Immune Response to Borrelia: Lessons from Lyme Disease Spirochetes.

The mammalian host responds to infection with Borrelia spirochetes through a highly orchestrated immune defense involving innate and adaptive effector functions aimed toward limiting pathogen burdens, minimizing tissue injury, and preventing subsequent reinfection. The evolutionary adaptation of Borrelia spirochetes to their reservoir mammalian hosts may allow for its persistence despite this immune defense. This review summarizes our current understanding of the host immune response to B. burgdorferi sensu lato, the most widely studied Borrelia spp. and etiologic agent of Lyme borreliosis. Pertinent literature will be reviewed with emphasis on in vitro, ex vivo and animal studies that influenced our understanding of both the earliest responses to B. burgdorferi as it enters the mammalian host and those that evolve as spirochetes disseminate and establish infection in multiple tissues. Our focus is on the immune response of inbred mice, the most commonly studied animal model of B. burgdorferi infection and surrogate for one of this pathogen's principle natural reservoir hosts, the white-footed deer mouse. Comparison will be made to the immune responses of humans with Lyme borreliosis. Our goal is to provide an understanding of the dynamics of the mammalian immune response during infection with B. burgdorferi and its relation to the outcomes in reservoir (mouse) and non-reservoir (human) hosts.

Rickettsia hoogstraalii and a Rickettsiella from the Bat Tick Argas transgariepinus, in Namibia.

Ectoparasites were collected from Eptesicus hottentotus, the long-tailed serotine bat, caught in Namibia as part of an ecological study. Larvae of Argas transgariepinus, a blood-feeding ectoparasite of bats in Africa, were removed from 3 of 18 bats. We present scanning electron microscope images of unengorged larvae. As with other ectoparasites, this bat tick might transmit pathogens such as Borrelia and Rickettsia to their hosts as has been reported for bat ticks in Europe and North America. We screened 3 pools (25 total) of larvae of A. transgariepinus removed from the long-tailed serotine bat Eptesicus hottentotus caught in Namibia. Two microbes of unknown pathogenicity, including Rickettsia hoogstraalii, a spotted fever group pathogen, and a Rickettsiella sp. were detected by molecular techniques.

Molecular detection of avian spirochete Borrelia anserina in Argas persicus ticks in Algeria.

Argasid ticks are one of the most important poultry ectoparasites. They affect poultry directly through blood meal and indirectly through the transmission of pathogens essentially Borrelia anserina, agent of avian borreliosis, one of the most widespread poultry diseases in the world, and is of great economic importance. This study was conducted between April 2014 and March 2015 in the region of Ksar El Boukhari, Algeria, in order to investigate the presence of soft ticks in laying hen farms and to detect B. anserina bacteria using molecular tools. DNA was extracted and screened for the presence of Borrelia spp. DNA by real-time polymerase chain reaction (qPCR). Borrelia spp. screening was performed using primers and probe targeting the 16S rRNA gene. A total of 83 traditional laying hen farms were visited, of which 39 (46.98 %) were found infested with A. persicus tick. Molecular analysis revealed that 2/34 (5.88 %) of ticks were infected by B. anserina. None of the ticks tested were positive for Rickettsia spp., and Coxiella burnetii. These results constitute the first report in Algeria of A. persicus harboring B. anserina.

Experimental Demonstration of Reservoir Competence of the White-Footed Mouse, Peromyscus leucopus (Rodentia: Cricetidae), for the Lyme Disease Spirochete, Borrelia mayonii (Spirochaetales: Spirochaetaceae).

The white-footed mouse, Peromyscus leucopus (Rafinesque), is a reservoir for the Lyme disease spirochete Borrelia burgdorferi sensu stricto in the eastern half of the United States, where the blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae), is the primary vector. In the Midwest, an additional Lyme disease spirochete, Borrelia mayonii, was recorded from naturally infected I. scapularis and P. leucopus. However, an experimental demonstration of reservoir competence was lacking for a natural tick host. We therefore experimentally infected P. leucopus with B. mayonii via I. scapularis nymphal bites and then fed uninfected larvae on the mice to demonstrate spirochete acquisition and passage to resulting nymphs. Of 23 mice fed on by B. mayonii-infected nymphs, 21 (91%) developed active infections. The infection prevalence for nymphs fed as larvae on these infected mice 4 wk post-infection ranged from 56 to 98%, and the overall infection prevalence for 842 nymphs across all 21 P. leucopus was 75% (95% confidence interval, 72-77%). To assess duration of infectivity, 10 of the P. leucopus were reinfested with uninfected larval ticks 12 wk after the mice were infected. The overall infection prevalence for 480 nymphs across all 10 P. leucopus at the 12-wk time point was 26% (95% confidence interval, 23-31%), when compared with 76% (95% confidence interval, 71-79%) for 474 nymphs from the same subset of 10 mice at the 4-wk time point. We conclude that P. leucopus is susceptible to infection with B. mayonii via bite by I. scapularis nymphs and an efficient reservoir for this Lyme disease spirochete.

Vertical transmission rates of Borrelia miyamotoi in Ixodes scapularis collected from white-tailed deer.

Borrelia miyamotoi is a relapsing fever spirochete transmitted by ticks in the Ixodes ricinus complex. In the eastern United States, B. miyamotoi is transmitted by I. scapularis, which also vectors several other pathogens including B. burgdorferi sensu stricto. In contrast to Lyme borreliae, B. miyamotoi can be transmitted vertically from infected female ticks to their progeny. Therefore, in addition to nymphs and adults, larvae can vector B. miyamotoi to wildlife and human hosts. Two widely varying filial infection prevalence (FIP) estimates - 6% and 73% - have been reported previously from two vertically infected larval clutches; to our knowledge, no other estimates of FIP or transovarial transmission (TOT) rates for B. miyamotoi have been described in the literature. Thus, we investigated TOT and FIP of larval clutches derived from engorged females collected from hunter-harvested white-tailed deer in 2015 (n = 664) and 2016 (n = 599) from Maine, New Hampshire, Tennessee, and Wisconsin. After engorged females oviposited in the lab, they (n = 492) were tested for B. miyamotoi infection by PCR. Subsequently, from each clutch produced by an infected female, larval pools, as well as 100 individual eggs or larvae, were tested. The TOT rate of the 11 infected females was 90.9% (95% CI; 57.1-99.5%) and the mean FIP of the resulting larval clutches was 84.4% (95% CI; 68.1-100%). Even though the overall observed vertical transmission rate (the product of TOT and FIP; 76.7%, 95% CI; 44.6-93.3%) was high, additional horizontal transmission may be required for enzootic maintenance of B. miyamotoi based on the results of a previously published deterministic model. Further investigation of TOT and FIP variability and the underlying mechanisms, both in nature and the laboratory, will be needed to resolve this question. Meanwhile, studies quantifying the acarological risk of Borrelia miyamotoi disease need to consider not only nymphs and adults, but larval I. scapularis as well.

Manifestações neuroftalmológicas associadas a doença de Lyme / Neuro-ophthalmological manifestations associated with Lyme disease

Resumo A doença de Lyme é uma infecção sistêmica causada pela espiroqueta Borrelia burgdorferi e transmitida pelo carrapato do gênero Ixodes sp. e espécie Amblyomma cajennense. A doença costuma se manifestar em três estágios clínicos distintos, que podem variar de acordo com as características de cada hospedeiro. O objetivo deste trabalho é relatar o caso de uma paciente de 33 anos com doença de Lyme que apresentou como manifestações neuroftalmológicas diplopia, lagoftalmo paralítico e ceratite punctata, com exames laboratoriais negativos. Embora a espiroqueta Borrelia burgdorferi tenha maior tropismo pelos tecidos da pele, sistema nervoso e articulações, o acometimento ocular não deve ter sua importância diminuída, sendo descrito neste relato de caso, que abordou os aspectos mais pertinentes à doença para auxiliar seu diagnóstico e tratamento.

Abstract Lyme disease is a systemic infection caused by the spirochete Borrelia burgdorferi and transmitted by the tick of the genus Ixodes sp. and species Amblyomma cajennense. The disease usually manifests itself in three distinct clinical stages, which may vary according to the characteristics of each host. The objective of this paper is to report the case of a 33-year-old patient with Lyme disease who presented as neuro-ophthalmological manifestations diplopia, paralytic lagophthalmos and punctate keratitis, with negative laboratory tests. Although the spirochete Borrelia burgdorferi has a greater tropism in the tissues of the skin, nervous system and joints, the ocular involvement should not be diminished, being described in this case report, which approached the most pertinent aspects to the disease to aid its diagnosis and treatment.

An immunocompromised mouse model to infect Ixodes scapularis ticks with the relapsing fever spirochete, Borrelia miyamotoi.

The hard tick-borne relapsing fever spirochete, Borrelia miyamotoi, has recently gained attention as a cause of human illness, but fundamental aspects of its enzootic maintenance are still poorly understood. Challenges to experimental studies with B. miyamotoi-infected vector ticks include low prevalence of infection in field-collected ticks and seemingly inefficient horizontal transmission from infected immunocompetent rodents to feeding ticks. To reliably produce large numbers of B. miyamotoi-infected ticks in support of experimental studies, we developed an animal model where immunocompromised Mus musculus SCID mice were used as a source of B. miyamotoi-infection for larval and nymphal Ixodes scapularis ticks. Following needle inoculation with 1 × 105 spirochetes, the SCID mice developed a high spirochetemia (greater than 1 × 107 copies of B. miyamotoi purB per mL of blood) that persisted for at least 30 d after inoculation. In comparison, immunocompetent M. musculus CD-1 mice developed transient infections, detectable for only 2-8 d within the first 16 d after needle inoculation, with a brief, lower peak spirochetemia (8.5 × 104 - 5.6 × 105purB copies per mL of blood). All larval or nymphal ticks fed on infected SCID mice acquired B. miyamotoi, but frequent loss of infection during the molt led to the proportion infected ticks of the resulting nymphal or adult stages declining to 22-29%. The ticks that remained infected after the molt had well-disseminated infections which then persisted through successive life stages, including transmission to larval offspring.

Role of mustelids in the life-cycle of ixodid ticks and transmission cycles of four tick-borne pathogens.

BACKGROUND: Elucidating which wildlife species significantly contribute to the maintenance of Ixodes ricinus populations and the enzootic cycles of the pathogens they transmit is imperative in understanding the driving forces behind the emergence of tick-borne diseases. Here, we aimed to quantify the relative contribution of four mustelid species in the life-cycles of I. ricinus and Borrelia burgdorferi (sensu lato) in forested areas and to investigate their role in the transmission of other tick-borne pathogens. Road-killed badgers, pine martens, stone martens and polecats were collected in Belgium and the Netherlands. Their organs and feeding ticks were tested for the presence of tick-borne pathogens. RESULTS: Ixodes hexagonus and I. ricinus were found on half of the screened animals (n = 637). Pine martens had the highest I. ricinus burden, whereas polecats had the highest I. hexagonus burden. We detected DNA from B. burgdorferi (s.l.) and Anaplasma phagocytophilum in organs of all four mustelid species (n = 789), and Neoehrlichia mikurensis DNA was detected in all species, except badgers. DNA from B. miyamotoi was not detected in any of the investigated mustelids. From the 15 larvae of I. ricinus feeding on pine martens (n = 44), only one was positive for B. miyamotoi DNA, and all tested negative for B. burgdorferi (s.l.), N. mikurensis and A. phagocytophilum. The two feeding larvae from the investigated polecats (n = 364) and stone martens (n = 39) were negative for all four pathogens. The infection rate of N. mikurensis was higher in feeding nymphs collected from mustelids compared to questing nymphs, but not for B. burgdorferi (s.l.), B. miyamotoi or A. phagocytophilum. CONCLUSIONS: Although all stages of I. ricinus can be found on badgers, polecats, pine and stone martens, their relative contribution to the life-cycle of I. ricinus in forested areas is less than 1%. Consequently, the relative contribution of mustelids to the enzootic cycles of I. ricinus-borne pathogens is negligible, despite the presence of these pathogens in organs and feeding ticks. Interestingly, all four mustelid species carried all stages of I. hexagonus, potentially maintaining enzootic cycles of this tick species apart from the cycle involving hedgehogs as main host species.

Evidence for Clinical Anaplasmosis and Borreliosis in Cats in Maine.

The objectives of this study were to use data from client-owned cats in an Ixodes scapularis endemic area to evaluate for clinical associations with diagnostic test results for Anaplasma phagocytophilum and Borrelia burgdorferi and to provide information from a group of cats with possible borreliosis as the cause of clinical manifestations of disease. All cases were evaluated at one clinic, medical records were evaluated, and sera from all cats were tested using one of two commercially available assays labeled for the use with dog sera (SNAP 4Dx or SNAP 4Dx Plus; IDEXX Laboratories, Westbrook, ME). Of the 159 cats evaluated, 42 cats (26.4%) had clinical signs consistent with A. phagocytophilum or B. burgdorferi infection and 117 cats (73.6%) were apparently healthy. Antibodies against B. burgdorferi or A. phagocytophilum were detected in sera of 18.2% and 6.3% of the 159 cats, respectively. Cats with clinical signs of disease were 4 times more likely to have antibodies against one or both agents than healthy cats (95% confidence interval [CI] 1.7928-8.9246; P = .0007), cats allowed outdoors were 5 times more likely to have antibodies against one or both agents than cats housed exclusively indoors (95% CI 2.0196-12.4497; P = .0005), and cats of owners who purchased acaricides were more likely (odds ratio = .3977) to have antibodies against one or both agents than cats of owners who did not purchase acaricides (95% CI .1659-.9534; P = .0387). The cats in the case series were believed to have B. burgdorferi infection as the most likely cause of illness based on serological testing, select clinical information, and apparent response to administration of doxycycline. The results suggest that both A. phagocytophilum and B. burgdorferi are associated with clinical illness in cats. Owners of cats allowed outdoors should be diligent in the use of acaricides.

[Borrelia miyamotoi: a recently identified human pathogenic tick-borne relapsing fever spirochete]. / Borrelia miyamotoi: egy újabb, humán patogén, kullancs által terjesztett, visszatéro lázat okozó baktérium.

Borrelia miyamotoi is a recently described relapsing fever spirochete transmitted by ticks of the Ixodes ricinus complex. This pathogen is different from Borrelia burgdorferi sensu lato (the Lyme borreliosis spirochetes) in its epidemiology, ecology and also genetics. Over 50 patients have been described worldwide with Borrelia miyamotoi disease, and three immunocompromised patients were reported with neurological symptoms. Our knowledge about Borrelia miyamotoi infection in ticks and its distribution in different habitats and also the mechanism of the infection is limited. The most common symptom is fever; thus it can be easily confused with other tick-borne diseases. Due to the intensive research in recent years, Borrelia miyamotoi infection in ticks and hosts has been reported from different regions and also the number of patients is increasing, thus this bacterium is considered as an emerging pathogen. In this literature review we would like to summarize the available knowledge about this spirochete. Orv Hetil. 2017, 158(29): 1124-1130.

What's eating you? lone star tick (Amblyomma americanum).

Amblyomma americanum, also known as the lone star tick, is found in much of the eastern United States. Since the mid-20th century, the lone star tick has been implicated in human disease. Today, A americanum remains an important vector for tick-borne illness. In addition to others, species of Rickettsia, Ehrlichia, and Borrelia are all transmitted by the lone star tick. Recently described conditions such as Southern tick-associated rash illness and anaphylaxis to red meat following tick bites have been attributed to the lone star tick. Impressive local reactions also can result after bites from A americanum. Early treatment of tick-borne illness is crucial to ensure good patient outcomes. Tick-control measures also are an important part of disease management in endemic areas. We discuss the tick's biology, human illnesses associated with A americanum, and methods to control tick numbers and eliminate disease in local reservoirs.

Borrelia miyamotoi - new etiologic agent of neuroborreliosis?

Borrelia miyamotoi spirochetes discovered in Ixodes persulcatus ticks in Japan, in 1994 and documented in ticks and rodents in moderate climate zone of northern hemisphere. They belong to tick-borne relapsing fever group spirochetes. Borrelia miyamotoi is an etiologic agent of B.miyamotoi disease with acute febrile illness, including fever, headache, dizziness, fatigue, chills, and muscle and joint pain. Recurrence of fever has been observer in more than 10% patients. In some patients meningoencephalitis, encephalitis and cranial neuritis were observed. Laboratory recognition is based mainly upon PCR testing. Serological testing is limited due to inaccessibility of tests.

Prevalence of Rickettsia spp. in Ticks and Serological and Clinical Outcomes in Tick-Bitten Individuals in Sweden and on the Åland Islands.

Tick-transmitted diseases are an emerging health problem, and the hard tick Ixodes ricinus is the main vector for Borrelia spp., tick-borne encephalitis virus and most of the spotted fever Rickettsiae in Europe. The aim of the present study was to examine the incidence of rickettsial infection in the southernmost and south central parts of Sweden and the Åland Islands in Finland, the risk of infection in humans and its correlation with a bite of a Rickettsia-infected tick, the self-reported symptoms of rickettsial disease, and the prevalence of co-infection between Rickettsia spp. and Borrelia spp. Persons with a recent tick bite were enrolled through public media and asked to answer a questionnaire, provide a blood sample and bring detached ticks at enlistment and at follow-up three months later. Blood samples were previously analysed for Borrelia spp. antibodies and, for this report, analysed for antibodies to Rickettsia spp. by immunofluorescence and in 16 cases also using Western Blot. Ninety-six (44.0%) of the 218 participants were seropositive for IgG antibodies to Rickettsia spp. Forty (18.3%) of the seropositive participants had increased titres at the follow-up, indicating recent/current infection, while four (1.8%) had titres indicating probable recent/current infection (≥1:256). Of 472 ticks, 39 (8.3%) were Rickettsia sp. positive. Five (31.3%) of 16 participants bitten by a Rickettsia-infected tick seroconverted. Experience of the self-reported symptoms nausea (p = 0.006) and radiating pain (p = 0.041) was more common among those with recent, current or probable infection compared to those who did not seroconvert. Participants who showed seroreactivity or seroconversion to Rickettsia spp. had more symptoms than those who were seronegative. Seven (3.2%) participants showed seroconversion to Borrelia spp., and three (1.4%) of these showed seroconversion to both Rickettsia spp. and Borrelia spp., in accordance with previous studies in Sweden. Symptoms of rickettsial disease were in most of the cases vague and general that were difficult to differentiate from other tick-borne diseases.

[POPULATION MORBIDITY BY INFECTIONS TRANSMITIED VIA IXODES PER- SULCATUS IN THE NORTH AND SOUTH OF IRKUTSK REGION].

AIM: Evaluate the degree of epidemic risk of emergence of tick-borne encephalitis (TBE) and ixodes ticks' borreliosis (ITB) in municipalities (MPs) of Irkutsk region with various natural- climate conditions. MATERIALS AND METHODS: Morbidity was compared for TBE and ITB during 2001 - 2015 in MPs of Irkutsk region located, to the north or south of the 55th parallel, i.e. in the con- ditions of different severity of sharply continental climate. 5-year average data were analyzed. RESULTS: ITB morbidity was 2 - 3 times higher than TBE for all the 5-year periods in the north of the region, whereas in the south -differences were not present. Moreover, in MPs located to the north of the 55th parallel in 2001 - 2015 a decrease of TBE and ITB morbidity did not occur, as in the south of the region. CONCLUSION: The lack of population morbidity reduction by TBE and ITB in the north and prevalence of the latter nosoform requires enhancement of a complex of prophylaxis measures in these MPs with an accent in development of non-specific means, includ- ing acaricidic treatment, enhancement of operations of centers of express diagnostics of transmissible infections, familiarizing of the population with the use of protective suits during periods of high activity of ticks.

Borrelia miyamotoi Disease: Neither Lyme Disease Nor Relapsing Fever.

Borrelia miyamotoi disease (BMD) is a newly recognized borreliosis globally transmitted by ticks of the Ixodes persulcatus species complex. Once considered to be a tick symbiont with no public health implications, B miyamotoi is increasingly recognized as the agent of a nonspecific febrile illness often misdiagnosed as acute Lyme disease without rash, or as ehrlichiosis. The frequency of its diagnosis in the northeastern United States is similar to that of human granulocytic ehrlichiosis. A diagnosis of BMD is confirmed by polymerase chain reaction analysis of acute blood samples, or by seroconversion using a recombinant glycerophosphodiester phosphodiesterase enzyme immunoassay. BMD is successfully treated with oral doxycycline or amoxicillin.

Borrelia infection in small mammals in West Africa and its relationship with tick occurrence inside burrows.

Tick-borne relapsing fever (TBRF) is a zoonotic disease caused by several Borrelia species transmitted to humans by Ornithodoros tick vectors. In West Africa, Borrelia crocidurae is a common cause of disease in many rural populations. Small mammals act as reservoirs of infection. We report here the results of surveys that investigated the occurrence of B. crocidurae infection in rodents and insectivores from eight countries of West and Central Africa. Animals were identified at the species level and tested for Borrelia either by examination of thick blood film, intra-peritoneal inoculation of blood or brain tissues into laboratory mice, or by molecular techniques. A total of 4358 small mammals belonging to 38 species and 7 families were collected, including 3225 specimens collected in areas where the occurrence of Ornithodoros sonrai tick in rodent burrows was documented, and 1133 in areas where this tick was absent. In areas with O. sonrai, Borrelia infection was demonstrated in 287 of 3109 (9.2%) small mammals tested, and none was documented in 1004 animals tested from other areas. There was no relationship between the occurrence of Rhipicephalus, Hyaloma and Argas ticks in burrows and the distribution of Borrelia infection in small mammals. The 287 specimens infected by Borrelia belonged to 15 rodent and shrew species, including three Saharo-Sahelian species - Gerbillus gerbillus, Gerbillus occiduus and Gerbillus tarabuli - identified as reservoirs for TBRF with a distribution restricted to this area. In Sudan and Sudano-Sahelian areas, Arvicanthis niloticus, Mastomys erythroleucus and Mastomys huberti were the main reservoir of infection. Although most small mammals species collected had a large distribution in West and Central Africa, the fact that only animals collected in areas with O. sonrai were found infected suggest that this tick is the only vector of TBRF in rodents and insectivores in this part of Africa.

Borrelia miyamotoi: a widespread tick-borne relapsing fever spirochete.

Borrelia miyamotoi is a relapsing fever spirochete that has only recently been identified as a human pathogen. Borrelia miyamotoi is genetically and ecologically distinct from Borrelia burgdorferi sensu lato, while both are present in Ixodes ticks. Over 50 patients with an acute febrile illness have been described with a B. miyamotoi infection, and two infected immunocompromised patients developed a meningoencephalitis. Seroprevalence studies indicate exposure in the general population and in specific risk groups, such as patients initially suspected of having human granulocytic anaplasmosis. Here, we review the available literature on B. miyamotoi, describing its presence in ticks, reservoir hosts, and humans, and discussing its potential impact on public health.

Transstadial transmission of Borrelia turcica in Hyalomma aegyptium ticks.

Borrelia turcica comprises the third major group of arthropod-transmitted borreliae and is phylogenetically divergent from other Borrelia groups. The novel group of Borrelia was initially isolated from Hyalomma aegyptium ticks in Turkey and it was recently found in blood and multiple organs of tortoises exported from Jordan to Japan. However, the ecology of these spirochetes and their development in ticks or the vertebrate hosts were not investigated in detail; our aims were to isolate the pathogen and to evaluate the possibility of transstadial transmission of Borrelia turcica by H. aegyptium ticks. Ticks were collected from Testudo graeca tortoises during the summer of 2013 from southeastern Romania. Engorged nymphs were successfully molted to the adult stage. Alive B. turcica was isolated from molted ticks by using Barbour-Stoenner-Kelly (BSK) II medium. Four pure cultures of spirochetes were obtained and analyzed by PCR and sequencing. Sequence analysis of glpQ, gyrB and flaB revealed 98%-100% similarities with B. turcica. H. aegyptium ticks collected from T. graeca tortoises were able to pass the infection with B. turcica via transstadial route, suggesting its vectorial capacity.