula. Ticks survived for greater than 6 mo after this procedure and were able to feed, molt to the next stage, or oviposit. In contrast, fewer ticks survived after intracelomic inoculation. The course of infection in ticks receiving anal infusions of Borrelia burgdorferi (the Lyme disease spirochete) was assessed. Such infections appear to differ from those established by feeding on infected hosts. Contents of the tick gut can be sampled nondestructively by anal perfusion to diagnose infection by this spirochete.

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Tick (Acari: Ixodidae) attachment cement and salivary gland cells contain similar immunoreactive polypeptides.

A specific antiserum (12C) raised to a 90-kDa immunogenic component of salivary glands of the tick Rhipicephalus appendiculatus recognized similar 90-kDa polypeptides from salivary glands of the American dog tick, Dermacentor variabilis, and the lone star tick, Amblyomma americanum, as well as 70-kDa polypeptides in the cement of D. variabilis, A. americanum, and R. sanguineus (brown dog tick). The reduction in size of the polypeptide for these ticks suggests that it is modified in some way during or after secretion. Immunostaining of salivary glands of unfed- and partially-fed female D. variabilis localized an immunoreactive protein in the d- and e-cells of the type III acini. The quantity of label in granules of glands from unfed ticks was visibly greater than in the granules of glands from partially fed ticks, suggesting that this component is secreted within the first 2 d of feeding. Collectively, these data support the conclusion that a 90-kDa polypeptide of saliva is conserved among ixodid tick genera and is a component of the attachment cement.

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African swine fever virus infection in the Iberian soft tick, Ornithodoros (Pavlovskyella) marocanus (Acari: Argasidae).

One thousand six hundred Ornithodoros (Pavlovskyella) marocanus Velu larvae were fed on a pig infected with African swine fever virus (titer: 10(7.4) HAd50/ml), and 1,600 larvae were fed on an uninfected pig. Ticks in each group were compared for mortality rates, mean time to death for ticks that died, mean time from feeding to either molting or eclosion, percentage of ticks that eclosed or molted, and the number of blood meals per nymph or instar. Cumulative virus-induced mortality for all immature stages (larvae to adult) of O. marocanus that had been fed as larvae on a pig infected with African swine fever was ca. 73% over a 390-d period. In contrast, less than 9% mortality was observed among ticks fed on uninfected pigs. Mean time to death for infected ticks was 15-87 d versus 10-17 d for uninfected ticks. Differences in the premolt period (number of days from blood meal to molt) between infected and control ticks were not observed. Mean premolt periods for larvae and first-, second-, third-, fourth-, and fifth-instar nymphs fed on pigs were 7, 9, 15, 11, 15, and 15 d, respectively. The majority of infected and all uninfected ticks required only one blood meal from pigs to molt. Mean weights for unfed second-, third-, and fourth-instar nymphs and males and females were 0.50, 0.67, 3.07, 3.63, and 5.91 mg, respectively.

African Swine Fever↗

Integumental glands of the tick Rhipicephalus appendiculatus (Acari:Ixodidae) as potential producers of semiochemicals.

The opisthosomal integument and associated secretory organs of Rhipicephalus appendiculatus Neumann larvae, nymphs, females and males, both unfed and fed were examined by light and electron microscopy. Type 1 dermal glands were found on the alloscutum and scutum of all ticks. They were undeveloped in unfed ticks and reached full development in engorging ticks. They produced secretory granules from 2 glandular cells but without accumulation of a reservoir of secreted material. Type 2 dermal glands were found in all ticks, with pores on the alloscutum, edge of scutum, and on anal plates. These glands produced secretion during feeding and accumulated large reservoirs of secreted material that were present in engorging, recently detached and questing ticks. Spiracular glands were found in all ticks below the spiracle plate. They produced small amounts of secretion and had pores to the exterior by way of spiracle goblets. No obvious cycle of secretory activity was recorded. Foveal glands were present and produced secretions in nymphs, females, and males. They were largest in females with an accumulation of secretory vesicles in feeding ticks. The potential function of these glands is discussed in the context of the chemical ecology of this tick.

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Phylogenetic analysis of spotted fever group rickettsiae based on gltA, 17-kDa, and rOmpA genes amplified by nested PCR from ticks in Japan.

In order to understand the natural situation of rickettsiae in the ticks in Japan, the rickettsial genes, gltA gene, rOmpA gene, and 17-kDa gene, were amplified from the ticks by nested PCR. The prevalences of rickettsial gltA genes among Haemaphysalis formosensis, H. longicornis, H. megaspinosa, Ixodes ovatus, H. flava, H. kitaokai, and I. persulcatus were 62, 57, 24, 24, 19, 13, and 10%, respectively; 26% (186/722) being the average. The gltA genes amplified from the ticks were classified into 9 genotypes (I to IX) by the difference in nucleotide sequences. Genotype I was detected from 7 species of ticks. Genotype II mainly was detected from H. longicornis and H. formosensis. Genotypes III and VII mainly were detected from H. flava and I. ovatus. The polarization in the distribution of genotypes among regions where the ticks were collected was not clear. Based on the phylogenetic analysis of the three genes presented here, genotypes I, III, and IV (detected from H. formosensis, H. hystricia, and I. ovatus ) are genetically close with each other, but rickettsiae of the same property still have not been isolated from ticks anywhere in the world. These genotypes should be considered as new species among SFG rickettsiae. Genotype II was identical with strain FUJ-98, genetically close to R. japonica which has been isolated from ticks in China. Genotype V was identical with R. felis and strain California 2 isolated from the cat flea. This is the first report on the detection of R. felis from ticks. Genotype VI detected from Ixodes sp. did not seem to belong to genus Rickettsia. Based on the previous antigenic data and the phylogenetic analysis presented here, Genotype VII should be considered a variant of R. helvetica and genotype VIII detected from I. ovatus and I. persulcatus were identical with R. helvetica. Genotype IX detected from I. nipponensis was genetically close to the strains IRS3, IRS4, and IrR/Munich isolated from I. ricinus in Slovakia and German.

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Tick bites in Korea.

BACKGROUND: Tick bites are dermatoses not commonly encountered in Korea. Recognizing their clinical signs as well as their histopathologic findings is important in making a diagnosis of tick-related dermatoses. The incidence and causative species are different depending on the geographic areas. The histopathologic findings of tick bites are known to be a variable depending on the species of ticks involved and the duration of their bloodsucking. METHODS: Five ticks were collected from five patients and three of them were identified as Ixodes (I.) nipponensis. RESULTS: Histopathologic findings of panniculitis were prominent in four of five cases; septal panniculitis in two cases, and lobular panniculitis in the other cases. CONCLUSIONS: Ixodes nipponensis was the most common causative species of ticks responsible for tick bites in Korea, and tick bite panniculitis must be considered in the differential diagnosis of panniculitis which is mainly composed of neutrophils.

Aged↗

Detection of Astrakhan fever rickettsia from ticks in Kosovo.

Astrakhan fever is a summer spotted fever resembling Mediterranean spotted fever, endemic in Astrakhan, a region of Russia located by the Caspian sea. Its agent is a spotted fever group rickettsia, member of the Rickettsia conorii complex, transmitted to humans by Rhipicephalus sanguineus and Rhipicephalus pumilio ticks. In Summer 2001, French United Nations troops in Kosovo collected 2 ticks on asymptomatic soldiers (1 R. sanguineus and 1 Hyalomma marginatum) and 10 ticks on dogs (7 R. sanguineus, 2 Ixodes ricinus, and 1 H. marginatum) in the Morina region. By PCR amplification of both the gltA and ompA genes, we detected a rickettsia in 4 R. sanguineus, i.e., 3 of those collected on dogs and those taken from military personnel. As ticks were preserved in alcohol, culture was not possible. The sequences obtained from these PCR products identified, with a 100% homology, Astrakhan fever rickettsia. None of the other collected tick species was positive. The patient with the positive tick remained asymptomatic. Our study demonstrates, for the first time, the presence of Astrakhan fever rickettsia in ticks outside Russia. We suspect that the area of distribution of this rickettsia could be wider than initially suspected. Moreover, as R. sanguineus ticks bite humans, Astrakhan fever might be a cause of spotted fever in Kosovo.

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Pheromone-mediation of host-selection in bont ticks (Amblyomma hebraeum koch).

The bont tick, Amblyomma hebraeum, is the principal vector to southern African ruminants of heartwater (Cowdria ruminantium infection). The role of feeding male ticks, which emit an aggregation-attachment pheromone, in attracting unfed ticks to cattle was investigated. Calves infested with feeding male ticks were more attractive to unfed adult ticks than were uninfested calves. The presence of the pheromone on previously infested cattle apparently allows unfed ticks to discriminate between hosts on which these parasites have fed successfully (suitable hosts) and those on which they have not (potentially unsuitable hosts). The use of acaricides is thus unlikely to reduce bont tick populations in areas where adequate numbers of alternate (wild) hosts are present. Also, cattle so treated may lose their resistance to heartwater through lessened exposure to infected ticks.

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Prevalence of Borrelia burgdorferi sensu lato-infected ticks on migrating birds.

The prevalence of Lyme disease Borrelia-infected ticks on migrating birds was studied in Scandinavia. A total of 22,998 birds were caught at eight different bird observatories and examined for ticks. Five different species of ticks were found infesting the birds. The dominant species, Ixodesricinus, constituted 98.3% of the ticks collected. The presence of spirochetes was determined by an immunofluorescence assay of tick larvae and DNA amplification by PCR on all ticks. To determine which Borrelia burgdorferi sensu lato species were present, a species classification was performed by DNA amplification with species-specific 16S rDNA primers and by DNA sequencing (rDNA is DNA coding for rRNA). Flagellin gene sequences of all species of B. burgdorferi sensu lato previously recorded in Europe were observed. Borrelia garinii was the most prevalent Lyme disease Borrelia species in ticks collected from birds arriving from the South or Southeast in the spring, whereas the distribution was more heterogeneous in ticks from birds migrating from the Southwest. These data support the notion that birds are partly responsible for the heterogeneous distribution of Lyme disease Borrelia spirochetes in Europe.

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Interaction and transmission of two Borrelia burgdorferi sensu stricto strains in a tick-rodent maintenance system.

In the northeastern United States, the Lyme disease agent, Borrelia burgdorferi sensu stricto, is maintained by enzoonotic transmission, cycling between white-footed mice (Peromyscus leucopus) and black-legged ticks (Ixodes scapularis). B. burgdorferi sensu stricto is genetically variable and has been divided into three major genotypes based on 16S-23S ribosomal DNA spacer (RST) analysis. To better understand how genetic differences in B. burgdorferi sensu stricto may influence transmission dynamics in nature, we investigated the interaction between an RST1 and an RST3 strain in a laboratory system with P. leucopus mice and I. scapularis ticks. Two groups of mice were infected with either BL206 (RST1) or B348 (RST3). Two weeks later, experimental mice were challenged with the opposite strain, while control mice were challenged with the same strain as that used for the primary infection. The transmission of BL206 and B348 from infected mice was then determined by xenodiagnosis with uninfected larval ticks at weekly intervals for 42 days. Mice in both experimental groups were permissive for infection with the second strain and were able to transmit both strains to the xenodiagnostic ticks. However, the overall transmission efficiencies of BL206 and B348 were significantly different. BL206 was more efficiently transmitted than B348 to xenodiagnostic ticks. Significantly fewer double infections than expected were detected in xenodiagnostic ticks. The results suggest that some B. burgdorferi sensu stricto strains, such as BL206, may be preferentially maintained in transmission cycles between ticks and white-footed mice. Other strains, such as B348, may be more effectively maintained in different tick-vertebrate transmission cycles.

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Transcript heterogeneity of the p44 multigene family in a human granulocytic ehrlichiosis agent transmitted by ticks.

Human granulocytic ehrlichiosis (HGE) is an emerging tick-borne zoonosis caused by a strain of Anaplasma phagocytophila called the HGE agent, an obligatory intracellular bacterium. The agent expresses immunodominant 44-kDa outer membrane proteins (P44s) encoded by a multigene family. The present study established an experimental process for transmission of the HGE agent from infected mice (a reservoir model) to nymphal Ixodes scapularis ticks (a biological vector) and subsequently to horses (a patient model) by the adult infected ticks. Overall, a total of 20 different p44 transcripts were detected in the mammals, ticks, and cell cultures. Among them, a transcript from a p44-18 gene was major at acute stage in mice and horses but minor in ticks. Both mRNA and protein produced from the p44-18 gene were detected in the HGE agent cultivated in HL-60 cells at 37 degrees C, but their expression levels decreased in the organisms cultivated at 24 degrees C, suggesting that temperature is one of the factors that influence the expression of members of the p44 multigene family. Several additional p44 transcripts that were not detected in the mammals at the acute stage of infection were detected in ticks. Phylogenetic analysis of the 20 different p44 transcripts revealed that the major transcripts found in mammals and ticks were distinct, suggesting a difference in surface properties between populations of the HGE agent in different host environments. The present study provides new information for understanding the role of the p44 multigene family in transmission of the HGE agent between mammals and ticks.

Amino Acid Sequence↗

Characterization of spotted fever group rickettsiae in flea and tick specimens from northern Peru.

Evidence of spotted fever group (SFG) rickettsiae was obtained from flea pools and individual ticks collected at three sites in northwestern Peru within the focus of an outbreak of febrile disease in humans attributed, in part, to SFG rickettsia infections. Molecular identification of the etiologic agents from these samples was determined after partial sequencing of the 17-kDa common antigen gene (htrA) as well as pairwise nucleotide sequence homology with one or more of the following genes: gltA, ompA, and ompB. Amplification and sequencing of portions of the htrA and ompA genes in pooled samples (2 of 59) taken from fleas identified the pathogen Rickettsia felis. Four tick samples yielded molecular evidence of SFG rickettsiae. Fragments of the ompA (540-bp) and ompB (2,484-bp) genes were amplified from a single Amblyomma maculatum tick (tick 124) and an Ixodes boliviensis tick (tick 163). The phylogenetic relationships between the rickettsiae in these samples and other rickettsiae were determined after comparison of their ompB sequences by the neighbor-joining method. The dendrograms generated showed that the isolates exhibited close homology (97%) to R. aeschlimannii and R. rhipicephali. Significant bootstrap values supported clustering adjacent to this nodule of the SFG rickettsiae. While the agents identified in the flea and tick samples have not been linked to human cases in the area, these results demonstrate for the first time that at least two SFG rickettsia agents were circulating in northern Peru at the time of the outbreak. Furthermore, molecular analysis of sequences derived from the two separate species of hard ticks identified a possibly novel member of the SFG rickettsiae.

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Ability of the vector tick Boophilus microplus to acquire and transmit Babesia equi following feeding on chronically infected horses with low-level parasitemia.

The protozoan parasite Babesia equi replicates within erythrocytes. During the acute phase of infection, B. equi can reach high levels of parasitemia, resulting in a hemolytic crisis. Horses that recover from the acute phase of the disease remain chronically infected. Subsequent transmission is dependent upon the ability of vector ticks to acquire B. equi and, following development and replication, establishment of B. equi in the salivary glands. Although restriction of the movement of chronically infected horses with B. equi is based on the presumption that ticks can acquire and transmit the parasite at low levels of long-term infection, parasitemia levels during the chronic phase of infection have never been quantified, nor has transmission been demonstrated. To address these epidemiologically significant questions, we established long-term B. equi infections (>1 year), measured parasitemia levels over time, and tested whether nymphal Boophilus microplus ticks could acquire and, after molting to the adult stage, transmit B. equi to naive horses. B. equi levels during the chronic phase of infection ranged from 10(3.3) to 10(6.0)/ml of blood, with fluctuation over time within individual horses. B. microplus ticks fed on chronically infected horses with mean parasite levels of 10(5.5) +/- 10(0.48)/ml of blood acquired B. equi, with detection of B. equi in the salivary glands of 7 to 50% of fed ticks, a range encompassing the percentage of positive ticks that had been identically fed on a horse in the acute phase of infection with high parasitemia levels. Ticks that acquired B. equi from chronically infected horses, as well as those fed during the acute phase of infection, successfully transmitted the parasite to naive horses. The results unequivocally demonstrated that chronically infected horses with low-level parasitemia are competent mammalian reservoirs for tick transmission of B. equi.

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