A population-based case-control study was conducted to evaluate the relative factors in the environments, agricultural works, outdoor activities, and the effectiveness of Lyme borreliosis (LB)- associated personal protective measures in Beijing. Thirty-four cases and 272 controls were personally interviewed by well-trained interviewers. Venous blood samples were taken from each subject. Sowing or harvesting in summer (OR=2.571, 95% CI: 1.109-5.962), living in house with weeding in the yard (OR=2.247, 95% CI: 1.062-4.755), and residence at the plain area (OR=2.630, 95% CI: 1.050-6.588) were the independent relative factors for seropositive LB. Wearing long pants and clothes with cuffs was the only protective behavior against tick bite (OR=0.186, 95% CI: 0.041-0.846). The findings showed that local farmers were easily infected with LB and almost no protective measure was taken against LB infection. Infection with LB was easier in residents of plain regions. Pets raising and outdoor activities were not the risk factors for infection with LB. Further studies are needed to fully understand the risk of infection with LB in China.
Adult ; Aged ; Agriculture ; Case-Control Studies ; China ; epidemiology ; Cities ; Environment ; Female ; Human Activities ; Humans ; Lyme Disease ; epidemiology ; microbiology ; prevention & control ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure.

METHODSFP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and immunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples (159 patients with Lyme disease and 292 controls), all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index.

RESULTSCriteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.

CONCLUSIONEstablishment of WB criteria for B. afzelii is important in validating the diagnostic assays for Lyme disease in China.

Blotting, Western ; methods ; Borrelia burgdorferi Group ; pathogenicity ; China ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Humans ; Lyme Disease ; diagnosis ; microbiology

Severe tick infestation was found in a hare in a suburban area of Nanchang, Jiangxi Province, China. We sampled ticks and identified them based on their morphologic characteristics. Three species, Ixodes sinensis, which is commonly found in China and can experimentally transmit Borrelia burgdorferi, Rhipicephalus haemaphysaloides, and Haemaphysalis longicornis which can transmit Lyme disease were detected with an optical microscope and a stereomicroscope. Risk of spreading ticks from suburban to urban areas exists due to human transportation and travel between the infested and non-infested areas around Nanchang.
Animals ; Arachnid Vectors/*classification/microbiology ; Borrelia burgdorferi/physiology ; China ; Fatal Outcome ; Female ; Hares/*parasitology ; Humans ; Ixodidae/*classification/microbiology ; Lyme Disease/microbiology/*transmission ; Male ; Risk ; Tick Infestations/parasitology/transmission/*veterinary

Animals ; Borrelia burgdorferi Group ; classification ; genetics ; isolation & purification ; China ; Genotype ; Lyme Disease ; microbiology ; transmission ; Rats ; Ticks ; microbiology

OBJECTIVETo explore the fact that the east border of Heilongjiang had been a lyme disease natural focus,we investigated the species and distribution of ticks and isolated bacteria from ticks and identified genomic species of Borrelia burdorferi sensu lato. This study provided evidence for prevention and control of lyme disease.

METHODSTicks were caught by flagging method and Direct immunofluorescence method was used to detect the rate of bacteria borne by the tick. BSK UI culture medium was used to isolate the agent and Specific McAbs were used to identify the bacteria. SDS-PAGE protein profile and PCR-RFLP method were also used to identify the species of Spirochetes.

RESULTSTicks, collected from China-Russia border of east Heilongiiang province were classified including Ixodes persulcatus Schulze, Dermacentor sivarum Olener, Haemaphysalis concinna Kock,and Haemaphysalis japonica Kock. We found that the distributon of ticks was different under different circumstances and the predominant species were also different in different ports. The rate of bacteria borne by Iodes persulaatus Schulze was 31.4% ,by Dermacentor sivarum Olener and Haemaphysalis concinna Kock were 2.2% and 3.8%, respectively. However,it was negative for Haenaphysalis japonica Kock. Spirochetes isolated from Ixodes persulcatus Schulze were collected from Dongning and Tongjiang while Genomic species of Spirochetes, isolated from ticks of the border belonged to B. garinii.

CONCLUSIONAll the results showed that the east border of Heilongjiang province was the natural focus of lyme disease.

Animals ; Arachnid Vectors ; classification ; microbiology ; Borrelia burgdorferi ; classification ; genetics ; isolation & purification ; China ; Humans ; Lyme Disease ; microbiology ; Russia ; Ticks ; classification ; microbiology

OBJECTIVETo detect and study the types of Borrelia burgdorferi sensu lato in ticks and rodents from Da Xing-An Mountains Forest areas of China.

METHODSNested PCR was performed to amplify 5S-23S rRNA intergenic spacer of B. burgdorferi. Positive products were analysed by restriction fragment length polymorphism (RFLP) and single strand conformation polymorphism (SSCP), specimens showing unique RFLP profile were sequenced and analysed.

RESULTS1336 Ixodes persulcatus, 144 Dermacento silvarum, 144 Haemaphysalis concinna and 145 rodents of 9 species were collected from 16 sections of Da Xing-An Mountains Forest areas of China. Specific fragments were amplified from 293 I. persulcatus and 6 D. silvarum and 5 rodents of 4 species. B. burgdorferi was not detected in H. concinna. Among the positively tested I. persulcatus, 209 contained B. garinii genospecies and 45 contained B.afzelii genospecies based on RFLP. Moreover, B.garinii genospecies consisted of B. garinii 20047 and B. garinii NT29. 17 adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29. Nine adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. afzelii. Four adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29 and B. afzelii. Two D. silvarum were infected with B. garinii 20047, 1 D. silvarum with B. garinii 20047, 2 D. silvarum with B. afzelii. 3 rodents were infected with B. garinii 20047 while 2 rodents were infected with B. garinii NT29. Mixed infection was not found in D. silvarum and rodents. In addition, nine I. persulcatus and one D. silvarum specimens showed unique RFLP pattern. Data from sequential analysis showed that they all belonged to B. garinii. PCR-SSCP profiles of 5S-23S rRNA intergenic spacer of B. burgdorferi in the positive specimens exceeded 36 types; B. garinii 20047 showed 16 types while B. garinii NT29 showing 11 types, B. afzelii showing 9 types. SSCP profiles of the specimens coinfected with multiple B. burgdorferi was relatively complex.

CONCLUSIONThe infection of B. burgdorferi was found in the ticks and rodents in Da Xing-An Mountains Forests areas. The infection rate of I. persulcatus was high. B. garinii was predominant genospecies, and the population of B. burgdorferi was heterogeneous in the area. Mixed infections of different B. burgdorferi genospecies in ticks were found. I. persulcatus and Clethrionomys rufocanus were possibly served as major vector and major host for B. burgdorferi, respectively, suggesting that further study is needed to confirm the coinfection in humans and animals in this region.

Animals ; Borrelia burgdorferi Group ; genetics ; isolation & purification ; China ; epidemiology ; Humans ; Lyme Disease ; epidemiology ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; RNA, Bacterial ; analysis ; Rodentia ; microbiology ; Ticks ; microbiology ; Trees

OBJECTIVETo understand the coinfection status of Borrelia burgdorferi sensu lato (B.b.s.l) and spotted fever group Rickettsia (SFGR) in Hunchun of Jilin province, China.

METHODSPolymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B. b. s. l and ompA of SFGR in ticks was collected in Hunchun,Jilin province. The amplification products of positive ticks were sequenced, and phylogenetic analysis was conducted by PHYLIP software package.

RESULTSThe infection rate of B. b. s. l was 36.0% in Ixodes persulcatus ticks and the SFGR was discovered in I. persulcatus ticks,with an infection rate of 2.0%. The coinfection rate of both agents was 2.0%. In 327 Dermacentor siltarum ticks, the positive rates of B. b. s. l and SFGR were 30.9% and 29.1% respectively. 55 ticks (16.8%) were coinfected with the two pathogens. The sequence analysis of B. b. s. l showed that the B. b. s. l in Jilin area, which were highly homologous, all belonged to B. garinii genotypes. The sequence analysis of SFGR positive products showed that the DNA secquence of the newly detected agent (JL-95) was close to the two previously described rickettsiae which were detected in I. ricinus from Slovakia (called IRS3 and IRS4). Phylogenetic relationships inferred from the comparison of these sequences with those of other genus Rickettsiae indicated that JL-95, IRS3 and IRS4 constituted a new rickettsial genotype and formed a separate cluster among the spotted fever group Rickettsiae.

CONCLUSIONCoinfection of B. b. s. l and SFGR existed in Hunchun, Jilin province. The sequencing of specific fragment confirmed a new SFGR which was different from other rickettsiae known in China.

Animals ; Borrelia burgdorferi Group ; genetics ; isolation & purification ; China ; DNA, Bacterial ; analysis ; Genotype ; Lyme Disease ; veterinary ; Phylogeny ; Polymerase Chain Reaction ; Rickettsia ; genetics ; isolation & purification ; Rickettsia Infections ; veterinary ; Ticks ; microbiology

Antimicrobial susceptibility testing was conducted with 6 different spirochetal strains (4 strains of Leptospira spp. and 2 strains of Borrelia burgdorferi) against 3 antimicrobial agents, commonly used in equine and bovine practice. The ranges of MIC and MBC of amoxicillin against Leptospira spp. were 0.05 - 6.25 microgram/ml and 6.25 - 25.0 microgram/ml, respectively. And the ranges of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of amoxicillin against B. burgdorferi were 0.05 - 0.39 microgram/ml and 0.20 - 0.78 microgram/ml, respectively. The ranges of MIC and MBC of enrofloxacin against Leptospira spp. were 0.05 - 0.39 microgram/ml and 0.05 - 0.39 microgram/ml, respectively. Two strains of B. burgdorferi were resistant to enrofloxacin at the highest concentration tested for MBC (> or = 100 microgram/ml). Therefore, the potential role of tilmicosin in the treatment of leptospirosis and borreliosis should be further evaluated in animal models to understand whether the in vivo studies will confirm in vitro results. All spirochetal isolates were inhibited (MIC) and were killed (MBC) by tilmicosin at concentrations below the limit of testing (< or = 0.01 microgram/ml).
Amoxicillin/pharmacology ; Anti-Bacterial Agents/*pharmacology ; Borrelia burgdorferi/*drug effects/growth & development/isolation & purification ; Fluoroquinolones/pharmacology ; Leptospira/*drug effects/growth & development/isolation & purification ; Leptospirosis/*microbiology ; Lyme Disease/*microbiology ; Macrolides/pharmacology ; Microbial Sensitivity Tests ; Reproducibility of Results ; Tylosin/analogs & derivatives/pharmacology

OBJECTIVETo study the existence of Ehrluichiosis, lyme disease and tick-borne spotted fever coinfection in some areas in China.

METHODSUsing polymerase chain reaction (PCR), B. burgdorferi sensu lato, spotted fever group (SFG) Rickettsiae and human granulocytic ehrlichia (HGE), Ehrlichia chaffeensis (EC) were detected in ticks and mouse samples collected from Inner Mogolia autonomous region, Heilongjiang province, Beijing and Fujian province.

RESULTS408 Ixodes persulcatus collected from Inner Mogolia autonomous region, HGE and B. burgdorferi sensu lato and SFG Rickettsiae were detected positive, with rates as 6.8%, 7.8%, 45.6%, respectively. 5 (5/408) were coinfection with HGE and B. burgdorferi sensu lato while 1 (1/408) was coinfection with HGE and SFG Rickettsiae. 46 Ixodes persulcatus collected from Helongjiang province were determined positive, with rates as 6.5%, 10.8% and 34.8%, respectively including 1 (1/46) coinfected with HGE and B. burgdorferi sensu lato. 2 of 922 ticks collected from Beijing were detected positive with B. burgdorferi sensu lato. Among 283 groups of Haemaphysalis yeni ticks (3/group) and from 38 rodent samples collected from Ninghua county of Fujian province HCE and B. burgdorferi sensu lato and SFG Rickettsiae were detected. Out of them, 25 groups were positive with EC and the minimal positive rate was 3.8% while 21 rodent samples were positive with EC with a positive rate of 56.4%. 2 ticks and 1 rodent sample were detected positive with EC and spotted fever group.

CONCLUSIONCoinfection of HGE and B. burgdorferi sensu lato or spotted fever group Richi did exist in Ixodes persulcatus collected from Inner Mogolia autonomous region and Heilongjiang province. Coinfection of EC and spotted fever group Richi was found in the ticks and rodents collected from Fujian province.

Animals ; Arachnid Vectors ; Borrelia burgdorferi Group ; isolation & purification ; China ; epidemiology ; DNA, Bacterial ; analysis ; Disease Vectors ; Ehrlichia ; isolation & purification ; Ehrlichiosis ; epidemiology ; Humans ; Ixodes ; microbiology ; Lyme Disease ; epidemiology ; Polymerase Chain Reaction ; Rats ; Rickettsia ; isolation & purification ; Rickettsia Infections ; epidemiology ; Rodentia ; microbiology ; Tick-Borne Diseases ; epidemiology ; Ticks ; microbiology

OBJECTIVETo study the cloning and expression of flagellin gene from Chinese Borrelia burgdorferi, PD91 strain and to evaluate the feasibility of using recombinant protein as diagnostic antigen when comparing the gene sequence with flagellin gene from North American Borrelia burgdorferi B31.

METHODSThe piece of genes coding flagellin from Chinese Borrelia burgdorferi PD91 by polymerase chain reaction (PCR) method was obtained, and constructed recombinant plasmid, before transformed into E. coli BL21 strain, and induced. The recombinant plasmid was identified with enzyme cutoff and gene sequence comparison. Efficient expression strain was selected and the expression product was analyzed with sodium amplified polymorphic-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot method.

RESULTSThe recombinant protein (r-flagellin) expressed in host bacteria was successful. By means of western-blot assay, the immunological response showed the same antigenicity between r-flagellin and PD91 flagellin. The piece of genes coding flagellin of PD91 was 1011 bp, but when comparing with that of North American Borrelia burgdorferi it showed 94.70% homology. Homology between the sequence of amino acid of the r-flagellin and that of B31 flagellin was 95.85%.

CONCLUSIONFlagellin gene of Borrelia garinii of Chinese Lyme disease spirochete was successfully cloned and expressed for the first time. It was proved that the immunoreactivity of r-flagellin was the same as the natural flagellin.

Amino Acid Sequence ; Base Sequence ; Borrelia burgdorferi ; genetics ; isolation & purification ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Flagellin ; biosynthesis ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics