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Detection of the agent of heartwater, Cowdria ruminantium, in Amblyomma ticks by PCR: validation and application of the assay to field ticks.

We have previously reported that the pCS20 PCR detection assay for Cowdria ruminantium, the causative agent of heartwater disease of ruminants, is more sensitive than xenodiagnosis and the pCS20 DNA probe for the detection of infection in the vector Amblyomma ticks. Here, we further assessed the reliability of the PCR assay and applied it to field ticks. The assay detected DNA of 37 isolates of C. ruminantium originating from sites throughout the distribution of heartwater and had a specificity of 98% when infected ticks were processed concurrently with uninfected ticks. The assay did not detect DNA of Ehrlichia chaffeensis, which is closely related to C. ruminantium. PCR sensitivity varied with tick infection intensity and was high (97 to 88%) with ticks bearing 10(7) to 10(4) organisms but dropped to 61 and 28%, respectively, with ticks bearing 10(3) and 10(2) organisms. The assay also detected C. ruminantium in collections of Amblyomma hebraeum and Amblyomma variegatum field ticks from 17 heartwater-endemic sites in four southern African countries. Attempts at tick transmission of infection to small ruminants failed with four of these collections. The pCS20 PCR assay is presently the most characterized and reliable test for C. ruminantium in ticks and thus is highly useful for field and laboratory epidemiological investigations of heartwater.

Animals↗

Study on the occurrence of Borrelia burgdorferi sensu lato and tick-borne encephalitis virus (TBEV) in ticks collected in Lublin region (eastern Poland).

In 114 Ixodes ricinus ticks from 1 district of Lublin region (eastern Poland) examined by dark field microscopy method, the presence of motile spirochetes, morphologically corresponding to Borrelia species was detected in 8.8% of the total examined ticks. The highest infection rate was noted among females (16.7%), much lower in males (7.1%) and nymphs (4.0%). Examination of 550 ticks collected from 3 districts of the Lublin region by polymerase chain reaction (PCR) showed that 5.3% of the total number of ticks were infected with Borrelia spirochetes. The highest rates of Borrelia infection were observed in Ixodes ricinus ticks from the Zamość and Lublin districts (9.6% and 4.7% respectively). In the Włodawa district, only 2.4% ticks showed the presence of Borrelia DNA. In contrast to the results obtained by dark field microscopy method, the highest infection rate was noted in males (11.2%), followed by females (6.9%) and nymphs (1.7%). 57 Ixodes ricinus ticks collected from 3 Lublin districts (Lubartów, Lublin, Radzyń Podlaski) were tested for the presence of tick-borne encephalitis virus (TBEV) by the inoculation of 5-week-old Swiss mice, followed by blind passages and inoculation of GMK cell culture with brain suspension of the infected mice. One strain of TBEV was isolated from a pool of 24 I. ricinus ticks collected from the Radzyń Podlaski district. The minimum infection rate of ticks from this district with TBEV was estimated as 4.2%, and in the total area of Lublin region estimated as 1.8%.

Animals↗

[Comparative data on the tick-borne encephalitis virus infectiousness of hungry and satiated taiga ticks (based on the results of an immunoenzyme analysis)].

The results of individual investigation of 25,500 Ixodes persulcatus ticks from east Siberia are presented. The ticks were collected from grass, men, and animals before their sticking and after feeding at different intervals. The quantity of positive specimens was 6-11 times higher among the fed ticks hungry ones and averaged 10.36 and 1.85%, respectively, at the first stage due to the higher aggression of the infected females at the first stage and due to viral replication when the tick had feed and its better recognition as the titers increased. The proportion of ticks having high levels of tick-borne encephalitis virus antigen among the fed ticks was also considerably higher. The content of virus antigen increased in proportion with the duration of feeding. There is a moderate correlation (r = 0.59) of the infection index of hungry and fed ticks in different years and in various areas. The findings suggest that the fed ticks should be used as an additional marker for the features of tick-borne encephalitis virus circulation.

Animals↗

Introduced ticks and tick-borne diseases: the threat and approaches to eradication.

Exotic tick species and tick-borne diseases are serious threats to live-stock, companion animals, and wildlife in the United States. Recurring introductions of exotic tick species into the United States are a significant indicator of the degree of risk. Successful tick-eradication campaigns, such as the national program that eradicated Boophilus annulatus and B. microplus from the United States, the Cattle Fever Tick Eradication Program of the US Department of Agriculture's Veterinary Services that protects against the re-entry and dissemination of Boophilus ticks from Mexico back into their former haunts in the southern states, and the eradication action that eliminated Rhipicephalus evertsi from a game park in Florida, are sources of useful information that aid in elucidating essential elements of successful eradication programs. Examples of failed eradication programs in places such as Puerto Rico and St. Croix also have heuristic value. Among the varieties of tick species and related infectious agents that threaten the United States, Boophilus ticks and bovine babesiosis, Amblyomma species (especially the tropical bont tick) and heartwater, and equine babesiosis, for which endemic vectors exist, are of special concern. Risk assessments to accumulate, evaluate, and synthesize information needed to appraise risks, consequences, and preparedness are necessary not just to inform federal, state, and local officials, as well as producers and stakeholders, but also to facilitate the creation of emergency response plans.

Animal Diseases↗

Acquired resistance to ixodid ticks induced by tick cement antigen.

Antisera from guinea pigs made resistant to infestation with an ixodid tick of east and central Africa, Rhipicephalus appendiculatus, were used to identify the tick antigens they recognized by immunoblotting. Most of the antigens were found in tick salivary glands and in tick attachment cement. Antisera from R. appendiculatus-resistant guinea pigs also recognized some salivary-gland antigens in ticks of other species (R. pulchellus, R. evertsi, Amblyomma variegatum and A. gemma). Antibodies against the most strongly recognized R. appendiculatus antigen, a 20-kDa molecule, were only poorly reactive with similar-sized molecules in the other ticks. A 94-kDa antigen, which appeared to have broader cross-reactivity, was purified from R. appendiculatus attachment cement, and a monospecific rabbit serum was raised against it. This antiserum clearly recognized a molecule of similar molecular weight in R. pulchellus and R. evertsi. Intravenous inoculation of rabbits with the purified molecule elicited delayed-type hypersensitivity to the antigen. The hypersensitive rabbits demonstrated resistance to feeding of R. appendiculatus ticks but slight enhanced feeding of R. pulchellus ticks. These results are discussed with respect to their relevance for artificial induction of tick-feeding resistance.

Animals↗

Enhanced immunity to Hyalomma anatolicum anatolicum ticks in cross-bred (Bos indicus x Bos taurus) calves using ascaris extract immunomodulator with the tick salivary gland extract antigens.

Development of acquired immunity in cross-bred (Bos indicus x Bos taurus) calves against Hyalomma anatolicum anatolicum, a tick vector of bovine tropical theileriosis was studied using ascaris extract (AE), an immunomodulator of IgE responses, along with the tick salivary gland extract (SGE) antigens in Freund's incomplete adjuvant (FIA) emulsion. Calves immunised with SGE + FIA showed significant rejection (47.4 +/- 2.8%) of larvae, whereas only marginal rejection (12.47 +/- 1.5%) of nymphs was observed. In contrast, calves immunised with the immunomodulator AE in addition to SGE + FIA showed significant enhanced rejection of nymphs (50.25 +/- 5.2%), while the rejection of larvae was only slightly higher (55.8 +/- 11.4%), and not statistically different. In addition, incorporation of the immunomodulator AE also resulted in significant enhancement in the percentage recovery of abnormally fed larvae and nymphs. This abnormal feeding was characterised by their white to pale-yellow colour instead of the dark-grey colour of normally fed ticks. It may possibly be attributed to their inability to gain access to the blood vessels owing to the host immunological reaction at the bite sites. Consequently, the ticks fed on extravascular fluid devoid of red blood cells resulting in their white to pale-yellow colour. Difference in the disease transmission potential of normally and abnormally fed ticks is of future interest. Sera from all the immunised calves after 2 weeks of immunisation were positive for anti-tick SGE antibodies in the dot enzyme immunoassay (DOT-EIA). The immunised calves were positive for immediate type hypersensitivity (ITH) skin reaction on intradermal inoculation of the tick SGE antigens. There was no apparent difference in the DOT-EIA antibody titres between the two immunised groups. However, ITH skin swelling was significantly higher in AE + SGE + FIA immunised calves. The study indicates that use of the AE as an immunomodulator along with the tick SGE antigens enhanced anti-tick immunity.

Adjuvants, Immunologic↗

Tick species and tick-borne infections identified in population from a rural area of Spain.

To determine the tick species that bite humans in the province of Soria (Spain) and ascertain the tick-borne pathogens that threaten people's health in that province, 185 tick specimens were collected from 179 patients who sought medical advice at health-care centres. The ticks were identified, and their DNA examined by PCR for pathogens. Most ticks were collected in autumn and spring (59 and 57 respectively). Nine species of ticks were identified, the most frequent being Dermacentor marginatus (55.7%), Ixodes ricinus (12.4%) and Rhipicephalus bursa (11.9%). Ninety-seven females, 66 males, 21 nymphs and one larva were identified. Twenty-six ticks carried DNA from Rickettsia spp. (11 Rickettsia slovaca, 6 Rickettsia spp. RpA4/DnS14, 1 Rickettsia massiliae/Bar29, and 8 unidentified); two ticks carried DNA from Borrelia burgdorferi sensu lato and seven ticks harboured DNA from Anaplasma phagocytophilum.

Anaplasma phagocytophilum↗

Efficient transmission of tick-borne encephalitis virus between cofeeding ticks.

Most of the data on oral infections of ticks with tick-borne encephalitis virus have been derived from experiments using animals infected by syringe inoculation. To mimic the natural conditions of virus transmission, tick-borne encephalitis virus-infected Ixodes ricinus (Linnaeus) or Rhipicephalus appendiculatus Neumann adults (donors) were cofed with uninfected nymphs (recipients) of either tick species on uninfected guinea pigs. Two tick-retaining cells were attached to each guinea pig: cell 1 contained uninfected nymphs and virus-infected adults, and cell 2 contained uninfected nymphs. Following engorgement, 55% of I. ricinus nymphs and 65% of R. appendiculatus nymphs were shown to have acquired the virus while cofeeding with I. ricinus donor ticks. Similarly, 66% of R. appendiculatus recipient nymphs that cofed with R. appendiculatus virus-infected adults were infected. Some of the guinea pigs on which the ticks cofed were apparently nonviremic. The results indicate that efficient transmission of tick-borne encephalitis virus can occur between cofeeding ticks even when the host on which they feed does not develop a detectable viremia.

Animals↗

Enhancement of tick-borne encephalitis virus transmission by tick salivary gland extracts.

To investigate the role of ticks in TBE virus transmission, salivary gland extract (SGE) was derived from partially fed female Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus appendiculatus ticks. Guinea-pigs were infested with uninfected R. appendiculatus nymphs and inoculated with a mixture of TBE virus and SGE or with virus alone. The number of ticks which on average acquired virus from feeding on animals inoculated with TBE virus and SGE from partially fed ticks was 4-fold greater than the number that became infected by feeding on animals inoculated with virus alone or virus plus SGE from unfed I. ricinus. Viraemia was detected in 67% of guinea-pigs inoculated with virus plus SGE compared to 30% of guinea-pigs inoculated with virus alone. Virus titres in the blood were similar for both groups of animals [range 2.0-2.8 log10 plaque-forming units (PFU)/ml of blood]; however, the number of ticks that became infected was significantly higher on animals inoculated with virus plus SGE from partially fed ticks. No significant difference was observed with respect to the tick species used to derive SGE. The results indicate that TBE virus transmission is enhanced by factor(s) associated with the salivary glands of feeding ticks, and that these factor(s) may facilitate efficient transmission of TBE virus between infected and uninfected ticks even when they feed on hosts that have no detectable viraemia.

Animals↗

Efficacy of orally administered powdered aloe juice (Aloe ferox) against ticks on cattle and ticks and fleas on dogs.

The efficacy of orally administered powdered aloe juice (Aloe ferox) was evaluated against ticks on cattle and against ticks and fleas on dogs. Twelve calves were each infested over a 25-day period with approximately 4000 larvae of Rhipicephalus (Boophilus) decoloratus and allocated to 3 groups of 4 calves each. Three days after the last larval infestation and daily for 22 days thereafter, the calves in 1 group were fed 5 mg/kg body weight and those in another 25 mg/kg body weight of powdered aloe juice incorporated in game maintenance pellets, while the animals in the 3rd group received only pellets. Detached female ticks were collected daily and counted and the weights and the fertility of groups of 50 engorged female ticks collected from the animals were ascertained. The powdered aloe juice in the game maintenance pellets had no effect on the tick burdens of the calves or on the fertility of the ticks. Six dogs, in each of 2 groups, were treated daily for 15 consecutive days, commencing on Day -5 before the 1st tick infestation, with either 0.39 g or 0.74 g of powdered aloe juice, administered orally in gelatin capsules, while a 3rd group of 6 dogs served as untreated controls. All the dogs were challenged with Haemaphysalis leachi on Days 0 and + 7, and with Ctenocephalides felis on Days + 1 and + 8, and efficacy assessments were made 1 day after flea and 2 days after tick challenge, respectively. Treatment was not effective against ticks or fleas on the dogs.

Administration, Oral↗

Probability models of the rate of infection with tick-borne encephalitis virus in Ixodes persulcatus ticks.

A total of 3,254 adults of Ixodes persulcatus tick were collected in a taiga forest habitat situated in the Amgun river basin (Khabarovsk region, the Far East, USSR) and examined individually for the presence and amount of tick-borne encephalitis virus. The over-all proportion of infected ticks was 6.6% and it varied between 3.4% and 9.4% in the years 1982 to 1985. The amount of virus per tick was approximated by the gamma distribution determining a probability that the number of plaque-forming units (PFU) per tick is not greater than a selected value. The frequency distribution of infected ticks followed a model of the negative binomial distribution, enabling the estimation of probability of the occurrence of a given number of infected ticks in the area. However, the parameters of both probability models (i.e., the PFU content per tick, and the frequency of infected ticks) varied for particular years.

Animals↗

Ticks and tick paralysis: imaging findings on cranial MR.

Tick paralysis is an acute, progressive, and potentially fatal muscle paralysis secondary to a toxin secreted by a pregnant tick during a bite. Although tick bites can occur anywhere on the body, ticks are frequently overlooked on the scalp because of overlying hair. Children with acute neurologic symptoms frequently undergo MR scanning that may incidentally reveal the offending tick. Timely identification and removal of the tick leads to rapid recovery from tick paralysis. We report the MRI findings at 1.5 T of tick paralysis with an attached tick.

Child, Preschool↗

The tick protective antigen, 4D8, is a conserved protein involved in modulation of tick blood ingestion and reproduction.

The gene that encodes the tick protective antigen, 4D8, was cloned from 10 species belonging to 6 genera, and the nucleotide and amino acid sequences were analyzed. 4D8 nucleotide and protein sequences were conserved among these tick species with identity/similarity between 65-98 and 60-98%, respectively. The function of 4D8 was characterized by RNA interference (RNAi) in five tick species. After the ticks were allowed to feed, degeneration of gut, salivary glands and reproductive tissues was observed, and tick survival, weight and oviposition were significantly reduced. 4D8 RNAi effected >90% reduction in oviposition in all tick species tested. Because of the critical role that 4D8 plays during tick feeding and oviposition, which ultimately results in the reduction of tick progeny, we proposed the generic name "subolesin" (Latin, suboles: offspring, progeny) for tick 4D8 proteins and subA for the subolesin-encoding gene.

Amino Acid Sequence↗

Field sampling of the tick Amblyomma variegatum (Acari: Ixodidae) on pastures in Guadeloupe; attraction of CO2 and/or tick pheromones and conditions of use.

Field experiments were conducted in Guadeloupe to evaluate the attraction of different chemicals on a Guadeloupean strain of Amblyomma variegatum. Such tick sampling techniques may help to assess the size of the tick population on pastures before, during and after an attempted tick eradication campaign. CO2 was absolutely necessary to activate ticks, including the larvae. In addition to activation, our data suggested that CO2 itself is attractive for ticks. The reactivity of the ticks varied greatly with the period of the year, the adults being insensitive to CO2 alone or combined with pheromones for a period extending from December to June. For the whole period, the total captures were equal for the males and females, but the males were relatively more sensitive at the beginning of the season of activity (June-July), while the females dominated, but not significantly, for the rest of the period. There was also a gradient of reactivity depending on the distance from the trap; 28% of the ticks were captured at 0.5 m from the CO2 traps and 1.2% at a distance of 6 m. The ticks arrived rapidly at the trap. After 1 h of trapping, 55, 68 and 81% of the active nymphs, males and females, respectively, had reached the trap. Synthetic pheromones (methyl salicylate, o-nitrophenol and nonanoic acid in the proportions 1:2:8) or freshly detached males added to CO2, greatly increased the efficacy of the CO2 traps. Depending on the concentration of the pheromones, the attraction was four to 70 times higher than with CO2 alone. The effect depended on the sex of the ticks, males being attracted in a greater proportion by small amounts of pheromones, while the females were activated by higher doses. From a practical point of view, sampling should be conducted from June to December, with CO2 and high concentrations of pheromones, the traps being in operation for approximately 1 h at least, the area concerned by effluents being located downwind and covering an area of approximately 36 m2.

Age Factors↗

Prefeeding Dermacentor andersoni (Acari: Ixodidae) on cattle with prior tick exposure may inhibit detection of tick paralysis by using hamster bioassay.

The effect of prefeeding Dermacentor andersoni Stiles on naive and previously challenged cattle before conducting virulence assays by using hamsters was examined. In an initial observational trial, ticks prefed on a previously challenged heifer did not cause paralysis after transfer to hamsters, whereas ticks from the same lot that were prefed on a naive heifer did cause paralysis. Subsequent paired trails were conducted to simultaneously compare paralysis after feeding on naive and previously challenged heifers. In the first trial, prefeeding ticks on a previously challenged heifer did not result in loss of virulence. In a second trial, ticks fed on a previously challenged heifer had greatly diminished virulence after transfer to hamsters. This also occurred in a third trial after the previously challenged animal had two prior exposures to virulent D. andersoni. Weight gains of ticks that did not cause paralysis after transfer to hamsters were higher after feeding on previously challenged cattle, indicating that loss of virulence was not due to failure of ticks to reattach and feed. Ticks gained little weight on paralyzed hamsters due to the reduced time spent feeding on hamsters when paralysis occurred. Results indicate that assaying ticks for virulence requires prefeeding on naive hosts. If these are not available, rabbits, a resistant host, can be used for prefeeding ticks.

Animals↗

Strategies for the control of one-host ticks and relationship with tick-borne diseases in South America.

A variety of hemoparasites occurs in ruminants of South America (12 degrees 00'N-56 degrees 00'S), but there is consensus on the significant economic impact of babesiosis (Babesia bovis and Babesia bigemina) and anaplasmosis (Anaplasma marginale). The tick vector for the Babesia spp. for the whole region is Boophilus microplus which has been the most important target in control/eradication programs. Boophilus microplus and many different hematophagous diptera species are considered vectors of A. marginale. In areas where babesiosis and anaplasmosis are present, the wide range of climates (tropical, subtropical, temperate), husbandry practices (subsistence, extensive, intensive), and tick control strategies has led to the concepts of enzootic stability and instability. Within this framework, six control strategies for Boophilus microplus can be identified: (1) absolute tick control--with the establishment of quarantine areas and the implementation of state legislation; (2) absolute tick control in marginal areas--using local epidemiological knowledge, strategic treatments, quarantine areas and the intervention of the state; (3) prophylactic treatment--with a predetermined acaricide treatment schedule aimed at reducing tick burdens to an acceptably low level; (4) threshold treatment--acaricides are applied by the farmer whenever the number of ticks on cattle exceed a threshold level; (5) opportunistic treatment--acaricides are applied when cattle are assembled for other purposes; (6) integrated tick control--principally involving the combination of acaricides with the use of tick-resistant crossbred cattle.

Anaplasmosis↗

[Fever after a tick bite: clinical manifestations and diagnosis of acute tick bite-associated infections in northeastern Switzerland].

BACKGROUND: Different tick-borne infections can cause an acute febrile illness. The study objectives were to investigate the clinical manifestations and diagnosis of infections among patients who presented with fever after a tick-bite, and to detect newly described pathogens, including Ehrlichia, Babesia and Rickettsia helvetica, in North-Eastern Switzerland. PATIENTS AND METHODS: : We studied 75 patients (41 male, 34 female, median age 38 years, among them 10 children) who had fever within 3 weeks after a tick-bite. Paired sera were tested for antibodies to Borrelia burgdorferi, tick-borne encephalitis virus, Anaplasma (Ehrlichia) phagocytophila, Babesia microti, B. divergens, and Rickettsia helvetica. In addition, microscopy and polymerase chain reaction was used to detect Ehrlichia. Clinical data were obtained at baseline and at 1 and 2 year follow-up. RESULTS: Tick-borne infections were confirmed or possible in 36 (48 %) patients: 7 (9 %) Erythema migrans, 6 (8 %) other specific manifestations of Lyme borreliosis, 6 (8 %) Lyme borreliosis presenting as non-specific febrile illness, 8 (11 %) tick-borne encephalitis, 7 (10 %) granulocytic ehrlichiosis, 1 B. microti infection in a traveler from the US and 6 (8 %) dual infections. In 8 (11 %) patients serological findings were suggesting possible acute or past R. helvetica infection. CONCLUSION: Among patients with fever after a tick-bite, Lyme borreliosis was most frequently found. There was no evidence for babesiosis among the resident population. Serologic data suggest that human granulocytic ehrlichiosis and R. helvetica infections may be endemic in Switzerland. Among 50 % of the patients no tick-borne infections could be diagnosed.

Acute Disease↗

[Tick-borne encephalitis and Lyme borreliosis. Spread of pathogens and risk of illness in a tick-borne encephalitis region].

A knowledge of the distribution of tick-borne agents and the related risks of contracting diseases are essential to ensure an appropriate response between hysteria and disregard, and to decide the appropriateness of vaccination. Illustrated by the German town of Lohr a. M. (Bavaria), the prevalence of TBE-virus and Borrelia burgdorferi in ticks was investigated and compared, and the respective risk of contracting disease from a bite assessed. Using the polymerase chain reaction (PCR), a total of 1657 ticks obtained from five different biotopes around Lohr were examined for the TBE-virus, and 408 ticks for Borrelia burgdorferi. The results were compared with earlier findings in other regions of Germany. The risk of contracting illness was estimated on the basis of transmission and manifestation rates, together with epidemiological data from the region. The prevalence of TBE-virus was 0.12% (95% CI: 0.05-0.44%) in the ticks investigated. This is comparable with that in 4 other regions rated as TBE-risk regions, but significantly lower than that in rated high-risk regions. Borrelia burgdorferi was detected in 14.9% (11.8-19.0%) of adult ticks, roughly twice the prevalence found in nymphs (7.2%, range 4.6-11.7%). On the basis of these prevalences, the risk of contracting meningitis/encephalitis from a tick bite is about 1:10,000, and the risk for Lyme Borreliosis is about 1:100, the latter requiring that the tick remains attached for at least 2-3 days.

Adult↗