Animals ; China ; epidemiology ; Epidemiologic Studies ; Molecular Epidemiology ; Rickettsia ; genetics ; isolation & purification ; Rickettsia Infections ; epidemiology ; transmission ; Ticks ; microbiology

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

OBJECTIVE: To investigate the infection in spotted fever group Rickettsiae (SFGR) in wild rodents from Heilongjiang, China. METHODS: Polymerase chain reaction (PCR) was used to detect the OmpA gene of SFGR in rodents collected in Heilongjiang. The PCR products amplified from rodent specimens were sequenced and compared with the corresponding part of the sequences deposited in the GenBank. Phylogenetic trees were constructed with Mega 5.0 software. RESULTS: A total of 514 rodents were collected from Heilongjiang during 2009-2011 and 11 species were included. The infection rate of SFGR in the rodents was 9.3% (95% CI: 7.1%-12.2%). Statistical analysis showed a significant difference in different areas of Heilongjiang (P=0.023). The highest prevalence was observed in Mudanjing area (12.42%). There were significant differences in different species of rodents (P=0.002). The infection rate of SFGR determined in Clethrionomys rufocanus was the highest (22.1%). Sequence analysis revealed SFGR belonged to R.heilongjiangensis and a new unknown rickettsia genotype. CONCLUSION R.heilongjiangensis has been presented in rodents in Heilongjiang, and a new SFGR genotype different from other rickettsiae genotypes may exist in this area.
Animals ; China ; DNA, Bacterial ; genetics ; isolation & purification ; Forests ; Phylogeny ; Polymerase Chain Reaction ; Rats ; Rickettsia ; classification ; genetics ; isolation & purification ; Rickettsia Infections ; microbiology ; veterinary ; Rodentia ; microbiology ; Sequence Analysis

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Rickettsia prowazekii.

METHODSPrimers and TaqMan-MGB probes designed based on ompB gene of R. prowazekii, were used to develop this method.

RESULTSFor the quantitative real-time PCR, the relationship between the values of threshold cycle (Ct) and the DNA copy number was linear (r = 0.999) and the sensitivity was about 100 times higher than that of the nested PCR for detecting the same DNA sample. The results of the genomic DNA samples of other rickettsial and bacterial agents detected by real-time PCR were all negative. DNAs extracted from blood samples of guinea pig infected with R. prowazekii were examined by real-time PCR and the positive results were obtained from some of these samples. However, the results of some samples in nested PCR assay were all negative.

CONCLUSIONThese results suggested that the real-time PCR was highly specific and sensitive for detection of R. prowazekii that was useful for the detection of tiny DNA of R. prowazekii in blood samples from patients suspected of having epidemic typhus.

DNA Primers ; DNA, Bacterial ; analysis ; Humans ; Polymerase Chain Reaction ; methods ; Rickettsia prowazekii ; genetics ; isolation & purification ; Sensitivity and Specificity ; Typhus, Epidemic Louse-Borne ; diagnosis

OBJECTIVETo study the existence of natural foci of Spotted Fever in Ninghua, Fujian province.

METHODSUsing DNA polymerase chain reaction and restriction endonuclease fragment length polymorphism analysis (PCR/RFLP) to detect spotted fever group rickettsiae (SFGR) in ticks and rodents.

RESULTSIt was found that H. wellingtoni, H. yeni, and Dermacentor auratus were infected with Rickettsia sibirica; the DNA fragments were cloned, the PCR products from isolated strain NH-97 were antigenically and genotypically identical to Rickettsia sibirica. Rattus flavipectus were found infected with R. conorii. One of the sequeuce analysis showed that the DNA sequence was different from other SFGR and close to R. japanic.

CONCLUSIONNatural foci of R. sibirica, R. sibrica, R. japanic and R.conorii are found in Ninghua, Fujian province of China.

Animals ; Boutonneuse Fever ; microbiology ; China ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Rats ; Rickettsia ; genetics ; isolation & purification ; Rodentia ; microbiology ; Ticks ; microbiology

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 understand the epidemic status of Rickettsia in Xinyang areas of Henan province.

METHODSSamples including liver, spleen, kidney from mouse and chigger mites from Xinyang areas and serum samples were detected by nested-polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA).

RESULTSIn 62 viscus samples from mice organs, the positive rates were 16.13%, 8.06% and 6.45% for Orientia tsutsugamushi, R. typhii and Spotted fever group rickettsiae respectively. In blood clots samples from mice, the positive rates were 8.06%, 6.45% and 1.61 % for O. tsutsugamushi, R. typhii and Spotted fever group rickettsiae respectively. Three out of 26 mouse serum samples were positive for the predicted fluorexcent intensity O. tsutsugamushi.

CONCLUSIONUsing nested-PCR and IFA methods, O. tsutsugamushi, R. typhii and Spotted fever group rickettsiae were detected in the captured mice living in Xinyang areas of Henan province. Results showed that there were intensive natural reserviors of Rickettsia in Henan province, suggesting that the risk of outbreak of Rickettsia in these areas was high.

Animals ; China ; Fluorescent Antibody Technique, Indirect ; Humans ; Kidney ; microbiology ; Liver ; microbiology ; Mice ; Orientia tsutsugamushi ; classification ; genetics ; isolation & purification ; pathogenicity ; Phylogeny ; Polymerase Chain Reaction ; Rickettsia ; classification ; genetics ; isolation & purification ; pathogenicity ; Scrub Typhus ; epidemiology ; microbiology ; Spleen ; microbiology

OBJECTIVETo explore the existence of spotted fever group Rickettsiae (SFGR) in Guangdong province.

METHODSSera were tested to find the SFGR in population and host animals. The target samples were screened by polymerase chain reaction (PCR), and Rickettsiae was isolated with embryonated hen eggs and identified by serological tests.

RESULTSEight hundred and sixty people in natural condition and 321 of mice were determined. The mean positive rate of healthy population was 3.84%. To compare results among elected places, Fisher's exact test was applied. The difference was suggestive (P < 0.01), and there was no significant difference between mountain and plain areas. There was also no significant difference between mountain and plain areas (P > 0.05). Positive rate of mice was 4.67%, with Rattus fulvescens, Rattus edwardsi, Bandicota indica 11.59%, 12.90%, 3.13% respectively. It was the first time that SFGR antibodies in Rattus fulvescens, Rattus edwardsi, Bandicota indica were reported. A total number of 321 mice spleens and 394 ticks from the surface of mice body were collected. Two strains of SFGR, GDFK58-2000 and GDFK59-2000, were isolated in the ticks from the body surface of 2 Rattus fulvescens. They were identified as Rickettsia sibirica by serological tests. Five hundred thirty-three bp OmpA gene fragments of the two strains were cloned and sequenced. Compared with other relevant strains in Genbank, the rates of homology of nucleotide sequences of GDFK58-2000 and GDFK59-2000 and other Rickettsia sibirica strains were from 99.6% to 100%, and the homology of amino acid speculated was 100%.

CONCLUSIONIt has been proved that epidemic areas of north Asia tick-transmitted SFGR, did exist in Guangdong province confirmed by hostanimals, transmission vectors and aetiology.

Adolescent ; Adult ; Aged ; Animals ; Child ; China ; epidemiology ; Disease Reservoirs ; Female ; Humans ; Male ; Mice ; Middle Aged ; Rats ; Rickettsia rickettsii ; classification ; genetics ; isolation & purification ; Rocky Mountain Spotted Fever ; epidemiology ; microbiology ; Rodentia ; microbiology ; Sequence Homology, Amino Acid ; Ticks ; microbiology

The objective of this study was to investigate the prevalence of tick-borne pathogens in the Korean water deer (Hydropotes inermis argyropus). Pathogens were identified using PCR which included Anaplasma, Ehrlichia, Rickettsia, and Theileria. Rickettsia was not detected, whereas Anaplasma, Ehrlichia, and Theileria infections were detected in 4, 2, and 8 animals, respectively. The most prevalent pathogen was Theileria. Of the 8 Theileria-positive animals, 2 were mixed-infected with 3 pathogens (Anaplasma, Ehrlichia, and Theileria) and another 2 animals showed mixed-infection with 2 pathogens (Anaplasma and Theileria). Sequencing analysis was used to verify the PCR results. The pathogens found in this study were identified as Anaplasma phagocytophilum, Ehrlichia canis, and Theileria sp. To the best of our knowledge, this is the first report identifying these 3 pathogens in the Korean water deer. Our results suggest that the Korean water deer may serve as a major reservoir for these tick-borne pathogens, leading to spread of tick-borne diseases to domestic animals, livestock, and humans. Further studies are needed to investigate their roles in this respect.
Anaplasma/isolation & purification ; Animals ; Bacterial Infections/epidemiology/microbiology/*veterinary ; Cluster Analysis ; Coinfection/epidemiology/microbiology/veterinary ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Ehrlichia/*isolation & purification ; Korea/epidemiology ; Molecular Sequence Data ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*isolation & purification ; Ruminants/*microbiology ; Sequence Analysis, DNA ; Theileria/*isolation & purification