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[New molecular biology detection methods for tick-borne infectious agents].

Tick-borne zoonotic pathogens are well known in many areas all over the world. Among the tick-borne transmitted diseases in Switzerland, Lyme disease caused by Borrelia burgdorferi, ehrlichiosis caused by various species of Ehrlichia and tick-borne encephalitis caused by the tick-borne encephalitis virus (TBEV) are the most important zoonotic diseases. Early diagnosis and treatment is necessary to prevent fatal infections and chronic damage to various tissues. Due to the variety of uncharacteristic clinical signs, tick-borne diseases are not easily recognized. Diagnosis is based on clinical findings, a record of tick exposure, and direct or indirect detection of the pathogen. Here we discuss briefly the most important tick-borne infections and their diagnosis with emphasis on a new molecular diagnostic tool--the real-time TaqMan PCR--and its importance for the diagnosis of tick-borne pathogens.

Animals↗

Effects of infection of the tick Ornithodoros moubata with African swine fever virus.

The effects of infection with African swine fever virus (ASFV) on adult and nymphal Ornithodoros moubata Murray (Ixodoidea, Argasidae) ticks were examined. Three groups of ticks were used, an uninfected control group, one group infected with the VIC T90/1 isolate of ASFV and another group infected with the LIV 13/33 isolate of ASFV. Infection with ASFV did not affect the oviposition rates of infected ticks when compared with uninfected ticks. There was no difference between infected and uninfected ticks in progeny hatching rates and first nymphal stage feeding rates. Feeding rates of infected adult ticks were also unaffected. However, a significant increase in mortality rates was observed amongst the adult ticks that fed on an infective bloodmeal compared to ticks fed on an unifected bloodmeal.

African Swine Fever↗

Relationship between tick bites and the seroprevalence of Babesia microti and Anaplasma phagocytophila (previously Ehrlichia sp.) in blood donors.

BACKGROUND: Tick-borne diseases, particularly babesiosis and ehrlichiosis, represent recently emerging infections. Despite an increased recognition of the threat tick-borne agents pose to blood safety, our understanding of the prevalence and transmissibility of these agents in blood donors is limited. STUDY DESIGN AND METHODS: Babesia microti and Anaplasma phagocytophila (previously Ehrlichia sp.) seroprevalence was determined in random Connecticut and Wisconsin donors, and subsequently in Connecticut donors reporting tick bites. In the interim, a postcard survey regarding tick bites during the previous 6 months was sent to 6,000 random donors in six geographically distinct collection regions. RESULTS: In total, 3 of 999 Wisconsin donors (0.3%) and 6 of 1,007 Connecticut donors (0.6%) had antibodies to B. microti. Of 992 donors tested for A. phagocytophila, 5 Wisconsin donors (0.5%) and 35 Connecticut donors (3.5%) were seropositive. A total of 2,482 donors (41.4%) completed the survey; 103 (4.1%) reported a tick bite. Of 848 Connecticut donors (0.4%) reporting tick bites, 3 had B. microti antibodies, while 8 (0.9%) had A. phagocytophila antibodies. These rates were not significantly different from control donors. CONCLUSION: Blood donors seropositive for B. microti and A. phagocytophila are present in Connecticut and Wisconsin. Donors readily recall previous tick bites, but self-reported bites are not reliable indicators of serologic status. The exposure of blood donors to tick-borne pathogens does suggest a need to better understand the transfusion transmission potential of these agents.

Anaplasma phagocytophilum↗

Ticks and tickborne bacterial diseases in humans: an emerging infectious threat.

Ticks are currently considered to be second only to mosquitoes as vectors of human infectious diseases in the world. Each tick species has preferred environmental conditions and biotopes that determine the geographic distribution of the ticks and, consequently, the risk areas for tickborne diseases. This is particularly the case when ticks are vectors and reservoirs of the pathogens. Since the identification of Borrelia burgdorferi as the agent of Lyme disease in 1982, 15 ixodid-borne bacterial pathogens have been described throughout the world, including 8 rickettsiae, 3 ehrlichiae, and 4 species of the Borrelia burgdorferi complex. This article reviews and illustrate various aspects of the biology of ticks and the tickborne bacterial diseases (rickettsioses, ehrlichioses, Lyme disease, relapsing fever borrelioses, tularemia, Q fever), particularly those regarded as emerging diseases. Methods are described for the detection and isolation of bacteria from ticks and advice is given on how tick bites may be prevented and how clinicians should deal with patients who have been bitten by ticks.

Animals↗

Tick infestations on Zebu cattle in western Kenya: individual host variation.

Monthly collections were made of all ticks from 25 mature East African Zebu cattle and 5 yearlings for 13 mo on Rusinga Island, Lake Victoria, Kenya, from September 1986 to September 1987. The most common species were Rhipicephalus appendiculatus Neumann and Amblyomma variegatum F.; Boophilus decoloratus Koch and Rhipicephalus evertsi evertsi Neumann also were present in fewer numbers. Cattle were ranked according to their degree of resistance to each of the four species and to each stage in the life cycle of the three-host ticks. Ranking according to infestations of females and nymphs of R. appendiculatus, nymphs and larvae of A. variegatum, or all three stages of these two species when compared with the total burden gave statistically significant correlations. Sampling difficulties prevented the ranking of cattle for infestations of larvae of R. appendiculatus, although the numbers on cattle were high. On the other hand, failure of attempts to rank the cattle on females of A. variegatum, B. decoloratus, or R. e. evertsi counts was attributed to the small differences between tick numbers on cattle. Highly resistant cattle showed little or no seasonal fluctuations in tick numbers for most of the period compared with animals of low resistance, which showed an up to sevenfold increase in the magnitude of the tick burden when the tick challenge was high. Cattle with low resistance were responsible for much of the tick multiplication in the field, whereas highly resistant animals tended to limit the populations. There were no distinct differences between the numbers of each tick species on yearlings and mature cattle.

Animals↗

Experiments on the relationship between feeding of the tick Amblyomma variegatum (Acari: Ixodidae) and dermatophilosis skin disease in sheep.

Experiments on the systemic effect of feeding adult Amblyomma variegatum (F.) on dermatophilosis caused by Dermatophilus congolenis in six sheep were performed by feeding ticks at sites separate from sites of experimental infection with D. congolensis. Comparisons were made with D. congolensis infections on six control sheep without ticks. Tick-infested sheep developed chronic dermatophilosis lesions; tick-free sheep healed rapidly. On two tick-infested and two control sheep a chronological sequence of D. congolensis infection sites were made for histological comparisons. Dermatophilosis lesions on tick-infested sheep were characterized by chronic cellular infiltration and accumulation of plasma cells. Lesions on control sheep healed in association with transient cellular infiltration including plasma cells. In neither test nor control sheep was there evidence of effective phagocytosis of D. congolensis. It was concluded that feeding of these ticks had a systemic effect on the host such that dermatophilosis was aggravated via an effect on immunological reactions in the skin. Control of dermatophilosis by tick control is advocated.

Actinomycetales Infections↗

Salivary gland changes and host antibody responses associated with feeding of male lone star ticks (Acari:Ixodidae).

The goal of this study was to demonstrate that male lone star ticks, Amblyomma americanum (L.), actively feed on rabbits during attachment and that the host is capable of mounting an immune response against male salivary gland proteins. During attachment, it was shown that male ticks salivary glands hypertrophy. An enzyme-linked immunosorbent assay was used to detect rabbit serum proteins in the midgut of previously attached male ticks. SDS-polyacrylamide gel electrophoresis showed that the proteins expressed in the male tick salivary gland changed during feeding, with several new proteins in the 15- to 50-kDa range synthesized during attachment. Rabbits mounted a detectable antibody response against male tick salivary gland proteins after 2 sequential feedings of male ticks. The antibodies were directed against a spectrum of male salivary gland proteins ranging from 18 to 160 kDa. Several of these proteins were not recognized by antibodies directed against female tick saliva proteins, and thus may be specific male salivary gland components. This evidence indicates that male A. americanum ticks actively feed during attachment, that their salivary gland proteins change during feeding, and that male salivary gland proteins are immunogenic.

Animals↗

The effects of the brown ear-tick, Rhipicephalus appendiculatus, on milk production of Sanga cattle.

Lactating Sanga cows of the Mashona breed from Zimbabwe, receiving either a low or high level of nutritional supplement, were exposed to two levels of infestation of adults of the brown ear-tick, Rhipicephalus appendiculatus in the highveld of Zimbabwe. The effect of the ticks on the milk yield was measured over an 11-week period during the rainy season from January to April 1986. A technique in which calves were weighed before and after suckling was used to estimate milk yield. There were significant treatment effects of ticks (P < 0.05) on milk production but no significant differences in liveweight gain between calves from tick-free and tick-infested dams. The loss in milk production was poorly related to the number of female ticks that engorged, being 9 g (SEM 4) per tick. A Friesian x Hereford (Bos taurus) reference group of cattle carried 50% more ticks than the Mashona cows, illustrating a difference in resistance between the breeds. Thirteen screw-worm (Chrysomya bezziana) strikes were recorded amongst the thirty-two Mashona cows compared with twenty-one amongst the ten Friesian x Hereford animals between January and the end of March.

Animals↗

Tick saliva: recent advances and implications for vector competence.

Secretions of the tick salivary glands are essential to the successful completion of the prolonged feeding of these ectoparasites as well as the conduit by which most tick-borne pathogens are transmitted to the host. In ixodid ticks the salivary glands are the organs of osmoregulation, and excess water from the bloodmeal is returned via saliva into the host. Host blood must continue to flow into the feeding lesion as well as remain fluid in the tick mouthparts and gut. The host's haemostatic mechanisms are thwarted by various anti-platelet aggregatory, anticoagulatory and anti-vasoconstrictory factors in tick saliva. Saliva components suppress the immune and inflammatory response of the host permitting the ticks to remain on the host for an extended period of time and, adventitiously, enhancing the transmission and establishment of tick-borne pathogens. Over the years much work has been done on the numerous enzyme and pharmacological activities found in the tick saliva. The present article reviews the most recent work on salivary gland secretions with special emphasis on how they favour pathogen transmission.

Animals↗

Seasonal occurrence of Rhipicephalus sanguineus in Okayama Prefecture, Japan and effect of temperature on development of the tick.

The seasonal occurrence of Rhipicephalus sanguineus on dogs was examined at a kennel in Okayama Prefecture, Japan. The number of ticks suddenly decreased after treatment with an acaricide in late August. Small numbers of adults and nymphs were detected in September and October, then ticks were not seen on the dogs early in November, when the mean temperature was below 15 degrees C. Then 3 dogs were found to be infested by some adult ticks toward the end of March, when the mean temperature was about 11 degrees C. The effects of temperature on the oviposition and the development of the tick were examined under laboratory conditions. The larval and nymphal post parasitic period, the pre-oviposition period and the oviposition period were prolonged when the temperature was decreased from 37 to 23 degrees C. The oviposition period was extremely long at 14 degrees C, but the tick could not develop below 14 degrees C. No eggs hatched below 14 degrees C. The ability to attach an engorge of adult ticks was examined under cold conditions. Unengorged adult ticks could attach to rabbits on the ear which were kept in an outdoor kennel in October, November and March, however they could not engorge completely in November. They could not attach on rabbits from December to February. The longevity of the tick was also examined under low temperatures. Unengorged adults could attach and engorge on rabbits after kept at 12 degrees C with 50% relative humidity (RH) for 140 days or 12 degrees C with 50% RH for 40 days followed by 4 degrees C with 50% RH for 100 days. These findings suggest that R. sanguineus could be established in Okayama Prefecture under optimum condition.

Animals↗

Tick-borne pulmonary disease: update on diagnosis and management.

Ticks are capable of transmitting viruses, bacteria, protozoa, and rickettsiae to man. Several of these tick-borne pathogens can lead to pulmonary disease. Characteristic clinical features, such as erythema migrans in Lyme disease, or spotted rash in a spotted fever group disease, may serve as important diagnostic clues. Successful management of tick-borne diseases depends on a high index of suspicion and recognition of their clinical features. Patients at risk for tick bites may be coinfected with two or more tick-borne pathogens. A Lyme vaccine has recently become available for use in the United States. Disease prevention depends on the avoidance of tick bites. When patients present with respiratory symptoms and a history of a recent tick bite or a characteristic skin rash, a differential diagnosis of a tick-borne pulmonary disease should be considered. Early diagnosis and appropriate antibiotic therapy for these disorders lead to greatly improved outcomes.

Animals↗

Effectiveness of a repellent containing DEET and EBAAP for preventing tick bites.

OBJECTIVE: Topical repellents can provide effective personal protection from tick-borne diseases by preventing the attachment of ticks. The goal of this study was to assess the effectiveness of a commercially available repellent spray containing both N,N-diethyl-3-methylbenzamide, previously known as N,N-diethyl-m-toluamide (DEET), and ethyl-butylacetylaminopropionate (EBAAP) against tick bites in a population at risk in Switzerland under real-life conditions. METHODS: The effectiveness of an insect repellent spray containing both DEET and EBAAP was evaluated in a randomized, double-blind, placebo-controlled field study. The study, requiring simple application of the repellent to exposed skin, was carried out on 276 forestry workers and orienteers under everyday conditions in Switzerland from May to September 1999. We measured total effectiveness of the repellent by the following formula: percentage effectiveness = 100 x (T(P) - T(R))/T(P), where T(P) and T(R) were the average number of ticks per hour spent in wooded areas for the repellent and placebo groups, respectively. RESULTS: The average number of tick bites per hour of exposure to wooded areas differed significantly between the placebo (n = 138) and repellent (n = 138) groups, 0.17 vs 0.10 (P < .05). Total repellent effectiveness against tick attachment was 41.1% (95% CI, 2.5-79.6). On the arms, an effectiveness of 66% (95% CI, 17.3-114.7) was observed. No significant difference in the average number of unattached ticks could be found. CONCLUSIONS: This study found that an easily applied repellent is moderately effective in reducing the risk of tick bites.

Administration, Cutaneous↗

Effects of pyriproxyfen on off-host water-balance and survival of adult lone star ticks (Acari: Ixodidae).

Newly engorged nymphs of the lone star tick, Amblyoma americanum (L.), were continuously exposed to 4 microg/cm2 of pyriproxyfen residues in glass vials. Treatment of engorged nymphs (n = 285) resulted in significant molting inhibition, with more than one-fourth (26.7%, n = 76) of nymphs dying before or during ecdysis. Treatment effects were evident among ticks that molted to the adult stage, with 26.7% (n = 76) of females, and 17.9% (n = 51) of males exhibiting moribund physical characteristics (i.e., lethargy; dull, discolored and desiccated cuticles; lacking full locomotor competency). A few molted adult ticks (10 males, four females) were dead upon inspection. Only 11.2% of pyriproxyfen treated, emergent females (n = 32), and 11.5% of treated emergent males (n = 25) from 285 ticks treated as engorged nymphs, exhibited normal physical appearance and possessed a full range of locomotor activity. Treated adult ticks maintained within a desiccating environmental chamber at 0% RH and 23 degrees C, had significantly accelerated whole-body water loss rates in comparison to untreated males and females maintained under the same environmental conditions. Additionally, treated adult ticks maintained under optimal environmental conditions (23 degrees C and >95% RH) sustained 100% mortality within 32 d following assignment to these conditions (or 79 d posttreatment as engorged nymphs), whereas untreated ticks had 0% mortality for the same duration of time. Results demonstrate that continuous exposure of nymphs to pyriproxyfen disrupted molting, and accelerated both whole-body water loss and subsequent mortality among emergent adult ticks.

Animals↗

Experimental infection of lone star ticks, Amblyomma americanum (L.), with Rickettsia parkeri and exposure of guinea pigs to the agent.

The maculatum agent, Rickettsia parkeri (a member of the spotted fever group rickettsiae), was inoculated into a colony of the lone star tick, Amblyomma americanum, and followed for two tick generations. In addition, guinea pigs were exposed to the agent by direct injection and by feeding infected ticks on them. Eighty (53%) of 150 nymphal A. americanum that were inoculated with suspensions of R. parkeri were positive by hemolymph test and fluorescent antibody test for rickettsial infection when examined as adults. One-month survival of R. parkeri-infected ticks was similar to that of control (noninfected) ticks. Transstadial and transovarial transmission of R. parkeri in the laboratory was demonstrated in A. americanum. When guinea pigs were exposed to the maculatum agent by either direct injection of Vero cell-grown R. parkeri, injection of homogenates of infected ticks, or by feeding of infected ticks, they developed mild fevers and occasional scrotal reactions. These data indicate that R. parkeri can remain viable in lone star ticks for two generations and suggest that guinea pigs may become infected, displaying mild clinical signs.

Animals↗

Influence of sheep breed and application site on the efficacy of a flumethrin pour-on formulation against ticks.

The objectives of this study were to determine the influence of application site and sheep breed on the efficacy of a flumethrin (1% m/v) solution for the control of 'bont'-legged (Hyalomma spp.) and red-legged ticks (Rhipicephalus evertsi evertsi). This study was conducted from November 1996 to February 1997 on 3 farms in the southwestern Free State Province. Two trials were conducted on Dorper sheep and 2 on Merino sheep. For each specific application 30 sheep were selected and allocated to 3 groups of 10 animals each using randomisation through minimisation, with pre-treatment total tick count as only criterion. Groups consisted of an untreated control group, a group treated with 3 ml of a flumethrin (1% m/v) solution applied only to the anogenital region, and a group treated at a dose rate of 1 mf flumethrin (1% m/v)/5 kg host body mass. The total dose volume for animals in the last group was divided into 3 equal parts and applied to the brisket/axillae, groin and anogenital regions respectively. Animals grazed under extensive farming conditions and were infested by ticks that occurred naturally in the environment. Ticks were counted and removed weekly over a 6-week period. In all 4 trials, Rhipicephalus e. evertsi was the dominant tick species, followed, in 3 of the trials, by Hyalomma spp. Efficacy (%) of control against ticks for Dorper sheep, treated only on the anogenital region, was variable, ranging between 29.5 and 97%. In Merino sheep the efficacy values ranged between 23.1 and 90 %. The site-spcific (anogenital region) efficacy of control against ticks infesting Merino sheep was in general 100% or almost 100%. In Dorper sheep the efficacy values were >80 % for 3-5 weeks. The efficacy (%) of control against ticks for sheep treated on the brisket/axillae, groin and anogenital regions was always higher compared to sheep treated only on the anogenital region. In Dorper sheep, efficacy of control was >80 % for up to 4 weeks and in Merino sheep >80 % for 6 weeks.

Administration, Topical↗

Antibody to a cDNA-derived calreticulin protein from Amblyomma americanum as a biomarker of tick exposure in humans.

The antibody responses of human and animal hosts were studied to determine the utility of antibody against recombinant tick calreticulin (rTC), a cDNA-derived protein isolated from salivary glands of Amblyomma americanum L., as a biologic marker of tick exposure. Rabbits fed upon by either A. americanum or Dermacentor variabilis Say developed significant anti-rTC antibody responses, as measured by both ELISA and immunoblot assay. In contrast, gerbils exposed to Aedes aegypti did not develop anti-rTC antibodies, as measured by ELISA or immunoblot assay. The utility of the assay was next evaluated in humans at high risk for tick exposure. During April through September 1990, 192 military personnel who originated from either Fort Chaffee, Arkansas or Fort Wainwright, Alaska were studied during maneuvers in tick infested areas at Fort Chaffee. Study subjects completed a questionnaire and had pre- and post-maneuvers serum specimens analyzed for antibodies to rTC. In adjusted analysis (controlling for age, fort of origin, attached tick during maneuvers, and bed netting use), the use of bed netting and home station were associated with post-maneuvers anti-rTC antibody seropositivity by ELISA. Subjects from Fort Wainwright were more likely to be seropositive for anti-rTC antibody (adjusted odds ratio = 5.3, 95% confidence interval [CI] = 1.1-25.6). Personnel who did not report the use of bed netting were more likely to be anti-rTC seropositive (adjusted odds ratio = 6.8, 95% CI = 1.4-32.4). Immunoblot assays showed that humans had specific anti-rTC antibody responses. The animal experiments demonstrate that hosts exposed to naturally feeding ticks develop anti-rTC antibodies. The data also indicate that hosts exposed to Ae. aegypti saliva may not develop antibodies against rTC. Observations in tick-exposed humans support the hypothesis that anti-rTC antibody seropositivity is a biologic marker of tick exposure.

Adult↗

Tick infestations of white-tailed deer in Alabama.

Four species of ticks were collected from 537 white-tailed deer (Odocoileus virginianus), examined during the hunting seasons (November to January) of 1988-89 and 1989-90 at selected locations in Alabama (USA). Ixodes scapularis was the most common tick recovered (2,060 specimens) and infested 54% of the deer. Dermacentor albipictus was the second most frequent tick (1,253 specimens) and infested 15% of the deer. Amblyomma americanum was the third most frequent tick (315 specimens) and infested 24% of the deer; this was the only species of tick collected from deer at all sampling locations. Amblyomma maculatum was an infrequent parasite (five specimens) and infested only 1% of the deer; this tick species was only recorded during the 1989-90 season. Year-to-year and geographical differences in tick infestation parameters were noted. The data are compared with those reported for previous surveys of ticks infesting white-tailed deer in Alabama and adjacent states.

Alabama↗

Borrelia sp. in ticks recovered from white-tailed deer in Alabama.

Six hundred sixty-five hunter-killed white-tailed deer (Odocoileus virginianus) from 18 counties in Alabama (USA) were examined for ticks. Most of the collections were made at state-operated wildlife management areas. Four species of ticks (n = 4,527) were recovered: the lone star tick Amblyomma americanum (n = 482); the Gulf Coast tick A. maculatum (n = 11); the winter tick Dermacentor albipictus (n = 1,242); and the black-legged tick Ixodes scapularis (n = 2,792). Fifty-six percent of the ticks (n = 2,555) were examined for Borrelia sp. spirochetes using an immunofluorescent, polyclonal antibody assay. Spirochetes were detected in I. scapularis (five females, seven males) from Barbour, Butler, Coosa, and Lee counties and A. americanum (four males, four nymphs) from Hale, Lee, and Wilcox counties. Area-specific prevalences in ticks were as high as 3.3% for I. scapularis and 3.8% for A. americanum.

Alabama↗