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Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 (Acari: Ixodidae): hosts, distribution and 16S rDNA sequences.

DNA sequences of Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 were obtained to determine genetic differences between these tick species. Collections of these species are discussed in relation to distribution and hosts. Seven ticks collections (four from Brazil, one from Argentina, one from Uruguay and one from USA) house a total of 1272 A. aureolatum (224 males, 251 females, 223 nymphs and 574 larvae) and 1164 A. ovale (535 males, 556 females, 66 nymphs and 7 larvae). The length of the sequenced mitochondrial 16S rRNA gene fragment for A. aureolatum was 370bp and for A. ovale was 373bp. The DNA sequence analysis showed a 13.1% difference between the two species. Apart from one male A. ovale found on a toad, all adult ticks were found on mammals. The majority of adult specimens of both tick species were removed from Carnivora (96.1 and 84.3% of A. aureolatum and A. ovale, respectively), especially from dogs (53.1% of A. aureolatum, and 46.4% of A. ovale). Collections on wild Canidae were higher for A. aureolatum (23.3%) than for A. ovale (7.1%). On the other hand, collections of A. ovale adults on wild Felidae were higher (18.3%) than findings of A. aureolatum (9.2%). The contribution of other mammalian orders as hosts for adults of A. aureolatum and A. ovale was irrelevant, with the exception of Perissodactyla because Tapiridae contributed with 13.0% of the total number of A. ovale adults. Adults of both tick species have been found occasionally on domestic hosts (apart of the dog) and humans. Most immature stages of A. aureolatum were found on Passeriformes birds, while rodents and carnivores were the most common hosts for nymphs and larvae of A. ovale. A. aureolatum has been found restricted to the Neotropical region, covering the eastern area of South America from Uruguay to Surinam, including northeastern Argentina, eastern Paraguay, southeastern Brazil and French Guiana. A. ovale showed a distribution that covers the Neotropical region from central-northern Argentina throughout the Neotropics into the Nearctic region of Mexico with a few records from the USA, also with collection sites in Paraguay, Bolivia, most Brazilian states, Peru, Ecuador, French Guiana, Surinam, Guyana, Trinidad & Tobago, Venezuela, Colombia, Panama, Costa Rica, Nicaragua, Belize, Guatemala and several states of Mexico.

Animals↗

Experiments on the transmission of Babesia major and Babesia bigemina by Haemaphysalis punctata.

Experiments on the transmission by Haemaphysalis punctata of three large Babesia strains were carried out. Three Babesia-free batches of laboratory reared H. punctata ticks were infected with two strains of Babesia major, B. major (Xingjiang strain), isolated with adult ticks of H punctata and B. major (Henan strain), isolated with H. longicornis) and a strain of Babesia bigemina by feeding them on the calves infected by inoculation of blood stabilates. H. punctata was shown to be capable of transmitting the B. major strains transovarially. The larvae, nymphs and adults developed from female ticks engorged on the calf infected with B. major (Xingjiang strain) transmitted the pathogen to splenectomised calves with prepatent periods of 15, 11 and 12 days, respectively. The calves infested with larvae and nymphs died of babesiosis with parasitemias of 400 and 710 per 1000 erythrocytes. The calf infested with adult ticks survived babesiosis, but the number of erythrocytes and the amount of haemoglobin were reduced greatly. H. punctata transmitted B. major (Henan strain) in the same way. The prepatent periods of the calves infested with larvae, nymph and adult ticks were 9, 10 and 12 days, respectively. Calves infested with larvae survived, but those infested with nymphal and adult ticks died of babesiosis with parasitemias of 410 and 100 per 1000 erythrocytes, respectively. H. punctata ticks did not transmit the B. bigemina strain to splenectomised calves. There were no clinical symptoms and no parasites were discovered in the blood films during a 2 month observation period after the calves were infested with larval, nymphal and adult ticks derived from female ticks engorged on calves inoculated with B. bigemina.

Animals↗

Overexpression of CYP4G19 associated with a pyrethroid-resistant strain of the German cockroach, Blattella germanica (L.).

A new cytochrome P450 gene, CYP4G19, was identified and isolated as a differentially expressed gene between insecticide susceptible ACY and resistant Apyr-R German cockroach strains using PCR-selected subtractive hybridization and cDNA array techniques. The cDNA sequence of CYP4G19 has an open ready frame of 1638 nucleotides encoding a putative protein of 546 amino acid residues. Sequence analysis shows that CYP4G19 putative protein contains (1) a highly hydrophobic N terminus, (2) a P450 protein signature motif, FXXGXRXCXG, known to be an important ligand for heme binding, and (3) an important characteristic motif, EVDTFMFEGHDTT, for the family 4. Northern blot analysis indicated that levels of CYP4G19 expression were low in eggs, nymphs, and adults of the susceptible ACY strain with a similar expression pattern. The expression of CYP4G19 in the resistant Apyr-R strain was developmentally regulated, with very low expression in eggs, increasing in nymphs, and reaching a maximum in both female and male adults. Comparison of CYP4G19 expression between ACY and Apyr-R strains indicated that there was no difference in their eggs, but expression was higher ( approximately 1.7-fold) in the nymphs and much higher ( approximately 5-fold) in the male and female adults of the Apyr-R strain. Levels of CYP4G19 mRNA were readily detectable in head+thorax tissues and increased ( approximately 5-fold) in the abdomens of the ACY strain. In the Apyr-R strain, however, levels of CYP4G19 mRNA were relatively low in head+thorax tissues and were about 7-fold increase in the abdomen. Although expression patterns of CYP4G19 in head+thorax and abdomen tissues were similar (i.e. lower in head+thorax tissues and higher in abdomen tissues) in both the ACY and Apyr-R strains, the overexpression was more evident in the Apyr-R strain in both head+thorax and abdomen tissues than in the ACY strain.

Amino Acid Sequence↗

A new case of trophobiosis between ants and Heteroptera.

Despite the large specific diversity of equatorial rainforests, Caternaultiella rugosa (Heteroptera; Plataspidae) was only noted on two euphorbiaceous trees (Bridelia micrantha and B. grandis) and was attended by two ant species, Camponotus brutus (Formicinae) and Myrmicaria opaciventris (Myrmicinae). We recorded semiochemical (attending workers palpated the dorsal abdominal glands of the nymphs) and semantic signals (nymphs ready to excrete honeydew raised their bodies; they alternated the extrusion and withdrawal of the first honeydew droplet when the workers did not immediately absorb the honeydew). Cat. rugosa was recorded in carton pavilions built by the ants at the base of the tree trunks. During proliferations of the population, clusters of nymphs and adults developed outside pavilions. In the latter case, M. opaciventris workers did not modify their rhythm of activity in order to attend these clusters, while Camp. brutus workers, normally nocturnal, attended them day and night.

Animals↗

Changes in salivary nitrophorin profile during the life cycle of the blood-sucking bug Rhodnius prolixus.

The insect Rhodnius prolixus is a hematophagous hemipteran that has five nymphal instars. Fifth instar nymphs contain, in their salivary glands, four nitrophorins which have already been described in the literature (NP1, NP2, NP3 and NP4). Two new hemeproteins were isolated and partially characterized from first instar nymphs. NP2, that shows an anticoagulant activity, was also identified, but NP1, NP3 and NP4 were not found. As these new hemeproteins have amino-terminal sequences clearly homologous to already described nitrophorins and were capable of binding nitric oxide, they were named nitrophorins 5 and 6, although they showed an unusual Soret band at 412 nm. In each subsequent nymphal stage, a new nitrophorin emerges. In the second instar, NP4 comes into view, in the third instar NP1 appears, and NP3 is only found in fifth instar nymphs and adults, showing that the nitrophorin profile of R. prolixus saliva is stage-specific.

Amino Acid Sequence↗

Simple epidemiological model predicts the relationships between prevalence and abundance in ixodid ticks.

We tested whether the prevalence of ticks can be predicted reliably from a simple epidemiological model that takes into account only mean abundance and its variance. We used data on the abundance and distribution of larvae and nymphs of 2 ixodid ticks parasitic on small mammals (Apodemus agrarius, Apodemus flavicollis, Apodemus uralensis, Clethrionomys glareolus and Microtus arvalis) in central Europe. Ixodes trianguliceps is active all year round, occurs in the study area in the mountain and sub-mountain habitats only and inhabits mainly host burrows and nests, whereas Ixodes ricinus occurs mainly during the warmer seasons, occupies a large variety of habitats and quests for hosts outside their shelters. In I. ricinus, the models with k values calculated from Taylor's power law overestimated prevalences. However, if moment estimates of k corrected for host number were used instead, expected prevalences of both larvae and nymphs I. ricinus in either host did not differ significantly from observed prevalences. In contrast, prevalences of larvae and nymphs of I. trianguliceps predicted by models using parameters of Taylor's power law did not differ significantly from observed prevalences, whereas the models with moment estimates of k corrected for host number in some cases under-estimated relatively lower larval prevalences and over-estimated relatively higher larval prevalences, but predicted nymphal prevalences well.

Animals↗

Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique.

Although Babesia divergens is the the principal confirmed zoonotic Babesia sp. in Europe, there are gaps in our knowledge of its biology and transmission by the tick Ixodes ricinus. In order to reproduce the part of the parasite cycle that occurs in the vector, an in vitro animal skin feeding technique on blood containing in vitro cultivated B. divergens was developed. Parasite DNA was detected in all samples of salivary glands of nymphs and adults that had fed on parasitized blood as larvae and nymphs, respectively, indicating acquisition as well as a transtadial persistence of B. divergens. PCR performed on eggs and larvae produced by females that had fed on parasitized blood demonstrated the existence of a transovarial transmission of the parasite. Gorging B. divergens infected larvae on non-infected gerbils showed persistance of the parasite over moulting into the resulting nymphs. These results indicate that the parasitic stages infective for the vector (i.e. the sexual stages) can be produced in vitro. To our knowledge, this is the first report of artificial feeding of I. ricinus via membrane as well as in vitro transmission of B. divergens to its vector. The opportunities offered by the use of such a transmission model of a pathogen by I. ricinus are discussed.

Animals↗

Seasonal synchrony: the key to tick-borne encephalitis foci identified by satellite data.

A previous analysis of tick infestation patterns on rodents in Slovakia suggested that the key to the focal distribution of western-type tick-borne encephalitis virus (TBEv) in Europe is the geographically variable degree of synchrony in the seasonal activity of larval and nymphal Ixodes ricinus ticks. This prediction is here tested by examining records, from 7 different countries, of the seasonal variation in the abundance of larvae and nymphs feeding on rodents or questing on the vegetation. Larvae consistently started feeding and questing earlier in the year at sites within TBEv foci than elsewhere, so that they appeared in the spring as soon as nymphs were active. Such larval nymphal synchrony is associated with a rapid fall in ground-level temperatures from August to October as revealed by the satellite-derived index of Land Surface Temperature (LST). Likewise, of 1992 pixels sampled on a grid across Europe, the 418 that fell within TBEv foci were characterized by a higher than average rate of autumnal cooling relative to the peak midsummer LST. It is proposed that such a seasonal temperature profile may cause unfed larvae to pass the winter in quiescence, from which they emerge synchronously with nymphs in the spring.

Animals↗

Prevalence of Cowdria ruminantium infection in Amblyomma hebraeum ticks from heartwater-endemic areas of Zimbabwe.

Analysis of the transmission dynamics of Cowdria ruminantium, the tick-borne rickettsial agent of heartwater in ruminants, requires accurate measures of infection in vector populations. To obtain these, Amblomnia hebraeum ticks were collected at two heartwater-endemic locations in the lowveld and highveld regions of Zimbabwe and assessed for C. ruminantium infection with specific polymerase chain reaction (PCR) and DNA probe detection assays. At the lowveld site, 11.2% (50/446) of adult ticks and 8.5% (23/271) of nymphs carried C. ruminantium, as detected by PCR. At the highveld site, the prevalence of infection in adult ticks was 10.2% (40/392). DNA probe analysis revealed that most infections at both sites were of low intensity; only 9% and 23% of all nymph and adult tick infections, respectively, were greater than 70000 organisms, the detection limit of the DNA probe. However, the majority (70%) of probe-detectable adult tick infections were high, between 10(7) and 10(9) organisms/tick, while those within nymphs were lower, between 10(5) and 10(6) organisms/tick.

Animals↗

Field evidence that roe deer (Capreolus capreolus) are a natural host for Ehrlichia phagocytophila.

Samples of blood, spleen and legs from 112 culled roe deer (Capreolus capreolus) were collected from nine sites widespread in the United Kingdom. The prevalence of infection with Ehrlichia phagocytophila was determined by serology and polymerase chain reaction. Means of 58% of 102 plasma or serum samples were seroreactive by IFA, 38% of 84 blood samples and 29% of 82 spleen samples were positive by PCR. Ticks on legs of 71 roe deer were Ixodes ricinus larvae, nymphs and adults and 83% of legs were infested. Numbers of ticks corresponded positively to the percentage of samples positive for E. phagocytophila by serology and PCR for different sampling sites. Ixodes ricinus nymphs collected from the vegetation at one site with infected deer were analysed for infection with E. phagocytophila by examination of Feulgen stained salivary glands. Of 135 nymphs 5% were infected. These results confirm that roe deer are commonly parasitized by both E. phagocytophila and its vector tick in such a way that it is likely to be an important natural mammalian reservoir of E. phagocytophila.

Animals↗

Cross-bred cattle protected against Hyalomma anatolicum anatolicum by larval antigens purified by immunoaffinity chromatography.

Antigens from larvae of Hyalomma anatolicum anatolicum were extracted and purified by immunoaffinity chromatography using immunoglobulin ligands from cross-bred animals immunized with soluble larval antigen. Affinity-purified antigen (Aff-TLE) and a total larval extract (TLE) were used to immunize cross-bred (Bos indicus x Bos taurus) cattle. The group immunized with Aff-TLE rejected 71.6% of larvae and 77.3% of nymphs. However, the rejection percentages were lower in the TLE-immunized group. No significant changes in the feeding period, moulting percentages or moulting period of engorged larvae and nymphs were recorded. There was, however, a significant decrease in the number of resultant nymphs p < 0.01) and adults (p < 0.01) in the ticks fed on the Aff-TLE-immunized group. The Aff-TLE antigen was 93.3% purified. SDS-PAGE analysis identified a 39 kDa protein, reported for the first time, as the antigen responsible for the induction of resistance in the host.

Animals↗

Immunoprotective efficacy of a purified 39 kDa nymphal antigen of Hyalomma anatolicum anatolicum.

Soluble nymphal antigens (HNAg) were purified by immunoaffinity chromatography using CNBr-activated Sepharose 4B coupled with immunoglobulin ligands from animals immunized with HNAg and 69-71% protected against challenge infestations, and 8% recovery of the purified protein (Aff-HNAg) was obtained. Following immunization of crossbred calves (Bos indicus x Bos taurus) with 1600 sg of Aff-HNAg in three divided doses, significant rejections of larvae (p<0.001, 84.2%), nymphs (p<0.05, 61.4%) and adults (p<0.05, 58.7%) were recorded. No significant changes were recorded in the engorgement weights of the larvae and nymphs, but there was a significant (p<0.05) reduction in the weight of the engorged adults. Immunization conferred a significant decrease in the numbers of resultant nymphs (p<0.001) and adults (p<0.001) that had fed on the immunized animals. SDS-PAGE analysis identified a 39 kDa protein, previously isolated from larvae of Hyalomma anatolicum anatolicum, as the antigen responsible for the induction of resistance against all the stages of the tick.

Animals↗

Vector competence of Ixodes angustus (Acari: Ixodidae) for Borrelia burgdorferi sensu stricto.

The vector competence of Ixodes angustus for Borrelia burgdorferi sensu stricto (s.s.) was investigated in the laboratory. The larval progeny of female ticks from Washington State were placed on Swiss-Webster mice that had been inoculated intravenously with 10(8) spirochetes each of a Californian isolate of B. burgdorferi. Spirochetes were detected in 6 (12%) of 50 nymphs derived from larvae that had fed on these animals. Ten nymphs from the same cohort of experimentally infected ticks were placed on each of 4 naive deer mice (Peromyscus maniculatus). One of the mice seroconverted to B. burgdorferi and spirochetes were isolated from its ear tissues 4 weeks after exposure to ticks. Further vector competence trials were conducted with I. angustus ticks from California. Larvae were fed on deer mice that had been inoculated intradermally with B. burgdoferi along with larvae of I. spinipalpis as a comparison group. There was no significant difference in the prevalence of infection in nymphs of I. angustus (8.2%) versus those of I. spinipalpis (12.1%). We conclude that I. angustus is a competent experimental vector of B. burgdorferi s.s. and its efficiency for acquiring and transstadially passing such spirochetes is similar to that of I. spinipalpis.

Animals↗

Low risk of Lyme borreliosis in a protected area on the Tyrrhenian coast, in central Italy.

A comprehensive Lyme borreliosis risk assessment process was applied in S. Rossore Estate, on the Tyrrhenian coast, near Pisa, Italy. Host-seeking Ixodes ricinus nymphs peaked in May in oak-dominated deciduous wood (median, Q1-Q3, number of nymphs/50 m dragging = 4.5, 2.5-8), whereas host-seeking larvae peaked in August in the same habitat type (6.0, 4-17/50 m dragging). Prevalence of I. ricinus infestation was 88.9% in wild rodents (n = 11), 64.3% in fallow deer (n = 28) and 0.0% in wild boars (n = 5). Borrelia burgdorferi sensu lato was not isolated from rodents' organs, and from 80 I. ricinus nymphs and 50 adults. Moreover, PCR for B. burgdorferi sl carried out on 110 nymphs and 12 adult ticks also gave negative results. Forest workers were at higher risk of tick bite than other Estate employees (relative risk (RR): 1.7, p = 0.02). In spite of high levels of tick exposure, B. burgdorferi sl specific antibodies were not detected in sera from Estate personnel (n = 30) and sentinel animals (dogs, n = 23, fallow deer, n = 61).

Animals↗

Tick bite and Lyme borreliosis risk at a recreational site in England.

The risk of tick bite and Lyme borreliosis in a forested area in England with public access was studied over a two-year period. Tick infestation levels were high with more than 1000 members of the public reporting for tick removal at a local clinic. Most of the attached ticks were nymphs (82%) and distinct differences in anatomical sites of attachment were observed in children and adults. Children sustained nymphal bites to the head, neck and axilla region much more frequently than adults (48 vs. 10%), whereas adults were bitten on the lower legs more frequently than children (46 vs. 9%). The vegetation was heavily infested with ticks and high numbers were particularly associated with areas used by deer. The average density of nymphs collected from the vegetation was 14.1 per 10 m2 (range 5.1-43.6). Infection rates of these nymphs determined by PCR and indirect IFA ranged from 5.2-17.0%, and the genospecies Borrelia valaisiana and B. garinii were detected, suggesting that birds may be important reservoir hosts in this area. It is estimated that, at the level of tick challenge observed here, at least 50 persons per year may be bitten by infected ticks at this site. However, no cases of Lyme borreliosis have been reported through the clinic follow-up procedure, and sera from 19 forest workers were negative for antibody to B. burgdorferi sensu lato. Despite the high challenge from tick bites, this particular recreational forest site poses a low risk of infection to the general public, and prophylactic antibiotic treatment or serological testing following a bite is not justified.

Adult↗

Transmission of Cowdria ruminantium by Amblyomma gemma from infected African buffalo (Syncerus caffer) and eland (Taurotragus oryx) to sheep.

Two African buffalo (Syncerus caffer), an eland (Taurotragus oryx) and a waterbuck (Kobus defassa) were intravenously inoculated with Cowdria ruminantium (Kiswani). Amblyomma gemma nymphs were fed on the animals at 3 weekly intervals. Jugular blood was also collected at 3 weekly intervals and inoculated into sheep. Nymphal ticks that fed on one buffalo on days 16 and 37 and on the other buffalo on day 58 after infection transmitted the disease as adults to sheep. Nymphs that were applied to the eland 16 days after infection also transmitted the disease to sheep. No nymphs that had fed on the waterbuck transmitted the disease. This is the first report of transmission of heartwater by Amblyomma gemma from infected wild ruminant species to a susceptible domestic ruminant species.

Animals↗

High prevalence of Borrelia lusitaniae in Ixodes ricinus ticks in Tunisia.

To investigate whether ticks of the genus Ixodes are infected by Borrelia burgdorferi complex, 490 unfed Ixodes ricinus ticks were collected by flagging in three different areas of Tunisia in 1998. DNAs extracted from 81 adults, 60 nymphs and 38 larvae were analysed after genic amplification of the non-coding spacer between the two copies of the rrl-rrf genes of B. burgdorferi sl. The prevalence of B. burgdorferi sl. in adults, nymphs and larvae was found to be 34, 33.3 and 2.6%, respectively. All DNAs (n = 61) but one were identified as belonging to different genotypes of B. lusitaniae by analysis of the restriction fragment length polymorphism of amplification products. In addition, 290 adults, 14 nymphs and 7 larvae were used to inoculate BSK-H medium to isolate spirochetes. Fifteen strains were isolated from adult ticks in the humid areas of Tunisia, whereas only one was obtained from larvae. Isolates were identified as B. lusitaniae (15/16) and B. garinii (1/16). These results provide new evidence for the existence of Lyme borreliosis in North Africa.

Animals↗

Infestation of mammals by Ixodes ricinus ticks (Acari: Ixodidae) in south-central Sweden.

Infestation by Ixodes ricinus ticks on rodents, hares and cervids was examined at Bogesund, 10 km north of Stockholm, in south-central Sweden during 1991-1994 and on varying hares (Lepus timidus) at Stora Karlsö and Gotska Sandön in the Baltic Sea during 1992-1993. At Bogesund, there were great differences between two consecutive years in the number of I. ricinus larvae infesting bank voles (Clethrionomys glareolus). The seasonal pattern of infestation by I. ricinus larvae and nymphs on bank voles was unimodal in 1991, with peaks in June-July and bimodal in 1992, with peaks in June and August. Male bank voles, compared to females and older voles, compared to young voles, harboured greater numbers of I. ricinus ticks. Apodemus mice, compared to bank voles, harboured greater numbers of I. ricinus ticks. Ixodes ricinus larvae engorged on Apodemus mice were heavier than larvae engorged on bank voles and resulted in larger nymphs. However, there was no difference in the proportions of viable nymphs resulting from larvae engorged on mice or voles. The ranges in the numbers of I. ricinus ticks infesting individual hosts were 1-451 for rodents, 16-2374 for hares and 428-2072 for roe deer (Capreolus capreolus). These ranges of tick numbers are estimated to represent potential blood losses from individual hosts of approximately 0.2-65% for rodents, 0.2-13% for hares and 0.3-9.0% for roe deer. Within the populations of all host species examined, the distributions of all stages of I. ricinus were clumped, with most host individuals harbouring few ticks and only a few individuals harbouring many ticks. The data suggest that, even though a small proportion of tick hosts may be severely affected, the direct effects of feeding by I. ricinus are unlikely to play an important role on mammal population dynamics.

Age Factors↗