Coexistence of antibodies to tick-borne agents of babesiosis and Lyme borreliosis in patients from Cotia county,State of São Paulo,Brazil

This paper reports a case of coinfection caused by pathogens of Lyme disease and babesiosis in brothers. This was the first case of borreliosis in Brazil, acquired in Cotia County, State of S o Paulo, Brazil. Both children had tick bite history, presented erythema migrans, fever, arthralgia, mialgia, and developed positive serology (ELISA and Western-blotting) directed to Borrelia burgdorferi G 39/40 and Babesia bovis antigens, mainly of IgM class antibodies, suggestive of acute disease. Also, high frequencies of antibodies to B. bovis was observed in a group of 59 Brazilian patients with Lyme borreliosis (25.4%), when compared with that obtained in a normal control group (10.2%) (chi-square = 5.6; p < 0.05). Interestingly, both children presented the highest titers for IgM antibodies directed to both infective diseases, among all patients with Lyme borreliosis.     

Molecular Detection of Tick-Borne Pathogens in Humans with Tick Bites and Erythema Migrans,in the Netherlands

BackgroundTick-borne diseases are the most prevalent vector-borne diseases in Europe. Knowledge on the incidence and clinical presentation of other tick-borne diseases than Lyme borreliosis and tick-borne encephalitis is minimal, despite the high human exposure to these pathogens through tick bites. Using molecular detection techniques, the frequency of tick-borne infections after exposure through tick bites was estimated.MethodsTicks, blood samples and questionnaires on health status were collected from patients that visited their general practitioner with a tick bite or erythema migrans in 2007 and 2008. The presence of several tick-borne pathogens in 314 ticks and 626 blood samples of this cohort were analyzed using PCR-based methods. Using multivariate logistic regression, associations were explored between pathogens detected in blood and self-reported symptoms at enrolment and during a three-month follow-up period.ResultsHalf of the ticks removed from humans tested positive for Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, Rickettsia monacensis, Borrelia miyamotoi and several Babesia species. Among 92 Borrelia burgdorferi s. l. positive ticks, 33% carried another pathogen from a different genus. In blood of sixteen out of 626 persons with tick bites or erythema migrans, DNA was detected from Candidatus Neoehrlichia mikurensis (n = 7), Anaplasma phagocytophilum (n = 5), Babesia divergens (n = 3), Borrelia miyamotoi (n = 1) and Borrelia burgdorferi s. l. (n = 1). None of these sixteen individuals reported any overt symptoms that would indicate a corresponding illness during the three-month follow-up period. No associations were found between the presence of pathogen DNA in blood and; self-reported symptoms, with pathogen DNA in the corresponding ticks (n = 8), reported tick attachment duration, tick engorgement, or antibiotic treatment at enrolment.ConclusionsBased on molecular detection techniques, the probability of infection with a tick-borne pathogen other than Lyme spirochetes after a tick bite is roughly 2.4%, in the Netherlands. Similarly, among patients with erythema migrans, the probability of a co-infection with another tick-borne pathogen is approximately 2.7%. How often these infections cause disease symptoms or to what extend co-infections affect the course of Lyme borreliosis needs further investigations.     

European erythema migrans disease and related disorders

European erythema migrans disease, lymphocytoma, and acrodermatitis chronica are a group of disorders associated with the bite of ixodid ticks. These disorders are now thought to be due to a single, or closely related, ixodid tick spirochetes. European erythema migrans disease closely resembles Lyme disease. Serological evaluation may help to separate spirochetal lymphocytoma from other pseudolymphomas of nonspirochetal origin and from lymphoma. Acrodermatitis chronica atrophicans, so far observed mainly in Europe, is presumably a late manifestation of this group of spirochetal disorders.     

Borrelia burgdorferi in ticks and dogs in the province of Vojvodina, Serbia

Lyme disease is a tick borne zoonotic infection, caused by Borrelia burgdorferi s.l. bacteria. For the transmission of the disease, the presence of ticks is a prerequisite. Lyme borreliosis mostly occurs in people and dogs, but it may occur in other animals. Ticks which carry B. burgdorferi s.l. in Serbia are of the Ixodes ricinus specis. In Serbia, Lyme disease was detected for the first time in the late '80-es. In dogs, clinical symptoms may occur even months after a tick bite, and include weakness, lymphadenopathy, fever, lameness, arthritis, etc. In our survey, we have observed tick and dog populations in the province of Vojvodina (northern part of Serbia). I. ricinus ticks were collected and examined for the presence of B. burgdorferi s.l. in several chosen locations. In addition, blood samples were collected from house dogs and pets from the same locations, and analyzed for the presence of antibodies specific for B. burgdorferi s.l. The results showed a mean infection of ticks of 22.12%, and a mean seroprevalence of Lyme disease in dogs of 25.81%. We conclude that in Vojvodina there is an actual risk of Lyme borreliosis for other animals and humans, because of the persistence of B. burgdorferi s.l. in both tick and dog populations.     

Lyme borreliosis agents and the genetics and sex of their vector, Ixodes ricinus

The tick Ixodes ricinus is responsible for the transmission and maintenance of a wide variety of pathogenic organisms in the Northern Hemisphere, among which Lyme disease represents a major threat to humans. Despite numerous studies, the epidemiology of the different bacterial species responsible for this disease remains unclear. Recent evidence for a sex-biased genetic structure of its European vector leads us to analyse the consequences of this pattern on Borrelia transmission. Here we show that male and female ticks are not equivalently infected by Borrelia burgdorferi, that Borrelia afzelii affects tick migration capabilities, especially for the most vagile sex (i.e., male) and that Lyme borreliosis agents are consequently vectorised in a much more complex way than usually thought. Such results change the epidemiological perception of Lyme borreliosis and suggest new co-evolutionary pathways between the ticks and the borrelia.     

Tick salivary secretion as a source of antihemostatics

Ticks are mostly obligatory blood feeding ectoparasites that have an impact on human and animal health. In addition to direct damage due to feeding, some tick species serve as the vectors for the causative agents of several diseases, such as the spirochetes of the genus Borrelia causing Lyme disease, the virus of tick-borne encephalitis, various Rickettsial pathogens or even protozoan parasites like Babesia spp. Hard ticks are unique among bloodfeeders because of their prolonged feeding period that may last up to two weeks. During such a long period of blood uptake, the host develops a wide range of mechanisms to prevent blood loss. The arthropod ectoparasite, in turn, secretes saliva in the sites of bite that assists blood feeding. Indeed, tick saliva represents a rich source of proteins with potent pharmacologic action that target different mechanisms of coagulation, platelet aggregation and vasoconstriction. Tick adaptation to their vertebrate hosts led to the inclusion of a powerful protein armamentarium in their salivary secretion that has been investigated by high-throughput methods. The resulting knowledge can be exploited for the isolation of novel antihemostatic agents. Here we review the tick salivary antihemostatics and their characterized functions at the molecular and cellular levels.     

Bannwarth's syndrome (lymphocytic meningoradiculitis) in Sweden

Lymphocytic meningoradiculitis of Bannwarth is often associated with a tick bite and erythema chronicum migrans, and therefore may be a European counterpart of Lyme disease in North America. Of nine patients with lymphocytic meningoradiculitis studied at the Neurologic Clinic in Lund, Sweden, six were found to have elevated antibody titers to the Lyme spirochete. These studies support the conclusion that the two diseases are related and may be overlapping sectors of a larger clinical spectrum caused by one infectious agent.     

Preliminary Observations on Ultrastructure of Borreliae in Tissues of Ixodes persulcatus

We observed Lyme borrelia by electron microscopy in the tissues of the ticks, Ixodes persulcatus, which were indicated positive for borreliae by BSK cultures of their internal organs. Borreliae (0.25 μm in diameter) were found only in the lumen of the midgut. They were closely associated with the microvilli on the midgut epithelium but never penetrated into the epithelial cells. Ultrastructural features common to Lyme borreliae., i.e., the three-layered membranes surrounding the cytoplasm and orientation of the flagella insertions, were obviously confirmed. The present results are useful to understand tick tissue-borrelia interface.     

Perspectives of creation of vaccines against tick bite for nonspecific prophylaxis of vector borne diseases

Prophylaxis of infectious diseases transferred by ticks is an important problem of contemporary medicine. One of the perspective approaches to solve this problem is the creation of vaccines against tickbite (anti-tickvaccines). Contemporary methods of the control of infectious diseases transferred by ticks are described in the review. Features of naturally and artificially acquired immunity against ticks are examined. Candidate tick antigens for the construction of vaccines against genus Ixodes tick bite are described. Perspectives of use of anti-tick vaccines against tick vector borne diseases are evaluated.     

Molecular interactions that enable movement of the Lyme disease agent from the tick gut into the hemolymph

Borrelia burgdorferi, the causative agent of Lyme disease, is transmitted to humans by bite of Ixodes scapularis ticks. The mechanisms by which the bacterium is transmitted from vector to host are poorly understood. In this study, we show that the F(ab)(2) fragments of BBE31, a B.burgdorferi outer-surface lipoprotein, interfere with the migration of the spirochete from tick gut into the hemolymph during tick feeding. The decreased hemolymph infection results in lower salivary glands infection, and consequently attenuates mouse infection by tick-transmitted B. burgdorferi. Using a yeast surface display approach, a tick gut protein named TRE31 was identified to interact with BBE31. Silencing tre31 also decreased the B. burgdorferi burden in the tick hemolymph. Delineating the specific spirochete and arthropod ligands required for B. burgdorferi movement in the tick may lead to new strategies to interrupt the life cycle of the Lyme disease agent.     

Vertical vs. horizontal transmission of the microbiome in a key disease vector,Ixodes pacificus

Vector‐borne pathogens are increasingly found to interact with the vector's microbiome, influencing disease transmission dynamics. However, the processes that regulate the formation and development of the microbiome are largely unexplored for most tick species, an emerging group of disease vectors. It is not known how much of the tick microbiome is acquired through vertical transmission vs. horizontally from the environment or interactions with bloodmeal sources. Using 16S rRNA sequencing, we examined the microbiome of Ixodes pacificus, the vector of Lyme disease in the western USA, across life stages and infection status. We also characterized microbiome diversity in field and laboratory‐collected nymphal ticks to determine how the surrounding environment affects microbiome diversity. We found a decrease in both species richness and evenness as the tick matures from larva to adult. When the dominant Rickettsial endosymbiont was computationally removed from the tick microbial community, we found that infected nymphs had lower species evenness than uninfected ticks, suggesting that lower microbiome diversity is associated with pathogen transmission in wild‐type ticks. Furthermore, laboratory‐reared nymph microbiome diversity was found to be compositionally distinct and significantly depauperate relative to field‐collected nymphs. These results highlight unique patterns in the microbial community of I. pacificus that is distinct from other tick species. We provide strong evidence that ticks acquire a significant portion of their microbiome through exposure to their environment despite a loss of overall diversity through life stages. We provide evidence that loss of microbial diversity is at least in part due to elimination of microbial diversity with bloodmeal feeding but other factors may also play a role.     

Presence and indigenous nature of Lyme disease in southern Italy

Lyme disease is very common in the countries of the northern hemisphere. In Italy it is endemic in some regions of the northern part of the country and it is more frequent during summer. In Calabria (south Italy) no cases have been reported. To document the presence and indigenous nature of Lyme disease in this territory we conducted a study from 1999 to 2002. We defined as indigenous cases those with erythema migrans with the following characteristics: dimensions equal to or greater than 5 cm; localization on an area of the skin where there was a tick bite; appearance between 4 and 30 days after the tick bite; appearance in patients who had not resided out of Calabria in the previous 3 months. We found 23 patients with the necessary characteristics to be defined indigenous cases. Since 15 of these cases (65.2%) were observed in the October - December trimester and no case was found in the July - September trimester, we suspect that in Calabria the disease follows a seasonal distribution which differs with respect to countries where it is historically endemic.     

Lyme disease in Minnesota: epidemiologic and serologic findings

During the four years, 1980 to 1983, 83 Minnesota residents have been diagnosed with Lyme disease. Sixty-five of the patients were male. The median age of patients was 39 years with a range from one to 77 years. Seventy-five (90 percent) had onset in 1982 and 1983. Of these latter cases, 56 (75 percent) recalled a tick bite three to 27 days prior to the development of erythema chronicum migrans. Patients experienced possible exposure to Ixodes dammini in at least 24 (28 percent) of the 87 Minnesota counties; however, over 50 percent had reported exposure in one of eight east-central counties near or immediately west of the Wisconsin border. Serologic studies for antibody against the Ixodes dammini spirochete were completed on 30 patients with onset in 1982 and 1983. Of 28 patients with paired acute and convalescent serum samples, only two (7 percent) had fourfold rises in antibody titers. Lyme disease is an emerging public health problem in Minnesota. Additional studies are needed to define the risk of disease by geographic area within the state. Physicians statewide should be alert to the possibility of Lyme disease among their patients, since only 39 percent of patients with onset in 1982 and 1983 were exposed in their county of residence.     

Epidemiologic features of Lyme disease in New York

During 1982, surveillance identified 207 cases of Lyme disease in New York State. Cases were clustered in two geographic areas, eastern Long Island and northern Westchester counties. Symptoms and signs of Lyme disease in cases were consistent with previous reports, with erythema chronicum migrans (ECM) being the most frequently (77 percent) reported sign of disease. Facial palsy was reported in a surprisingly high 18 percent of cases. Of 160 cases whose sera were submitted for Lyme spirochete specific IgG antibody testing, 112 (70 percent) had titers greater than or equal to 64, while 88 (55 percent) had titers greater than or equal to 128. Positive titers were not associated with any single sign or symptom of disease, but were significantly associated with symptom onset or tick bite occurring during the three-month period of June, July, and August. We conclude that the incidence of Lyme disease in New York is much higher than previously recognized. In addition, our data suggest that a serologic test for Lyme-spirochete IgG antibody lacks sensitivity, but can be useful in confirming the diagnosis of Lyme disease when antibody titers are high.     

Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the attachment and blood feeding of Ixodes species ticks on mammalian hosts. In nature, this zoonotic bacterial pathogen may use a variety of reservoir hosts, but the white-footed mouse (Peromyscus leucopus) is the primary reservoir for larval and nymphal ticks in North America. Humans are incidental hosts most frequently infected with B. burgdorferi by the bite of ticks in the nymphal stage. B. burgdorferi adapts to its hosts throughout the enzootic cycle, so the ability to explore the functions of these spirochetes and their effects on mammalian hosts requires the use of tick feeding. In addition, the technique of xenodiagnosis (using the natural vector for detection and recovery of an infectious agent) has been useful in studies of cryptic infection. In order to obtain nymphal ticks that harbor B. burgdorferi, ticks are fed live spirochetes in culture through capillary tubes. Two animal models, mice and nonhuman primates, are most commonly used for Lyme disease studies involving tick feeding. We demonstrate the methods by which these ticks can be fed upon, and recovered from animals for either infection or xenodiagnosis.     

Human perception and awareness of ticks in a South Australian rural community and implications for management of Amblyomma triguttatum triguttatum

The ornate kangaroo tick, Amblyomma triguttatum triguttatum, is invasive on Yorke Peninsula, South Australia. To assess the potential for developing comprehensive management strategies for the species, we mailed 1,000 surveys to Yorke Peninsula residents to investigate community perceptions of ticks, gather information on tick-host ecology, and assess whether or not occupation and length of residence affected whether residents had seen ticks. The response rate was 32%; most respondents (63%) had not seen ticks on Yorke Peninsula. Of those who had (n = 104), 56% had seen ticks on animals, especially lizards (31%) and kangaroos (20%); 50% had seen ticks on humans. Of the 64 respondents who believed places on Yorke Peninsula had many ticks, 56% identified Innes National Park and 75% areas on southern Yorke Peninsula. The likelihood of seeing ticks was not related to occupation. Whether respondents had seen ticks was related to length of residence on Yorke Peninsula (<10 years = 23.6%, 10-30 years = 44.0%, 31-50 years = 47.6%, 51-70 years = 30.9%, 71+ years = 21.4%). The number of respondents who had seen ticks on humans suggests that tick-borne diseases could become a public health issue on Yorke Peninsula. This finding is of particular concern because A. t. triguttatum is implicated in the epidemiology of Q fever in Queensland. Finally, respondents were unaware of the responsibility for tick management, which demonstrates a need for collaboration amongst stakeholders.     

Abundance and Borrelia burgdorferi-infection Prevalence of Nymphal Ixodes scapularis Ticks along Forest–Field Edges

More than 19,000 human cases of Lyme disease (LD) are reported each year in the United States. Lyme disease cases occur when humans are exposed to the bacterium Borrelia burgdorferi through the bite of an infected ixodid tick. The probability of human exposure to infected ticks results from a combination of human behaviors and entomological risk. Human behaviors include use of tick habitats, use of protective clothing, and grooming for tick removal. Entomological risks include the density of ticks in a habitat and the proportion of these that are infected with B. burgdorferi. Recent studies have suggested that humans are at higher risk of exposure to B. burgdorferi near edges between forests and herbaceous communities, including lawns and old fields, but whether this increased risk is a function of human behaviors, entomological risk, or both, is unknown. We assessed entomological risk across forest–old field edges in Dutchess County, NY. Densities of ticks and of infected ticks were considerably higher within forests than at forest–field edges, and were lowest within fields. Thus, edges between forests and fields do not pose a higher entomological risk than do the forests themselves, although risk at the edge is higher than in herbaceous habitat. Landscapes with abundant edges between forested and herbaceous habitat, and roughly even proportions of both, might attract both heavy human use and pose moderately high entomological risk, and thus could be targeted for mitigation. We suggest that determining appropriate methods for reducing human exposure to LD requires differentiating entomological risk from human behaviors.     

Functional analysis of the Borrelia burgdorferi bba64 gene product in murine infection via tick infestation

Borrelia burgdorferi, the causative agent of Lyme borreliosis, is transmitted to humans from the bite of Ixodes spp. ticks. During the borrelial tick-to-mammal life cycle, B. burgdorferi must adapt to many environmental changes by regulating several genes, including bba64. Our laboratory recently demonstrated that the bba64 gene product is necessary for mouse infectivity when B. burgdorferi is transmitted by an infected tick bite, but not via needle inoculation. In this study we investigated the phenotypic properties of a bba64 mutant strain, including 1) replication during tick engorgement, 2) migration into the nymphal salivary glands, 3) host transmission, and 4) susceptibility to the MyD88-dependent innate immune response. Results revealed that the bba64 mutant's attenuated infectivity by tick bite was not due to a growth defect inside an actively feeding nymphal tick, or failure to invade the salivary glands. These findings suggested there was either a lack of spirochete transmission to the host dermis or increased susceptibility to the host's innate immune response. Further experiments showed the bba64 mutant was not culturable from mouse skin taken at the nymphal bite site and was unable to establish infection in MyD88-deficient mice via tick infestation. Collectively, the results of this study indicate that BBA64 functions at the salivary gland-to-host delivery interface of vector transmission and is not involved in resistance to MyD88-mediated innate immunity.     

Tick-borne meningopolyneuritis (Garin-Bujadoux, Bannwarth)

We studied 100 patients with tick-borne meningopolyneuritis (Garin-Bujadoux, Bannwarth), the neurologic component of European erythema chronicum migrans disease. They had intensive radicular pain, asymmetric polyneuritis combined often with uni- or bilateral facial palsy, lymphocytic meningitis without or with only slight meningismus, and a course lasting three to five months. Neurologic abnormalities were preceded by the bite of a tick or an insect in 37 percent of patients or by an erythema in 41 percent. In addition, many patients had extraneural involvement, such as fever or fatigue. The outcome was favorable in all cases, and occurred faster with antibiotic treatment, but a few patients had slight residual peripheral nervous system deficits.