Borrelia burgdorferi sensu lato infecting Ixodes auritulus ticks in Uruguay.

In the southern cone of South America different haplotypes of Borrelia burgdorferi sensu lato (Bbsl) have been detected in Ixodes spp. from Argentina, southern Brazil, Chile, and Uruguay. So far, Lyme borreliosis has not been diagnosed in Uruguay and the medical relevance of the genus Ixodes in South America is uncertain. However, the growing number of new genospecies of Bbsl in the southern cone region and the scarce information about its pathogenicity, reservoirs and vectors, highlights the importance of further studies about spirochetes present in Uruguay and the region. The aim of this study was to determine the presence of Bbsl in Ixodes auritulus ticks collected from birds and vegetation in two localities of southeastern Uruguay. In total 306 I. auritulus were collected from 392 passerine birds sampled and 1110 ticks were collected by flagging in vegetation. Nymphs and females were analyzed for Borrelia spp. by PCR targeting the flagellin (fla) gene and the rrfA-rrlB intergenic spacer region (IGS). The phylogenetic analysis of Borrelia spp. positive samples from passerine birds and vegetation revealed the presence of four fla haplotypes that form a clade within the Bbsl complex. They were closely related to isolates of Borrelia sp. detected in I. auritulus from Argentina and Canada.

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.

Wild birds as host of Borrelia burgdorferi sensu lato in northwestern Argentina.

Borrelia burgdorferi sensu lato (s. l.) spirochetes are associated with a wide range of vectors and hosts. Birds are important hosts in the ecology of some hard ticks (Ixodidae) in northwestern Argentina, where B. burgdorferi s.l. have been detected in Ixodes pararicinus. We evaluated Borrelia infection in ticks collected from wild birds by molecular analysis through the presence of Borrelia DNA (by nested-PCR targeting the fla gene). A total of 381 ticks (357 larvae and 24 nymphs) belonging to four species (I. pararicinus, Haemaphysalis juxtakochi, Haemaphysalis leporispalustris and Amblyomma sp.) were collected. Partial sequences of the fla gene of Borrelia (100% identical to Borrelia sp. haplotype I from Argentina) were detected in 9 of 70 tick pools (6 pools of larvae and 1 pool of nymphs of I. pararicinus, and in 2 pools of H. juxtakochi larvae) collected on Turdus rufiventris, Syndactila rufosuperciliata and Troglodytes aedon. The results of this study suggest that resident birds have reservoir capacity for Borrelia sp. haplotype I.

Serological Evidence of Borrelia Burgdorferi Infection in Mexican Patients with Facial Palsy.

BACKGROUND: Facial palsy is the most frequent manifestation of neuroborreliosis in the United States, Europe, and Asia, whereas in Mexico, its frequency is unknown. OBJECTIVE: We aimed to determine the frequency of Borrelia spp. infection in patients with acute facial palsy in Mexico. MATERIALS AND METHODS: In this cross-sectional, referral hospital-based survey, 191 patients with facial palsy were selected and clinical and epidemiologic data recorded. IgM and IgG serum antibodies to Borrelia burgdorferi were tested by enzyme-linked immunosorbent assay (ELISA) and confirmed by Western-Blot (WB). IgM and IgG antibodies against the herpes viruses HSV-1, HSV-2, cytomegalovirus, and Epstein-Barr virus were tested by ELISA. RESULTS: 71 patients (37%) tested positive by ELISA to either Borrelia spp. or the herpes viruses. Of 25 patients (13%) who tested positive for B. burgdorferi by ELISA, 23 (12%) were confirmed by WB; 14 had IgM and 9 had IgG antibodies. Among the 14 IgM-WB positive patients, two cases recognized antigens of B. burgdorferi sensu stricto (s.s.), 10 of Borrelia garinii and 2 of B. afzelii, whereas all 9 IgG-WB positive were reactive against B. burgdorferi s.s. 14 patients had facial palsy in addition to other clinical data compatible with Lyme borreliosis. Patients infected with B. burgdorferi s.s. had a longer recovery time and a significantly higher risk (odds ratio 4.4, 95% confidence interval 1.5-12.9) of recurrent facial palsy than patients infected with other Borrelia genospecies. CONCLUSIONS: Borrelia infection is frequent in facial palsy patients in Mexico, with B. burgdorferi s.s. and B. garinii being the most frequent causative species.

Borrelia burgdorferi sensu lato in Ixodes longiscutatus ticks from Brazilian Pampa.

Borrelia burgdorferi sensu lato (s.l.) complex includes the agents of Lyme disease/borreliosis in North America, Europe, and Asia, such Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia bavariensis, Borrelia spielmanii, Borrelia bissettiae, and Borrelia mayonii. In 2013 B. burgdorferi s.l. was reported for the first time in the Neotropical region, from Ixodes aragaoi ticks in Uruguayan Pampa. In addition, from 2011 to 2016, 17 suspected human cases of borreliosis-like syndrome were reported in Rio Grande do Sul (RS) state, Brazil, which contains only part of country in the Pampa biome. The goal of this work is to report the results of a state surveillance program conducted in order to investigate the presence of B. burgdorferi s.l. in its classic vector, Ixodes spp. ticks, from the Brazilian Pampa. For this, we searched for Ixodes spp. ticks in 307 rodents from 11 municipalities of RS state. We then tested the ticks for the presence of B. burgdorferi s.l. DNA using PCR analysis. Of 35 Ixodes spp. ticks tested, one larva and one nymph of Ixodes longiscutatus ticks tested positive for Borrelia sp. DNA. The phylogenetic analysis of the flaB fragment grouped our samples (referred as Borrelia sp. haplotype Pampa) into B. burgdorferi s.l. group in a particular branch with other South American haplotypes, and this group was close to Borrelia carolinensis, B. bissettiae, and Borrelia californiensis. This is the first evidence of B. burgdorferi s.l. circulation in ticks of the genus Ixodes in Brazil. These results highlight the need for the implementation of public health policies for the diagnosis and prevention of potential cases of human borreliosis in Brazil. Further studies are needed to fill the gaps in our knowledge of the distribution, pathogenicity, reservoirs, and vectors of these emerging South American B. burgdorferi s.l. haplotypes.

Presence of Borrelia in different populations of Ixodes pararicinus from northwestern Argentina.

This work was performed to evaluate the presence of Borrelia in different populations of Ixodes pararicinus from northwestern Argentina (Jujuy, Salta and Tucumán provinces). Questing adults and nymphs of I. pararicinus were collected from vegetation, and I. pararicinus nymphs were also collected on birds. Eighty-two ticks were tested for Borrelia presence by PCR targeting the gene flagellin and the rrfA-rrlB intergenic spacer region. Pools of ticks positive to Borrelia were formed by two nymphs collected on Turdus rufiventris in Tucumán, one nymph collected on Syndactyla rufosuperciliata in Jujuy, one nymph collected on Turdus nigriceps in Tucumán, three nymphs collected on T. nigriceps in Tucumán, and two females collected from vegetation in Salta. Two haplotypes of Borrelia sp. belonging to the Borrelia burgdorferi sensu lato complex were found. One of them is closely related to the haplotypes of Borrelia genospecies previously reported in I. aragaoi from Uruguay (haplotypes D and E) and in I. pararicinus from Jujuy Province in Argentina. The second haplotype (detected in the sample of Salta) is closely related to the haplotypes A, B and C associated with I. aragaoi from Uruguay. All these results suggest that the presence of B. burgdorferi s.l. genospecies in I. pararicinus ticks is widespread along the entire distribution of this tick species in northwestern Argentina. However, the Borrelia presence in I. pararicinus cannot be directly assumed as a phenomenon of medical relevance, because Ixodes ticks are not relevant as human parasites in South America, and none of the two Borrelia genospecies detected in this work is related to any of the Borrelia genospecies currently known to be pathogenic to humans.

Serological detection of antibodies to Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen in dogs from Costa Rica.

In a study in Costa Rica 314 serum samples from dogs throughout all seven provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis, and of circulating antigen of Dirofilaria immitis. A total of 6.4% (20/314) and 38.2% (120/314) were positive for Anaplasma spp. (An) and E. canis (Ec) antibodies. Overall, 8.0% (25/314) were positive for D. immitis (Di) antigen. One single dog reacted positive with B. burgdorferi s.l. (Bb) antigen (0.3%, 1/314). E. canis positive dogs were detected in all provinces (highest percentages in Guanacaste, Puntarenas [both significantly different compared to the overall] and Limón). Guanacaste and Puntarenas also showed the highest prevalences of Anaplasma spp. (both significantly different compared to the overall). The highest prevalence of D. immitis was detected in Puntarenas (significantly different compared to the overall). Double pathogen exposure (Ec plus An; Ec plus Di; Ec plus Bb) were recorded in 8.9% (28/314). Two dogs showed a triple pathogen exposure (0.6%, 2/314; An, Ec and Di). There was a significant difference between male (11.5%, 18/156) and female (4.4%, 7/158) animals for D. immitis positive results. There was also a significant difference between breed and no breed dogs regarding the characteristics of a general positive test, as well as seropositivity to the single pathogens of Anaplasma spp., E. canis and D. immitis. Finally there was a significant difference in the presence of clinical signs again regarding the characteristics of a general positive test, as well as seropositivity to Anaplasma spp., E. canis and D. immitis. Practitioners in Costa Rica should be aware of the canine vector-borne diseases mentioned as dogs are at risk of becoming infected. Concerning the positive B. burgdorferi s.l. dog, an autochthonous occurrence cannot be confirmed due to a history of adoption and an unusual tattoo number. Veterinary advice to protect dogs and limit transmission of vector-borne pathogens, also to humans, by using prophylactic measures is strongly recommended.

Multiplex PCR for molecular screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp.

INTRODUCTION: Ticks transmit a great variety of pathogenic microorganisms to humans and animals. The detection of tick-borne pathogens (TBP) is mainly by molecular techniques based on polymerase chain reactions (PCR). OBJECTIVE: To design and evaluate a multiplex PCR for the molecular screening of zoonotic TBP for exploratory studies. MATERIAL AND METHODS: Control DNA from reference strains, DNA from experimentally-infected biological specimens, and from Rhipicephalus sanguineus ticks collected from domestic and homeless dogs were used. A multiplex PCR assay to detect the presence of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. was designed and optimized using primers previously reported for B. burgdorferi sensu lato and Anaplasma spp., while for Babesia spp. they were designed in silico. The multiplex PCR was evaluated on the DNA from biological samples. RESULTS: A new set of specific primers for Babesia spp. was designed. Adjustment of the master mix reactive concentrations and amplification conditions for the multiplex PCR allowed the successful amplification of the specific amplicons for each microbial group from the control DNA and experimentally-infected biological specimens. The efficiency of the multiplex PCR amplifying three DNA targets was confirmed. Individual and co-infection of Anaplasma spp. and Babesia spp. were detected in the R. sanguineus ticks from dogs. CONCLUSIONS: A multiplex PCR assay for the screening of three TBP is available. By using it, B. burgdorferi sensu lato, Anaplasma spp. and Babesia spp. can be detected accurately in one PCR reaction.

Borrelia burgdorferi sensu lato in humans in a rural area of Paraná State, Brazil.

This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s.) in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.

Borrelia burgdorferi sensu lato in humans in a rural area of Paraná State, Brazil

This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s.) in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.

Borrelia burgdorferi sensu lato in humans in a rural area of Paraná State, Brazil

This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s.) in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.(AU)

Borrelia infection in Ixodes pararicinus ticks (Acari: Ixodidae) from northwestern Argentina.

The aim of this work was to describe for the first time the presence of Borrelia burgdorferi sensu lato infecting ticks in Argentina. Unfed specimens of Ixodes pararicinus collected from vegetation in Jujuy Province were tested for Borrelia infection by PCR targeting the gene flagellin (fla), the rrfA-rrlB intergenic spacer region (IGS) and the 16S rDNA (rrs) gene. One male and one female of I. pararicinus collected in Jujuy were found to be positive to Borrelia infection with the three molecular markers tested. Phylogenetically, the Borrelia found in I. pararicinus from Jujuy belongs to the B. burgdorferi s.l complex, and it was similar to one of the genospecies detected in I. aragaoi from Uruguay. Also, this genospecies is closely related to two genospecies known from USA, Borrelia americana and the Borrelia sp. genospecies 1. The epidemiological risk that implies the infection with Borrelia in I. paracinus ticks from Argentina appears to be low because the genospecies detected is not suspected of having clinical relevance and there are no records of Ixodes ticks biting humans in the southern cone of South America. Further studies are needed to assess accurately if there is risk of borreliosis transmitted by ticks in South America.

Borrelia chilensis, a new member of the Borrelia burgdorferi sensu lato complex that extends the range of this genospecies in the Southern Hemisphere.

Borrelia burgdorferi sensu lato (s.l.), transmitted by Ixodes spp. ticks, is the causative agent of Lyme disease. Although Ixodes spp. ticks are distributed in both Northern and Southern Hemispheres, evidence for the presence of B. burgdorferi s.l. in South America apart from Uruguay is lacking. We now report the presence of culturable spirochetes with flat-wave morphology and borrelial DNA in endemic Ixodes stilesi ticks collected in Chile from environmental vegetation and long-tailed rice rats (Oligoryzomys longicaudatus). Cultured spirochetes and borrelial DNA in ticks were characterized by multilocus sequence typing and by sequencing five other loci (16S and 23S ribosomal genes, 5S-23S intergenic spacer, flaB, ospC). Phylogenetic analysis placed this spirochete as a new genospecies within the Lyme borreliosis group. Its plasmid profile determined by polymerase chain reaction and pulsed-field gel electrophoresis differed from that of B. burgdorferi B31A3. We propose naming this new South American member of the Lyme borreliosis group B. chilensis VA1 in honor of its country of origin.

Borrelia burgdorferi sensu lato infecting ticks of the Ixodes ricinus complex in Uruguay: first report for the Southern Hemisphere.

The Borrelia burgdorferi sensu lato (s.l.) group comprises genetically related spirochetes, mostly associated with tick species belonging to the Ixodes ricinus complex in the Northern Hemisphere. The present study evaluated borrelial infection in the tick Ixodes pararicinus, which is the only representative species of the I. ricinus complex in Uruguay. A total of 137 I. pararicinus ticks were collected from deer, cattle, or vegetation in 2 Uruguayan Departments. A part of these ticks was tested directly by PCR targeting the borrelial gene flagellin (fla), whereas another part of the ticks was inoculated into Barbour-Stoenner-Kelly (BSK)-H medium in an attempt to isolate Borrelia. Overall, Borrelia infection was detected in 9 males and 1 nymphal tick pool. These ticks were found to be infected by unique fla haplotypes, which were shown through phylogenetic analysis to represent possibly 2 new B. burgdorferi s.l. genospecies, 1 associated with B. bissettii, the other phylogenetically closest to B. americana. These results were reinforced by PCR and DNA sequencing analyses of portions of 2 additional borrelial genes, rrfA-rrlB intergenic spacer region (IGS) and 16S rDNA (rrs). Weekly examinations of BSK cultures by dark-field microscopy failed to demonstrate live Borrelia through a 100-day incubation period. However, Borrelia DNA was detected by fla-PCR in culture media from 2 vials up to 90 days after inoculation. To the best of our knowledge, this is the first report of B. burgdorferi s.l. infecting ticks in South America.

Presence of Borrelia burgdorferi "Sensu Lato" in patients with morphea from the Amazonic region in Brazil.

BACKGROUND: In the present study, Borrelia spirochetes were found in four (26.6%) out of 15 patients with Atrophoderma of Pasini and Pierini (IAPP) and lichen sclerosis et atrophicans (LSA) from the Brazilian Amazon Region. MATERIAL AND METHODS: Borreliosis was investigated by immunohistochemistry and focus floating microscopy for Borrelia burgdorferi in skin biopsy samples from 15 patients with both clinical and histopathology evidences compatible with Morphea, LSA, and IAPP. RESULTS: Spirochetes were detected by specific immunohistochemistry and focus floating microscopy for B. burgdorferi in samples from three patients. A limitation of our study was the fact that we were not able to isolate and culture these organisms. CONCLUSION: Our data confirm the presence of borreliosis cases in the Amazon.

Borrelia Burgdorferi "sensu lato" in Brazil: Occurrence confirmed by immunohistochemistry and focus floating microscopy.

In the present study, we report the occurrence of Lyme's borreliosis in patients from the Brazilian Amazon Region. Borreliosis was investigated by immunohistochemistry and focus floating microscopy for Borrelia burgdorferi in skin biopsy samples from 22 patients with both clinical and histopathology evidences compatible with Erythema Migrans. Spirochetes were detected by specific immunohistochemistry and focus floating microscopy for B. burgdorferi in samples from five patients. Clinical cure of the cutaneous lesions was observed in all the patients after treatment with doxycycline regimen as proposed by the Center Disease Control guidelines. A limitation of our study was the fact that we were not able to isolate and culture these organisms. These are the first known Brazilian cases of borreliosis to have Focus Floating Microscopy confirmation.

[Search for the spirochete Borrelia burgdorferi sensu lato by polymerase chain reaction in wild Chilean ticks]. / Búsqueda de la espiroqueta Borrelia burgdorferi sensu lato mediante PCR en garrapatas ixoideas chilenas silvestres.

BACKGROUND: Lyme disease is a tick-borne human disease caused by the spirochete Borrelia burgdorferi. Main vectors of Lyme disease are ticks of the Ixodes and Amblyomma genera. Cases with clinical manifestations of Lyme disease and favorable responses to antimicrobial agents have been reported in Chile, some of them with erythema migrans, the hallmark of B burgdorferi infection. AIM: To detect the presence of B burgdorferi in Chilean ticks. MATERIAL AND METHODS: A total of 62 ticks were recollected from wild rodents and cervidae in the Southern region of Chile. Infected and non infected ticks of the species Ixodes ricimus, were used as controls. Insects were homogenised and B burgdorferi was detected using classical and nested polymerase chain reactions. RESULTS: B burgdorferi was not detected in the studied ticks. CONCLUSIONS: Although all the elements required for the enzootic cycle of B burgdorferi are present in Chile, its direct detection in Chilean ticks using the nested polymerase chain reaction assay was negative.

Serosurvey for selected disease agents in white-tailed deer from Mexico.

Serum samples from 350 white-tailed deer (Odocoileus virginianus texanus) collected in March 1994 from northeastern Mexico were tested for the prevalence of antibody activity against five infectious diseases of ruminants. The prevalence rate was 81% for bluetongue virus (BTV) of all serotypes, 72% for epizootic hemorrhagic disease virus (EHDV), 3% for Borrelia burgdorferi, 69% for Anaplasma marginale, and 0% for Brucella abortus, B. melitensis, and B. ovis. These are diseases that affect domestic ruminants, and deer may act as a reservoir of infection. In addition, if deer are translocated, they may introduce pathogens to formerly disease-free areas. The high seroprevalence of BTV and EHDV cannot be related to the presence of hemorrhagic disease in the deer in this region. This is the first report to indicate the presence of B. burgdorferi infection of deer in Mexico. Despite the high prevalence of A. marginale titers, it is uncertain that deer play a role in the epizootiology of cattle anaplasmosis in the region. Apparently, white-tailed deer are unimportant in the epizootiology of brucellosis of both cattle and goats in northeastern Mexico.

[Laboratory diagnosis of Lyme borreliosis]. / Borreliosis de Lyme: aproximaciones al diagnóstico de laboratorio.

Some direct methods that can be used for the diagnosis of Lyme borreliosis are the culture, direct visualization or the detection of microbial DNA using polymerase chain reactions, in body tissues or fluids. Unfortunately, all these methods have a low sensitivity. There is a wide assortment of tests and antigens for indirect diagnosis, and the most recommended are ELISA tests and Western blot. The main inconvenient of these tests are the existence of shared serologic reactions, the variability of immune response and the difficult interpretation of results. Therefore, we propose the following guidelines for the diagnosis of Lyme borreliosis: For sero-epidemiological studies and to diagnose infection, antibodies should be determined in subjects with a compatible clinical picture, using an ELISA test that must be positive in at least two separate samples. All positive ELISA results should be confirmed with Western blot analysis, that must be interpreted using established criteria. Polymerase chain reactions should be used when they are available.