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PubMed2001

Morphology and structural organization of gené's organ in Dermacentor reticulatus (Acari: Ixodidae).

Scanning and transmission electron microscopical investigations revealed that Genés organ in unfed and ovipositing females of Dermacentor reticulatus is formed as a double-sac-structure consisting of an outer epithelial and an inner cuticular sac. In ovipositing ticks the latter emerges through the camerostomal aperture to the exterior. Genés organ in unfed ticks consists of a corpus, two posterior horns and a pair of undeveloped glands at each side, which differentiate in ovipositing ticks to compound, branched tubular glands with a main efferent duct for each gland opening into the lumen between the epithelial and the cuticular sac. Genés organ of egg-laying females corresponds basically in morphology and structural organization to that of unfed ticks. Compared with unfed ticks, however, in ovipositing ticks the corpus and horns are longer and broader, the glands are fully developed and the cuticular sac is evertable. The epithelial sac as the outermost part of Genés organ is continuous with the hypodermis of the basis capituli and the scutum, arises at the camerostomal aperture, forms the corpus and the two blind-ending horns, passes into the epithelium of the main excretory ducts of the glands and envelops the cuticular sac. The cuticular sac passes into the cuticle of the basis capituli and the scutum, arises at the camerostomal aperture, is folded, expands into the horn tips and consists inwards of a smooth epicuticula and outwards of a fibrous endocuticula. Muscles originating from the scutum pass caudomedially through the epithelial sac and are inserted into the cuticular sac. The entire surface of the maximally everted cuticular sac is covered with an amorphous mass. In cleaned samples, ledge-like structures appear on the lateral surface. These ledges turn into balloon-like structures which extend over the medial and dorsal surface. The entire surface including the balloon-like structures and the ledges are provided with numerous cribrate pits.

Animals↗
PubMed2001

Cockroach allatostatin-like immunoreactivity in the synganglion of the American dog tick Dermacentor variabilis (Acari: Ixodidae).

Using immunocytochemistry based on a monoclonal antibody against Diploptera punctata allatostatin I and horseradish peroxidase-diaminobenzidine reaction, the presence of allatostatin-like immunoreactivity is demonstrated in the synganglion of Dermacentor variabilis females. The immunoreactive cells are located in the protocerebral, cheliceral, palpal, stomodeal, postesophageal, and opisthosomal regions of the synganglion. Strongly immunoreactive granules accumulate in the boundary area of the subganglia in the preesophageal part of the synganglion. This suggests that the immunoreactive materials may be released directly from there. In addition, a putative neurohemal area is found in the anterior area of the opisthosomal ganglion, where abundant immunoreactive materials are stored. Weak immunoreactivity and fewer immunoreactive cells are seen in newly molted females compared with one month old, unfed females. Thus, the immunoreactive products may be depleted during molting and synthesized in females before feeding.

Animals↗
PubMed2000

Host utilization and seasonal occurrence of Dermacentor species (Acari:Ixodidae) in Missouri, USA.

A total of 3,235 Dermacentor variabilis (Say) specimens were collected from birds, mammals, and by dragging vegetation, and 2,683 D. albipictus (Packard) ticks were collected from deer from 1993 to 1996. Peak seasonal occurrence of adult D. variabilis was from May through July with a precipitous decrease in August. Nymphal D. variabilis populations peaked in June. Peak activity of larvae was bimodal, with one activity peak during late summer (September) and a second peak in winter or early spring. The raccoon, Procyon lotor (L.), was the principal host of adults followed by the Virginia opossum, Didelphis virginiana Kerr. Rodents and the eastern cottontail rabbit, Sylvilagus floridanus (J. A. Allen), were the primary hosts of nymphs. The marsh rice rat, Oryzomys palustris (Harlan), was the principal host of larvae followed by the pine vole, Microtus pinetorum (Le Conte), and white-footed mouse, Peromyscus leucopus (Rafinesque). All stages of D. albipictus were found only on white-tailed deer, Odocoileus virginianus (Zimmermann). Numbers of adult and nymphal D. albipictus peaked in November, whereas larvae peaked in September.

Animals↗
PubMed2003

Ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum ticks to acquire and transstadially transmit Borrelia garinii.

In China, the ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum to transmit Borrelia spirochetes was determined under laboratory conditions. Results showed that all three tick species can acquire spirochetes by feeding on infected mice. However, the capability of the three species to maintain spirochetes was different. Only I. persulcatus is able to maintain spirochetes alive during molting, and subsequent tick stages transmitted the infection to naive mice. In H. concinna and D. silvarum ticks, spirochetes could not survive after the digestion period of blood and after the molting process was finished, spirochetes were no longer detected. Contrary to H. concinna and D. silvarum, I. persulcatus should be considered the principal vector of Lyme disease in north China.

Animal Feed↗
PubMed2001

Seasonal activity and spatial distribution of host-seeking adults of the tick Dermacentor variabilis.

The seasonal temporal and spatial distribution and emergence of overwintering and summer cohorts of host-seeking American dog tick, Dermacentor variabilis (Say) (Acari: Ixodidae), adults were examined for 5 years in the Bluegrass region of Kentucky. Weekly drag cloth sampling was conducted from early March until late September 1992-1995 and early March until the end of May 1996. Overwintering adult cohorts became active in early to mid-April followed by bimodal host-seeking activity each year, with the duration and intensity of the first activity period being related to the previous year's summer cohort activity. The first activity peak extended from mid-April through late May and the second activity peak occurred in July. All host-seeking activity ceased by early August, except in 1995, when activity ended on 22 August. Laboratory survival of adults, collected weekly during 1993-1996, was used to estimate the emergence of summer adult cohorts. Summer adult cohorts became active in late May or early June and overlapped with overwintering adult host-seeking activity. The termination of host-seeking activity in overwintering adults could not be determined using the methods employed in this study and is most likely to have continued for the entire season. Host-seeking adults displayed contagious distribution patterns along trails bordering ecotone habitats, as determined by fitting a negative binomial frequency distribution to weekly sampling data. These patterns did not vary from year to year. In addition, the location of D. variabilis activity did not change from year to year, which suggests that microhabitats within apparently homogeneous environments enhance adult survival and lead to 'hot spots' of tick activity despite the ability of vertebrate hosts to disperse D. variabilis populations more widely. These data better define our understanding of D. variabilis adult seasonal and spatial host-seeking activity, estimate for the first time the intensity of summer cohort host-seeking activity, and relate summer cohort activity with spring activity the following year.

Animals↗
PubMed2003

Dynamics of Rickettsia-tick interactions: identification and characterization of differentially expressed mRNAs in uninfected and infected Dermacentor variabilis.

To begin to explore the molecular dynamics of rickettsial tick interaction, subtractive hybridization was used to screen mRNAs in Rickettsia montanensis-infected and uninfected Dermacentor variabilis. We isolated 30 cDNA fragments, 22 of which were up-regulated and eight were down-regulated in response to rickettsial infection. Based on a putative identity of 11 cDNA fragments with similarity to known protein families, the tick genetic factors have been assigned into three groups including, the tick immune response factors (alpha-2 macroglobulin and IgE-dependent histamine release factor), the receptor/adhesion molecules (the signal transducer and activator of transcription-1/3 protein inhibitor, the clathrin adaptor protein and tetraspanin) and the stress response proteins (aldose reductase, glutathione-S transferase, ferritin, nucleosome assembly protein and cyclin A protein). Density analyses of semiquantitative RT-PCR amplified products demonstrated differential expression for 18 of the 23 tested genetic factors, apparently representing a 78% agreement with results obtained by subtractive hybridization. Additionally, mRNA transcripts of 17 of the 23 tested genetic factors were amplified from tick haemocytes/circulatory cells demonstrating that their expression is not restricted to the ovaries and suggesting a potential involvement in the immune response.

Animals↗
PubMed2003

Characterization of a recombinant immunomodulatory protein from the salivary glands of Dermacentor andersoni.

The gene encoding a 36-kDa (p36) immunomodulatory protein present in saliva of Dermacentor andersoni was cloned in prokaryotic and eukaryotic expression vectors. A polymerase chain reaction (PCR)-generated cDNA lacking signal peptide was cloned into the Escherichia coli expression vector pET28 and a similar sequence was cloned into pIB/V5-His-TOPO expression vector for stable transfection of insect cells, High 5 trade mark. The 26-kDa molecular mass of p36 expressed by bacteria is in agreement with that predicted from the translated full-length cDNA sequence. Eukaryotic-cell-expressed p36 consisted of multiple forms with molecular masses between 34 and 36 kDa. These multiple forms were attributed to differences in post-translational modifications. N-linked mannose was detected on insect-cell-expressed and tick-derived p36. Multiple bands remained after endoglycosidase removal of N-linked sugars, indicating the presence of other modifications. Both bacterial- and insect-cell-expressed p36 reacted on immunoblots with polyclonal antibodies raised against tick-derived p36. Insect-cell-expressed p36 suppressed T-lymphocyte-mitogen-driven in vitro proliferation of splenocytes from tick-naïve mice in a dose-dependent manner. Bacterial-cell-expressed p36 lacked immunomodulatory activity.

Animals↗
PubMedDate not supplied

The ultrastructure of Rickettsia slovaca in naturally infected females of the tick Dermacentor marginatus.

The ultrastructure of Rickettsia slovaca in naturally infected females of the tick Dermacentor marginatus is similar to that of other members of the spotted fever group rickettsiae. R. slovaca was found to multiply in all organs, tissues and cells examined. It is primarily found free in the cytoplasm; occasionally it may also invade the cell nucleus. Special attention was paid to the rickettsiae in the salivary gland. They multiply in all cells of the three acini types I-III and also in the duct cells. The amount of rickettsiae eliminated via the saliva reached values of about 10(4)EID50/ml.

Animals↗
PubMed1996

Variabilin, a novel RGD-containing antagonist of glycoprotein IIb-IIIa and platelet aggregation inhibitor from the hard tick Dermacentor variabilis.

A novel inhibitor of human platelet aggregation, named variabilin, was isolated from salivary glands of the hard tick Dermacentor variabilis using a combination of gel filtration and high pressure liquid chromatography. Variabilin was a potent antagonist of the fibrinogen receptor glycoprotein IIb-IIIa (GPIIb-IIIa; alphaIIbbeta3) and the vitronectin receptor alphavbeta3. Amino acid sequence analysis by Edman degradation revealed that it has 47 residues, with a molecular weight of 4968.5. Like many other naturally occurring antagonists of GPIIb-IIIa, variabilin contains the RGD (Arg-Gly-Asp) motif. However, unlike the RGD-containing antagonists of GPIIb-IIIa, the RGD sequence of variabilin is not positioned in a loop bracketed by cysteine residues. It has little sequence homology to the other known naturally occurring antagonists of GPIIb-IIIa, including the disintegrins from snakes, decorsin and ornatin from leeches, and disagregin from soft ticks. Variabilin is the first RGD-containing antagonist isolated from ticks.

Amino Acid Sequence↗
PubMed2001

Dermacentor andersoni: effects of repeated infestations on lymphocyte proliferation, cytokine production, and adhesion-molecule expression by BALB/c mice.

The effects of repeated infestations with Dermacentor andersoni nymphs on the lymphocyte functions of BALB/c mice were investigated. The in-vitro proliferation responses to concanavalin-A or salivary-gland molecules, the production of cytokines, and the expression of two adhesion molecules-leucocyte function-associated antigen-1 (LFA-1) and very late activation-4 (VLA-4)-were all studied. In addition, the ability of salivary-gland extract or saliva from D. andersoni to modulate expression of lymphocyte adhesion molecules in vitro was determined. The proliferative responses of T-lymphocytes to concanavalin-A were significantly suppressed after first and second infestations, and significant increases in lymphocyte proliferation in the presence of tick salivary-gland antigen were observed in infested mice. After two infestations, production of interleukin-2 was significantly decreased but that of interferon-gamma remained unchanged. Production of interleukin-4 and interleukin-10 was significantly enhanced in infested mice after both the first and second infestations. Expression of LFA-1 and VLA-4 by lymphocytes from infested mice was suppressed. Furthermore, both a salivary-gland extract and the saliva of D. andersoni reduced the in-vitro expression of both of these adhesion molecules by lymphocytes from tick-naive mice.

Analysis of Variance↗
PubMed2001

Infection and transovarial transmission of rickettsiae in Dermacentor variabilis ticks acquired by artificial feeding.

In this study we examined the efficiency of an in vitro feeding technique using glass microcapillaries as a method of establishing rickettsiae-infected lines of ticks. To quantify the volume ingested by ticks during microcapillary feeding, the incorporation of radiolabeled amino acids in tick gut and hemolymph was calculated. Fifteen of 18 ticks consumed between 0.06 and 6.77 microl. However, ingestion of fluid was not correlated to weight gain during capillary feeding. Uninfected and partially fed laboratory-reared female Dermacentor variabilis ticks were exposed to either Rickettsia montana- or Rickettsia rhipicephali-infected Vero cells via microcapillary tubes, returned to rabbit hosts, and allowed to feed to repletion. All tissues collected from ticks allowed to feed overnight on rickettsiae-infected fluids were found to be infected when examined by IFA. When rickettsiae-infected and uninfected capillary-fed ticks were allowed to feed to repletion and lay eggs, no significant differences in mean engorgement weight or fecundity was observed. When we assessed the efficiency of transovarial transmission of rickettsiae by ticks that imbibed rickettsiae-infected cells by polymerase chain reaction (PCR) and IFA, infection was detected by PCR in the eggs from 85% of the ticks exposed to R. montana and 69% of the ticks exposed to R. rhipicephali. Rickettsial genes were not amplified in samples of the uninfected controls. Examination by IFA of egg samples from females exposed to rickettsiae-infected cells identified rickettsiae in 100% of the samples tested, while the uninfected controls were negative.

Amino Acids↗
PubMed2005

Babesia DNA detection in canine blood and Dermacentor reticulatus ticks in southwestern Siberia, Russia.

Babesia infection was studied in 21 blood samples of dogs with symptoms of babesiosis and among 72 Dermacentor reticulatus and 70 Ixodes persulcatus ticks from southwestern Siberia, Russia. Babesia DNA was detected by hemi-nested PCR based on the 18S rRNA gene with subsequent direct sequencing. All of the analyzed canine blood samples and three D. reticulatus, but none from I. persulcatus ticks studied were shown to contain Babesia DNA. Nucleotide sequences of the Babesia 18S rRNA gene fragment of 354 bp long for all 24 positive samples appeared to belong to the subspecies Babesia canis canis and differed only at three positions. The Babesia nucleotide sequences from 17 canine blood samples and from one D. reticulatus tick were identical to each other and to previously known B. canis canis from canine blood in Slovenia. Four canine blood samples and the second tick sample contained a mixture of two nucleotide sequences previously found in canine blood. B. canis canis nucleotide sequence from the third tick differed in the unique nucleotide transition and could correspond to a new genetic variant. Thus, the main etiological agent of canine babesiosis in Novosibirsk region is B. canis canis, and D. reticulatus, but not I. persulcatus, ticks could serve as a vector of this infectious agent. To our knowledge, this is the first report of the B. canis canis nucleotide sequences from ticks.

Animals↗
PubMed1989

Species recognition elicited by differences in composition of the genital sex pheromone in Dermacentor variabilis and D. andersoni (Acari: Ixodidae).

The chemical basis for the ability of fed males of Dermacentor variabilis (Say) and D. andersoni (Stiles) to differentiate conspecific and heterospecific females was investigated using a neutered female bioassay. Male D. variabilis responded strongly in a dose-response manner to increasing concentrations of stearic and palmitic acids and an extract of the anterior reproductive tract (ART) of fed D. variabilis females. Mating responses were greatest at 1 microgram of fatty acid and 1 female equivalent (FE) of ART extract. D. variabilis males also responded strongly to D. variabilis females treated with a conspecific ART extract and a mixture of fatty acids simulating the extract; responses were moderate to D. andersoni females treated with the same extract or fatty acid mixture. They also responded moderately to conspecific females treated with D. andersoni extract. Male D. andersoni displayed moderate increases in mating behavior over a range of concentrations of stearic and palmitic acids and conspecific ART extract. D. andersoni males mated only with D. andersoni females treated with a conspecific extract or a mixture of fatty acids simulating the extract. Mating by D. andersoni did not occur in response to D. variabilis females treated with D. andersoni or D. variabilis ART extracts. Stearic acid alone or in combination with 20-hydroxyecdysone, oleic, or myristic acid did not account for the specificity of response of either species. D. andersoni males were much less responsive to fatty acids, alone or in mixtures, than D. variabilis males. These findings indicate that fatty acids are important in mate recognition by D. variabilis, but explain only part of that by D. andersoni. The possibility is raised that another compound is required for mate recognition by D. andersoni.

Animals↗
PubMed1990

Metabolism of juvenile hormone during adult development of Dermacentor variabilis (Acari: Ixodidae).

Juvenile hormone (JH)-I and -III were used as model substrates to study the in vitro metabolism of JH in the hemolymph and body homogenates of the American dog tick, Dermacentor variabilis (Say). Ester hydrolysis was the principal pathway of JH metabolism in hemolymph and homogenates. JH also was converted into JH-diol primarily by body homogenates, indicating the presence of JH epoxide hydrolase activity. JH epoxide hydrolase activity, alpha-naphthyl acetate esterase activity, and protein concentration per milligram wet weight were significantly lower (t test, alpha = 0.05) in homogenates of partially fed, virgin and replete, mated females of D. variabilis compared with unfed, virgin females. The decline in these factors was probably because of the influx of water into the tissues caused by the blood meal. In addition, the epoxide hydrolase and alpha-naphthyl acetate esterase activity per milligram tissue protein decreased significantly during this time. Mating of fed females rather than feeding alone caused a significant decline in the tissue JH esterase activity per milligram wet weight but not per milligram protein. The JH esterase activity per milligram protein was significantly higher in partially fed, virgin and replete, mated females compared with unfed females, indicating that feeding may actually increase JH esterase activity on a protein basis. JH-III was metabolized 1.4 times faster than JH-I by the hemolymph of partially fed, virgin females. The inhibitors O,O-diisopropyl phosphorofluoridate and octylthio-1,1,1-trifluoropropan-2-one at 10(-4) M inhibited JH and alpha-naphthyl acetate esterase activity in hemolymph and body homogenate.

Animals↗
PubMed1990

Fecundity of partially engorged female Dermacentor albipictus (Acari: Ixodidae) removed by cattle grooming.

Eight Hereford heifer calves were infested 28 times over a 64-d period with approximately 49,560 Dermacentor albipictus (Packard) larvae per calf. The fecundity of partially engorged and small, fully engorged female ticks, prematurely removed by grooming behavior of the calves, were compared. A total of 3,532 females of various engorged weights was collected from all calves, resulting in a mean female tick yield of 1.78% based on the number of larvae used for all infestations. The ratio of partially to fully engorged females was 1:16. Female oviposition success was 96%. The mean (+/- SE) values for female weight, number of larvae, and percentage larval eclosion of 50 partially engorged females that ranged in weight from 54 to 397 mg was 174 +/- 99, 1,272 +/- 931 and 85 +/- 14, respectively. The correlation between preoviposition weight and number of larvae from partially engorged females was highly significant (r = 0.95, P less than 0.0001). A prediction model (Y = -354.6 + 9.07X) showed that the preoviposition weight of partially engorged females and larval fecundity from a separate study was highly correlated (r = 0.92, P less than 0.0001). There was no significant difference between mean values of larval numbers and percentage eclosion for partially engorged and small, fully engorged females of similar weight.

Animals↗
PubMed1990

Micrometeorologic factors affecting field host-seeking activity of adult Dermacentor variabilis (Acari: Ixodidae).

Fifteen micrometeorologic or microenvironmental parameters, including temperature, moisture, wind, and solar radiation, were repeatedly measured at 1 m and 2.5 cm aboveground, in the litter layer, and in the soil concurrently with drag samples of questing American dog tick, Dermacentor variabilis (Say), in five plots (1 by 10 m) in Delaware County, Ohio. Multivariate statistical analyses of the resultant data showed that ambient temperature was the best general predictor of adult tick host seeking under the observed ranges of the observed parameters. Multivariate procedures included forward stepwise multiple regressions and principal components analyses. Solar radiation was covariant with ambient temperature but was much less predictive. A suppression of questing activity with increased ambient temperature was evident at the highest observed temperatures, implying an upper temperature limit for this activity.

Analysis of Variance↗
PubMed1990

Hispid cotton rats (Sigmodon hispidus) as a source for infecting immature Dermacentor variabilis (Acari: Ixodidae) with Rickettsia rickettsii.

Hispid cotton rats (Sigmodon hispidus Say and Ord) were susceptible to infection with Rickettsia rickettsii Wolbach under laboratory conditions and were capable of serving as sources for infecting ticks with rickettsiae. Cotton rats developed rickettsemias that could be detected for as long as 6 h following intraperitoneal inoculation of 10(5) plaque-forming units (PFUs) of R. rickettsii (Morgan strain). An estimate of the median infectious dose (ID50) was made by inoculating six groups (eight animals per group) with serial dilutions of 10(1) to 10(6) PFUs. In this experiment, cotton rats became infected after exposure to relatively few organisms (ID50 = 37 PFUs). None of the 48 cotton rats succumbed to infection, and only 6 of the 48 rats failed to seroconvert. Dermacentor variabilis (Say) larvae were fed on rickettsemic cotton rats to determine whether this species could serve as a source for infecting ticks with R. rickettsii. A small percentage (0.9-3.0%) of larval ticks that fed on three of the four cotton rats had R. rickettsii in their tissues when examined as nymphs. A fourth cotton rat died 7 d after inoculation with R. rickettsii and infected a much higher percentage of ticks (64.0%) than the other three animals.

Animals↗
PubMed1990

Cytosystematics of five North American Dermacentor (Acari: Ixodidae) species.

The karyotypes of five species of Dermacentor ticks found in North America were examined for the presence and location of constitutive heterochromatin (C-bands) and the location of the nucleolar organizing regions (NORs). All specimens examined had 20 autosomes and an XX:XO sex-determining system; the X was the largest element in the karyotype. Except as noted, the autosomes of all species were acrocentric, and heterochromatin was restricted to the centromeric region. The one-host species D. nitens and D. albipictus were distinguished from the three-host species by a lack of additional bands of heterochromatin and by having a subtelocentric X chromosome. However, some individuals of D. albipictus were observed to have acrocentric X chromosomes. D. nitens differed from D. albipictus in having two subtelocentric autosomal pairs (pairs 1 and 8). A single pair of NORs was noted for the one-host species; however, heterochromatin was not associated with the NORs. Among the three-host species, D. occidentalis showed a band of heterochromatin proximal to the centromere in autosomal pairs 1 and 8, and D. andersoni had similar bands in autosomal pairs 1, 3, 7, and 9. D. variabilis displayed heterochromatic bands in autosomal pairs 3, 7, and 9. All three-host species displayed a single pair of NORs, which was associated with the band of heterochromatin on pair 9. Indications of some genetic incompatibility between the two recognized forms of D. albipictus are discussed.

Animals↗