Olaf Kahl

Borrelia burgdorferi sensu lato in Ixodes ricinus ticks and rodents in a recreational park in south-western Ireland

Ixodes ricinus ticks infected with Borrelia burgdorferi sensu lato were numerous on the edges of paths and roads in a recreational park in south-western Ireland. The abundance of ticks at different sites was related to the presence of deer, but a negative relationship was shown between tick abundance and tick infection rates. This is thought to be due to the deposition of large numbers of uninfected ticks by deer, which are apparently not good reservoir hosts of B. burgdorferi s.l. Blood meal analysis only detected deer DNA in uninfected nymphs. Reservoir competent rodents, Apodemus sylvaticus and Clethrionomys glareolus, were abundant at all sites and a high proportion of captured specimens were infested with larval ticks. However, very few rodents were infected with B. burgdorferi s.l. and none of the unfed infected nymphs analysed for the identity of their larval blood meal had fed on rodents. The spirochaetes detected in I. ricinus in the study area may be poorly adapted to rodents or are not transmitted readily because of the absence of nymphal infestation. The majority of spirochaetes in these ticks were apparently acquired from non-rodent hosts, such as birds.

Water vapour uptake from subsaturated atmospheres by engorged immature ixodid ticks

Contrary to current opinion, fully engorged and detached larvae and nymphs of some ixodid ticks consistently take up substantial amounts of atmospheric water vapour and thereby display their regulative capacity for maintaining water balance in subsaturated air. Net uptake of vapour generally begins some days after detachment and the capability persists until shortly after initiation of apolysis, a period which in diapausing specimens may extend up to several months. This was shown forIxodes ricinus, Haemaphysalis punctata, and the North AmericanI. dammini. Apparently, some other engorged ixodid immatures fail to exhibit net vapour uptake, as was shown for both larvae and nymphs ofDermacentor marginatus and nymphs ofHyalomma anatolicum excavatum. But there is some evidence for engorged nymphs ofD. marginatus that active uptake of vapour does occur, masked by spiracular transpiration. Net uptake of vapour is apparently not possible during the pharate phases. InI. ricinus both teneral nymphs and adults are capable of achieving net water gains by active vapour uptake on the first day following ecdysis. There is new evidence from fully engorgedI. ricinus immatures for the decisive role of agranular alveoli in the production of the salivary secretion involved in vapour uptake.

Bodies of liquid water as a source of water gain for Ixodes ricinus ticks (Acari: Ixodidae)

It is well established that free-living unfed ticks can compensate for their inevitable body water losses by active water vapour absorption in periods of ambient relative humidity (RH) of greater than 75-90%. Whilst many species of terrestrial arthropods are known to be capable of locating liquid water and drinking when in need, the existing knowledge concerning the ecological significance of bulk water for rehydration in ticks is scarce. In the present laboratory study batches of unfed Ixodes ricinus (larvae, nymphs, and adults) in varying states of (de)hydration were supplied with tapwater either as droplets for 45 min in a Petri dish or in a small trough for 24 h. The body mass of each tick was gravimetrically determined before and after exposure. Though fully hydrated ticks were not usually attracted to liquid water, the response became increasingly positive with a growing body water deficit (p > 0.05). No tick was ever found with its mouthparts inserted into the water, nor had any tick residues of orange G in its alimentary tract when supplied with orange G-coloured water. Linear regression analysis demonstrated that water intake took place in a regulated manner in adult ticks, i.e. the more severely the ticks had been dehydrated the more net water they subsequently gained (p > 0.01). The maximum uptake rates were <20% of the original body mass per day. These findings indicate that unfed I. ricinus do not drink liquid water but are well able to use the high RH in the immediate vicinity of bulk water to actively take up vapour.

Tick infection rates with Borrelia Ixodes ricinus versus Haemaphysalis concinna and Dermacentor reticulatus in two locations in eastern Germany

Unfed nymphal Ixodes ricinus, Haemaphysalis concinna, and adult Dermacentor reticulatus were collected in two locations of Saxony in July and September 1991 by flagging. In July, the abundance of nymphal I. ricinus was about 2–3 times higher than that of nymphal H.concinna, a time of the year when nymphs of both species are reported to have a seasonal peak of activity. No D.reticulatus were flagged concurrently. In September, host‐seeking activity of nymphal I.ricinus was again quite high as was that of adult D.reticulatus but only low numbers of nymphal H.concinna were collected. The flagged ticks were individually examined for Borrelia by an indirect immunofluorescence assay {I.ricinus: n = 414; H.concinna: n = 96; D.reticulatus: n= 116). The prevalence of Borrelia (probably B.burgdorferi) in I.ricinus varied from 12.1% to 21.0%. No borreliae were found in H. concinna. Of the examined D. reticulatus from one site (n= 97) 11.3% contained either B. burgdorferi or a related Borrelia. This may be the first finding of Borrelia in an Eurasian Dermacentor species.

Acquisition of Borrelia burgdorferi by Ixodes ricinus ticks fed on the European hedgehog, Erinaceus europaeus L.

A hedgehog, Erinaceus europaeus, was found to be heavily infested with larval and nymphal Ixodes ricinus in a forest park in Co. Galway, Ireland. A large proportion of the ticks that engorged and detached were infected with the spirochacte, Borrelia burgdorferi, the causative agent of human Lyme borreliosis. The identity of these spirochaetes was confirmed by immunofluorescent assay with B. burgdorferi-specific monoclonal antibody and by polymerase chain reaction test and they were transmitted from the hedgehog to laboratory-reared ticks and from the ticks obtained from the hedgehog to gerbils (Meriones unguiculatus). The high infection rate of the larvae that fed on the hedgehog in comparison with unfed larvae from the same habitat was interpreted as strong evidence that this host species is reservoir competent. Since hedgehogs can evidently feed adult ticks as well as many immature stages, they may well have an important role in the ecology of Lyme borreliosis in some habitats.

Prevalence of borrelia burgdorferi in Ixodes ricinus ticks in Berlin (West)

In 1986, 1711 nymphal and adult Ixodes ricinus orginating from Berlin (West) forests were examined individually or in pools of up to 10 ticks for the presence of Borrelia burgdorferi, the etiologic agent of Lyme borreliosis. Detection of borreliae was carried out by means of a culture method using modified Barbour-Stoenner-Kelly-Medium (BSK II). Tick populations from 14 out of 15 locations contained positive specimens. The calculated minimal infection rate of pooled ticks was 2.5% in nymphs (n = 1365), 10.2% in females (n = 59), and 5.3% in males (n = 114). Among those ticks examined individually, none of the nymphs (n = 49) proved to be positive but B. burgdorferi was isolated from 8.2% of the females (n = 73) and 7.8% of the males (n = 51). Fifty-five out of 56 isolates were identified as B. burgdorferi by means of an indirect immunofluorescence test (IFT) using monoclonal antibody H 5332. From these results B. burgdorferi must be considered as being present in the Berlin area.

Dynamics of Dissemination and Outer Surface Protein Expression of Different European Borrelia burgdorferi Sensu Lato Strains in Artificially Infected Ixodes ricinus Nymphs

ABSTRACT Unfed Ixodes ricinus nymphs were infected with eight different strains and clones of Borrelia afzelii and B. garinii by capillary feeding. Except one B. afzelii clone, all expressed OspC in culture. Tick midguts and salivary glands were investigated at different time intervals for the presence of borreliae and for OspA and OspC phenotypes by immunofluorescence with simultaneous staining of OspA and OspC with monoclonal antibodies. Both species were transmittable to I. ricinus. All OspC-expressing strains and clones were able to disseminate into the salivary glands. In contrast, the OspC-negative B. afzelii clone was not detectable in the salivary glands, an indication that OspC plays an important role in dissemination. OspA-positive borreliae prevailed in the midgut. OspC positives were more frequent in the salivary glands than in the midgut. Notably, simultaneously OspA- and OspC-negative borreliae were detected in both organs. Kinetics of dissemination varied with the strains. The OspC-positive B. afzelii clone and all B. garinii OspA type 4 strains were detectable in the salivary glands right after feeding, while one B. garinii OspA type 6 strain invaded the salivary glands with a delay of 24 h. These findings support the hypothesis that OspA is abundantly expressed in unfed ticks while upregulation of OspC is also a prerequisite for dissemination in the vector for the Eurasian species B. afzelii and B. garinii. However, we found strain-specific dynamics of Osp expression and strain-specific kinetics of systemic infection in the vector tick and it appears that additional factors are involved in the initiation and regulation of the dissemination process.

Variability in cuticular hydrocarbons and phenotypic discrimination of Ixodes ricinus populations (Acarina: Ixodidae) from Europe

The cuticular hydrocarbon composition and a stepwise discriminant analysis are used to elucidate the phenotypic relationships of 66 populations of Ixodes ricinus in Europe. The method correctly allocates Ixodes persulcatus (outgroup) populations away from the main cluster of I. ricinus samples and separates the samples into ten relatively defined clusters of specimens. Populations from Poland are inseparable from samples collected in Germany, Switzerland and the Italian Alps, while individuals from Slovakia and the Czech Republic come into separate groups of phenotypic similarity. Irish and British specimens are separated but highly related and Spanish populations show an unexpectedly high distance from the remaining clusters.

Tick-borne encephalitis in the age of general mobility

ZusammenfassungDie 11. Konferenz der International Scientific Working Group on Tick-borne Encephalitis (ISW TBE) stand unter dem Motto Von der Kindheit bis zum Seniorenalter: erhöhte Mobilität – gesteigertes Risiko für FSME-Infektionen?. Teilnehmer aus 26 Ländern einschließlich den USA und China berichteten über die jüngsten Entwicklungen und Trends im Hinblick auf lokale FSME-Infektionen, Impfschutz und Risikofaktoren. Im Speziellen wurde die Situation der Kinder und der Senioren diskutiert. Der aktuellen Datenlage zufolge haben sich Lokalisation und Ausdehnung endemischer FSME-Areale in den letzten Jahren im vermuteten Zusammenhang mit der Klimaerwärmung und der Ausbreitung der Zecken in höher gelegene Gebiete geändert. Die gesteigerte Mobilität der Menschen verstärkt die Exposition weiter; Aktivitäten im Freien und internationale Reisetätigkeit nehmen zu, auch und vor allem bei Personen über 50 Jahren, bei denen bereits per se ein höheres Risiko für Krankheitsmanifestationen, Komplikationen und Tod besteht. Die meisten Europäer verreisen innerhalb von Europa und wissen dabei oft nicht über endemische Gebiete Bescheid. Auf lange Sicht kann nur eine hohe Durchimpfungsrate eine niedrige Morbidität sicherstellen. Die Information der Öffentlichkeit ist die einzige effektive Maßnahme, mit der ein entsprechendes Bewusstsein geschaffen und dieses Ziel umgesetzt werden kann. Insgesamt sollte das Risiko der einzelnen Person für eine FSME-Infektion nicht als fixe Größe angesehen werden, sondern muss im individuellen Fall mithilfe des Wissens um endemische Gebiete und Risikofaktoren abgeschätzt werden.SummaryThe 11th meeting of the International Scientific Working Group on Tick-borne Encephalitis (ISW-TBE) was conducted under the title of, From childhood to golden age: increased mobility – increased risk of contracting TBE? Participants from 26 countries, including the United States of America and China, presented reports on the latest developments and trends in local TBE cases, vaccination coverage and risk factors. In particular, the situation of children and the elderly (the golden agers) was discussed. As the current evidence suggests, the location and extension of endemic areas for TBE have changed over the last few years, along with global warming and the shift of infected ticks to higher altitudes. The increased mobility of the human population adds to the heightened exposure; outdoor activities and international travel are on the rise also, and especially, amongst the 50+ generation, who are already per se at higher risk of disease manifestation, complications and case fatality. Most Europeans travel within Europe, often without sufficient awareness of endemic areas. Only high immunization rates can ensure low disease rates in the long run. To achieve this goal, public education is the sole effective approach for raising the level of awareness. Overall, the risk of any given person to contract TBE should not be regarded as a fixed entity, but rather it must be estimated individually, on the basis of knowledge of the TBE virus endemic areas and risk factors.

Visualization of Borrelia burgdorferi sensu lato by fluorescence in situ hybridization (FISH) on whole-body sections of Ixodes ricinus ticks and gerbil skin biopsies.

The objective of this study was to visualize borreliae directly in whole-body sections of Ixodes ricinus by fluorescence in situ hybridization (FISH). Borrelia afzelii mono-infected or Borrelia burgdorferi sensu stricto (ss)/B. afzelii double-infected nymphs were fixed, embedded in cold polymerizing resin and sectioned. The same sample processing was applied to skin biopsies taken from a Mongolian gerbil after an infectious tick-bite. FISH was carried out using 16S-rRNA-directed, fluorescence-labelled oligonucleotide probes specific for the genus Borrelia and specific within the group of Lyme borreliosis-associated genospecies B. afzelii, B. burgdorferi ss, Borrelia garinii and Borrelia valaisiana. Sensitivity and specificity of the newly designed probes were evaluated using PCR, dot-blot hybridizations and FISH. Despite significant autofluorescence of certain tick tissues (which allowed good histological orientation within the sections), borreliae showing the typical spirochaetal morphotype were clearly visible in five out of six putatively infected ticks. These findings were confirmed by electron microscopy of ticks from the same infected batch as used for FISH. Attempts to produce ticks infected by two different Borrelia genospecies were not successful. FISH on whole-body sections of resin-embedded ticks offers the possibility of visualizing and identifying borreliae within tick tissues. This technique has great potential for the investigation of the transmission of bacteria or other micro-organisms by arthropod vectors. Furthermore, clear visualization of single spirochaetes distributed along subcutaneous fat cell membranes in gerbil skin biopsies suggests that FISH might also be suitable for the detection of borreliae in clinical tissue specimens.