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<p><b>OBJECTIVE</b>To study the pathogenic ecology characteristics of plague in Qinghai plateau.</p><p><b>METHODS</b>Applied molecular biology techniques, conventional technologies and geographic information system (GIS) to study phenotypic traits, plasmid spectrum, genotype, infected host and media spectrum etc.of 952 Yersinia pestis strains in Qinghai plateau plague foci, which were separated from different host and media in different regions during 1954 to 2012.</p><p><b>RESULTS</b>The ecotypes of these strains were Qingzang plateau (91.49%, 871/952),Qilian mountain (6.41%, 61/952) and Microtus fuscus (1.26%, 12/952).83.6% (796/952) of these strains contained all the 4 virulence factors (Fr1, Pesticin1,Virulence antigen, and Pigmentation), 93.26% (367/392) were velogenic strains confirmed by virulence test.725 Yersinia pestis strains were separated from Qinghai plateau plague foci carried 9 kinds of plasmid, among which 713 strains from Marmot himalayan plague foci carried 9 kinds of plasmid, the Mr were 6×10(6), 7×10(6), 23×10(6), 27×10(6), 30×10(6), 45×10(6), 52×10(6), 65×10(6) and 92×10(6) respectively. 12 Yersinia pestis strains were separated from Microtus fuscus plague foci carried only 3 kinds of plasmid, the Mr were 6×10(6), 45×10(6), 65×10(6). Meanwhile, the strains carrying large plasmid (52×10(6), 65×10(6) and 92×10(6)) were only distributed in particular geographical location, which had the category property. The research also confirmed that 841 Yersinia pestis strains from two kinds of plague foci in Qinghai plateau had 11 genomovars. The strains of Marmot himalayan plague foci were given priority to genomovar 5 and 8, amounted to 611 strains, genomovar 8 accounted for 56.00% (471/841), genomovar 5 accounted for 23.07% (194/841). Besides, 3 new genomovars, including new 1(62 strains), new 2(52 strains), new 3(48 strains) were newly founded, and 12 strains of Microtus fuscus plague foci were genomovar 14.</p><p><b>CONCLUSION</b>The main host and media of Qinghai plateau plague foci directly affected the spatial distribution regularities of plague epidemic and the pathogens characteristics, meanwhile the polymorphism of plague ecological geographic landscape leds to the complexity of Yersinia pestis' genotype.</p>
Subject(s)
Animals , Arvicolinae , Microbiology , China , Epidemiology , Disease Reservoirs , Microbiology , Ecology , Genotype , Marmota , Microbiology , Plague , Epidemiology , Microbiology , Virulence , Genetics , Yersinia pestis , Genetics , VirulenceABSTRACT
Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.
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ObjectiveTo establish a comprehensive evaluation method on sensitivity of Yersinia pestis to different antibiotics, then use the method to evaluate the sensitivity of Yersinia pestis to 6 antibiotics. Methods Through literature review, related articles were consulted and sensitivity testing data to antibiotics were collected,and the sensitivity of Yersinia pestis to the 6 antibiotics was assessed with analytic hierarchy process. The sensitivity of Yersinia pestis to the 6 antibiotics(Cefotaxime penicillin, Ampicillin, Cefazolin, Ciprofloxacin, Ciprofloxacin Buddha's, and Streptomycin) was evaluated comprehensively with analytic hierarchy process from the antibacterial situation of the antibiotics to Yersinia pestis of 141 strains of virulent, attenuated EV76paris strains and isolates of the bacteria of Yunnan. Results The evaluation showed that the sensitivity of Yersinia pestis to antibiotics was different. The rank of the sensitivity of Yersinia pestis to the 6 antibiotics was: Ceftazidime, Ampicillin, Cefazolin,Ciprofloxacin, Norfloxacin and Streptomycin. Composite score index were 1.730 77, 1.631 77, 1.581 95, 1.567 80,1.449 48, and 0.999 99, respectively. Conclusions Analytic hierarchy process evaluation can be a reasonable,objective and more accurate evaluation on the sensitivity of Yersinia pestis to different antibiotics. This method could provide some references for screening feasible pharmaceuticals against plague.
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Objective To explore the plague epidemical trend of nearly a 10 years data in Qinghai province to provide basis for making the prevention and control measures. Method The regional distribution and time distribution of animal and human plague, monitoring and plague foci of survey data in Qinghai from 2001 to 2010 were analyzed with Excel software 2003. Results In Qinghai province, a total of 167 strains of Yersinia pestis were isolated from infected animals and insects in 10 years. Yersinia pestis was mainly distributed in Wulan,Delinha, Geermu, and Tianjun, along the Qinghai-Xizang railway. Human plague was occurred every year from 2001 to 2010 except 2002, 2007, 2008, and 2010. In the 10 years, there were 37 plague cases and 16 of these cases died, the mortality was 43.24％. The plague cases were mainly distributed in Nangqian, Qumalai, Chenduo,Zhiduo, Xinghai, Tongde, Tianjun, Wulan and Qilian. And these cases were found mostly in the period from May to October, especially in the period from August to October. Major clinical type of the plague cases was lung-type (62.16％,23/37). Conclusions The plague epidemic situation in Qinghai province is still severe, animal plague occurred year after year, and human plague outbreaks occasionally. Monitoring and early warning in the key areas should be strengthened, and the comprehensive measures of plague prevention and control should be carried out to reduce the incidence and prevalence of plague.
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Objective To study the biological characteristics of Yersinia pestis and to develop prevention and control program on plague in Sanjiangyuan areas,Qinghai province.Methods To identify the biologic types and molecular biological features of Y.pestis isolated in Sanjiangyuan area from 1954-2007.Results Among the 411 strains ofY.pestis,12 strains belonged to the microtus type Y.pestis with denitrification (-) and donkey-bible gelatin carbohydrate(-) and glycerine(+).399 strains belonged to classic type Y.pestis with denitrification (+) and donkey-hide gelatin carbohydrate (+) and glycerine (+).411 Y.pestis strains had factor F I and Pst I.Among them,VW + strains of Y.pestis accounted for 95.13%(391/411),VW accounted for 4.87%(20/411),Pgm+accounted for 80.78%(332/411 ),Pgm±accounted for 9% (37/411 ) and Pgm-accounted for 10.22% (42/411) respectively.96.82% (213/220) of the Y.pestis strains showed strong virulence to laboratory mice while 3.18% (7/220) of the strains carried medium virulence.90.02% of the tested Y.pestis (370/411) strains had 6×106,45×106,65×106 plasmids.8 types of genome were found among 80 strains of Y.pestis,with 6 of them resembling ZHOU Dongsheng's classification.Two new genome types were found.Conclusion The Y.pestis in the Sanjiangyuan area had the characteristics of plague pathogen,identified in Qinghai-Tibet plateau.It is estimated that human beings are highly susceptible to the disease which spread fast,causing serious signs and symptoms with high death rate.
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Objective Throush identify biochemical characteristics and virulence factors of 2 strains suspected Yersinia pestis(Y.pestis)isolated from the dead Marmota himalayana(M.himalayana)to confirm the nature epidemic focus in Dege County,Sichuan Province.Methods Y.pestis was analyzed by specific staining and shape,culturing characteristics,splitting-test by bacteriophage,test of biochemical characteristics and glycolysis ability,virulence factors,virulence,nutritional requirement,plasmid,genetic test and genetic type. Results The tested strains were Gram staining bacilus.The main biochemical characteristics were Arabinose(+)、 Rhamnose(-),Maltose(+),Melibiose(-),Glycerol(+),Denitrification(+).The virulence factors with FI+.VW+, Pgm+,Pst I+;and with the common 6.0×106,45.0×106,65.0×106 plasmids,also with the virulence-relative plasmid gene.Both their absolutely lethal dose(LD100)in mice were 50 bacteria.The nutritional requirement appeared which were depended on Phenylalanine and Methionine.With the Genomovar 5 genotype characteristics of M.himalayana plague foci of Qinghai-Tibet plateau.The difference between tested strains and Yersinia pseudotubercuosis on the 3 different culture medium was obvious.The tested strains had a Y.pestis' specific 3a fragment,Pst I and FI-Ag,at 22 ℃,the strains could be split by bacteriophage completely.Conclusions According to the diagnostic criteria of plague in China,the 2 suspected strains isolated from Dege County,Sichuan Province ale confirmed as Y.pestis.both with powerful virulenceand with the characteristics of the Y.pestis of M.himahtyana in Qinghai-Tibet plateau plague natural focus.
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Objective To investigate the space structure of plague natural foci in the area of Lantsang, Yellow and Yangtse River in Qinghai Province to provide references for making decisions to eontrol the occurrence of human plague. Methods Data was collected from the survey on natural foci and surveillance of plague from 1954 to 2006 and descriptive epidemiological method was used to analyze the data. Results Marmata hirnalayana and Microtus fuscua natural foci were known in Sanjiangyuan area. Callopsylla dolabris, Oropsylla silantiewi, Citellophilus sparsilis and Amphipsylla tuta were vectors; Microtus fuscus plague natural foci was in a range of about 9500 km2, distributing in Zhenqin Town, Chengduo County. Marmata himalayana plague natural foci distributed over 13 countries, a range nearly 107 000 km2. By the end of 2006, 450 strains of Yersinia pestis were detected and separated from 6 kinds of rodents, 6 kinds of carnivora, 3 kinds of artiodactyls and 9 insects vectors. Between 1960 and 2006, 238 cases and 134 deaths from plague were reported. Most human plague cases occurred in the months from May to November and usually presented as one of three primary forms-bubonic 17.23%(41/238), septicemic 16.81% (40/238), pneumonic 61.34% (146/238) and other types 4.62% (11/238). However, the first epidemic plague case was mainly the glandular plague. Conclusions Date suggested that plague is still a critical public health problem in Sanjiangyuan area, against which countermaeasure needs to be strengthened in the main epidemic areas. More scientific researches on plague should be carried out. Surveillance networks of reporting suspected plague have been established and reduce the number of human plague cases.
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Objective To study the features of Yersinia pestis(Y.pestis)in areas along Qinghai-Tibet Railroad in Qinghai Province.Methods To identify the biologic types and the molecular biological feathers of Y.pestis isolated from areas along Qinghai-Tibet Railroad in Qinghai from 2001-2006.Results All the tested Y.pestis was biologically of classical type and ecologically of Qinghai-Tibet plateau type.The Y.pestis had high virulence.The Y.pestis of 65×106 plasmids was distributed in the Tanggula area,the Y.pestis of 52×106plasmids,in Tianjun and Delingha areas.The Y.pestis srains carried 52 × 106 plasmids.except the two containing 65 X 106 plasmids in Wulan County.The genetic type of Y.pestis in Tanggula was type 5 and that in Zongwulong of Delingha,Saishike,Keke,Tongpu of Wulan was type 8 except 2 strains of Y.pestis isolated from woodchuck and the patients in Dananwan of Tongpu,Wulan County were type 15.Conclusion The Y.pestis in the area along Qinghai-Tibet Railroad in Qinghai belongs to Qinghai-Tibet plateau type with high virulence.