Coinfection of Babesia and Borrelia in the Tick Ixodes ricinus-A Neglected Public Health Issue in Europe?

Ixodes ricinus nymphs and adults removed from humans, and larvae and nymphs from birds, have been analysed for infection with Babesia species and Borrelia species previously in separately published studies. Here, we use the same data set to explore the coinfection pattern of Babesia and Borrelia species in the ticks. We also provide an overview of the ecology and potential public health importance in Sweden of I. ricinus infected both with zoonotic Babesia and Borrelia species. Among 1952 nymphs and adult ticks removed from humans, 3.1% were PCR-positive for Babesia spp. Of these Babesia-positive ticks, 43% were simultaneously Borrelia-positive. Among 1046 immatures of I. ricinus removed from birds, 2.5% were Babesia-positive, of which 38% were coinfected with Borrelia species. This study shows that in I. ricinus infesting humans or birds in Sweden, potentially zoonotic Babesia protozoa sometimes co-occur with human-pathogenic Borrelia spp. Diagnostic tests for Babesia spp. infection are rarely performed in Europe, and the medical significance of this pathogen in Europe could be underestimated.

The biology of Ixodes ricinus with emphasis on its ecology.

Prior to its identification as the vector of Lyme borreliosis spirochaetes in Europe in 1983, interest in Ixodes ricinus (L.) was moderate and mainly concerned the transmission of pathogens to farm animals and of tick-borne encephalitis virus to humans. The situation now is very different, and more papers have been published on I. ricinus than on any other ixodid tick species. However, this large literature is scattered and in recent years has become dominated by the molecular detection and characterization of the many pathogens that I. ricinus transmits. Several decades have now elapsed since a review addressing its basic biology and ecology appeared, and the present publication seeks to present basic aspects of its biology and ecology that are related to its role as a vector of disease agents, including its life cycle, feeding behaviour, host relations, survival off the host, and the impact of weather and climate.

Ticks, Human Babesiosis and Climate Change.

The effects of current and future global warming on the distribution and activity of the primary ixodid vectors of human babesiosis (caused by Babesia divergens, B. venatorum and B. microti) are discussed. There is clear evidence that the distributions of both Ixodes ricinus, the vector in Europe, and I. scapularis in North America have been impacted by the changing climate, with increasing temperatures resulting in the northwards expansion of tick populations and the occurrence of I. ricinus at higher altitudes. Ixodes persulcatus, which replaces I. ricinus in Eurasia and temperate Asia, is presumed to be the babesiosis vector in China and Japan, but this tick species has not yet been confirmed as the vector of either human or animal babesiosis. There is no definite evidence, as yet, of global warming having an effect on the occurrence of human babesiosis, but models suggest that it is only a matter of time before cases occur further north than they do at present.

Vectors of Babesiosis.

Babesiosis, caused by piroplasmid protozoans in the genus Babesia, is arguably the most important vector-borne disease of livestock and companion animals and is growing in importance as a zoonosis. Ixodid ticks were identified as vectors more than a hundred years ago, but the particular tick species transmitting some significant pathogens are still unknown. Moreover, it is only recently that the complexity of the pathogen-tick relationship has been revealed as a result of studies enabled by gene expression and RNA interference methodology. In this article, we provide details of demonstrated and incriminated vectors, maps of the current knowledge of vector distribution, a summary of established features of the pathogen life cycle in the vector, and an outline of molecular research on pathogen-tick relationships. The article concludes with a discussion of vector ecology and disease epidemiology in a global-change context and with suggestions for future research.

Ecosystem change and zoonoses in the Anthropocene.

Changes in land use, animal populations and climate, primarily due to increasing human populations, drive the emergence of zoonoses. Force of infection (FOI), which for these diseases is a measure of the ease with which a pathogen reaches the human population, can change with specific zoonoses and context. Here, we outline three ecosystem categories-domestic, peridomestic and sylvatic, where disease ecology alters the FOI of specific zoonoses. Human intervention is an overriding effect in the emergence of zoonoses; therefore, we need to understand the disease ecology and other influencing factors of pathogens and parasites that are likely to interact differently within ecological and cultural contexts. Planning for One Health and community ecology, such as an ecological impact assessment, is required to prepare and manage the emergence and impact of zoonoses in the Anthropocene.

Blood donor Borrelia burgdorferi sensu lato seroprevalence and history of tick bites at a northern limit of the vector distribution.

In order to study the antibody seroprevalence of the causal agent of Lyme borreliosis, Borrelia burgdorferi sensu lato (s.l.), and the history of tick bites at a geographical distribution limit of Ixodes ricinus, we compared healthy blood donors in geographically extreme regions: the borreliosis-endemic Vestfold County (59°N) and the region of northern Norway. Blood samples were screened using IgG/VlsE ELISA, and positive/borderline samples were confirmed using C6 ELISA and immunoblot assays. Also, donors completed a questionnaire consisting of several items including the places they have lived, and whether they owned any pets. The seroprevalence was 0.48% (5/1048) in northern Norway and 9.25% (48/519) in Vestfold County. Seven donors (of 1048) had experienced a single tick bite in the southern part of Nordland County (65°N) in northern Norway. This first study on B. burgdorferi s.l. antibody seroprevalence and tick bites on humans and pets in northern Norway showed that the seroprevalence of B. burgdorferi s.l. infection and the risk of tick bite in northern Norway are insignificant; the fact that only five positive IgG samples were detected underscores the very low background seroprevalence. These results suggest that so far I. ricinus has not expanded north of the previously established geographical distribution limit.

Diapause in ticks of the medically important Ixodes ricinus species complex.

Four members of the Ixodes ricinus species complex, Ixodes pacificus, Ixodes persulcatus, Ixodes ricinus and Ixodes scapularis, have, between them, a worldwide distribution within the northern hemisphere. They are responsible for the transmission of several animal and human pathogens, including the causal agents of Lyme borreliosis, tick-borne encephalitis, human granulocytic anaplasmosis and human babesiosis. Despite the importance of these ticks as vectors, the knowledge and understanding of the role that diapause plays in their complex life cycles are confused and incomplete. In view of the continuing geographic spread of these tick species, as well as the effects of climate change on vector-borne diseases, it is timely to encourage research on diapause phenomena to improve understanding of their biology and of pathogen transmission dynamics. In our review we seek to clarify thinking on the topic and to address gaps in our knowledge that require the attention of researchers.

Changing incidence of bovine babesiosis in Ireland.

BACKGROUND: In Ireland bovine babesiosis is caused by the tick-borne blood parasite, Babesia divergens. A survey of veterinary practitioners and farmers in the 1980's revealed an annual incidence of 1.7% associated with considerable economic losses. However, two subsequent surveys in the 1990's indicated a decline in clinical babesiosis. Recent evidence from continental Europe suggests that, probably due to climate change, the distribution of the tick vector of B. divergens, Ixodes ricinus is extending to more northerly regions and higher altitudes. In addition, milder winters are thought to widen the window of tick activity. In order to determine whether any such changes have affected the incidence of bovine babesiosis in Ireland, a questionnaire survey of farmers and veterinarians was carried out and compared with data from previous surveys. RESULTS: Our survey indicates that while the incidence of clinical disease has continued to decline, cases can occur at any time of year. In contrast to previous surveys, affected farms were the same size as unaffected ones. There was no correlation between disease risk and the presence of deer on the land. Disease severity and mortality rates were increased because many infections were advanced by the time they were detected and treated. CONCLUSION: While the precise reasons for the decline in the incidence of redwater are unknown, changes in agricultural practice are likely to be of importance. A reversal of the trend could be devastating, as vigilance among farmers and veterinarians is flagging and the national herd is losing its protective immunity to disease.

Research on the ecology of ticks and tick-borne pathogens--methodological principles and caveats.

Interest in tick-transmitted pathogens has experienced an upsurge in the past few decades. Routine application of tools for the detection of fragments of foreign DNA in ticks, together with a high degree of interest in the quantification of disease risk for humans, has led to a marked increase in the number of reports on the eco-epidemiology of tick-borne diseases. However, procedural errors continue to accumulate in the scientific literature, resulting in misleading information. For example, unreliable identification of ticks and pathogens, erroneous interpretations of short-term field studies, and the hasty acceptance of some tick species as vectors have led to ambiguities regarding the vector role of these arthropods. In this review, we focus on the ecological features driving the life cycle of ticks and the resulting effects on the eco-epidemiology of tick-transmitted pathogens. We review the factors affecting field collections of ticks, and we describe the biologically and ecologically appropriate procedures for describing tick host-seeking activity and its correlation with environmental traits. We detail the climatic variables that have biological importance on ticks and explain how they should be properly measured and analyzed. We also provide evidence to critically reject the use of some environmental traits that are being increasingly reported as the drivers of the behavior of ticks. With the aim of standardization, we propose unambiguous definitions of the status of hosts and ticks regarding their ability to maintain and spread a given pathogen. We also describe laboratory procedures and standards for evaluating the vectorial capacity of a tick or the reservoir role of a host. This approach should provide a coherent framework for the reporting of research findings concerning ticks and tick-borne diseases.

Natural isotope signatures of host blood are replicated in moulted ticks.

This proof-of-concept study demonstrates that stable isotope ratios of nitrogen and carbon (expressed as δ(13)C and δ(15)N) of host blood are faithfully reproduced in unfed nymphal Ixodes ricinus that developed from larvae fed on that host. Measured isotopic discrimination (i.e. the tick-blood spacing) was between -0.1 and 0.7‰ for δ(13)C and 3.8 and 3.9‰ for δ(15)N. Both δ(13)C and δ(15)N increased significantly with tick ageing. The isotopic analysis of unfed ticks has potential for determining the physiological age of unfed ticks, for identifying the season in which the previous stage had fed and for identifying the main hosts utilized by ticks.

Babesias of red deer (Cervus elaphus) in Ireland.

Blood samples were obtained from 38 wild red deer (Cervus elaphus) at two sites in Ireland and subjected to PCR analysis of the 18S rRNA gene followed by sequencing. Two fragments of the 18S rRNA gene were generated by two different PCR protocols and subsequent sequencing suggested that at least six of the deer were infected by a babesia that, in those loci, is indistinguishable from Babesia divergens, an important tick-borne pathogen of cattle and of zoonotic significance. Additionally, a B. odocoilei-like parasite was detected in three samples and a babesia that did not match any sequences in the GenBank database was found in five samples. Neither B. capreoli nor B. venatorum (EU1) were found. There have been several reports of B. divergens occurring in deer species, including red deer, roe deer (Capreolus capreolus) and reindeer (Rangifer tarandus). However, in view of recent re-sequencing of bovine-origin samples deposited previously in GenBank, it is unlikely that any of these sequences from deer are B. divergens. The present study describes the only deer piroplasm detected so far that shows complete identity with B. divergens, in just over half of the 18S rRNA gene. The entire gene of this deer parasite should be analysed and transmission experiments undertaken before the infectivity of B. divergens for red deer can be confirmed.

Identity of the causal agents of human babesiosis in Europe.

Most cases of human babesiosis are caused either by Babesia divergens in Europe or Babesia microti in America. B. microti, once regarded as a single species, occurs as a world-wide species complex and although both phenotypic and genotypic features lend support to suggestions that zoonotic B. microti may occur in Europe, convincing medical evidence is lacking. Several B. divergens-like parasites have emerged in the last few years, but 18S rRNA gene analysis suggests that B. divergens 'sensu stricto' is restricted to European (and North African) cattle. Some of the B. divergens-like parasites only differ from the bovine type by a few bases, and it remains to be determined whether this is sufficient to accord them separate species status. Comparative biology should support genetic data in taxonomic studies of both B. divergens and B. microti.

Possible mechanisms underlying age-related resistance to bovine babesiosis.

Calves infected with the tick-borne parasites Babesia spp. do not develop severe clinical babesiosis. Instead they display persistent low parasitaemias without any apparent ill-effects. This age-related resistance not only benefits the host, but also furthers parasite transmission. Both calves and adult animals respond with a Th I immune response to primary infection. Here we hypothesize that the difference in the outcome of infection may at least partly be explained by the localization and timing of the inflammatory response: in calves NO production occurs early and appears to be concentrated in the spleen. On the other hand, there is evidence that a delayed and systemic inflammatory response occurs in adult animals that is ineffectual and probably contributes to the pathogenesis. An improved understanding of the possible mechanisms that underlie this phenomenon may lead to new approaches for the treatment and immune prophylaxis of the disease.

Babesia sp.: emerging intracellular parasites in Europe.

The emergence of Lyme borreliosis as the most prevalent arthropod disease of humans in the temperate northern hemisphere has resulted in renewed interest in human babesiosis, transmitted by the same tick vectors. The advent of new molecular tools has made possible a reappraisal of the main parasites involved (Babesia divergens in Europe and Babesia microti in the USA). B. divergens is probably restricted to European cattle, though there are several nearly identical species. B. microti occurs as a world-wide species complex rather than as a single species, and although both phenotypic and genotypic features lend support to suggestions that zoonotic B. microti may occur in Europe, convincing medical evidence is lacking. Comparative biology should support genetic data in taxonomic studies of these parasites.

Babesia divergens, a bovine blood parasite of veterinary and zoonotic importance.

Babesia divergens is an intraerythrocytic protozoan parasite, transmitted by the tick Ixodes ricinus, and is the main agent of bovine babesiosis in Europe. It is not only a cause of significant loss to the cattle industry; it can also infect immunocompromised humans, causing medical emergencies characterized by rapid fulmination and parasitemias that may exceed 70%. The current emphasis in Europe on sustainable agriculture and extensification is likely to lead to an increase in vector tick populations with increased risk of infection. Despite the veterinary and zoonotic importance of this parasite, relatively little research has been carried out on B. divergens, and many questions regarding the parasite's epidemiology and the host's response remain unanswered. A better understanding of the species' biology and host-parasite interactions may lead to improved control mechanisms and new trends in vaccine and antibabesial drug development. This review provides the first comprehensive summary of B. divergens biology, including its morphology, life cycle, and host specificity, and the current state of knowledge of both human and bovine infections.

Biology of Ixodes species ticks in relation to tick-borne zoonoses.

At least six tick-borne zoonoses are transmitted by members of the Ixodes ricinus species complex in northern hemisphere temperate regions, and include the relatively recently emerged diseases, Lyme borreliosis and ehrlichiosis. An understanding of the biology of these ticks is essential for the prevention and control of the zoonoses they transmit and this review summarises established knowledge in addition to addressing recent work on host specificity of ticks, inter- and intra-specific biological variation and factors affecting tick distribution and abundance.