2. Suurin osa punkeista kantaa enemmän kuin yhtä borrelia-bakteerin alalajia. B.lusitaniae on dokumentoitu nyt ensimmäistä kertaa Tanskassa.
3. Borrelia bavariensis sp. nov. esiintyy laajasti sekä Euroopassa että Aasiassa
1. FEMS Microbiol Lett. 2009 Jan 28; [Epub ahead of print]
Molecular detection of Borrelia bissettii DNA in serum samples from patients inthe Czech Republic with suspected borreliosis. Rudenko N, Golovchenko M, Ruzoek D, Piskunova N, Mallatova N, Grubhoffer L. Biology Centre of the Academy of Sciences of the Czech Republic, Institute ofParasitology AS CR, Faculty of Science, University of South Bohemia in CeskeBudejovice, Ceske Budejovice, Czech Republic. Abstract
Until recently, three spirochete genospecies were considered to be thecausative agents of Lyme borreliosis (LB) in Europe: Borrelia burgdorferi sensustricto, Borrelia afzelii and Borrelia garinii. However, the DNA of Borreliavalaisiana, Borrelia lusitaniae, Borrelia spielmanii and Borrelia bissettii hasalready been detected in samples of human origin, or the spirochetes wereisolated from the patients with symptoms of LB.
Molecular analysis of 12selected serum samples collected in the regional hospital confirmed the presenceof B. bissettii DNA in cases of single and multiple infection in patients withsymptomatic borreliosis or chronic borrelial infection.
The presence of B.bissettii as a single strain in patients provides strong support of the factthat B. bissettii might be a causative agent of the disease. After the firstisolation of B. bissettii from the samples of human origin in Slovenia,following the detection of this species in cardiac valve tissue of the patientwith endocarditis and aortic valve stenosis in the Czech Republic, here we present additional molecular data supporting the involvement of B. bissettii in LB in Europe. PMID: 19187198 [PubMed - as supplied by publisher]
2. Parasitol Int. 2007 Aug 1;
Occurrence of multiple infections with different Borrelia burgdorferi genospecies in Danish Ixodes ricinus nymphs.
Vennestrøm J, Egholm H, Jensen PM.
Section for Genetics and Microbiology, Department of Ecology, The Royal Veterinary and Agricultural University, Frederiksberg, Denmark.
The pathogen Borrelia burgdorferi causes Lyme Borreliosis in human and animals world-wide. In Europe the pathogen is transmitted to the host by the vector Ixodes ricinus. The nymph is the primary instar for transmission to humans. We here study the infection rate of five Borrelia genospecies: B. burgdorferi sensu stricto, B. afzelii, B. garinii, B. valaisiana, B. lusitaniae in nymphs, by IFA and PCR. 600 nymphs were collected in North Zealand of Denmark. Each nymph was first analysed by IFA. If positive for spirochaetal infection, the genospecies was determined by PCR. The infection rate of B. burgdorferi sensu lato was 15.5%, with the primary genospecies being B. afzelii (64.3%), B. garinii (57.1%), and B. lusitaniae (26.8%). It is the first time B. lusitaniae is documented in Denmark. Even though, the highest infection rate was discovered for B. afzelii and B. garinii, mixed infections are more common than single infections. Fifty-one percent (29/56) of these were infected with two genospecies, 7.1% (4/56) with three, and 5.3% (3/56) with four. We try to explain the high infection rate and the peculiar number of multiple infections, with a discussion of changes host abundance and occurrence of different transmission patterns.
PMID: 17804280 [PubMed - as supplied by publisher]
Borrelia bavariensis sp. nov. is widely distributed in Europe and Asia.
Authors: Margos G, Wilske B, Sing A, Hizo-Teufel C, Cao WC, Chu C, Scholz H, Straubinger RK, Fingerle V
Citation: Int. J. Syst. Evol. Microbiol. 2013(Jul)
Location: Bavarian Health and Food Safety Authority, Oberschleissheim, Germany;
Since the original description of Borrelia bavariensis sp. nov. in 2009, additional samples available from humans and ticks from Europe and Mongolia, respectively, have been used to further characterize Borrelia strains belonging to this group of spirochetes that utilize rodents as reservoir hosts. These investigations suggested the presence of related strains in Europe and Asia and confirmed their status as a distinct species. Furthermore, samples that were investigated by researchers from China and Japan confirm the ecological relationship of members of this proposed species with rodents and suggest that it has a wide distribution in Eurasia. Here, we use phylogenetic and genetic distance analyses to validate B. bavariensis sp. nov. as species within the Borrelia burgdorferi sensu lato species complex.
Vector Borne Zoonotic Dis. 2007 Sep 3; [Epub ahead of print] Links
First Record of Lyme Disease Borrelia in the Arctic.
Larsson C, Comstedt P, Olsen B, Bergström S.
Department of Molecular Biology, Umeå University, Umeå, Sweden.
The epidemiology and ecology of Lyme disease is very complex, and its reported geographical distribution is constantly increasing. Furthermore, the involvement of birds in long distance dispersal and their role as reservoir hosts is now well established. In this study, we have shown that sea birds in the Arctic region of Norway carry Ixodes uriae ticks infected with Lyme disease Borrelia garinii spirochetes. Interestingly, DNA sequencing showed that these isolates are closely related to B. garinii previously isolated from birds, as well as from clinical specimens in northern Europe.
PMID: 17767412 [PubMed - as supplied by publisher]
Suom. huom. Englantilaisen ystäväni mukaan kyseinen alalaji on tyypillinen Briteissä ja Irlannissa. Heille on kerrottu että kyseinen alalaji ei aiheuta sairautta!
Am J Trop Med Hyg. 2007 Dec;77(6):1124-1127.
Borrelia valaisiana Infection in a Japanese Man Associated with Traveling to Foreign Countries.
Saito K, Ito T, Asashima N, Ohno M, Nagai R, Fujita H, Koizumi N, Takano A, Watanabe H, Kawabata H.
Department of Cardiovascular Medicine, and Department of Dermatology, University of Tokyo Graduate School of Medicine, Tokyo, Japan; Ohara Research Laboratory, Ohara General Hospital, Fukushima, Japan; Department of Bacteriology, National Institute of Infectious Diseases, Tokyo, Japan; Doctoral Course of the United Graduate School of Veterinary Sciences, Gifu University, National University Corporation, Gifu, Japan.
A 78-year-old Japanese man who had clinical symptoms and a flu-like illness with fever, chills, diarrhea, and arthralgia had traveled to Cambodia and Khabarovsk, Russia, before the onset of symptoms and illness. He had been bitten by an Ixodes persulcatus tick in which the DNA of Borrelia valaisiana was detected.
The patient's symptoms improved rapidly after treatment with minocycline. Serologic examination detected antibodies to Lyme disease Borrelia. An flaB polymerase chain reaction with the patient's plasma amplified a DNA fragment similar to that of B. valaisiana.
PMID: 18165534 [PubMed - as supplied by publisher]
Parasitol Int. 2008 Mar;57(1):32-7. Epub 2007 Aug 1. Links
Occurrence of multiple infections with different Borrelia burgdorferi genospecies in Danish Ixodes ricinus nymphs.
Vennestrøm J, Egholm H, Jensen PM.
Section for Genetics and Microbiology, Department of Ecology, The Royal Veterinary and Agricultural University, Frederiksberg, Denmark.
The pathogen Borrelia burgdorferi causes Lyme Borreliosis in human and animals world-wide. In Europe the pathogen is transmitted to the host by the vector Ixodes ricinus. The nymph is the primary instar for transmission to humans.
We here study the infection rate of five Borrelia genospecies: B. burgdorferi sensu stricto, B. afzelii, B. garinii, B. valaisiana, B. lusitaniae in nymphs, by IFA and PCR. 600 nymphs were collected in North Zealand of Denmark. Each nymph was first analysed by IFA. If positive for spirochaetal infection, the genospecies was determined by PCR.
The infection rate of B. burgdorferi sensu lato was 15.5%, with the primary genospecies being B. afzelii (64.3%), B. garinii (57.1%), and B. lusitaniae (26.8%). It is the first time B. lusitaniae is documented in Denmark.
Even though, the highest infection rate was discovered for B. afzelii and B. garinii, mixed infections are more common than single infections. Fifty-one percent (29/56) of these were infected with two genospecies, 7.1% (4/56) with three, and 5.3% (3/56) with four.
We try to explain the high infection rate and the peculiar number of multiple infections, with a discussion of changes host abundance and occurrence of different transmission patterns.
PMID: 17804280 [PubMed - in process]
Arch Dermatol. 1999 Nov;135(11):1317-26.
Arch Dermatol. 1999 Nov;135(11):1398-400.
Solitary erythema migrans in Georgia and South Carolina.
Felz MW, Chandler FW Jr, Oliver JH Jr, Rahn DW, Schriefer ME.
Department of Family Medicine, Medical College of Georgia, Augusta 30912-3500, USA.
OBJECTIVE: To evaluate the incidence of Borrelia burgdorferi infection in humans with erythema migrans (EM) in 2 southeastern states.
DESIGN: Prospective case series. SETTING: Family medicine practice at academic center.
PATIENTS: Twenty-three patients with solitary EM lesions meeting Centers for Disease Control and Prevention (CDC) criteria for Lyme disease.
INTERVENTIONS: Patients underwent clinical and serologic evaluation for evidence of B burgdorferi infection. All lesions underwent photography, biopsy, culture and histopathologic and polymerase chain reaction analysis for B burgdorferi infection. Patients were treated with doxycycline hyclate and followed up clinically and serologically.
MAIN OUTCOME MEASURES: Disappearance of EM lesions and associated clinical symptoms in response to antibiotic therapy; short-term and follow-up serologic assays for diagnostic antibody; growth of spirochetes from tissue biopsy specimens in Barbour-Stoenner-Kelly II media; special histopathologic stains of tissue for spirochetes; and polymerase chain reaction assays of tissue biopsy specimens for established DNA sequences of B burgdorferi.
RESULTS: The EM lesions ranged from 5 to 20 cm (average, 9.6 cm). Five patients (22%) had mild systemic symptoms. All lesions and associated symptoms resolved with antibiotic therapy. Overall, 7 patients (30%) had some evidence of B burgdorferi infection. Cultures from 1 patient (4%) yielded spirochetes, characterized as Borrelia garinii, a European strain not known to occur in the United States; 3 patients (13%) demonstrated spirochetallike forms on special histologic stains; 5 patients (22%) had positive polymerase chain reaction findings with primers for flagellin DNA sequences; and 2 patients (9%) were seropositive for B burgdorferi infection using recommended 2-step CDC methods. No late clinical sequelae were observed after treatment.
CONCLUSIONS: The EM lesions we observed are consistent with early Lyme disease occurring elsewhere, but laboratory confirmation of B burgdorferi infection is lacking in at least 16 cases (70%) analyzed using available methods. Genetically variable strains of B burgdorferi, alternative Borrelia species, or novel, uncharacterized infectious agents may account for most of the observed EM lesions.
PMID: 10566829 [PubMed - indexed for MEDLINE]
Alalajeista puheenollen... ja niihin liittyvistä testeistä. Ilkka Seppälän sanoin " Huomattavia eroja antigeenirakenteissa. Optimaalista yhtä testiä ei ole." Tästä huolimatta esim. Seppälän analysoimien borrelioositestien perässä lukee useimmiten: löydös sopii vanhaan immuniteettiin, tulos on raja-arvo tms.
Esitelmä löytyy googlettamalla esim.
Borrelia burgdorferi-ryhmä Seitsemän tunnistettua ...
Tiedostomuoto: PDF/Adobe Acrobat - HTML-versio
Borrelia burgdorferi-ryhmä. Seitsemän tunnistettua humaanipatogeenia. Euroopassa . B. burgdorferi sensu stricto. B. afzelii. B. garinii. B. valaisiana ...
http://www.google.fi/url?sa=t&rct=j&q=& ... 4661,d.bGE
1. Parasite. 2008 Sep;15(3):244-7.
Borrelia burgdorferi sensu lato, the agent of lyme borreliosis: life in the wilds.
Gern L.Institute of Biology, University of Neuchâtel, Switzerland, Emile Argand 11, Case postale 158, 2009 Neuchâtel, Switzerland. email@example.com
In Europe, Borrelia burgdorferi sensu lato (sl) the agent of Lyme
borreliosis circulates in endemic areas between Ixodes ricinus ticks and a large number of vertebrate hosts upon which ticks feed. Currently, at least 12 different Borrelia species belonging to the complex B. burgdorferi sl have been identified among which seven have been detected in I. ricinus: B. burgdorferi sensu stricto (ss), B. garinii, B. afzelii, B. valaisiana, B. spielmanii and B. bissettii.
A few dozens of vertebrate hosts have been
identified as reservoirs for these Borrelia species. Specific associations
were rather early observed between hosts, ticks and borrelia species, like for example between rodents and B. afzelii and B. burgdorferi ss, and between birds and B. garinii and B. valaisiana. The complement present in the blood of the hosts is the active component in the Borrelia host specificity. Recent studies confirmed trends toward specific association between Borrelia species and particular host, but also suggested that loose associations may be more frequent in transmission cycles in nature than previously thought.
PMID: 18814688 PubMed - in process
B.bissettii -alalajia esiintyy mm. Itä-Euroopan maissa. Myös se näyttäisi aiheuttavan Borrelioosia. Hiirillä kudosmuutoksia esiintyi esim. virtsarakossa, sydämessä ja nivelissä.
Vector-Borne and Zoonotic Diseases
Borrelia bissettii Isolates Induce Pathology in a Murine Model of Disease
To cite this paper:
Bradley S. Schneider, Martin E. Schriefer, Gabrielle Dietrich, Marc C. Dolan, Muhammad G. Morshed, Nordin S. Zeidner. Vector-Borne and Zoonotic Diseases. ahead of print. doi:10.1089/vbz.2007.0251.
Full Text PDF: ? HiRes for printing (384.7 KB) ? PDF Plus w/ links (191.6 KB)
Bradley S. Schneider
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO.
Institut Pasteur, Département de Parasitologie, Unités de Réponses précoces aux Parasites et Immunopathologie, 25 Rue du Docteur Roux, 75724 Paris Cedex 15, France.
Martin E. Schriefer
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO.
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO.
Marc C. Dolan
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO.
Muhammad G. Morshed
Zoonotic Diseases and Emerging Pathogens Section, Laboratory Services, BC Centre for Disease Control, and Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V5Z 4R4 Canada.
Nordin S. Zeidner
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO.
The spirochete Borrelia burgdorferi is a tick-borne pathogen that causes Lyme disease. Although B. burgdorferi sensu lato is a diverse group of bacteria, only three genospecies, B. burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii, are known to be pathogenic and commonly recognized to cause human disease. To assess the potential of another common genospecies, Borrelia bissettii, to induce disease, a mouse model was employed.
Two Colorado isolates of B. bissettii (CO-Bb) induced lesions of the bladder, heart, and femorotibial joint 8 weeks after inoculation into mice. In contrast, two British Columbia (BC-Bb) isolates, could not be cultured or amplified by PCR from target organs, and did not induce lesions. Consistent with pathology and culture results, the antibody response in mice to BC-Bb was minimal compared to CO-Bb, indicating either transient localized infection or rapid immune clearance of BC-Bb. Although sequence analysis of the rrf (5S)?rrl (23S) intergenic spacer region indicated 99% homology between CO-Bb and BC-Bb, polyacrylamide gel electrophoresis (PAGE) analysis indicated five distinct protein differences between these low-passage isolates.
These studies support the prospect that B. bissettii may indeed be the causative agent of Lyme borreliosis cases in Eastern Europe, associated with the atypical Borrelia strain 25015, and in other regions. To our knowledge, this is the first evidence that B. bissettii can induce pathology in a vertebrate host.
Diversity of Borrelia burgdorferi sensu lato in natural foci of Novosibirsk region.
Fomenko NV, Livanova NN, Chernousova NY.
Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, Lavrentyev's Avenue 8, 630090 Novosibirsk, Russia.
Ixodes persulcatus ticks, wild small mammals, and infected people in Novosibirsk region of Russia were investigated to analyze the local distribution and diversity of Borrelia burgdorferi sensu lato. The molecular typing was based on the partial sequencing of the 16S rRNA gene and the 5S-23S rRNA intergenic spacer region. The analysis of our data revealed the presence of two Borrelia species, B. garinii and B. afzelii. B. garinii DNA was found much more often, namely in 74% of all PCR-positive human blood samples, in 72% of all samples from small mammals (including blood and tissues), and in 76% of Borrelia-positive unfed adult I. persulcatus. B. afzelii DNA was found in 21% of the PCR-positive human blood samples, in 20% of the PCR-positive samples from the small mammals (including blood and tissues) and in 19% of the PCR-positive ticks. Mixed infection with B. garinii and B. afzelii was observed in both clinical and environmental samples. Interestingly, four samples isolated from I. persulcatus were closely related to the relapsing fever species B. miyamotoi.
PMID: 18249150 [PubMed - as supplied by publisher]
http://www.webmd.com/news/20080609/bad- ... RSS_PUBLIC
Bad Lyme Bug Spreading
Virulent Strain of Lyme Disease Spreading in U.S., Europe
By Daniel J. DeNoon
WebMD Health News
Reviewed by Louise Chang, MD
June 9, 2008 -- A virulent strain of Lyme disease germ is spreading in the U.S. and in Europe, a new study shows.
It's not a new strain of Borrelia burgdorferi, the spirochete or spiral-shaped bacterium that causes Lyme disease. In fact, it was one of the first strains ever identified -- found in the cerebrospinal fluid of a patient with severe Lyme meningitis.
But now Wei-Gang Qiu, PhD, Benjamin Luft, MD, and colleagues find that the particularly nasty ospC type A strain appears to be the most common of the 20 or so B. burgdorferi strains found in the U.S. The spread of this virulent strain, they suggest, could be part of the reason for the increase in Lyme disease cases seen over the past two decades.
"OspC type A is the type most widely distributed in the U.S. -- and, as others have shown, this is the most virulent strain," Qiu, an assistant professor at New York's Hunter College, tells WebMD. "If this is widespread, it is not good. You don't want to see this thing increase."
Luft, professor of medicine and former chief of infectious diseases at SUNY Stony Brook, says the spread of the ospC type A strain may explain part of the U.S. Lyme disease epidemic.
"Perhaps part of the Lyme disease story is not that B. burgdorferi just emerged in the 1970s, but that this group of strains that cause more disease became more dominant in our environment," Luft tells WebMD. "So instead of people getting infected with less virulent strains, they got infected with a more virulent strain and got more disease."
Qiu and Luft note that the rise of a more virulent Lyme spirochete isn't the whole story. People get the infection from the bite of a deer tick. As suburbs encroach on rural areas, and as more homes are built near forests, more people are at risk of tick bites. Increased exposure to ticks accounts for most of the increase in Lyme disease.
Qiu and colleagues found that the ospC type A strain is the most widespread strain in the U.S. It's also widespread in Europe. That was a surprise, as Lyme disease spirochetes in Europe are spread by different ticks and harbored by different animal hosts than in the U.S.
"What is surprising is these ospC type A strains in the U.S. and in Europe are genetically almost identical," Qiu says. "So this type is quite unusual in its ability to colonize new habitats. ... This is very strong evidence for this type having a very broad ecological niche in terms of the species that can carry it."
"This means it went from one continent to another continent relatively recently," adds Luft. "And it means that, as Europe and U.S. have very different ecosystems, this strain is highly adaptable to new environments. ... This makes it a formidable foe -- and it causes significant disease."
It's not clear whether this bad Lyme germ traveled from North America to Europe or vice versa. And it's not clear when this happened, although it seems to have occurred in the last 200 years, possibly when a tick-infested bird crossed the Atlantic.
It's likely, Qiu says, that this strain will continue to become more prominent in areas where Lyme disease is established.
CDC medical epidemiologist Kevin Griffith, MD, MPH, says that while Lyme disease has been reported in nearly every state, 10 mostly Northeastern states account for 92% of cases.
Although the 20,000 cases reported to the CDC in 2006 were fewer than the 23,000 cases reported in 2005, Griffith says the true number of cases is probably larger.
"There is probably a true increase in the number of cases," Griffith tells WebMD.
The good news, he says, is that there's been a drop in the most severe, late-stage manifestations of Lyme disease. He attributes this to doctors identifying the disease -- and beginning treatment -- sooner now than in the earlier years of the epidemic.
Qiu and colleagues report their findings in the July issue of the CDC's Emerging Infectious Diseases
http://www.newsday.com/news/printeditio ... 5964.story
Clue to Lyme disease seen in single bacterial strain
BY DELTHIA RICKS | firstname.lastname@example.org
July 2, 2008
Medical investigators have identified a single bacterial strain that causes Lyme disease in patients on two continents and may be responsible for a growing number of severe, invasive cases in the United States and Europe since the 1980s.
Dr. Ben Luft and colleagues at Stony Brook University Medical Center have isolated a strain of Borrelia burgdorferi - the bacterium that causes Lyme - and declared it prevalent not only in the United States, but also Europe. The finding, considered unusual, is reported in the journal Emerging Infectious Diseases. "We studied a large number of strains from throughout the United States and Europe, well over 100 strains," Luft said yesterday. "But there was only one that was identical. And when you consider that we are a world apart separated by an ocean where there is no common interaction ... it is surprising to find that [the American and European strains] were the same."
He believes migratory birds may have transported the strain across continents. Luft said the strain is responsible for 30 percent to 40 percent of all cases of Lyme.
Lyme is the most common tick-transmitted disorder in the United States. It may start as a rash, but an invasive form, caused by a bacterium like the one identified by Luft and his colleagues, can enter the bloodstream and attack major organs. Invasive Lyme has become more prevalent over the last 20 years.
Long Island is endemic for the disease, particularly Suffolk County, where the majority of cases occur. Suffolk Health Commissioner David Graham hailed the new research yesterday, saying answers are needed to help keep pace with the growing incidence of Lyme.
Last year, federal investigators reported cases of Lyme had more than doubled since 1991, with 93 percent of all cases concentrated in New York and nine other states.
"Lyme disease is a serious, chronic problem, especially when it is not detected right away," Graham said. "I am pleased that they're looking into the virulence of these strains from throughout the world. That's very important basic research on the disease pathogen."
Graham estimates 585 cases occurred in Suffolk last year, up dramatically from 2006, when there were 190. However, there were 542 in 2005. In Nassau, there were 123 last year, 109 in 2006 and 135 in 2005.
Advocates for patients applauded the finding, saying the discovery helps explain cases of long-term Lyme disease.
Eva Haughie, president of the Empire State Lyme Disease Association, said 15 members of her immediate and extended family developed Lyme disease. Even her dog got Lyme and died of the disorder. "We've known this all along that there is a difference in virulence," said Haughie, of Manorville, referring to the differing strains of Lyme bacteria.
Some patients say their bout with Lyme was short-lived; others have suffered a variety of symptoms for years. In Haughie's case, she suffered a form of memory impairment comparable to Alzheimer's. Her daughters have what many patients refer to as persistent Lyme.
Int J Med Microbiol. 2002 Jun;291 Suppl 33:80-7.
Horses and Borrelia: immunoblot patterns with five Borrelia burgdorferi sensu lato strains and sera from horses of various stud farms in Austria and from the Spanish Riding School in Vienna.
Müller I, Khanakah G, Kundi M, Stanek G.
Institute for Hygiene and Medical Microbiology of the University Vienna, Wien, Austria.
Grazing animals are continuously exposed to tick bites. Consequently, one may expect that horses will become infected with the various pathogens carried by ticks including Borrelia burgdorferi sensu lato. Whether horses may develop clinical disease due to this pathogen is controversially discussed. We were interested to learn about the infection of horses with Borrelia burgdorferi sensu lato within one season by studying the dynamics of the humoral immune response in paired blood samples. The majority of horses examined were Lipizzaner from the stud farm in Piber/Steiermark, and from the Spanish Riding School in Vienna. Smaller groups of animals of different breeds were from stud farms in Kärnten, Niederösterreich, Salzburg and Steiermark. Clinical status and medical history were obtained and blood was drawn at the beginning of the highest tick activity and nine months later in 1998. Immunoblot technique (Western blot) was used in order to determine the dynamics in the immune response patterns. As antigens served the genospecies Borrelia afzelii, Borrelia burgdorferi sensu stricto, Borrelia garinii, Borrelia lusitaniae, and Borrelia valaisiana. 309 horses (age median 7 years, range 1/12 to 33 years) were seen at the first round. 186 of these animals (60.2%; median age 6 years, range 4/12 to 33 years) were re-examined in the second round. All animals were in normal health condition during both rounds of examination and blood sampling. Analysis of the immunoblot patterns was based on in-house-, Pko-, Pka2-, Pbi-, and European Union Concerted Action on Lyme Borreliosis (EUCALB) 2 & 3-criteria; analyses revealed a variety of positive results with different strains and criteria. Positive immunoblot results with 186 paired samples and B. afzelii as antigen, for example, ranged from 52 to about 91% in the first, and 53 to 93% in the second round. The age dependency analyses showed that the first infection with B. burgdorferi sensu lato occurs in the first year. Re-infection is characterised by appearance of additional bands. Continuously tick-exposed horses show a stable pattern of bands whilst in unexposed horses the number of bands decreases with age. In this study horses became repeatedly infected with B. burgdorferi sensu lato but, apparently, developed only rarely, if at all, clinical diseases. The infectious agent is predominantly B. afzelii. Antibodies to other borrelia genospecies are predominantly due to cross reactivity.
PMID: 12141766 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/1902 ... rom=pubmed
Borrelia carolinensis sp.nov. - a new (14th) member of Borrelia burgdorferi sensu lato complex from the southeastern United States
Nataliia Rudenko, Maryna Golovchenko, Libor Grubhoffer, and James H. Oliver Jr.
Journal of Clinical Microbiology Accepts, published online ahead of print on 19 November 2008
Pol Merkur Lekarski. 2008 Sep;25(147):254-6. [Re-infection with Borrelia burgdorferi s.l in a patient with a history ofneuroborreliosis--case report] [Article in Polish] Grygorczuk S, Pancewicz S, Zajkowska J, Kondrusik M, Swierzbinska R, MoniuszkoA, Pawlak-Zalewska W. Uniwersytet Medyczny w Bialymstoku, Klinika Chorob Zakaznych i Neuroinfekcji.email@example.com
Reinfection with Borrelia burgdorferi s.l., which is likely in highly exposedpersons, has not been described in Poland so far. Symptoms of Lyme arthritis,preceded by typical skin lesion (erythema migrans) appeared in 46 years oldwomen 5 years after successful treatment of borrelial meningitis. Re-appearanceof symptoms of Lyme borreliosis following localized skin lesion, after a longasymptomatic period, as well as accompanying increase in specific antibodies,point to reinfection with Borrelia burgdorferi sensu lato. Differentlocalization of systemic symptoms during two episodes of Lyme disease suggestsinfection with distinct genospecies of B. burgdorferi s.l. This case confirmsrisk of recurrent infections with different B. burgdorferi s.l. genospecies ininhabitants of highly endemic areas in the north-east of Poland, which may posenecessity of repeated antibiotic treatment.
Publication Types:English Abstract PMID: 19112843 [PubMed - in process]
Am J Trop Med Hyg. 2009 Feb;80(2):252-8.
Arthritogenicity of Borrelia burgdorferi and Borrelia garinii: comparison ofinfection in mice.
Craig-Mylius KA, Lee M, Jones KL, Glickstein LJ. Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital,Charlestown, Massachusetts 02129, USA. firstname.lastname@example.org
Arthritogenicity, as determined by joint swelling and synovial histology, wascompared between or within two Borrelia genospecies that cause Lyme arthritis inhumans. The spirochete burden in bladder tissue (a site of spirochetepersistence) was documented by quantitative polymerase chain reaction, andimmune responses were analyzed. In C3H/HeJ mice, three B. burgdorferi isolatesand two of the three B. garinii isolates induced severe arthritis and swelling.Previous designation as invasive or noninvasive B. garinii, or RNA spacer typeof B. burgdorferi did not determine arthritis severity induced by isolates.Compared with the other five isolates, the B. garinii PBi isolate inducedsignificantly less arthritis, a lower humoral immune response, and persisted ata much lower level in bladder tissue. However, B. garinii PBi isolates inducedsimilar Borrelia antigen-specific inflammatory T cell responses from the localdraining lymph node. Thus, diverse B. burgdorferi and B. garinii isolates werehighly arthritogenic in immune competent mice.
Publication Types:Research Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov't PMID: 19190223 [PubMed - in process]
Br J Dermatol. 2009 Mar 19; [Epub ahead of print]
Species of Borrelia burgdorferi complex that cause borrelial lymphocytoma inFrance.
Lenormand C, Jaulhac B, De Martino S, Barthel C, Lipsker D. Clinique Dermatologique, Universite Louis Pasteur, 67091 Strasbourg, France. Background Only about 30 cases of borrelial lymphocytoma (BL) withidentification of the causative species of Borrelia have been published to date,mainly from Eastern or Central European countries. Objectives To identify thespecies of B. burgdorferi complex responsible for BL in France. Methods Ninepatients with BL acquired in France and for whom skin samples were sent to thenational reference centre laboratory between 1994 and 2007 were included in thisretrospective study. Direct detection of Borrelia in skin samples was made bypolymerase chain reaction targeting the fla gene. Culture was performed whentechnically possible, and identification of each species was made byhybridization of a fragment of the fla gene with a panel of species-specificoligonucleotides. Results Borrelia afzelii was identified in three cases, B.garinii and B. burgdorferi sensu stricto in one case each. Culture was positivein only one case (B. garinii).
Conclusions Borrelia afzelii seems to be the predominant species of Borrelia responsible for BL in France, as alreadyreported in other European countries.
PMID: 19302068 [PubMed - as supplied by publisher]
Zh Mikrobiol Epidemiol Immunobiol. 2009 Jan-Feb;(1):63-6.
[Isolation of tick-borne borreliosis agent from blood of patients] [Article in Russian] [No authors listed]
During spring-autumn period of 2006 Borrelia were isolated for the first time inRussia from blood of 79 patients treated in Perm City Clinical Hospital forInfectious Diseases No. 1 with diagnosis "tickborne borreliosis, manifestiveform with migrating erythema, localized stage". Ten primary isolates (12.7% oftotal seeded samples) were obtained by seeding plasma samples on the BSK medium.Their subsequent identification by polymerase chain reaction-restrictionfragments length polymorphism revealed presence of Borrelia garinii NT29 in allpatients. Length of sequenced fragment of rrfA-rrlB region was 253 b.p. Sevenisolates had 100% and 3 - 99.6% similarity with typical strain NT29 (L30130).Nucleotide sequences of 4 obtained isolates were deposited in GenBank database(No. AM932199 - AM932202). It was proposed that B. garinii NT29 more frequentlythan other Borrelia species can be an etiologic agent of tick-borne borreliosisnot only in Perm region but also in whole Russia. Publication Types:English Abstract PMID: 19340967 [PubMed - in process]
Curr Probl Dermatol. 2009;37:1-17. Epub 2009 Apr 8. Borrelia burgdorferi sensu lato Diversity and Its Influence on Pathogenicity inHumans. Baranton G, De Martino SJ. Centre National de Reference Borrelia, Institut Pasteur, Paris, France. Among the Spirochaetes, the
Borrelia burgdorferi sensu lato complex isresponsible for Lyme borreliosis. This complex comprises more than 13 Borreliaspecies. Four of them are clearly pathogenic for humans: B. burgdorferi sensustricto, B. afzelii, B. garinii and B. spielmanii. They can generate erythemamigrans, an initial skin lesion, and can then spread deeply into the host toinvade distant tissues, especially the nervous system, the joints or the skin.In humans, Borrelia pathogenicity seems to be linked with taxonomic position, butin vitro studies show the role of plasmids in B. burgdorferi s.l. pathogenesis.The inter- and intraspecies genetic diversity of B. burgdorferi s.l. evidences aclonal evolution of the chromosome, while plasmid genes are quite variable,suggesting their major role in Borrelia adaptability. The plasmid-encodedadhesins and vlse, crasps and osp genes determine invasiveness and host immuneevasion of B. burgdorferi s.l., and select the bacterial host spectrum. Thegeographic distribution of B. burgdorferi s.l. is closely related to its vectorsand competent hosts, and its development within these influences its diversity,taxonomy and pathogenesis, primarily via genetic lateral transfer. Copyright (c)2009 S. Karger AG, Basel. PMID: 19367094 [PubMed - as supplied by publisher]
Curr Probl Dermatol. 2009;37:18-30. Epub 2009 Apr 8.
Life Cycle of Borrelia burgdorferi sensu lato and Transmission to Humans. Gern L. Institute of Biology, University of Neuchatel, Neuchatel, Switzerland.
Lyme borreliosis is a zoonosis: its causative agent, Borrelia burgdorferi sensulato, circulates between Ixodes ricinus ticks and a large variety ofvertebrates. I. ricinus has a wide geographical distribution throughout Europewithin the latitudes of 65 degrees and 39 degrees and from Portugal into Russia.Enzootic cycles in Europe involve at least 7 Borrelia species. Apparently,associations exist in nature between Borrelia species and hosts. B. afzelii andB. burgdorferi sensu stricto are associated with rodents, and B. garinii and B.valaisiana with birds. B. lusitaniae may be transmitted to ticks by some lizardspecies and birds. B. spielmanii appears to be associated with dormice and hedgehogs.
Less strict associations also exist. Transmission of Borreliainfection by I. ricinus to their hosts, including humans, does not occurimmediately when ticks attach to host skin. A delay is observed, which maydepend on the Borrelia species infecting the tick.
B. afzelii can be transmittedduring the first 24 h, whereas B. burgdorferi needs 48 h of tick attachmentbefore its transmission begins. Nothing is known about the other Borreliaspecies; however, success of transmission always increases with tick attachmentduration. Therefore, careful visual examinations of the body for at least 2successive days are recommended after visiting an endemic area.
Copyright (c)2009 S. Karger AG, Basel. PMID: 19367095 [PubMed - as supplied by publisher]
Przegl Epidemiol. 2009;63(1):19-22. [STARI--a new tick borne spirochetosis] [Article in Polish] Zajkowska J, Moniuszko AM, Czupryna P, Pancewicz SA, Grygorczuk S, Kondrusik M. Klinika Chorob Zakaznych i Neuroinfekcji Uniwersytetu Medycznego w Bialymstoku.email@example.com
Southern Tick Associated Rash Illness (STARI), also known as Masters disease iscaused by Borrelia lonestari spirochetes and it is transmitted by Amblyommaamericanum tick. Early symptoms of STARI are similar to early Lyme borreliosis(skin rash resembling Erythema migrans). In diagnostic of STARI traditionalmethods are not effective. The decision of antibiotic treatment (similar to Lymeborreliosis) still remain based on clinical picture.
Publication Types:English Abstract PMID: 19522220 [PubMed - in process]
1. Borrelia valaisiana, previously of unknown pathogenic significance, has now been detected by PCR in the cerebrospinal fluid of a patient from Greece showing clinical evidence of advanced neuroborreliosis.
Diza E, Papa A, Vezyri E, Tsounis S, Milonas I, Antoniadis A. Borrelia valaisiana in cerebrospinal fluid.Emerg Infect Dis. 2004 Sep;10(9):1692-3.
2. Borrelia lusitaniae, also previously of unknown pathogenic significance, has been isoolated from a human for the first time. The patient, a Portuguese woman from the Lisbon area, presented with skin lesions that had persisted for 10 years, and B. lusitaniae was isolated from skin biopsies cultured in BSK medium. The identity of the spirochaetes was determined by RFLP-PCR and confirmed by sequence analysis of the intergenic spacer region.
Collares-Pereira M, Couceiro S, Franca I, Kurtenbach K, Schafer SM, Vitorino L, Goncalves L, Baptista S, Vieira ML, Cunha C. First isolation of Borrelia lusitaniae from a human patient. J Clin Microbiol. 2004 Mar;42(3):1316-8
3. Borrelia spielmani (Hollanti). Correctly possierlich - borrelia spielmani - the name of a new Borrelienart, which was discovered by the Parasitologen team around Professor Dr. Franz Franz-Rainer Matuschka, Charité Berlin, sounds.
Under the designation the Borrelie found in a Netherlands patient already strewed A14S before five years suspicion on a new species. One found it in three-landhits a corner to Germany, France and Switzerland, in the elsaessischen Petite in such a way specified Camargue. Also the natural landlords are exposed. The Haselmaus concerns, in Central Europe of common rodents and in exceptions. "spielmani" after that Zeckenfor Andrew Spielman, Harvard University of, was designated the USA.
Frankfurt general newspaper, 15,12,2004, NR. 293/page 38
New research results and studies
http://www.borreliose-magazin.de/cms/mo ... cle&sid=23
Articles by Richter, D.
Articles by Matuschka, F.-R.
Applied and Environmental Microbiology, November 2004, p. 6414-6419, Vol. 70, No. 11
Relationships of a Novel Lyme Disease Spirochete, Borrelia spielmani sp.nov., with Its Hosts in Central Europe
Bakteerin alalajien ja sitä kautta oireiden tunnistaminen parantaa hoitotuloksia. Esim. jotkin alalajit tunkeutuvat toisia nopeammin keskushermostoon. Alalajin tunnistaminen auttaa tunnistamaan tilanteen varhaisessa vaiheessa ja esim. keskushermostoon paremmin vaikuttava suonensisäinen antibioottihoito aloitamaan aiemmin." (2010)
Genotypic Variation and Mixtures of Lyme Borrelia in Ixodes Ticks from North America and Europe
http://www.plosone.org/article/info%3Ad ... ne.0010650
"The presence of multiple Borrelia genotypes in ticks increases the probability that a person will be infected with more than one genotype of B. burgdorferi, potentially increasing the risks of disseminated Lyme disease. Our study indicates that the genotypic diversity of Borrelia in ticks in both North America and Europe is higher then previously reported and can have potential clinical consequences"....
"The ability to quickly identify theBorrelia species and associate particular genotypes with certain symptoms can have important patient-management implications. For example, the clinical observation of a single erythema migrans lesion where at least one of the recovered spirochetes is associated with an invasive phenotype might alert the treating physician to consider the use of an antibiotic, such as an intravenously administered one, that will reliably penetrate the central nervous system ,  or, at the very least, the physician would monitor the patient for development of neurologic symptoms even with administration of oral antibiotics."
J Bacteriol. 2011 Jan 7; [Epub ahead of print]
Whole genome sequence of an unusual Borrelia burgdorferi sensu lato isolate.
Casjens SR, Fraser-Liggett CM, Mongodin EF, Qiu WG, Dunn JJ, Luft BJ, Schutzer
Department of Pathology, Division of Microbiology and Immunology, University of
Utah Medical School, Salt Lake City, UT 84112; Institute for Genome Sciences,
University of Maryland, School of Medicine, Department of Microbiology and
Immunology, Baltimore, MD 21201; Department of Biological Sciences, Hunter
College of the City University of New York, New York, NY 10065; Biology
Department, Brookhaven National Laboratory, Upton, NY 11793; Department of
Medicine, Health Science Center, Stony Brook University, Stony Brook, NY 11794;
Department of Medicine, University of Medicine and Dentistry of New Jersey-New
Jersey Medical School, Newark, NJ 07103.
Human Lyme disease is caused by a number of related Borrelia burgdorferi sensu
We report here the complete genome sequence of Borrelia sp.
isolate SV1 from Finland. It is a so far the closest known relative of B.
burgdorferi sensu stricto, but it is sufficiently genetically distinct from that
species that it and its close relatives warrant its candidacy as a new species
status. We suggest that it should be named Borrelia finlandensis.
http://eutils.ncbi.nlm.nih.gov/entrez/e ... md=prlinks
PMID: 21217002 [PubMed - as supplied by publisher]
1. sensu stricto
Argas ja Ornitodorus suvun punkkien valittama, joita esiintyy linnuissa (mm. kanat, kalkkunat, ankat jne). Oireet: anemia, ripuli, neurologiset ongelmat
?h recurrentis (Toisintokuume)
Paaasiallinen levittaja on tai. Oireet: relapsoiva kuume.
Esiintyy lahinna Lansi-Afrikassa. Oireet: relapsoiva kuume.
Esiintyy lahinna Espanjassa, Portugalissa ja Luoteis-Afrikassa. Oireet: relapsoiva kuume.
Oireet: relapsoiva kuume. Aiheuttaa enemman kuolemantapauksia kuin Borrelia recurrentis.
¡¨Mykoplasmat ovat pienimpia (halkaisijaltaan noin 0,2 £gm) ja yksinkertaisimpia itsenaisesti lisaantyvia prokaryootteja, jotka on tunnistettu yli sata vuotta sitten.¡¨
¡¨Pienen genomikoon vuoksi mykoplasmat elavat usein loisina, joilla on tarkka isanta- ja kudosspesifisyys. Paastyaan sopivaan isantaorganismiin mykoplasmat elavat ja lisaantyvat huomattavan pitkiakin aikoja. Ne suojautuvat isantaorganismin immuunivasteelta mm. jaljittelemalla isannan omia antigeeneja¡¨
¡¨ Mykoplasmoja tunnetaan yli 190 lajia, jotka infektoivat ihmisia, elaimia, hyonteisia ja kasveja.¡¨
Itioelaimiin kuuluvia alkuelaimia, jotka aiheuttavat malariaa muistuttavan taudin (babesiaasin) loisimalla punasolujen sisalla.
?h Bartonella (kissanraapimatauti, juoksuhautakuume)
Aiheuttaa kaksi erilaista tautia ihmisessa. Oroyan kuume joka on vakava ja iho-oireita aiheuttava Verruga peruana.
?« symbiont of Trichobius major
Riketsiat jaetaan Rickettsia-, Coxiella- ja Ehrlichia-sukuihin
¡¨Riketsiat ovat solunsisaisia parasiitteja, jotka ovat riippuvaisia isantasolusta kasvaakseen ja lisaantyakseen. Riketsiat ovat pleiomorfisia bakteereita eli niiden muoto voi vaihdella.¡¨
Riketsiat aiheuttavat elaimille ja ihmisille monia sairauksia, lahinna erilaisia pilkkukuumeita. R.Akari aiheuttaa riketsiarokkoa. Jotkut riketsiat aiheuttavat kasvitauteja.
Bakteeri sarkoidoosin takana ?
?« sibirica subgroup
Rotan valittama pilkkukuume. Ihmisella vakava tauti, mutta ei
?h Anaplasma (entinen Ehrlichia)
Ehrlichiat aiheuttavat sairauksia seka ihmisella etta elaimilla.
?h TBE (Puutiaisaivokuume, Kumlingentauti)
Taudin aiheuttaa arbovirus. Suomessa punkeista alle 1 % levittaa virusta. Maassamme sairastuu vuosittain noin 20 ihmista. Virus jattaa pysyvan immuniteetin. Tautiin on tehokas rokote.
Ticks Tick Borne Dis. 2011 Sep;2(3):123-8. Epub 2011 May 27.
Updates on Borrelia burgdorferi sensu lato complex with respect to public health.
Rudenko N, Golovchenko M, Grubhoffer L, Oliver JH Jr.
Biology Centre AS CR v.v.i. AS CR, Institute of Parasitology, Branisovska 31, 37005 Ceske Budejovice, Czech Republic; Faculty of Sciences, University of South Bohemia, 37005 České Budějovice, Czech Republic; Georgia Southern University, Institute of Arthropodology and Parasitology, Statesboro, GA 30460-8056, USA.
Borrelia burgdorferi sensu lato (s.l.) complex is a diverse group of worldwide distributed bacteria that includes 18 named spirochete species and a still not named group proposed as genomospecies 2. Descriptions of new species and variants continue to be recognized, so the current number of described species is probably not final. Most of known spirochete species are considered to have a limited distribution.
Eleven species from the B. burgdorferi s.l. complex were identified in and strictly associated with Eurasia (B. afzelii, B. bavariensis, B. garinii, B. japonica, B. lusitaniae, B. sinica, B. spielmanii, B. tanukii, B. turdi, B. valaisiana, and B. yangtze), while another 5 (B. americana, B. andersonii, B. californiensis, B. carolinensis, and B. kurtenbachii) were previously believed to be restricted to the USA only.
B. burgdorferi sensu stricto (s.s.), B. bissettii, and B. carolinensis share the distinction of being present in both the Old and the New World.
Out of the 18 genospecies, 3 commonly and 4 occasionally infect humans, causing Lyme borreliosis (LB) - a multisystem disease that is often referred to as the 'great imitator' due to diversity of its clinical manifestations. Among the genospecies that commonly infect people, i.e. B. burgdorferi s.s., B. afzelii, and B. garinii, only B. burgdorferi s.s. causes LB both in the USA and in Europe, with a wide spectrum of clinical conditions ranging from minor cutaneous erythema migrans (EM) to severe arthritis or neurological manifestations.
The epidemiological data from many European countries and the USA show a dramatic increase of the diagnosed cases of LB due to the development of new progressive diagnostic methods during the last decades (Hubálek, 2009). Recently, the definition of the disease has also changed. What was not considered Lymeborreliosis before might be now.
Copyright © 2011 Elsevier GmbH. All rights reserved.
New York Medical College, Yale look for new tick-borne disease in region
1:17 AM, Sep. 29, 2011 |
Westchester County, New York
VALHALLA ? New York Medical College plans to help Yale University study whether a new deer-tick-borne disease that has reportedly infected people in Russia also has caused illness here.
Yale researchers received a grant from the National Institutes of Health to develop a diagnostic test for Borrelia miyamotoi, a bacterium that causes high fever and is a distant relative of the pathogen that causes Lyme disease. The medical college will provide blood specimens from patients for the new test.
"Our role is to see if patients in our region who had tick bites or were exposed to ticks are, in fact, coming down with this infection that up until now we may have attributed to Lyme disease or something else," said Dr. Gary Wormser, a tick-borne-disease expert at New York Medical College and chief of infectious diseases at Westchester Medical Center.
Though one study showed that up to 4 percent of adult deer ticks in Westchester have B. miyamotoi, "what hasn't been addressed is whether it causes an illness at all in humans in our area," Wormser said.
If it is discovered in local people, antibiotic treatment likely would be similar to what's prescribed for Lyme disease, he said.
While there is currently no test in the United States to check for it, researchers have reported finding B. miyamotoi in patients in Russia, according to a study to be published in next month's Emerging Infectious Diseases journal.
Yale and Russian researchers said it was discovered in 46 patients using a diagnostic test that picked up the organism's DNA in their blood.
Symptoms included headache, chills and a high fever that sometimes subsided and recurred. The disease does not seem to be life-threatening, and the infection appeared to clear with antibiotics.
Durland Fish, an epidemiology professor at Yale School of Public Health and first director of the Lyme Disease Research Center at New York Medical College, discovered B. miyamotoi in deer ticks in Connecticut 10 years ago.
"We've known for a long time that this organism occurs in ticks, and we've since found it in ticks wherever there is Lyme disease," said Fish, who worked on the Russian study.
Before Yale was awarded the grant, researchers had tried unsuccessfully for years to get funding to study the organism's possible human connection, he said.
"The main concern is if people have this infection, and if they aren't treated, we don't know what the outcome of that will be, so certainly we would like it to be appropriately diagnosed," he said.
Yale researchers are developing a blood test to detect antibodies produced in response to the infection. The DNA of the organism is not always detected in a patient's blood.
There are currently three deer-tick-borne diseases identified in humans in the Northeast: Lyme disease, babesiosis and human granulocytic anaplasmosis, also known as ehrlichiosis.
Multilocus sequence analysis of Borrelia bissettii strains from North America reveals a new Borrelia species, Borrelia kurtenbachii
Margos, G., Hojgaard, A., Lane, R. S., Cornet, M., Fingerle, V., Rudenko, N., Ogden, N., Aanensen, D. M., Fish, D. and Piesman, J., 2010. Multilocus sequence analysis of Borrelia bissettii strains from North America reveals a new Borrelia species, Borrelia kurtenbachii. Ticks and Tick-Borne Diseases, 1 (4), pp. 151-158.
Using multilocus sequence analysis (MLSA), we investigated the phylogenetic relationship of spirochaete strains from North America previously assigned to the genospecies Borrelia bissettii. We amplified internal fragments of 8 housekeeping genes (clpA, clpX, nifS, pepX, pyrG, recG, rplB, and uvrA) located on the main linear chromosome by polymerase chain reaction. Phylogenetic analysis of concatenated sequences of the 8 lad showed that the B. bissettii clade consisted of 4 closely related clusters which included strains from California (including the type strain DN127-C19-2/p7) and Colorado that were isolated from Ixodes pacificus, I. spinipalpis, or infected reservoir hosts. Several strains isolated from I. scapularis clustered distantly from B. bissettii. Genetic distance analyses confirmed that these strains are more distant to B. bissettii than B. carolinensis, a recently described Borrelia species, which suggests that they constitute a new Borrelia genospecies.
**We propose that it be named Borrelia kurtenbachii sp. nov. in honour of the late Klaus Kurtenbach.**
The data suggest that ecological differences between B. bissettii and the new Borrelia genospecies reflect different transmission cycles. In view of these findings, the distinct vertebrate host-tick vector associations and the distributions of B. bissettii and B. kurtenbachii require further investigation.
BORRELIA KURTENBACHII IS WIDELY DISTRIBUTED IN SOUTHEASTERN UNITED
Corresponding Author: Nataliia Rudenko, Biology Centre, Branisovska 31, Ceske Budejovice
Authors: Rudenko, Nataliia; Golovchenko, Maryna ; Grubhoffer, Libor ; Oliver Jr., James
Borrelia kurtenbachii, B. bissettii, southeastern US, tick vectors, hosts
Borrelia burgdorferi sensu lato (s.l.) complex is a diverse group of worldwide distributed bacteria that includes 18
named spirochete species and not named yet group proposed as genomospecies 2. B. bissettii is considered to
be the second major North American spirochete genospecies widely distributed in southeastern and far western
United States and in Colorado. Until recently it was believed that B. bissettii is restricted to U.S.A. only, but
newest publication confirmed that B. bissettii shares with B. burgdorferi sensu stricto the distinction of being
present in both the Old and New World.
Recent reinvestigation of the phylogenetic relatedness of several North American strains from California,
Colorado, New York and Illinois previously included into DN127 group split B. bissettii genospecies into two
separate species showing that strain 25015 and few other strains from Illinois represent a distinct group
proposed as B. kurtenbachii sp. nov. Until now it was believed that B. kurtenbachii is restricted to the east of the
Analysis of 118 Borrelia isolates cultured from tick, rodents and birds in southeastern part of the United States
(GA, SC and FL) revealed the presence of rather divergent group of B. bissettii strains (35 isolates) with high
similarity to strains DN127 and 25015. Using the reverse line blotting and two newly designed probes that are
able to distinguish B. bissettii and B. kurtenbachii we found that the majority of strains previously defined as B.
bissettii represents in reality B. kurtenbachii. While the RLB probe for detection of B. kurtenbachii was 100%
identical to the selected region of 5S-23S IGS of 21 southeastern isolates, the sequence similarity between this
group and strain 25015 was not too high. The presence of the 6 nt gap was revealed at the 5? end of 5S-23S
IGS of 15 out of 21 southeastern B. kurtenbachii strains showing the major differences between type strain of B.
kurtenbachii and southeastern isolates.
Out of 21 southeastern B. kurtenbachii strains 14 were cultivated from eastern woodrat (Neotoma floridana) ear
biopsy, 3 from cotton mouse (Peromyscus gossypinus) ear biopsy, 2 from Ixodes minor, 1 from I. affinis and 1
from I. scapularis.
It is interesting to note that among 23 randomly picked B. bissettii samples analyzed by reverse line blotting, 7
samples were confirmed to contain a single B. kurtenbachii species while the rest of the isolates (16) represent
co-infection of B. kurtenbachii and B. bissettii. Newly described B. kurtenbachii was detected in 4 distinct
locations of South Carolina and in Bulloch County, Georgia. Further molecular and phylogenetic analysis of
southeastern B. bissettii and B. kurtenbachii strains is in progress.
J Clin Microbiol. 2002 Sep;40(9):3308-12.
Determination of novel Borrelia genospecies in Swedish Ixodes ricinus ticks.
Fraenkel CJ, Garpmo U, Berglund J.
Department of Infectious Diseases, Blekinge Hospital, S-371 85 Karlskrona, Sweden. firstname.lastname@example.org
A total of 301 adult questing Ixodes ricinus ticks were collected at 15 different locations along the south and east coasts of Sweden to determine the Borrelia genospecies diversity. Thirty-two ticks (11%) were found to be positive by nested PCR with Borrelia burgdorferi sensu lato-specific primers. Species determination was based on partial sequencing of the 16S rRNA gene and the flagellin gene. Five different Borrelia species were found. The nucleotide sequence of the Borrelia DNA found in two ticks differed extensively from the nucleotide sequences of the Borrelia DNA found in the other ticks, and analysis revealed that they were closely related to the relapsing fever borrelia species Borrelia miyamotoi. This is the first report of a B. miyamotoi-like borrelia in I. ricinus and in Europe.
Moreover, the Borrelia DNA of two ticks (6%) clustered within the B. valaisiana complex. B. valaisiana has not previously been reported in Sweden. B. afzelii DNA was found in 14 ticks (44%), and B. garinii DNA was found in 10 ticks (31%). B. burgdorferi sensu stricto DNA was found in four ticks (13%). We conclude that all of the known human-pathogenic species (B. garinii, B. afzelii, and B. burgdorferi sensu stricto) and B. valaisiana found elsewhere in Europe are also present in the Swedish host-seeking tick population and that a B. miyamotoi-like Borrelia species seems to be present in I. ricinus ticks in Europe.
PMID: 12202571 [PubMed - indexed for MEDLINE] PMCID: PMC130762 Free PMC Article
http://www.ncbi.nlm.nih.gov/pmc/article ... figure/F1/
Assessment of the frequency of different Borrelia burgdorferi sensu lato species in patients with Lyme borreliosis from north-east Poland by studying preferential serologic response and DNA isolates.
Authors: Grygorczuk S, Peter O, Kondrusik M, Moniuszko A, Zajkowska J, Dunaj J, Zukiewicz-Sobczak W, Pancewicz S
Citation: Ann Agric Environ Med 2013(Mar); 20(1): 21-9.
Location: Department of Infectious Diseases and Neuroinfections, Medical University in Białystok, Poland.
Introduction and objective.
Several Borrelia burgdorferi sensu lato species cause Lyme borreliosis throughout Europe and their geographic distribution may influence clinical manifestations of the disease. In Poland, Lyme borreliosis presents mainly with neurologic and cutaneous symptoms, while clinically overt arthritis is rare. The presented study investigates the prevalence of B. burgdorferi s.l. genospecies in a group of patients with different clinical forms and stages of Lyme borreliosis in north-east of Poland. This has not previously been studied.
Material and methods.
Preferential reactivity towards different B.burgdorferi s.l. species was investigated with a previously validated immunoblot assay in sera of 33 patients with disseminated Lyme borreliosis: 10 with neuroborreliosis, 6 with acrodermatitis chronica atrophicans and 17 with osteoarticular symptoms. Also typed were B.burgdorferi s.l. DNA isolated from the skin and synovial fluid of 7 patients with erythema migrans, acrodermatitis chronic atrophicans and arthritis.
Preferential reactivity was detected in 30 out of 33 serum samples. Of these, 25 reacted preferentially with B.afzelii, 3 with B. garinii and 2 with B. burgdorferi ss. B.burgdorferi DNA was isolated from all studied samples and typed as B.afzelii in 5. In a patient with acrodermatitis chronica atrophicans studied with both methods simultaneously, B.afzelii was identified by both genotyping and serotyping.
Both methods gave consistent results, indicating B.afzelii as the main agent of all the clinical forms of the Lyme borreliosis in the study area.
The Lancet, Volume 382, Issue 9892, Page 658, 17 August 2013
doi:10.1016/S0140-6736(13)61644-XCite or Link Using DOI
Copyright © 2013 Elsevier
A case of meningoencephalitis by the relapsing fever spirochaete Borrelia miyamotoi in Europe
Joppe W R Hovius MD a Corresponding AuthorEmail Address, Bob de Wever MSc b, Maaike Sohne MD c, Matthijs C Brouwer MD d, Jeroen Coumou MSc e, Alex Wagemakers MD e, Anneke Oei b, Henrike Knol b, Sukanya Narasimhan PhD g, Caspar J Hodiamont MD b, Setareh Jahfari MSc h, Prof Steven T Pals MD f, Hugo M Horlings MD f, Prof Erol Fikrig MD g, Hein Sprong PhD h, Prof Marinus H J van Oers MD c
On April 1 2012, a 70-year-old patient came to our clinic reporting slow cognitive processing, memory deficits, and a disturbed gait, all of which had gradually developed over several months and progressed during the last few weeks before the patient's initial visit. He did not report fever, and he had not been outside the country for several years. He had recently been treated with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone), polychemotherapy, and rituximab (last dose on ...
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 PCR 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 Borrelia chilensis VA1, in honor of its country of origin.
http://www.lakartidningen.se/Klinik-och ... fastingar/
Sju Borreliaarter i svenska fästingar
Thomas G T Jaenson, professor, medicinsk entomologi, Uppsala universitet
Flera betydelsefulla upptäckter om de fästingöverförda infektionerna Lyme-borrelios och fästingöverförd hjärninflammation (TBE) har under de senaste åren gjorts inom det så kallade Stingprojektet vid Linköpings universitet. En av gruppens forskare, Peter Wilhelmsson, presenterar i dagarna i sin doktorsavhandling flera nya betydelsefulla upptäckter om Borreliabakteriernas biologi, speciellt rörande deras överföring från fästingen till människan. Resultaten är viktiga eftersom de ger vägledning om hur vi lämpligast kan minska risken att drabbas av fästingöverförda sjukdomsorsakande mikrober.
Avhandlingen baseras på mikrobiologiska analyser av drygt 2 000 fästingar, som ca 1 500 fästingbitna personer lämnade in till totalt 34 olika vårdcentraler i främst södra Sverige och på Åland under 2007 och 2008. Betydligt fler kvinnor än män lämnade in fästingar.
Dessutom analyserades från varje fästingbiten person två blodprov avseende antikroppar mot Borrelia. Det första blodprovet togs strax efter det att en person tagit bort en fästing från sin hud och lämnat in fästingen till en vårdcentral, det andra blodprovet togs tre månader senare.
Information om eventuella sjukdomssymtom till följd av fästingbetten erhölls genom enkäter och patientjournaler.
Sju arter av Borreliaspiroketer påvisades i fästingarna. Sex av dessa Borreliaarter tillhör artkomplexet Borrelia burgdorferi sensu lato, som kan ge upphov till det sjukdomssyndrom som brukar benämnas Lyme-borrelios, eller i vardagligt tal borrelia. Den sjunde arten, Borrelia miyamotoi, orsakar fästingburen återfallsfeber.
Borreliabakterier påvisades i genomsnitt i var fjärde avlägsnad fästing. Trots detta utvecklade endast 2 procent av de bitna personerna symtom eller tecken på Lyme-borrelios. Av de fästingbitna personerna utvecklade 2,5 procent antikroppar mot Borrelia, trots att man inte fann några sjukdomssymtom av infektionen. Majoriteten av fästingarna, 72 procent, var nymfer, dvs i stadiet mellan larv och vuxen fästing, och hade vanligtvis bitit fast sig på benen.
Genom att i mikroskop mäta avstånden mellan speciella strukturer på en fästing går det att avgöra hur länge den sugit blod. Tiden som fästingen suttit förankrad i huden är starkt positivt korrelerad till sannolikheten att spiroketer överförts från fästing till människa, dvs att en infektion ägt rum. De Borreliainfekterade fästingar som suttit länge i huden (>36 timmar) innehöll färre spiroketer än de som suttit fast en kortare tid. Och de personer som utvecklade antikroppar mot Borreliabakterierna hade tagit bort »sina« fästingar från huden betydligt senare (efter i genomsnitt 58 timmar) jämfört med dem som inte serokonverterade (29 timmar). Yngre personer och kvinnor avlägsnade fästingarna snabbast, äldre personer och män långsammast. Fästingar som fäst sig i huden på huvudet eller i underlivet upptäcktes i allmänhet betydligt senare än de som fäst sig på andra områden på kroppen.
Borrelia miyamotoi sensu lato Seroreactivity and Seroprevalence in the Northeastern United States
Peter J. KrauseComments to Author , Sukanya Narasimhan, Gary P. Wormser, Alan G. Barbour, Alexander E. Platonov, Janna Brancato, Timothy Lepore, Kenneth Dardick, Mark Mamula, Lindsay Rollend, Tanner K. Steeves, Maria Diuk-Wasser, Sahar Usmani-Brown, Phillip Williamson, Denis S. Sarksyan, Erol Fikrig, Durland Fish, and the Tick Borne Diseases Group
Author affiliations: Yale School of Public Health, New Haven, Connecticut, USA (P.J. Krause, J. Brancato, L. Rollend, T.K. Steeves, M. Diuk-Wasser, D. Fish); Yale School of Medicine, New Haven (P.J. Krause, S. Narasimhan, M. Mamula, E. Fikrig); New York Medical College, Valhalla, New York, USA (G.P. Wormser); University of California, Irvine, California, USA (A.G. Barbour); Central Research Institute of Epidemiology, Moscow, Russia (A.E. Platonov); Nantucket Cottage Hospital, Nantucket, Massachusetts, USA (T. Lepore); Mansfield Family Practice, Mansfield, Connecticut, USA (K. Dardick); L2 Diagnostics, LLC, New Haven (S. Usmani-Brown); Creative Testing Solutions, Tempe, Arizona, USA (P. Williamson); State Medical Academy, Izhevsk, Russia (D.S. Sarksyan)
Suggested citation for this article
Borrelia miyamotoi sensu lato, a relapsing fever Borrelia sp., is transmitted by the same ticks that transmit B. burgdorferi (the Lyme disease pathogen) and occurs in all Lyme disease–endemic areas of the United States. To determine the seroprevalence of IgG against B. miyamotoi sensu lato in the northeastern United States and assess whether serum from B. miyamotoi sensu lato–infected persons is reactive to B. burgdorferi antigens, we tested archived serum samples from area residents during 1991–2012. Of 639 samples from healthy persons, 25 were positive for B. miyamotoi sensu lato and 60 for B. burgdorferi. Samples from ≈10% of B. miyamotoi sensu lato–seropositive persons without a recent history of Lyme disease were seropositive for B. burgdorferi. Our resultsA suggest thatA human B. miyamotoiA sensu latoA infection may be common in southern New England and that B. burgdorferi antibody testing is not an effective surrogate for detecting B. miyamotoi sensu lato infection.
Relapsing fever, an arthropod-borne infection caused by several Borrelia spp. spirochetes, is transmitted by ticks and lice (1,2). In 1995, Fukunaga et al. (3) discovered a novel relapsing fever spirochete in the hard-bodied (ixodid) tick Ixodes persulcatus and named it Borrelia miyamotoi. This discovery greatly expanded the potential geographic range of relapsing fever borreliae for humans. Before this finding, only soft-bodied ticks were known to transmit tick-borne relapsing fever spirochetes to humans. In 2001, a related spirochete was detected in I. scapularis ticks in the northeastern United States (4); this and similar organisms have been designated B. miyamotoi sensu lato to distinguish them from the B. miyamotoi sensu stricto isolates from Japan (5). A subsequent study showed that ticks in 15 states in the northeastern and northern midwestern regions of the United States are infected with B. miyamotoi sensu lato and have an average prevalence of infection of 1.9% (range 0–10.5%) (6). B. miyamotoi sensu lato has now been found in all tick species known to be vectors of Lyme disease, including I. pacificus in the western United States, I. ricinus in Europe, and I. persulcatus and I. ricinus in Russia (7–9). The first human cases of B. miyamotoi sensu lato infection were reported from central Russia in 2011 (9). Several reports of B. miyamotoi sensu lato infection in humans have subsequently been published, including 3 in the United States, 1 in Europe, and 1 in Russia (10–14). Some of these reports suggest that B. miyamotoi sensu lato infection causes a nonspecific, virus-like illness. B. miyamotoi sensu lato and B. burgdorferi, the agent of Lyme disease, share several antigens that might cause cross-reactivity during serologic testing, which could lead to a misdiagnosis.
There are few data on the seroprevalence of B. miyamotoi sensu lato infection. To increase knowledge of the seroprevalence of this infection, we used assays for antibodies against B. miyamotoi sensu lato glycerophosphodiester phosphodiesterase (GlpQ), a protein that is absent from all Lyme disease Borrelia species (15), for evaluation of >1,000 archived serum samples from persons living in a Lyme disease–endemic region of the United States. We also performed standard 2-tiered testing for B. burgdorferi antibodies (16). Our aim was to compare the seroprevalence of B. miyamotoi sensu lato with that of B. burgdorferi. We also sought to determine whether persons seropositive for B. miyamotoi sensu lato would also have positive results for standard B. burgdorferi antibody testing.
Materials and Methods
The serum samples evaluated in our study were obtained during 1991–2012 from 3 groups of persons living in areas of the northeastern United States where Lyme disease is endemic. Group 1 consisted of 639 persons from Block Island and Prudence Island, Rhode Island, and from Brimfield, Massachusetts, who participated in serosurveys for tick-borne infections. Persons participating in the serosurvey were healthy at the time of blood sampling and were enrolled during the spring and autumn of each year (16). All participants were asked to respond to a questionnaire and to provide a blood sample for serologic analyses of tick-borne infections.
Group 2 consisted of 194 patients from Block Island; Nantucket, Massachusetts; Mansfield, Connecticut; and the Lower Hudson Valley, New York, who were enrolled in studies of tick-borne diseases. At or near the time of sample collection, persons in this group were treated with doxycycline, amoxicillin, or amoxicillin/clavulanic acid for acute Lyme disease.
Group 3 consisted of 221 adult patients who experienced a febrile illness in the late spring or summer without features suggestive of an upper respiratory tract infection or gastroenteritis. A subgroup of group 3 consisted of 17 patients from the Lower Hudson Valley who were enrolled in a study during 1992–2009 to better characterize the clinical and laboratory features of human granulocytic anaplasmosis as a single infection or as a co-infection with early Lyme disease. Results for serologic testing, culture, buffy coat examination for morulae, and/or PCR showed that none of the patients was infected with Anaplasma phagocytophilum (17). All these patients resided in an area where I. scapularis–transmitted infection is highly endemic and, thus, had possible exposure to I. scapularis ticks. A second subgroup of group 3 consisted of 204 adult patients from Block Island, Mansfield, or Nantucket who had suspected Lyme disease or babesiosis. Testing showed that 25 of these patients had babesiosis but none had Lyme disease or anaplasmosis.
Serum samples were also obtained from 2 patients from the Udmurtia Republic, Russia, who had PCR-confirmed acute B. miyamotoi sensu lato infection. In addition, Creative Testing Solutions (Tempe, AZ, USA) provided an aliquot of residual serum used for blood screening from 300 blood donors who lived in Tempe or in Miami, Florida.
De-identified serum samples were used in this study. The study was approved by the Yale School of Public Health Human Investigation Committee, the New York Medical College Institutional Review Board, and the University of Connecticut Institutional Review Board.
Production of B. miyamotoi sensu lato GlpQ Antigen
Thumbnail of Polyacrylamide gel electrophoresis purification (A) and Western blot analysis (B) of recombinant glycerophosphodiester phosphodiesterase (rGlpQ). A) Coomassie blue staining of purified Borrelia miyamotoi sensu lato rGlpQ (lane 1) and of Precision Plus Protein Prestained Standards (Bio-Rad, Laboratories, Hercules, CA, USA) (lane 2). B) Western blot analysis of B. miyamotoi sensu lato–positive control mouse serum shows 39-kDa rGlpQ-specific band (arrow).
Figure 1. Polyacrylamide gel electrophoresis purification (A) and Western blot analysis (B) of recombinant glycerophosphodiester phosphodiesterase (rGlpQ)A) Coomassie blue staining of purified Borrelia miyamotoi sensu lato rGlpQ (lane 1) and of Precision Plus...
B. miyamotoi sensu lato glpQ from strain LB-2001 cloned into the prokaryotic expression vector pXT7 (18), a derivative of pGEM4Z and pSP64T (Promega, Madison, WI, USA), was transformed into BL21 Star (DE3)/pLysS cells (Invitrogen, Carlsbad, CA, USA), and transformants were used for protein production (6). The chromosome sequence for the protein is in GenBank (accession no. CP006647) (19). The 39.1-kDa recombinant GlpQ (rGlpQ) containing an N-terminal His tag was purified over an Ni-NTA Superflow affinity column (QIAGEN, Valencia, CA, USA) as described by the manufacturer. Purity was assessed by sodium dodecyl sulfate electrophoresis of ≈1 I1/4g of rGlpQ on a 4%–20% polyacrylamide gel and by Coomassie blue staining (Figure 1).
GlpQ Antibody ELISA
We developed a B. miyamotoi sensu lato IgG ELISA by using 20 C3H/HeJ mice (Jackson Laboratory, Bar Harbor, ME, USA). Ten of the mice were not infected. The other 10 age-matched mice were infected by using B. miyamotoi sensu lato–infected I. scapularis nymphal ticks. A month after the mice were infected, blood was obtained from all 20 mice for testing. Titrating concentrations of GlpQ protein and secondary antibody were tested in a checkerboard assay to determine the optimal concentrations for detecting B. miyamotoi sensu lato antibody. Results for the B. miyamotoi sensu lato ELISA were positive for all 10 B. miyamotoi sensu lato–infected mice and negative for all 10 uninfected mice.
To test the human serum samples, we coated ELISA plates with 100 I1/4L of 1 I1/4g/mL GlpQ protein in phosphate-buffered saline (PBS) and incubated the plates at 4° for 18 h. We then added 300 I1/4L of 1% bovine serum albumin in PBS buffer to the plates and incubated them for 2 h at room temperature. The plates were then emptied, and serum was added at a 1:320 dilution and incubated for 1 h. If acute- and convalescent-phase serum samples were available for a study participant, the initial dilution of the acute-phase sample was 1:80, and convalescent-phase samples were diluted to endpoint. The plates were then washed 3 times with wash buffer, and 100 I1/4L of goat antihuman IgG secondary antibody was added at 0.002 mg/mL, incubated for 1 h, and then washed 3 times. BluPhos substrate (Kirkegaard & Perry, Gaithersburg, MD, USA) was added and allowed to react for 20 min before absorbance at 630 nm was determined. B. miyamotoi sensu lato–infected mouse serum was used as a positive control. As a negative control for each plate, we used serum samples that were negative for B. miyamotoi sensu lato antibody, as determined by ELISA and Western blot. The serum was obtained from 3 healthy participants who had no history of tick bite or tick-borne disease and who lived in an area where Lyme disease is endemic. The serum samples were tested by PCR for amplifiable B. miyamotoi sensu lato DNA and were negative. For mouse and human serum samples, a signal >3 SD above the mean of 3 noninfected serum controls was considered positive for B. miyamotoi sensu lato infection.
GlpQ Western Blot Antibody Assay
Purified GlpQ (500 ng) was electrophoresed on each replicate lane of a precast mini 4%–20% sodium dodecyl sulfate–polyacrylamide gel electrophoresis gel (Bio-Rad Laboratories, Hercules, CA, USA) and transferred to a nitrocellulose membrane using the Bio-Rad MiniTrans Blot Cell (Bio-Rad Laboratories). Replicate strips containing rGlpQ were blocked overnight at 4° in PBS (pH 7.2)/5% dried milk/0.05% Tween 20. The blocked strips were then individually incubated with human serum at a 1:250 dilution at room temperature in PBS (pH 7.2)/2.5% dried milk/0.05% Tween 20 for 1 h. The strips were then washed 3 times and incubated for 1 h with horseradish peroxidase–conjugated rabbit anti-human IgG (Sigma-Aldrich, St. Louis, MO, USA) or with horseradish peroxidase–conjugated goat anti-human IgM (Invitrogen) at a 1:5,000 dilution in PBS (pH 7.2)/2.5% dried milk/0.05% Tween 20. Bound antibodies were detected by using Thermo Scientific SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific, Inc., Rockford, IL, USA). Serum from ≈10% of the study participants reacted to a ≈55-kDa band, presumably a trace contaminant copurified with the rGlpQ generated in a bacterial expression system. Samples with a 39-kDa band corresponding to GlpQ on positive control mouse serum samples were considered GlpQ antibody–positive (Figure 1).
PCR DNA Amplification
We used a B. miyamotoi sensu lato PCR as described (4) to amplify B. miyamotoi sensu lato DNA in serum samples. B. burgdorferi DNA was amplified by using a standard PCR assay (16).
B. burgdorferi Antibody Detection
We detected serologic evidence of exposure to B. burgdorferi by using a whole-cell sonicate ELISA, C6 ELISA, or Western blot assay as described (16,20–22). Specimens were considered positive according to the criteria of the US Centers for Disease Control and Prevention (http://www.cdc.gov/lyme/diagnosistestin ... index.html).
B. miyamotoi sensu lato–seropositive serum samples were defined by the presence of B. miyamotoi sensu lato antibody as determined by using ELISA and confirmatory Western blot assays for IgG alone or IgG plus IgM antibody. B. burgdorferi seropositive serum samples were defined by the presence of B. burgdorferi antibody as determined by ELISA and supplemental Western blot IgM or IgG assays.
Study participants were considered to have B. miyamotoi sensu lato infection if they had exhibited a fever >37.5° and a >4-fold rise in antibody to B. miyamotoi sensu lato GlpQ protein between acute- and convalescent-phase serum samples, as determined by ELISA and confirmatory Western blot assays for IgG alone or IgG plus IgM. The time between acute- and convalescent-phase samples ranged from 2 wk to 2 mo. Study participants were considered to have Lyme disease if they had a physician-diagnosed erythema migrans skin lesion or a virus-like illness plus a test result that showed either PCR amplification of B. burgdorferi DNA in blood or B. burgdorferi seroconversion from negative to positive between acute- and convalescent-phase serum samples.
A 2-tailed Fisher exact test was used to compare the frequency of B. miyamotoi sensu lato–seropositive and –seronegative study participants in groups 1, 2, and 3. The McNemar χ2 test was used to compare the seroprevalence of B. miyamotoi sensu lato and B. burgdorferi among group 1 participants.
Seroprevalence of B. miyamotoi sensu lato Infection
Serum samples from 52 of the 1,054 study participants were seroreactive to B. miyamotoi sensu lato antigen by rGlpQ ELISA and Western blot assay (Table 1). The percentage of B. miyamotoi sensu lato–seropositive persons was greater among participants with Lyme disease (group 2; 19/194 [9.8%]) than among those who were healthy (group 1; 25/639 [3.9%], p<0.01 by Fisher exact test, odds ratio [OR] 2.66 [range 1.35–5.16]) or those who had a febrile illness in the late spring or summer (group 3; 8/221 [3.6%], p<0.05 by Fisher exact test, OR 2.89 [range 1.17–7.81]). B. miyamotoi sensu lato DNA could not be amplified from any serum samples (including 27 acute-phase serum samples) from the 52 participants who had test results positive for B. miyamotoi sensu lato antibody.
Of the 639 serum samples from group 1 participants, 25 (3.9%) were seroreactive to B. miyamotoi sensu lato antigen and 60 (9.4%) were seroreactive to B. burgdorferi antigen, as determined by using the standard 2-step ELISA and Western blot procedure (McNemar χ2 test, p<0.0001, OR 10.23 [range 7.84–13.57]). About half (51%) of group 1 participants were male, and the mean age of group 1 participants was 55 years (range 2–102). There was no significant difference in the sex of the group 1 study participants who were seropositive for B. miyamotoi sensu lato (40% male) and those who were seropositive for B. burgdorferi (53% male; p = 0.34). The mean age also did not differ significantly between participants who were seropositive for B. miyamotoi sensu lato (59 years [+15]) and those who were seropositive for B. burgdorferi (61 years [+15]; p = 0.62).
Of the participants from Brimfield, Massachusetts, 9.3% (10/107) were seropositive for B. miyamotoi sensu lato and 7.5% (8/107) were seropositive for B. burgdorferi, compared with 3.2% (15/474) and 11% (52/474), respectively, of the participants from Block Island, Rhode Island. None of the 58 participants from Prudence Island, Rhode Island, were seropositive for B. miyamotoi sensu lato or B. burgdorferi.
Serodiagnosis of B. miyamotoi sensu lato Infection
To assess B. miyamotoi sensu lato ELISA and Western blot assay accuracy in patients with confirmed B. miyamotoi infection, we tested acute- and convalescent-phase serum samples from 2 patients in Russia with B. miyamotoi sensu lato infection confirmed by real-time PCR–(9). Both patients had a >4-fold rise in B. miyamotoi sensu lato GlpQ antibody between acute- and convalescent-phase serum samples (1:80 and 1:2,560, respectively, for 1 patient and 1:640 and 1:2,560, respectively, for the other), as determined by ELISA and confirmed by Western blot.
To assess B. miyamotoi sensu lato ELISA and Western blot assay performance in persons at low risk for Lyme disease or B. miyamotoi sensu lato infection, we performed the GlpQ ELISA on 300 serum samples from healthy blood donors living in Tempe or Miami. For the 9 microtiter plates used for this serosurvey, the mean and standard deviation of the ELISA optical density values for 3 negative control serum samples ranged from 0.108 to 0.136 and from 0.03 to 0.07, respectively. Of the 300 samples, 19 (6.3%) exceeded the mean of the negative control serum by >3 SDs, but none was reactive by Western blot.
We determined whether B. miyamotoi sensu lato infection might be misdiagnosed as Lyme disease in persons whose serum was reactive by B. burgdorferi antibody testing. Of the 36 B. miyamotoi sensu lato–seropositive study participants without a clinical history of Lyme disease within the previous 2 years, 7 (19.4%) had test results positive for B. burgdorferi by IgG and/or IgM ELISA, 6 (16.7%) had test results positive for C6 ELISA, and 4 (11.1%) had test results positive for standard 2-tier ELISA plus confirmatory Western blot (Table 2). The 2-tier B. burgdorferi ELISA and Western blot assay combination used in our laboratory has a 2% false-positive rate.
Clinical Manifestations among Patients with B. miyamotoi sensu lato Seroconversion
Thumbnail of Western blot reactivity to recombinant Borrelia miyamotoi glycerophosphodiester phosphodiesterase in serum samples from 5 Borrelia miyamotoi sensu lato–seropositive patients in the northeastern United States, 1991–2012. Numbers at the top of rows are patient numbers and correspond to patients 26–29 and 45 in (Table 1). The letters a and c that follow patient numbers indicate acute- and convalescent-phase serum samples, respectively. Western blot results that show no seroreacti
Figure 2. Western blot reactivity to recombinant Borrelia miyamotoi glycerophosphodiester phosphodiesterase in serum samples from 5 Borrelia miyamotoi sensu lato–seropositive patients in the northeastern United States, 1991–2012Numbers at the top of rows are...
A clinical description of illness was available for 5 symptomatic patients who experienced a ≥4 fold rise in B. miyamotoi sensu lato IgG and/or IgM antibody between acute- and convalescent-phase serum samples, as determined by ELISA and confirmatory Western blot assays (Western blot data shown in Figure 2). Of the 5 patients, 4 were co-infected with Lyme disease, 1 of whom was also co-infected with babesiosis (determined by blood smear). The 4 patients all had an erythema migrans skin lesion, and 2 had culture results positive for B. burgdorferi. The fifth patient had no evidence of co-infection and was the only 1 of 17 (5.9%) participants with a febrile summertime illness who had acute- and convalescent-phase serum tested for B. miyamotoi sensu lato antibody and who seroconverted. Three of these 5 patients have been reported previously (11). All 5 patients had fever, but a relapsing fever pattern was not reported. Symptoms resolved in 4 of the patients after treatment with doxycycline for 7–14 days, and symptoms resolved in the fifth patient after treatment with amoxicillin/clavulanic acid for 14 days.
We found evidence of human infection with the spirochete B. miyamotoi sensu lato in 52 residents residing in southern New England or New York State during 1991–2012. Among healthy study participants from southern New England, the seroprevalence of B. miyamotoi sensu lato infection was about one third that of B. burgdorferi infection (3.9% vs. 9.4%, respectively). This finding is consistent with the higher rate of B. burgdorferi infection in I. scapularis ticks in the region (range 2:1–20:1) (4–6). As expected, the seroprevalence of B. miyamotoi sensu lato infection was higher in serum samples from patients with acute Lyme disease and recent I. scapularis tick bites than in serum samples from patients whose tick-bite status was unclear. B. miyamotoi sensu lato seroprevalence rates were similar among study participants with a febrile late spring or summertime illness and healthy participants, probably because B. miyamotoi sensu lato infection is unlikely to be a common cause of nonspecific febrile illness in the late spring or summer. The seroprevalence of B. miyamotoi sensu lato was less than that of B. burgdorferi but similar to that of Babesia microti among residents of the same southern New England and New York region (16,26–28).
Approximately 10 percent of the B. miyamotoi sensu lato–seropositive patients without a recent history of Lyme disease reacted to B. burgdorferi antigen by 2-tier testing. The reactivity could have represented a prior B. burgdorferi infection, a false-positive test reaction, and/or cross-reactivity of B. miyamotoi sensu lato antibody against >1 B. burgdorferi antigens. The frequency of antibody reactivity to B. burgdorferi in patients with relapsing fever is shown in Table 2 (23–25,29). Several proteins are found in common between B. burgdorferi and B. miyamotoi sensu lato, including the flagellin FlaB protein, the GroEL heat shock proteins, and the BmpA (P39) protein (19,25). Misdiagnosis of B. miyamotoi sensu lato infection as Lyme disease is therefore possible. Results of B. burgdorferi testing may be positive for B. miyamotoi sensu lato–infected patients who are co-infected with B. burgdorferi (as was the case for some persons in this study). Our findings suggest, however, that testing for antibodies against B. burgdorferi is not an appropriate surrogate for testing for antibodies against B. miyamotoi sensu lato; B. burgdorferi antibody testing should not be used in place of an assay for antibody against B. miyamotoi sensu lato GlpQ or another B. miyamotoi sensu lato–specific antigen.
Our study had several limitations. First, laboratory evidence for acute B. miyamotoi sensu lato infection was based on ELISA and Western blot antibody assay rather than on culture, blood smear, or B. miyamotoi sensu lato PCR. However, in agreement with the case definition commonly used for many infectious diseases by the US Centers for Disease Control and Prevention (30), we considered results positive if a >4-fold rise in antibody occurred between acute- and convalescent-phase serum samples.
Second, B. miyamotoi sensu lato from North American has not been cultured, and blood smears were not available from the patients in our study. We were unable to detect B. miyamotoi sensu lato DNA in frozen, archived serum samples; however, the process of preparing serum from whole blood likely removed some spirochetes from the samples, and freeze–thaw cycles may have destroyed bacterial DNA. Furthermore, almost half of the serum samples that we tested were obtained after the period of acute illness, when the bacteremia may have cleared.
Third, our seroprevalence rates presumably would have been higher if we had tested for both IgM and IgG antibody by ELISA and included patients with IgM antibody alone as seropositive patients; however, we chose a more stringent definition of seropositivity by requiring the presence of IgG antibody. On the other hand, our seroprevalence data may have been inflated as a result of cross-reactivity of antibodies from other infections reacting against B. miyamotoi sensu lato GlpQ antigen. Although all other relapsing fever species have the glpQ gene, no other relapsing fever Borrelia sp. has been identified in I. scapularis ticks or humans in the northeastern United States (2,4–6,15,17,31).
Last, we do not have travel histories for the B. miyamotoi sensu lato–seroreactive patients included in the study, but the probability that many of our patients would have had exposure to other relapsing fever Borrelia spp. in the United States seems highly unlikely because these infections are infrequent and occur in the western states (2). Cross-reactivity against other tick-borne infections in the Northeast also appears unlikely because the agents of Lyme disease, human granulocytic anaplasmosis, and Powassan virus disease lack a glpQ gene (15). Proteins homologous to the GlpQ protein of relapsing fever borreliae are found in some gram-negative bacteria, including Escherichia coli, but they are so distant in sequence that antibody cross-reactivity is not expected (15).
The determination of B. miyamotoi sensu lato seroprevalence in our population is important because it indicates that this pathogen may infect persons at a rate that is similar to that of B. microti in the northeastern United States (16,23,24). Our data suggest that acute B. miyamotoi sensu lato infection in some persons may be misdiagnosed as Lyme disease because of the presence of antibody to B. burgdorferi from a previous B. burgdorferi infection, a false-positive test reaction, and/or cross-reactivity. Antibody testing for B. burgdorferi, however, is not adequate to detect infection with B. miyamotoi sensu lato in the United States. The potential for misdiagnosis may be greater in locations like northern California, were the prevalence of B. miyamotoi sensu lato in ticks equals or exceeds the prevalence of B. burgdorferi in ticks (32). Further studies are needed to better characterize the epidemiology and improve the serodiagnosis of human B. miyamotoi sensu lato infection.
Parasit Vectors. 2014 Sep 4;7(1):418. doi: 10.1186/1756-3305-7-418.
The relapsing fever spirochete Borrelia miyamotoi is cultivable in a modified Kelly-Pettenkofer medium, and is resistant to human complement.
Wagemakers A, Oei A, Fikrig MM, Miellet WR, Hovius JW1.
1Center for Experimental and Molecular Medicine, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. email@example.com.
Borrelia miyamotoi is a relapsing fever spirochete found in Ixodes ticks in North America, Europe, and Asia, and has recently been found to be invasive in humans. Cultivation of this spirochete has not yet been described, but is important for patient diagnostics and scientific purposes. Host specificity of Borrelia species is dependent on resistance to host complement (serum resistance), and since B. miyamotoi has been identified as a human pathogen we were interested whether B. miyamotoi is resistant to human complement.
We inoculated B. miyamotoi strains LB-2001 and HT31 in modified-Kelly-Pettenkofer medium with 10% fetal calf serum (MKP-F), and used standard non-laborious Borrelia culture methods to culture the spirochetes. Next, we assessed serum sensitivity by a direct killing assay and a growth inhibition assay.
We were able to passage B. miyamotoi over 10 times using a standard culture method in MKP-F medium, and found B. miyamotoi to be resistant to human complement. In contrast to B. miyamotoi, Borrelia anserina - a relapsing fever spirochete unrelated to human infection- was serum sensitive.
Using a variation on MKP medium we were able to culture B. miyamotoi, opening the door to in vitro research into this spirochete. In addition, we describe that B. miyamotoi is resistant to human complement, which might play an important role in pathogenesis. We have also found B. anserina to be sensitive to human complement, which might explain why it is not related to human infection. Summarizing, we describe a novel culture method for B. miyamotoi and show it is resistant to human complement.
[PubMed - in process]
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Suomen puutiaisilta löydetty uusi infektioita aiheuttava bakteeri
Suomessa elävien puutiaisten on havaittu kantavan toisintokuumeborrelioiden ryhmään kuuluvaa Borrelia miyamotoi -bakteeria. Bakteeria on havaittu aiemmin Ahvenanmaan puutiaisista, Turun yliopiston tutkimuksissa sitä tavattiin Turunmaan sisäsaaristosta. Tutkijoiden mukaan löytö ennakoi uuden puutiaisvälitteisen infektiosairauden esiintymistä myös Suomessa.
Saaristomerellä sijaitsevalla Seilin saarella toteutetussa tutkimuksessa puutiaisista löydettiin Borrelia afzelii, B. burgdorferi, B. garinii, B. valaisiana ja B. miyamotoi -bakteerilajit. Neljän ensiksi mainitun tiedetään aiheuttavan Lymen borrelioosia, mutta toisintokuumeborrelioiden ryhmään kuuluva B. miyamotoi voi aiheuttaa infektion, johon liittyy influenssan kaltainen korkeakuumeinen yleisinfektio ja keskushermoston tulehdus.
Suomesta ei ole vielä diagnosoitu potilaita, joilla olisi ollut juuri tämä puutiaisvälitteinen infektiosairaus.
– B. miyamotoi:n aiheuttamaan infektioon ei liity tyypillistä Lymen borrelioosin ensioiretta eli rengasihottumaa (Erythema migrans), mikä hankaloittaa tämän infektion tunnistamista. Venäjältä ja muutamasta muusta maasta näitä tapauksia on jo diagnosoitu, joten potilaita voi löytyä Suomestakin, puutiaistutkimuksessa mukana ollut tutkijalääkäri, dosentti Jukka Hytönen sanoo.
B. miyamotoi -bakteerin tiedetään siirtyvän aikuisista naaraspuutiaisista transovariaalisesti eli munien kautta toukkavaiheen puutiaisiin. Lymen borrelioosia aiheuttavat bakteerit eivät nykytietämyksen mukaan siirry tätä reittiä, joten puutiaisten toukkamuodot eivät levitä Lymen borrelioosia.
– B. miyamotoi:n esiintymistä on nyt ryhdytty tutkimaan ja sitä on viime vuosina löydetty useasta Euroopan maasta. On käytännössä varmaa, että tätä bakteeria löytyy Suomessa muualtakin kuin Seilin saarelta. Löytöjä tehtäneen sekä tavallisista puutiaisista että taigapunkeista. Nämä asiat selviävät tulevissa tutkimuksissamme, tutkimuksen kenttätyöt tehnyt ja puutiaisten ekologiasta väitöskirjaansa valmisteleva tohtorikoulutettava Jani Sormunen sanoo.
B. miyamotoi voi oleellisesti muuttaa Borrelia-bakteerien aiheuttamien infektioiden luonnetta tulevaisuudessa, joten sen esiintymisen kartoittaminen ja seuraaminen on nyt tärkeää. Lääkäreillä tulisi olla valmius tunnistaa B. miyamotoi -infektio mahdollisena puutiaisvälitteisenä sairautena. Laboratoriodiagnostiikka tämän infektion tunnistamiseksi on kehitteillä.
Joka neljäs aikuinen kantaa Borrelia-bakteeria
Tutkimus paljasti havumetsän puutiaisen (Ixodes ricinus) suosituimmaksi elinympäristöksi.
– Vastoin vanhoja uskomuksia puutiaisia esiintyi avoimilla alueilla, kuten niityillä ja laitumilla, vähemmän kuin havu- tai sekametsässä. Puutiaisten viihtyminen erityisesti mustikkatyypin havumetsässä on huono uutinen esimerkiksi marjastajille ja sienestäjille, Sormunen sanoo.
Kaikki tutkimuksessa vuonna 2012 tai sen jälkeen Saaristomereltä pyydetyt puutiaiset ovat olleet ns. tavallisia puutiaisia. Siperian puutiaisia eli taigapunkkeja (Ixodes persulcatus) ei ole vielä tavattu Varsinais-Suomessa.
– Puutiaisia pyydettiin Seilissä viikoittain toukokuusta syyskuuhun. Yhteensä saaliiksi tuli noin 2000 puutiaista. Laboratoriossa niistä määritettiin taudinaiheuttajia kuten Borrelia-suvun bakteereita ja puutiaisvälitteistä aivokuumetta aiheuttavaa TBE-virusta. Virusta ei näistä näytteistä kuitenkaan löytynyt, Sormunen kertaa tutkimuksen vaiheita.
Borrelia-bakteerin esiintyvyys (eli prevalenssi) oli aikuisvaiheen puutiaisilla noin 25 prosenttia ja nymfeillä eli esiaikuisilla noin 15 prosenttia. Puutiaisten toukista bakteereita ei löytynyt. Nämä luvut vastaavat muiden eurooppalaisten tutkimusten tuloksia. Yleisesti voidaan todeta, että noin joka neljäs aikuisvaiheen puutiainen kantaa Borrelia-bakteereita.
– Borrelia-bakteerien kirjo on moninainen. Eri bakteerikantojen yhteys Lymen borrelioosin eri oireisiin ja ilmenemismuotoihin on huonosti tunnettua, Hytönen sanoo.
Tutkimus on julkaistu kansainvälisessä Ticks and Tick-borne Diseases –julkaisusarjassa (doi:10.1016/j.ttbdis.2015.10.011 )
http://www.ncbi.nlm.nih.gov/pubmed/2654 ... t=Abstract
Ticks Tick Borne Dis. 2016 Feb;7(1):208-15. doi: 10.1016/j.ttbdis.2015.10.011. Epub 2015 Oct 24.
Assessing the abundance, seasonal questing activity, and Borrelia and tick-borne encephalitis virus (TBEV) prevalence of Ixodes ricinus ticks in a Lyme borreliosis endemic area in Southwest Finland.
Sormunen JJ1, Klemola T2, Vesterinen EJ3, Vuorinen I4, Hytönen J5, Hänninen J4, Ruohomäki K2, Sääksjärvi IE6, Tonteri E7, Penttinen R6.
Studies have revealed that Ixodes ricinus (Acari: Ixodidae) have become more abundant and their geographical distribution extended northwards in some Nordic countries during the past few decades. However, ecological data of tick populations in Finland are sparse. In the current study, I. ricinus abundance, seasonal questing activity, and their Borrelia spp. and tick-borne encephalitis virus (TBEV) prevalence were evaluated in a Lyme borreliosis endemic area in Southwest Finland, Seili Island, where a previous study mapping tick densities was conducted 12 years earlier. A total of 1940 ticks were collected from five different biotopes by cloth dragging during May-September 2012. The overall tick density observed was 5.2 ticks/100m(2) for nymphs and adults. Seasonal questing activity of ticks differed between biotopes and life stages: bimodal occurrences were observed especially for nymphal and adult ticks in forested biotopes, while larvae in pastures exhibited mostly unimodal occurrence. Prevalence of Borrelia and TBEV in ticks was evaluated using conventional and real-time PCR. All samples were negative for TBEV. Borrelia prevalence was 25.0% for adults (n=44) and the minimum infection rate (MIR) 5.6% for pooled nymph samples (191 samples, 1-14 individuals per sample; 30/191 positive). No Borrelia were detected in pooled larval samples (63 samples, 1-139 individuals per sample). Five species of Borrelia were identified from the samples: B. afzelii, B. burgdorferi s.s., B. garinii, B. valaisiana and B. miyamotoi. In Finland, B. valaisiana and B. miyamotoi have previously been reported from the Åland Islands but not from the mainland or inner archipelago. The results of the present study suggest an increase in I. ricinus abundance on the island.
Copyright © 2015 Elsevier GmbH. All rights reserved.
PMID: 26548608 [PubMed - in process]
Lyme borreliosis (LB) on tick-välitteinen tarttuva tauti, jonka aiheuttaa Borrelia burgdorferi sensu lato (sl). Euroopassa kolme eri Borrelia- lajia ovat LB: n tärkeimmät aiheuttajat: B. burgdorferi sensu stricto (ss), Borrelia afzelii ja Borrelia garinii . Jälkimmäinen riippuu voimakkaasti linnuista, jotka ovat sen tärkeimmät säiliö isännät. Itse asiassa linnut voivat toimia sekä Borrelian biologisina kantajina että tartunnan saaneiden punkkien kuljettajina. Monien lintulajien kausiluonteinen muuttoliike ei ainoastaan auta B. gariniin leviämiseen uusiin fokaaleihin, vaan vaikuttaa myös tämän lajin monimuotoisuuteen. B. garinii on eristetty paitsi maanpäällisistä linnuista Euroopassa myös merilinnuista maailmanlaajuisesti, ja näiden kahden eri infektiokerroksen isolaattien homologia viittaa päällekkäisyyksien ja kantojen vaihtoon. Lisäksi on osoitettu, että linnut voivat ylläpitää ja levittää B. gariniin genotyyppejä, jotka liittyvät LB: hen ihmisillä. Tässä tarkastelussa käsitellään lintujen merkitystä B. garinii spirochetesin ekologiassa ja epidemiologiassa .
Avainsanat: Borrelia garinii , Lyme borreliosis, linnut, säiliöisännät, punkit, Ixodes uriae , Ixodes ricinus