OBJECTIVETo investigate rodents' natural infection of Orientia tsutsugamushi (Ot) in some areas of Inner Mongolia and Xinjiang, China.

METHODSDNAs were extracted from spleens of the captured mice and nested-polymerase chain reaction (nPCR) technique was used to detect the Ot-Sta56 gene. Six positive samples were sequenced and analyzed by Clustal X (5.0) and DNA Club software.

RESULTSA total of 90 rodents were captured in Inner Mongolia, and the overall prevalence of Ot was 6.67%. There was no significant difference in infection rates among the positive rodents species. 20 rodents were captured in Xinjiang, and the prevalence of Ot was 5.00%. The geographical difference in infection rates was not statistically significant between Inner Mongolia and Xinjiang. 9 rodents were captured in farmlands of Inner Mongolia and Xinjiang but there was no positive samples found. 101 rodents were captured in grasslands, and the prevalence of Ot was 6.93%. The Sta56 gene nucleotide sequence homology to Karp strain of N59 (from Microtus maximowiczii), N69 (from Cricetulus barabensis) and X33(from Cricetus cricetus) was 99%. The sequence homology to Taitung-2 strain and TW461 strain of N65 (from C. barabensis) was 94%, and the sequence homology to Taitung-2 strain and TW461 strain of N88(from Apodemus agrarius) was also 94%. The sequence homology to Oishi strain of N90 (from A. agrarius) was 96.00%.

CONCLUSIONOur findings indicated that infections of Ot did exist in rodents captured from Inner Mongolia and Xinjiang. The genotypes of Ot in Inner Mongolia and Xinjiang were quite complex, with some of them belonged to Karp type, and the others belonged to Taitung-2, TW461 and Oishi types which providing evidence for further investigation on the scrub typhus fuci in the two areas.

Animals ; China ; Geography ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Rodentia ; microbiology ; Scrub Typhus

A total of 9,281 larval chigger mites were collected from small mammals captured at Hwaseong-gun, Gyeonggi-do (Province) (2,754 mites from 30 small mammals), Asan city, Chungcheongnam-do (3,358 mites from 48 mammals), and Jangseong-gun, Jeollanam-do (3,169 for 62 mammals) from April-November 2009 in the Republic of Korea (= Korea) and were identified to species. Leptotrombidium pallidum was the predominant species in Hwaseong (95.8%) and Asan (61.2%), while Leptotrombidium scutellare was the predominant species collected from Jangseong (80.1%). Overall, larval chigger mite indices decreased from April (27.3) to June (4.9), then increased in September (95.2) and to a high level in November (169.3). These data suggest that L. pallidum and L. scutellare are the primary vectors of scrub typhus throughout their range in Korea. While other species of larval chigger mites were also collected with some implications in the transmission of Orientia tsutsugamushi, they only accounted for 11.2% of all larval chigger mites collected from small mammals.
Animals ; Arachnid Vectors ; Larva/*microbiology ; Orientia tsutsugamushi/*isolation & purification ; Republic of Korea ; Rodentia ; Scrub Typhus/*microbiology ; Trombiculidae/*classification/*microbiology

OBJECTIVETo determine genotype, nucleotide sequence homology and phylogenesis of Orientia tsutsugamushi isolated from Shandong, China.

METHODSOrientia tsutsugamushi isolated from patients, Apodemus agrarius and Leptotrombidium scutellare in Shandong area were identified by nested-PCR. On the basis of the nucleotide sequence of the gene that encoding the Ot M, 56 x 10(3) antigen, the primers were frequently used in Japan and Korea. Nucleotide sequences of three isolates were determined. The DNA sequences were compared with nucleotide sequences of Orientia tsutsugamushi registered in GenBank for sequence homology analysis. Phylogenetic analysis of the isolates and'some published sequences was carried out with Neighbor-joining method by MEGA 3.1 software.

RESULTS481- 507 bp DNA fragments encoding Orientia tsutsugamushi M, 56 x 10(3) protein were amplified successfully in the samples of Gilliam, Karp, Kato and Shandong isolates by group-specific primers. The corresponding target fragments of the three international reference strains of Gilliam, Karp, and Kato were amplified successfully with each of their own type specific primers. 523 bp DNA fragments were amplified successfully from Shandong isolates by the nPCR with Kawasaki-specific primer, and no DNA fragment was amplified by the nPCR with Gilliam, Karp, Kato, Kuroki and Saitama-specific primer. Comparing with the sequences of Orientia tsutsugamushi registered in GenBank, all the Shandong isolates shared higher than 95% nucleotide sequence homology with Kawasaki strain founded in Japan. Data from phylogenetic analysis showed that Shandong isolates belonged to the same branch with Kawasaki strain.

CONCLUSIONTo facilitate international comparison and communication, the primers should be employed in the Orientia tsutsugamushi research in China. Orientia tsutsugamushi isolated in China were similar to Kawasaki strain

Base Sequence ; China ; Genotype ; Humans ; Molecular Sequence Data ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA

A survey to determine the geographical distribution and relative abundance of potential vectors of scrub typhus was conducted from October to November 2006 at 13 localities throughout the Republic of Korea. Apodemus agrarius accounted for 97.6% (80/82) of all rodents, while only 2 Myodes regulus (2/82) were collected. A total of 10,860 chiggers were collected from A. agrarius belonging to 4 genera and 8 species, while only Walchia fragilis (40) was collected from Myodes regulus. Leptotrombidium pallidum (8,137; 74.9%), a vector of scrub typhus, was the predominant species collected from A. agrarius followed by Leptotrombidium scutellare (2,057, 18.9%), Leptotrombidium palpale (279; 2.7%), Leptotrombidium orientale (232; 2.1%), and Leptotrombidium zetum (79; 0.7%), Neotrombicula tamiyai (58; 0.5%), Euschoengastica koreaensis (16; 0.1%), and Cheladonta ikaoensis (2; < 0.1%). L. pallidum was the predominant chigger collected at collection sites in Gangwon (100%), Gyeonggi (87.2%), Chungnam (100%), Chungbuk (100%), Jeonbuk (73.9%), Jeonnam (77.0%), and Gyeongbuk (66.1%) provinces, whereas L. scutellare was the predominant chigger collected in Gyeongnam province (77.9%) and Jeju Island (62.3%). Data suggest a correlation between chigger population abundance and human cases of scrub typhus in Korea.
Animals ; Arvicolinae/*parasitology ; *Disease Vectors ; Geography ; Mites/*microbiology ; Murinae/*parasitology ; Orientia tsutsugamushi/*isolation & purification ; Republic of Korea ; Scrub Typhus/transmission

OBJECTIVETo investigate whether HV and Ot can coexist in their host (Leptotrombidium scutellare).

METHODSCollecting the separate Leptotrombidium scutellare and the ones from mice in epidemic area. The cells of mites at larva, nymph, and adult stages were cultured and made into smear. In situ RT-PCR and PCR were used to detect and locate HV RNA and Ot DNA in the primary cultured cells.

RESULTSPositive signals of HV RNA and Ot DNA distributed mostly in epithelial cells of digestive system and ovary cells of larva and nymph. The positive rate increased by the generation of passages.

CONCLUSIONCoinfection of HV and Ot did exist in wild Leptotrombidium scutellare.

Animals ; Cells, Cultured ; DNA, Bacterial ; analysis ; Female ; Hantavirus ; isolation & purification ; Mice ; Mites ; microbiology ; virology ; Orientia tsutsugamushi ; isolation & purification ; RNA, Viral ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the increasing incidence and the characteristics of Tsutsugamushi disease in the areas of Nan Peng Lie islands, Nan Ao island, Wan Shan archipelago, Nao Zhou island and Lei Zhou peninsula, located in the southern part of China and to develop strategies for preventive measures.

METHODSBoth epidemiological investigation, isolation and gene identification of Orientia tsutsugamushi, as well as pilot preventive measures were carried out.

RESULTSThese islands belonged to the epidemic area of south subtropical zone of Tsutsugamushi disease. The main host was Rattus norvegicu and the overall rates of infection on Orientia tsutsugamushi were 22.78%-33.75%. The main biological vector was Leptotrombidium (Leptotrombidium) deliens and the rates of infection on Orientia tsutsugamushi were 40.00%-75.00%. 25 strains of Orientia tsutsugamushi had been isolated from Rattus norvegicu and Leptotrombidium (Leptotrombidium) deliens. Results showed that the isolated strains of Orientia tsutsugamushi were 15 Karp, 8 Kato, 2 Yonchon. Results from serological studies showed that the positive rate of anti-Orientia tsutsugamushi antibodies was high, in both residents and soldiers stationed in these islands. On these islands, rats and biological vectors were killed. Results showed that these measures had positive impact in reducing the incidence.

CONCLUSIONIslands from the southern part of the country belonged to the epidemic area of Tsutsugamushi disease. People visiting this areas should be under protection.

Animals ; Antibodies, Bacterial ; blood ; Bacterial Typing Techniques ; China ; epidemiology ; Disease Outbreaks ; Disease Reservoirs ; microbiology ; Geography ; Humans ; Incidence ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Rats ; Scrub Typhus ; epidemiology ; Trombiculidae ; microbiology

OBJECTIVEScrub typhus is an infectious disease due to Orientia tsutsugamushi transmitted by infected chigger mites. Scrub typhus has long been recognized to occur in southern areas of China, but has recently been increasingly often reported from the north since the first case was reported in Mengyin County, Shandong Province in 1986. The key objectives of the present study were to investigate the clinical manifestations and epidemic factors of scrub typhus in children from the northern new natural foci.

METHODSThe case records of 56 children with scrub typhus who were admitted to the 5 hospitals of Fei County from September 1993 to January 2004 were reviewed. Orientia tsutsugamushi (Ot) was isolated from the cases. Based on ecological observations on the composition, seasonal fluctuation of animal hosts and chigger mites, Ot was isolated from rodents and chiggers. IgG antibodies to Ot was detected by IFA. Genotypes of the Ot isolates were also identified by nested PCR.

RESULTSAmong 56 children scrub typhus cases, 46 were male, 10 were female; 96% exhibited typical eschars or ulcers, 100% cases had high fever, skin rashes were observed in 55 cases (98%), and regional lymphadenopathy occurred in 48 cases (86%). All cases came from countryside, and all had histories of exposure to the crop field. fifty-one serum samples of suspected patients with scrub typhus were collected, 48 were positive for antibodies to Ot. The serotypes were Gilliam types. The cases only appeared in September to December with the peak at mid and late October. Leptotrombidium (L.) scutellare was the most important vector causing scrub typhus in the foci. Apodemus (A.) agrarius was the main host animals of Ot in the crop field. Totally 26 strains were isolated from patients, rodents, and chigger mites. The serotypes of 24 out of the 26 isolates were Gilliam types, while the genotypes of these isolates were Kawasaki types. The serotypes of the other 2 isolates were identical and both were Karp types.

CONCLUSIONChildren scrub typhus patients were frequently seen in the new natural foci of Shandong province. Exposure history, typical eschars or ulcers, and presence of IgG antibody were the important indexes to diagnose the disease.

Adolescent ; Animals ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mice ; parasitology ; Orientia tsutsugamushi ; isolation & purification ; Scrub Typhus ; epidemiology ; Seasons ; Trombiculidae ; 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

Comprehensive quarterly serosurveillance on scrub typhus in small mammals collected from military training sites located near the Demilitarized Zone (DMZ), northern Gyeonggi-do (Province), ROK was conducted to determine the potential rodent-borne and associated ectoparasite disease risks to military personnel. A total of 1,196 rodents and insectivores representing 8 species, Apodemus agrarius (87.3%, n = 1,044), Mus musculus (5.4%, n = 65), Crocidura lasiura (3.3%, n = 40), Microtus fortis (2.6%, n = 31), Micromys minutus (0.3%, n = 4), Tscherskia triton (0.3%, n = 4), Rattus norvegicus (0.3%, n = 4), and Myodes regulus (0.3%, n = 4) were assayed for the presence of antibodies to Orientia tsutsugamushi. O. tsutsugamushi antibodies were detected in 6 of 8 species and seroprevalence determined; A. agrarius (45.6%), M. musculus (23.1%), M. fortis (48.4%), M. minutus (50.0%), T. triton (50.0%), and R. norvegicus (25.0%). A total of 31,184 chigger mites collected from 508 rodents and insectivores were slide-mounted and 10 species belonging to 4 genera were identified. Leptotrombidium pallidum (53.4%) was the most frequently collected, followed by L. palpale (15.7%), Neotrombicula tamiyai (14.3%), L. orientale (10.7%), L. zetum (3.1%), Walchia fragilis (2.1%), and L. gemiticulum (0.8%), while the remaining 3 species, L. subintermedium, N. gardellai, and Euschoengastia koreaensis were rarely observed (prevalence < 10%). In contrast to previous surveys, higher chigger indices of the primary scrub typhus vectors, L. pallidum (165.4), L. orientale (45.0), and L. palpale (21.4), were observed during the spring season.
Animals ; Antibodies, Bacterial/immunology ; Arachnid Vectors/classification/*microbiology ; Disease Reservoirs/classification/microbiology/*parasitology ; Humans ; Military Facilities ; Orientia tsutsugamushi/*immunology/isolation & purification ; Republic of Korea ; Rodentia/classification/*immunology/microbiology/*parasitology ; Scrub Typhus/microbiology/*transmission/*veterinary ; Seroepidemiologic Studies ; Trombiculidae/classification/*microbiology

OBJECTIVETo analyze the genetic differences of Orientia tsutsugamushi (Ot) Sta56 gene between Shandong isolates and other strains deposited in GenBank.

METHODSPCR and restriction fragment length polymorphism (RFLP) were used to amplify the complete sequence of Ot-Sta56 gene. RFLP profiles of Ot were predicted by a computer program according to their complete sequences of Ot-Sta56 gene. PCR amplicon from XDM2 strain was sequenced and analyzed by Clustal X (1.8) and PHYLIP software.

RESULTSThe complete sequences (about 1.6 kbp) of Ot-Sta56 gene were amplified from B16 strain (isolated from patients), FXS2 strain (isolated from A. agrarius) and XDM2 strain. Four species of restriction endonucleases (Hha I, Hinf I, Hae III, Pst I) were used to digest the PCR amplicons from the 3 isolates. When comparing with the RFLP profiles of prototype Ot, the RFLP profiles of PCR amplicons from the 3 isolates were similar to those of Japan Kawasaki strain, but were quite different from the international reference strains Gilliam, Karp, Kato. Results from DNA sequence analysis showed that the complete sequence of Ot-Sta56 gene homology to Japan Kawasaki strain of XDM2 strain was 97%, and deduced amino acid sequence was 92%.

CONCLUSIONData from the complete sequence of Sta56 gene indicated that the genotypes of Ot isolates in Shandong province were similar, but with distinction from the Kawasaki strain.

Amplified Fragment Length Polymorphism Analysis ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Bacterial Typing Techniques ; classification ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Membrane Proteins ; genetics ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA