Co-feeding Ticks Research Articles

Interspecies co-feeding transmission of Powassan virus between a native tick, Ixodes scapularis, and the invasive East Asian tick, Haemaphysalis longicornis

BackgroundPowassan virus, a North American tick-borne flavivirus, can cause severe neuroinvasive disease in humans. While Ixodes scapularis are the primary vectors of Powassan virus lineage II (POWV II), also known as deer tick virus, recent laboratory vector competence studies showed that other genera of ticks can horizontally and vertically transmit POWV II. One such tick is the Haemaphysalis longicornis, an invasive species from East Asia that recently established populations in the eastern USA and already shares overlapping geographic range with native vector species such as I. scapularis. Reports of invasive H. longicornis feeding concurrently with native I. scapularis on multiple sampled hosts highlight the potential for interspecies co-feeding transmission of POWV II. Given the absence of a clearly defined vertebrate reservoir host for POWV II, it is possible that this virus is sustained in transmission foci via nonviremic transmission between ticks co-feeding on the same vertebrate host. The objective of this study was to evaluate whether uninfected H. longicornis co-feeding in close proximity to POWV II-infected I. scapularis can acquire POWV independent of host viremia.MethodsUsing an in vivo tick transmission model, I. scapularis females infected with POWV II (“donors”) were co-fed on mice with uninfected H. longicornis larvae and nymphs (“recipients”). The donor and recipient ticks were infested on mice in various sequences, and mouse infection status was monitored by temporal screening of blood for POWV II RNA via quantitative reverse transcription polymerase chain reaction (q-RT-PCR).ResultsThe prevalence of POWV II RNA was highest in recipient H. longicornis that fed on viremic mice. However, nonviremic mice were also able to support co-feeding transmission of POWV, as demonstrated by the detection of viral RNA in multiple H. longicornis dispersed across different mice. Detection of viral RNA at the skin site of tick feeding but not at distal skin sites indicates that a localized skin infection facilitates transmission of POWV between donor and recipient ticks co-feeding in close proximity.ConclusionsThis is the first report examining transmission of POWV between co-feeding ticks. Against the backdrop of multiple unknowns related to POWV ecology, findings from this study provide insight on possible mechanisms by which POWV could be maintained in nature.Graphical

Changes in host densities and co-feeding pattern efficiently predict tick-borne encephalitis hazard in an endemic focus in northern Italy

Tick-borne encephalitis is an important zoonosis in many parts of north-western, central and eastern Europe, Russia and the Far East, with considerable altitudinal and latitudinal shifts described during recent decades. The reported routes of transmission for TBE virus include the saliva-activated non-viraemic transmission between co-feeding ticks taking place on rodent hosts. During the period 2001–2014, a population of the yellow-necked mouse (Apodemus flavicollis), which is considered among the most efficient TBE competent host, especially in central and western Europe, was intensively live-trapped in a known TBE focus in the Province of Trento, Italy. Individual live-trapped mice were checked for the number and position of feeding ticks and serologically screened for TBEv antibodies. A combined effect of climatic conditions and density of both roe deer and mice on the number of co-feeding tick groups was observed. Specifically, the occurrence of co-feeding ticks on mice during the questing season was affected by autumnal cooling in the previous season. On the other hand, co-feeding occurrence was also positively associated with roe deer abundance, while mouse density showed a hump-shaped pattern. Individual features of A. flavicollis such as weight and sex also affected co-feeding occurrence with the heaviest (breeding adult) males carrying more co-feeding ticks. We also found that the overall number of co-feeding ticks on mice positively affected TBEv antibody detection in this species the following year. In conclusion, a specific combination of climatic conditions in conjunction with certain rodent and roe deer densities are the principal determinants of the number of co-feeding ticks on A. flavicollis and, consequently, TBEv circulation. These variables can be used to provide an early warning signal for a TBE hazard, thus representing a useful tool for Public Health authorities to prepare action for prevention and control within TBEv circulation areas.

Assessing systemic and non-systemic transmission risk of tick-borne encephalitis virus in Hungary.

Estimating the tick-borne encephalitis (TBE) infection risk under substantial uncertainties of the vector abundance, environmental condition and human-tick interaction is important for evidence-informed public health intervention strategies. Estimating this risk is computationally challenging since the data we observe, i.e., the human incidence of TBE, is only the final outcome of the tick-host transmission and tick-human contact processes. The challenge also increases since the complex TBE virus (TBEV) transmission cycle involves the non-systemic route of transmission between co-feeding ticks. Here, we describe the hidden Markov transition process, using a novel TBEV transmission-human case reporting cascade model that couples the susceptible-infected compartmental model describing the TBEV transmission dynamics among ticks, animal hosts and humans, with the stochastic observation process of human TBE reporting given infection. By fitting human incidence data in Hungary to the transmission model, we estimate key parameters relevant to the tick-host interaction and tick-human transmission. We then use the parametrized cascade model to assess the transmission potential of TBEV in the enzootic cycle with respect to the climate change, and to evaluate the contribution of non-systemic transmission. We show that the TBEV transmission potential in the enzootic cycle has been increasing along with the increased temperature though the TBE human incidence has dropped since 1990s, emphasizing the importance of persistent public health interventions. By demonstrating that non-systemic transmission pathway is a significant factor in the transmission of TBEV in Hungary, we conclude that the risk of TBE infection will be highly underestimated if the non-systemic transmission route is neglected in the risk assessment.

A simple model for the establishment of tick-borne pathogens of Ixodes scapularis: A global sensitivity analysis of R0

The basic reproduction number of a pathogen, R0, determines whether a pathogen will spread (R0>1), when introduced into a fully susceptible population or fade out (R0<1), because infected hosts do not, on average, replace themselves. In this paper we develop a simple mechanistic model for the basic reproduction number for a group of tick-borne pathogens that wholly, or almost wholly, depend on horizontal transmission to and from vertebrate hosts. This group includes the causative agent of Lyme disease, Borrelia burgdorferi, and the causative agent of human babesiosis, Babesia microti, for which transmission between co-feeding ticks and vertical transmission from adult female ticks are both negligible. The model has only 19 parameters, all of which have a clear biological interpretation and can be estimated from laboratory or field data. The model takes into account the transmission efficiency from the vertebrate host as a function of the days since infection, in part because of the potential for this dynamic to interact with tick phenology, which is also included in the model. This sets the model apart from previous, similar models for R0 for tick-borne pathogens. We then define parameter ranges for the 19 parameters using estimates from the literature, as well as laboratory and field data, and perform a global sensitivity analysis of the model. This enables us to rank the importance of the parameters in terms of their contribution to the observed variation in R0. We conclude that the transmission efficiency from the vertebrate host to Ixodes scapularis ticks, the survival rate of Ixodes scapularis from fed larva to feeding nymph, and the fraction of nymphs finding a competent host, are the most influential factors for R0. This contrasts with other vector borne pathogens where it is usually the abundance of the vector or host, or the vector-to-host ratio, that determine conditions for emergence. These results are a step towards a better understanding of the geographical expansion of currently emerging horizontally transmitted tick-borne pathogens such as Babesia microti, as well as providing a firmer scientific basis for targeted use of acaricide or the application of wildlife vaccines that are currently in development.

Non-viraemic transmission of tick-borne viruses

Tick-borne viruses are causative agents of several important human diseases. Tick-borne encephalitis virus (TBEV) is the most prominent representative considered medically to be the most important arbovirus (arthropod-borne virus) in Europe and northern Asia. Tick-borne virus transmission cycles are determined by the interactions between viruses, vectors, and their vertebrate hosts. Several mechanisms of tick-borne virus circulation in nature are currently considered to include transovarial transmission via the eggs from an infected female tick to its offspring, "viraemic" transmission between host and tick via feeding on a viraemic, infectious vertebrate hosts, and the virus transmission between co-feeding ticks, termed non-viraemic transmission (NVT). For NVT, the local skin site where ticks aggregately feed is an important focus of viral replication where migratory immune cells provide a vehicle for virus transmission from infected to uninfected co-feeding ticks. For TBEV at least, NVT is an important mechanism of virus maintenance in nature and offers explanations for some specific aspects of tick-borne virus ecology such as focal virus distribution and vector competency of particular tick species.

Transmission of tick-borne pathogens between co-feeding ticks: Milan Labuda's enduring paradigm

During the 1990s, Milan Labuda's experimental results established a new paradigm for the study of tick-borne viruses that has since been strengthened by its demonstrated effectiveness in explaining the epidemiology of tick-borne encephalitis (TBE). This brief review summarizes the essential features of the transmission of tick-borne pathogens such as TBE virus. Leukocytes migrate between tick feeding sites, bearing infective virions and providing a transport route for the virus between co-feeding ticks independent of a systemic viraemia. Such tick-borne pathogens are thus transmitted from tick to tick via vertebrates; the ticks are the reservoirs as well as the vectors, while the vertebrate is the transient bridge. The aim is to bring the related but non-synonymous terms (co-feeding and non-systemic) to the attention of workers who use simple PCR screening to identify additional vertebrate reservoir hosts of vector-borne pathogens that are not in fact maintained in nature through systemic transmission.

Borrelia afzelii ospC genotype diversity in Ixodes ricinus questing ticks and ticks from rodents in two Lyme borreliosis endemic areas: Contribution of co-feeding ticks

In Europe, the Lyme borreliosis (LB) agents like Borrelia burgdorferi sensu stricto (ss), B. afzelii, and B. garinii are maintained in nature by enzoonotic transmission cycles between vertebrate hosts and Ixodes ricinus ticks. The outer surface protein C is a highly antigenic protein expressed by spirochaetes during transmission from ticks to mammals as well as during dissemination in the vertebrate hosts. Previous studies based on analysis of ospC gene sequences have led to the classification of ospC genotypes into ospC groups. The aim of this study was to analyse and compare ospC group distribution among isolates of the rodent-associated genospecies, B. afzelii, at 3 levels (questing ticks, ticks feeding on rodents, and xenodiagnostic ticks). Isolates were obtained during a study carried out in 2 LB endemic areas located on the Swiss Plateau [Portes-Rouges (PR) and Staatswald (SW)], where rodents were differently infested by co-feeding ticks (Pérez et al., unpublished data). Overall, we identified 10 different ospC groups with different distributions among isolates from questing ticks, ticks that detached from rodents, and xenodiagnostic ticks at the 2 sites. We observed a higher ospC diversity among isolates from ticks that fed on rodents at SW, and mixed infections with 2 ospC groups were also more frequent among isolates from ticks that fed on rodents at SW ( n = 18) than at PR ( n = 1). At both sites, B. afzelii isolates obtained from larvae that were feeding on the rodents simultaneously with nymphs displayed a higher diversity of ospC groups (mean number of ospC groups: 2.25 for PR and 1.75 for SW) than isolates from larvae feeding without nymphs (mean number of ospC groups: 1.17 for PR and 1 for SW). We suggest that co-feeding transmission of Borrelia, previously described in laboratory models, contributes in nature in promoting and maintaining ospC diversity within local tick populations.

Co-feeding as a route for transmission of Rickettsia conorii israelensis between Rhipicephalus sanguineus ticks

Rickettsia conorii is widely distributed in Europe, Asia, and Africa. The brown dog tick, Rhipicephalus sanguineus, is the recognized vector of R. conorii. In this study, we assessed the efficiency of R. conorii israelensis transmission between co-feeding Rh. sanguineus ticks. Infected Rh. sanguineus adults and uninfected nymphs were fed simultaneously upon either naïve dogs or a dog previously exposed to this agent. When ticks were placed upon naïve dogs, 92-100% of nymphs acquired the infection and 80-88% of infected engorged nymphs transmitted it transstadially. When ticks were placed upon a seropositive dog, only 8-28.5% of recipient nymphs became infected. Our results establish the first evidence for efficient natural transmission of R. conorii israelensis between co-feeding ticks upon both naïve and seropositive dogs. This route of transmission can ensure continuous circulation of R. conorii israelensis in tick vectors even in the absence of naïve reservoir hosts.

To what extent has climate change contributed to the recent epidemiology of tick-borne diseases?

There is no doubt that all vector-borne diseases are very sensitive to climatic conditions. Many such diseases have shown marked increases in both distribution and incidence during the past few decades, just as human-induced climate change is thought to have exceeded random fluctuations. This coincidence has led to the general perception that climate change has driven disease emergence, but climate change is the inevitable backdrop for all recent events, without implying causality. Coincidence and causality can be disentangled using tick-borne encephalitis (TBE) as a test case, based on the excellent long-term data for this medically significant European disease system. Detailed analysis of climate records since 1970 has revealed abrupt temperature increases just prior to the dramatic upsurge in TBE incidence in many parts of central and eastern Europe. Furthermore, the seasonal patterns of this temperature change are such as might have favoured the transmission of TBE virus between co-feeding ticks. Nevertheless, the pattern of climate change is too uniform to explain the marked heterogeneity in the timing and degree of TBE upsurge, for example in different counties within each of the Baltic countries. Recent decreases as well as increases in TBE incidence must also be taken into account. Instead of a single cause, a network of interacting factors, acting synergistically but with differential force in space and time, would generate this epidemiological heterogeneity. From analysis of past and present events, it appears that human behavioural factors have played a more significant role than purely biological enzootic factors, although there is an explicit causal linkage from one to the other. This includes a range of abiotic and biotic environmental factors, together with human behaviour determined by socio-economic conditions. Many of the abrupt changes followed from the shift from planned to market economies with the fall of Soviet rule. Comparisons between eight countries have indeed revealed a remarkable correlation between poverty indicators and the relative degree of upsurge in TBE from 1993. Against this background of longer-term shifts in TBE incidence, sudden spikes in incidence appear to be due to exceptional weather conditions affecting people's behaviour, which have a differential impact depending on socio-economic factors. This new perspective may also help explain the epidemiology of Crimean-Congo haemorrhagic fever around the eastern Mediterranean region, including the current exceptional epidemic in Turkey.

Tick-borne disease systems emerge from the shadows: the beauty lies in molecular detail, the message in epidemiology

This review focuses on some of the more ground-shifting advances of recent decades, particularly those at the molecular and cellular level that illuminate mechanisms underpinning the natural ecology of tick-host-pathogen interactions and the consequent epidemiology of zoonotic infections in humans. Knowledge of components of tick saliva, now recognized as the central pillar in the tick's ability to complete its blood meal and the pathogen's differential ability to use particular hosts for transmission, has burgeoned with new molecular techniques. Functional studies have linked a few of them to saliva-assisted transmission of non-systemic infections between co-feeding ticks, the quantitative key to persistent cycles of the most significant tick-borne pathogen in Europe. Human activities, however, may be equally important in determining dynamic patterns of infection incidence in humans.

Vector seasonality, host infection dynamics and fitness of pathogens transmitted by the tickIxodes scapularis

Fitness of tick-borne pathogens may be determined by the degree to which their infection dynamics in vertebrate hosts permits transmission cycles if infective and uninfected tick stages are active at different times of the year. To investigate this hypothesis we developed a simulation model that integrates the transmission pattern imposed by seasonally asynchronous nymphal and larval Ixodes scapularis ticks in northeastern North America, with a model of infection in white-footed mice (Peromyscus leucopus) reservoir hosts, using the bacteria Borrelia burgdorferi and Anaplasma phagocytophilum as examples. In simulations, survival of microparasites, their sensitivity to reduced rodent and tick abundance, and to 'dilution' by a reservoir-incompetent host depended on traits that allowed (i) highly efficient transmission from acutely-infected hosts, (ii) long-lived acute or 'carrier' host infections, and/or (iii) transmission amongst co-feeding ticks. Minimum values for transmission efficiency to ticks, and duration of host infectivity, necessary for microparasite persistence, were always higher when nymphal and larval ticks were seasonally asynchronous than when these instars were synchronous. Thus, traits influencing duration of host infectivity, transmission efficiency to ticks and co-feeding transmission are likely to be dominant determinants of fitness in I. scapularis-borne microparasites in northeastern North America due to abiotic forcings influencing I. scapularis seasonality.

The basic reproductive number of tick–borne encephalitis virus

Tick-borne encephalitis virus (TBEV) is reciprocally transmitted between Ixodes ricinus ticks and small mammals. Recently, transmission between co-feeding ticks has been postulated as an epidemiological by important mechanism of perpetuating the agent. To empirically examine the question whether the "traditional" mode of transmission is sufficient to maintain enzootic TBEV transmission, the basic reproductive number R(0) of TBEV could be estimated under this model for sites in which TBEV is enzootic. I propose an empirical estimator of R(0) for TBEV which is based on longitudinal stage-specific local tick infestation densities assessed by live trapping of small mammals. A Gibbs sampler-based 95%-credibility interval is presented. When applied to published field data from TBEV enzootic sites sub-critical R(0) estimates are obtained for both sites. I discuss potential shortcomings of this method and possible implications of these findings on the discussion of supplemental mechanisms of transmission.

Co-feeding Transmission and Its Contribution to the Perpetuation of the Lyme Disease SpirocheteBorrelia afzelii(In Reply to Randolph and Gern)

To the Editor: Although transmission between co-feeding vector ticks may perpetuate particular tick-borne viruses, this mode of transmission plays no role in the epizootiology of Lyme disease spirochetes (1,2). In their letter, Randolph and Gern defend their suggestion that tick-borne pathogens perpetuate effectively by direct passage from one feeding tick to another by criticizing our analysis (3). These researchers mainly address our comparison of the transmission efficiency between simultaneously feeding ticks with that between ticks feeding sequentially on a persistently infected rodent. Our experiments demonstrate that approximately six times as many larvae (85.4%) acquire Borrelia afzelii spirochetes from a systemically infected mouse than from a mouse on which an infected nymph is feeding simultaneously (13.6%) (1). In nature, however, larval ticks rarely co-feed with nymphs on mice or voles; only approximately one fifth (18.8%) of these hosts harbor both subadult stages simultaneously. And of the nymphs, only approximately one quarter (26.4%) are infected by Lyme disease spirochetes. As a result, the natural transmission efficiency between simultaneously feeding ticks would be only one twentieth (5%) of that observed in the laboratory. Multiplying the experimentally observed efficiency of co-feeding transmission (13.6%) by the likelihood of larval and nymphal ticks co-infesting small rodents, as well as by the prevalence of infected nymphal ticks, reduces the efficiency of co-feeding transmission in nature to <1%. Although Randolph and Gern commit several minor mathematical errors, their calculations support our argument that few larval vector ticks would acquire spirochetal infection directly from an infected nymph (3). Randolph and Gern err, however, by applying the same mathematical modifications to the transmission efficiency by which larvae acquire spirochetes from a persistently infected host (3). Whereas the efficiency of co-feeding transmission observed in the laboratory must be modified to pay tribute to the rare event when larvae co-feed with an infected nymph in nature, the efficiency by which larvae acquire infection from a persistently infected host is independent of such limiting parameters. Because a competent rodent host remains infectious to larval ticks throughout its life, the proportion of hosts infested by particular subadult stages of the vector is irrelevant. Thus, the transmission efficiency on a persistently infected host is unchanged in the laboratory and the field. Almost 85.4% of larvae feeding on mice or voles in nature would, therefore, acquire spirochetal infection—far more than by co-feeding. We are correct in stating that natural transmission by sequentially feeding ticks is more efficient than transmission between co-feeding ticks. Randolph and Gern suggest that we could have recorded the distance between the feeding ticks to clarify whether the increase from a 13.6% transmission efficiency between co-feeding ticks to a transmission efficiency of 85.4% from a persistently infected host is associated with the development of a systemic infection. Our experimental observation (Table 1 in our article [1]), as well as a study on the movement of spirochetes through their host’s skin (4), conclusively demonstrates that the increase in transmission efficiency is due to the progressive dissemination of spirochetes from the site of inoculation. The likelihood of a larva’s acquiring spirochetes from any site of its host’s skin increases with the passage of time since the infected nymph attached. To compare the two modes of transmission in terms of efficiency (Table 2 in our article [1]), we permitted the larvae to attach randomly to their rodent hosts, just as they would do in nature. Table 1 Spirochetal infection in larval Ixodes ricinus ticks that fed on mice during the period of attachment of a single Borrelia afzelii–infected nymph and that fed at specified distances from the infecting nympha In the epizootiology of Lyme disease spirochetes, “simultaneous” transmission between co-feeding ticks (<1%) is some two orders of magnitude less efficient than sequential transmission between ticks feeding on persistently infected reservoir rodents (85.4%). The two studies that relied on natural infestation densities and refrained from using artificial feeding capsules conclusively demonstrated that transmission of Lyme disease spirochetes between ticks feeding simultaneously and in close proximity contributes little to the perpetuation of this pathogen, either in North America or in Europe (1,2). We are correct in concluding that Lyme disease spirochetes are maintained in nature mainly by sequential attachment of ticks to persistently infected reservoir hosts. Table 2. Spirochetal infection in larval Ixodes ricinus ticks that fed randomly on bodies of mice beginning at 3 days and 14 days after a single Borrelia afzelii–infected nymph had begun to feed

Evidence of ehrlichiosis agents found in ticks (Acari: Ixodidae) collected from migratory birds.

Two Ehrlichia pathogens were found in immature Ixodes ricinus (L.) ticks collected from migratory passerine birds in the Curonian Spit area of the Baltic Region of Russia (Kaliningrad enclave). During the spring and fall of 2000, 1,606 passerine birds (eight species) were collected; 6.8% of them (110/1,606) were infested by ticks, and 51.8% (57/110) of tick clusters contained various human pathogenic microorganisms. Human monocytic ehrlichiosis (HME) and human granulocytic ehrlichiosis (HGE) agents were found in 14% (8/57) of cases. Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi sensu stricto were found in 92.9% (53/57) of the ticks. In five out of eight cases, infection of both Ehrlichia and Borrelia were obtained. In one case, a single nymph contained HME, B. afzelii, and B. garinii. Borrelia burgdorferi s.s. and B. afzelii were found together in one pool of four nymphs and one larva. All agents were identified using polymerase chain reaction and species-specific primers. In 8.8% of the ticks collected from birds in the fall and 22% in the spring, pathogens were isolated from attached co-feeding nymphs and larvae. These data demonstrate that Ehrlichia exchange could occur between co-feeding ticks on animals without systemic infection.

Global warming and the spread of disease: the debate heats up

Many recent studies have suggested that as a consequence of global climate change there will be an increase in the incidence of vector-borne diseases, such as malaria, yellow fever, dengue fever and tick-borne encephalitis. This is based principally on the idea that, as the planet warms and parasite and vector development rates speed up, transmission will be possible at increasingly higher latitudes. However, two new papers by Sarah Randolph and David Rogers challenge this notion and suggest that due to the complex nature of most host–vector–parasite interactions it may be extremely difficult to make good generalizations about the future distributions of vector-transmitted diseases.In a paper published in Science, Rogers and Randolph explore the effects of global climate change on the future incidence of cerebral malaria, a life-threatening disease caused by Plasmodium falciparum and transmitted by anopheline mosquitoes 1xThe global spread of malaria in a future, warmer world. Rogers, D.J. and Randolph, S.E. Science. 2000; 289: 1763–1766Crossref | PubMedSee all

Survival strategy of tick-borne encephalitis virus: Cellular basis and environmental determinants

Although TBE virus can be transmitted in the laboratory by a wide variety of ixodid tick species to a wide variety of vertebrate host species, nevertheless in nature endemic cycles of TBE virus depend principally on just two tick species, Ixodes ricinus in the western and I. persulcatus in the eastern Palaearctic. A complete transmission cycle, from tick to tick via vertebrates, occurs most efficiently between co-feeding ticks in the absence of a systemic viraemia. This non-systemic route depends on TBE virus replication within particular immunocompetent cells in the skin, and only certain vertebrate species, notably Apodemus mice, are susceptible to this. Amongst the potential tick vectors in Europe, only I. ricinus has the correct host relationships and appropriate natural life cycle to support such non-systemic transmission cycles. Within the wide European distribution of this tick-host relationship, only in certain places do larval and nymphal ticks feed together on the same hosts with sufficient coincidence to ensure TBE virus survival. The environmental factors that determine this seasonal coincidence are being identified with the help of remotely-sensed meteorological satellite imagery to create predictive risk maps of TBE foci.

Tick-Borne Encephalitis Virus Transmission between Ticks Cofeeding on Specific Immune Natural Rodent Hosts

To determine whether the portion of a vertebrate host population having specific immunity to tick-borne encephalitis (TBE) virus can participate in the TBE virus transmission cycle, natural hosts immunized against TBE virus were challenged with infected and uninfected ticks. Yellow-necked field mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus) were either immunized with TBE virus by subcutaneous inoculation of the virus, or they were exposed to virus-infectedIxodes ricinusticks. One month later, when serum neutralizing antibody was detectable, the animals were infested with infected (donor) adult female ticks and uninfected (recipient) nymphal ticks; recipients were allowed to feed either in close contact (chamber 1) or physically separated (chamber 2) from the infected donor ticks. Following challenge with infected (and uninfected) ticks, viremia developed in all the control, nonimmune animals, whereas viremia was undetectable in all those animals naturally immunized by previous exposure to infected ticks. Despite the presence of neutralizing antibodies in all the immunized animals, 89% (24/27) immune animals supported virus transmission between infected and uninfected cofeeding ticks. Most transmission was localized, occurring within chamber 1; disseminated transmission from chamber 1 to chamber 2 was reduced. Immunization by tick bite was more effective than immunization by syringe inoculation in blocking cofeeding virus transmission. Nevertheless 75% (9/12) animals with “natural” immunity still supported transmission. The results demonstrate that natural hosts having neutralizing antibodies to TBE virus (and no detectable viremia) can still support virus transmission between infected and uninfected ticks feeding closely together on the same animal. These observations have important epidemiological implications relating to the survival of TBE virus in Nature.

Transmission of louping ill virus between infected and uninfected ticks co-feeding on mountain hares.

Most of the data on oral infection of ticks by louping ill virus have been obtained from experiments in which animals were infected by syringe inoculation with infectious material. Using infected ticks to mimic the natural situation, we have demonstrated that louping ill (LI) virus transmission can occur from infected to uninfected Ixodes ricinus feeding in close proximity on mountain hares (Lepus timidus). Under these conditions the hares developed either low or undetectable viraemias. Highest prevalence of LI virus infection was observed in recipient nymphs which had fed to repletion between days 3 and 7 post-attachment of virus-infected adults; following engorgement, 56% of nymphs acquired virus. These results demonstrate the efficient transmission of LI virus between co-feeding ticks on naive mountain hares. However, when ticks were allowed to co-feed on virus-immune hares a significant reduction in the frequency of infection was observed. Neither red deer (Cervus elaphus) nor New Zealand White rabbits supported transmission of LI virus. The significance of virus transmission between cofeeding ticks on LI virus epidemiology is discussed.