RESUMO
OBJECTIVE: To analyze the plasmid features and geographical distribution characteristics of Yersinia pestis of different plague foci in China. METHODS: A total of 2 213 Yersinia pestis strains were colected from 11 Chinese plague foci separated during 1943 to 2012, and plasmid DNA according to alkali cracking method, and measured the relative molecular mass (Mr) of plasmid DNA based on the standard plasmid contrast method, then analyzed the plasmid profiles by agar gel electrophoresis. RESULTS: A total of 2 213 strains had 16 kinds of plasmids with different Mr, including 4×10(6), 6×10(6), 7×10(6), 13×10(6), 16×10(6), 20×10(6), 22×10(6), 23×10(6), 27×10(6), 30×10(6), 36×10(6), 45×10(6), 52×10(6), 65×10(6), 72×10(6) and 90×10(6). Plasmid were classified into 26 kinds of plasmid profiles. A total of 2 213 Yersinia pestis strains contained 4 large plasmids, 52×10(6), 65×10(6), 72×10(6) and 90×10(6), whose ratio was 22.10% (589/2 213), 75.60% (1 672/2 213), 0.17% (4/2 213), 2.12% (47/2 213), respectively. Among which, strains with plasmid 52×10(6), 65×10(6), 90×10(6) distributed in Qinghai-Tibet plateau Himalayan Marmot natural plague foci, strains with 72×10(6) plasmid only distributed in Inner Mongolia Meriones unguiculatus natural plague foci and Junggar Basin R. opimus natural plague foci, and 65×10(6) plasmid distributed in all the other foci. CONCLUSION: Strains in Chinese 11 plague foci contained 4 kinds of large plasmid, the Mr respectively were 52×10(6), 65×10(6), 72×10(6), 90×10(6), which were classified into 26 kinds of plasmid profiles with other plasmid. These plasmid profiles distributed in relatively independent epidemic focus.
Assuntos
Genótipo , Plasmídeos , Yersinia pestis , Animais , China , PesteRESUMO
OBJECTIVE: To study the pathogenic ecology characteristics of plague in Qinghai plateau. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
Assuntos
Ecologia , Peste/microbiologia , Yersinia pestis , Animais , Arvicolinae/microbiologia , China/epidemiologia , Reservatórios de Doenças/microbiologia , Genótipo , Marmota/microbiologia , Peste/epidemiologia , Virulência/genética , Yersinia pestis/genética , Yersinia pestis/patogenicidadeRESUMO
Plague caused by Yersinia pestis remains a public health threat worldwide. Because multidrug-resistant Y. pestis strains have been found in both humans and animals, phage therapy has attracted increasing attention as an alternative strategy against plague. However, phage resistance is a potential drawback of phage therapies, and the mechanism of phage resistance in Y. pestis is yet to be investigated. In this study, we obtained a bacteriophage-resistant strain of Y. pestis (S56) by continuously challenging Y. pestis 614F with the bacteriophage Yep-phi. Genome analysis identified three mutations in strain S56: waaA* (9-bp in-frame deletion 249GTCATCGTG257), cmk* (10-bp frameshift deletion 15CCGGTGATAA24), and ail* (1-bp frameshift deletion A538). WaaA (3-deoxy-D-manno-octulosonic acid transferase) is a key enzyme in lipopolysaccharide biosynthesis. The waaA* mutation leads to decreased phage adsorption because of the failure to synthesize the lipopolysaccharide core. The mutation in cmk (encoding cytidine monophosphate kinase) increased phage resistance, independent of phage adsorption, and caused in vitro growth defects in Y. pestis. The mutation in ail inhibited phage adsorption while restoring the growth of the waaA null mutant and accelerating the growth of the cmk null mutant. Our results confirmed that mutations in the WaaA-Cmk-Ail cascade in Y. pestis contribute to resistance against bacteriophage. Our findings help in understanding the interactions between Y. pestis and its phages.
Assuntos
Bacteriófagos , Peste , Yersinia pestis , Animais , Humanos , Yersinia pestis/genética , Lipopolissacarídeos , Mutação , Bacteriófagos/genéticaRESUMO
Bacteriophages (phages) have been successfully used as disinfectors to kill bacteria in food and the environment and have been used medically for curing human diseases. The objective of this research was to elucidate the morphological and genomic characteristics of two novel Yersinia pestis phages, vB_YpeM_ MHS112 (MHS112) and vB_YpeM_GMS130 (GMS130), belonging to the genus Gaprivervirus, subfamily Tevenvirinae, family Myoviridae. Genome sequencing showed that the sizes of MHS112 and GMS130 were 170507 and 168552 bp, respectively. A total of 303 and 292 open reading frames with 2 tRNA and 3 tRNA were predicted in MHS112 and GMS130, respectively. The phylogenetic relationships were analysed among the two novel Y. pestis phages, phages in the genus Gaprivervirus, and several T4-like phages infecting the Yersinia genus. The bacteriophage MHS112 and GMS130 exhibited a wider lytic host spectrum and exhibited comparative temperature and pH stability. Such features signify that these phages do not need to rely on Y. pestis as their host bacteria in the ecological environment, while they could be based on more massive Enterobacteriales species to propagate and form ecological barriers against Y. pestis pathogens colonised in plague foci. Such characteristics indicated that the two phages have potential as biocontrol agents for eliminating the endemics of animal plague in natural plague foci.
Assuntos
Bacteriófagos , Peste , Yersinia pestis , Animais , Humanos , Peste/prevenção & controle , Bacteriófagos/genética , Filogenia , Bactérias , Myoviridae/genéticaRESUMO
The plague, which is caused by the Gram-negative coccobacillus bacterium Yersinia pestis, has been classified as a reemerging infectious disease by the World Health Organization. The Qinghai-Tibet Plateau natural plague focus is the largest plague focus in China, and Marmota himalayana is the primary host of the plague. Tibetan sheep (Ovis aries) were first identified as naturally infected hosts of Y. pestis based on etiological evidence in 1975, and activities such as slaughtering or skinning Tibetan sheep that have been infected by Y. pestis or died from Y. pestis infection had caused severe human plague in Qinghai. Tibetan sheep are important domestic livestock in the Qinghai-Tibet Plateau. Knowledge regarding the infection rate of Y. pestis in Tibetan sheep is important for understanding the range of infection and improving measures to control plague epidemics in this area. In this study, a serological survey involving 12,710 Tibetan sheep in all 44 counties in Qinghai Province was conducted. The total positive rate of indirect hemagglutination assay for Y. pestis in Tibetan sheep in Qinghai was 0.68% (86/12,710). Serological positivity to the Y. pestis F1 antibody was found in Tibetan sheep in all prefectures, except the Haidong and Haibei prefectures in Qinghai, with the seropositive rate in different counties ranging from 0.33% to 5.2% and the titers in the positive sera ranging from 1:20 to 1:5120. In addition, the seropositive rates in animal plague focus counties were higher than the rates in non-animal plague counties. Such results indicated a widespread infection of Tibetan sheep with Y. pestis in Qinghai, even though only sporadic epidemics of Tibetan sheep plague have been reported in Qinghai.
Assuntos
Peste/veterinária , Doenças dos Ovinos/microbiologia , Animais , Carnívoros , China/epidemiologia , Peste/epidemiologia , Roedores , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/epidemiologiaRESUMO
BACKGROUND: Plague, caused by the bacterium Yersinia pestis, is a highly infectious, zoonotic disease. Hundreds of human plague cases are reported across the world annually. Qinghai Plateau is one of the most severely affected plague regions in China, with more than 240 fatal cases of Y. pestis in the last 60 years. Conventional epidemiologic analysis has effectively guided the prevention and control of local plague transmission; however, molecular genetic analysis is more effective for investigating population diversity and transmission. In this report, we employed different genetic markers to analyze the population structure of Y. pestis in Qinghai Plateau. METHODOLOGY/PRINCIPAL FINDING: We employed a two-step hierarchical strategy to analyze the phylogeny of 102 Qinghai Plateau isolates of Y. pestis, collected between 1954 and 2011. First, we defined the genealogy of Y. pestis by constructed minimum spanning tree based on 25 key SNPs. Seven groups were identified, with group 1.IN2 being identified as the dominant population. Second, two methods, MLVA and CRISPR, were applied to examine the phylogenetic detail of group 1.IN2, which was further divided into three subgroups. Subgroups of 1.IN2 revealed a clear geographic cluster, possibly associated with interaction between bacteriophage and Y. pestis. More recently, Y. pestis populations appear to have shifted from the east toward the center and west of Qinghai Plateau. This shift could be related to destruction of the local niche of the original plague focus through human activities. Additionally, we found that the abundance and relative proportion of 1.IN2 subgroups varied by decade and might be responsible for the fluctuations of plague epidemics in Qinghai Plateau. CONCLUSION/SIGNIFICANCE: Molecular genotyping methods provided us with detailed information on population diversity and the spatial-temporal distribution of dominant populations of Y. pestis, which will facilitate future surveillance, prevention, and control of plague in Qinghai Plateau.
Assuntos
Variação Genética , Peste/microbiologia , Yersinia pestis/genética , Zoonoses/microbiologia , Animais , China/epidemiologia , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Epidemias , Genótipo , Técnicas de Genotipagem , Humanos , Filogeografia , Peste/epidemiologia , Polimorfismo de Nucleotídeo Único/genética , Análise Espaço-Temporal , Yersinia pestis/isolamento & purificação , Zoonoses/epidemiologiaRESUMO
Compared with high tibial osteotomy and total knee arthroplasty, the authors found a simpler surgical procedure, partial fibular osteotomy, could effectively relieve knee pain and also correct the varus deformity for patients with medial compartment knee osteoarthritis (OA). From January 1996 to April 2012, a total of 156 patients with medial compartment OA were treated by proximal fibular osteotomy in the authors' hospital. A 2-cm-long section of fibula was resected 6 to 10 cm below the fibular head. A total of 110 patients with follow-up of more than 2 years were included in the study, including 34 males and 76 females with an average age of 59.2 years. Anteroposterior and lateral weight-bearing radiographs, the femorotibial angle (FTA) and lateral joint space, and the American Knee Society Score (KSS) and the visual analog scale (VAS) score of the knee joint were evaluated preoperatively and at final follow-up, respectively. At final follow-up, mean FTA and lateral joint space were 179.4°±1.8° and 6.9±0.7 mm, respectively, which were significantly smaller than those measured preoperatively (182.7°±2.0° and 12.2±1.1 mm, respectively; both P<.001). Mean KSS at final follow-up was 92.3±31.7, significantly higher than the mean preoperative score of 45.0±21.3 (P<.001). Mean VAS score and interquartile range were 2.0 and 2.0, significantly lower than the preoperative data (7 and 1.0, respectively; P<.001). The authors found that proximal fibular osteotomy can significantly improve both the radiographic appearance and function of the affected knee joint and also achieve long-term pain relief. This procedure may be an alternative treatment option for medial compartment OA.
Assuntos
Descompressão Cirúrgica/métodos , Osteoartrite do Joelho/cirurgia , Osteotomia/métodos , Artroplastia do Joelho/métodos , Feminino , Fíbula/cirurgia , Humanos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Dor Musculoesquelética/prevenção & controle , Dor Musculoesquelética/cirurgia , Osteoartrite do Joelho/diagnóstico por imagem , Osteoartrite do Joelho/fisiopatologia , Projetos Piloto , Radiografia , Tíbia/cirurgia , Suporte de Carga/fisiologiaRESUMO
OBJECTIVE: To type Yersinia (Y.) pestis isolates under different regions (DFR) and to observe their geographical distributions in China. METHODS: 23 DFRs primers and PMT1 (plasmid) primer were used to verify the DFR genomovars of Y. pestiss strains from 11 plague foci in China. A total of 3 044 Y. pestis isolates were involved for analysis on DFR profiles with the characteristics of geographical distribution. RESULTS: 52 genomovars were verified in 3 044 Y. pestis strains in China in which 19 genomovars as major and 33 genomovars as minor genomovar. 21 new genomovars, namely genomovar 32 to genomovar 52 were described on the basis of 31 genomovars previously confirmed. Three new genomovars belonged to new major genomovars, namely Himalayan marmot natural plague foci of the Qinghai-Tibet plateau newly added genomovar 32 and genomovar 44 as major genomovars. Mongolian gerbil natural plague foci of Inner Mongolia plateau were newly added genomovar 50 as one of the major genomovars. CONCLUSION: Among 21 new genomovars, 3 were major genomovars, with Chinese Y. pestis DFR as the major genomovars which had obvious distribution characteristics.
Assuntos
Yersinia pestis/classificação , China , Genótipo , Geografia , Yersinia pestis/genética , Yersinia pestis/isolamento & purificaçãoRESUMO
OBJECTIVE: To study the biological and genetic characteristics of 119 strains of Yersinia (Y.) pestis isolated from plague patients in Qinghai province, from 1958-2012. METHODS: Both regular methods and different region(DFR)molecular typing techniques were used to study the epidemiological characteristics on 119 strains of Y. pesticin Qinghai during 1958-2012. Sources of Y. pestis from two outbreaks, in Nangqian county in 2004 and in Xinghai county in 2009,Qinghai province were also analyzed. RESULTS: 105 strains of Y. pestis were identified as Qinghai-Tibet Plateau Ecotype while the other 6 strains as Qilian Mountains Ecotype. 84.03% (100/119) of the tested strains carried 4 virulence factors F1(+), Pst I(+), VW(+) and Pgm(+)). 97.30% (72/74) of the tested strains showed high virulence. Strains that carrying 52×10(6), 65×10(6), 92×10(6) plasmids were distributed in Hainan, Haibei, Haixi,Yushu,Guoluo, Huangnan and Huangyuan counties. Genomovar 5 and 8 were the main gene types that circling around Qinghai Lake. Genomovar 10 was found in strains of Y. pesticin Nangqian county while Genomovar 8 was found in the strains isolated from human plague patient during the epidemics in Xinghai county in Qinghai. CONCLUSION: Data from biological and genetic analyses on the epidemics of human plague in Nangqian county in 2004 and in Xinghai county in 2009 demonstrated that methods as DFR genotyping and virulence factors profiles, as well as plasmids profiles were powerful tools in confirming the human plague epidemics and sources of infection.
Assuntos
Peste/epidemiologia , Peste/microbiologia , Yersinia pestis/genética , China/epidemiologia , Genótipo , Humanos , Yersinia pestis/isolamento & purificaçãoRESUMO
Artificially passive immunization has been demonstrated to be effective against Yersinia pestis infection in animals. However, maternal antibodies' protective efficacy against plague has not yet been demonstrated. Here, we evaluated the kinetics, protective efficacy, and transmission modes of maternal antibodies, using mice immunized with plague subunit vaccine SV1 (20 µg of F1 and 10 µg of rV270). The results showed that the rV270- and F1-specific antibodies could be detected in the sera of newborn mice (NM) until 10 and 14 weeks of age, respectively. There was no antibody titer difference between the parturient mice immunized with SV1 (PM-S) and the caesarean-section newborns (CSN) from the PM-S or between the lactating mice immunized by SV1 (LM-S) and the cross-fostered mice (CFM) during 3 weeks of lactation. The NM had a 72% protection against 4,800 CFU Y. pestis strain 141 challenge at 6 weeks of age, whereas at 14 weeks of age, NM all succumbed to 5,700 CFU of Y. pestis challenge. After 7 weeks of age, CFM had an 84% protection against 5,000 CFU of Y. pestis challenge. These results indicated that maternal antibodies induced by the plague subunit vaccine in mother mice can be transferred to NM by both placenta and lactation. Passive antibodies from the immunized mothers could persist for 3 months and provide early protection for NM. The degree of early protection is dependent on levels of the passively acquired antibody. The results indicate that passive immunization should be an effective countermeasure against plague during its epidemics.
Assuntos
Anticorpos Antibacterianos/sangue , Imunidade Materno-Adquirida , Vacina contra a Peste/imunologia , Peste/prevenção & controle , Yersinia pestis/imunologia , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Feminino , Humanos , Lactação/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Placenta/imunologia , Peste/imunologia , Vacina contra a Peste/administração & dosagem , Gravidez , Análise de Sobrevida , Fatores de TempoRESUMO
In our previous study, complete protection was observed in Chinese-origin rhesus macaques immunized with SV1 (20 µg F1 and 10 µg rV270) and SV2 (200 µg F1 and 100 µg rV270) subunit vaccines and with EV76 live attenuated vaccine against subcutaneous challenge with 6×10(6) CFU of Y. pestis. In the present study, we investigated whether the vaccines can effectively protect immunized animals from any pathologic changes using histological and immunohistochemical techniques. In addition, the glomerular basement membranes (GBMs) of the immunized animals and control animals were checked by electron microscopy. The results show no signs of histopathological lesions in the lungs, livers, kidneys, lymph nodes, spleens and hearts of the immunized animals at Day 14 after the challenge, whereas pathological alterations were seen in the corresponding tissues of the control animals. Giemsa staining, ultrastructural examination, and immunohistochemical staining revealed bacteria in some of the organs of the control animals, whereas no bacterium was observed among the immunized animals. Ultrastructural observation revealed that no glomerular immune deposits on the GBM. These observations suggest that the vaccines can effectively protect animals from any pathologic changes and eliminate Y. pestis from the immunized animals. The control animals died from multi-organ lesions specifically caused by the Y. pestis infection. We also found that subcutaneous infection of animals with Y. pestis results in bubonic plague, followed by pneumonic and septicemic plagues. The histopathologic features of plague in rhesus macaques closely resemble those of rodent and human plagues. Thus, Chinese-origin rhesus macaques serve as useful models in studying Y. pestis pathogenesis, host response and the efficacy of new medical countermeasures against plague.
Assuntos
Macaca mulatta/imunologia , Vacina contra a Peste/imunologia , Yersinia pestis/imunologia , Animais , Antígenos de Bactérias/imunologia , Membrana Basal/metabolismo , Feminino , Imunização , Imuno-Histoquímica/métodos , Injeções Subcutâneas , Masculino , Microscopia Eletrônica , Células-Tronco , Distribuição Tecidual , Vacinas de Subunidades Antigênicas/imunologiaRESUMO
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 of Y. pestis, 12 strains belonged to the microtus type Y. pestis with denitrification (-) and donkey-hide 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 x10(6), 45 x 10(6), 65 x 10(6) 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.
Assuntos
Peste/microbiologia , Roedores/microbiologia , Yersinia pestis/patogenicidade , Animais , China , Genoma Bacteriano , Genótipo , Camundongos , Plasmídeos , Virulência , Yersinia pestis/genética , Yersinia pestis/isolamento & purificaçãoRESUMO
OBJECTIVE: To study the distribution of genomovars and microevolution of Yersinia pestis in the Qinghai-Tibet Plateau. METHODS: Primer pairs targeting the twenty-two different regions(DFRs) were designed for detecting the presence or deletion of each DFR in 297 strains isolated from the Qinghai-Tibet Plateau. RESULTS: 9 genomovars, i. e. Genomovar 1, 5, 6, 7, 8, 10, 11, new type and Ype-ancestor were identified in the Marmota himalayana plague focus of the Qinghai-Tibet Plateau. Among these genomovars, genomovar 5,8 and 10 were dominant types. The total rate of the three genomovars was 80.6% (204/253) and the genomovars in different regions were different. All of 44 strains of Y. pestis in the Microtus fuscus plague focus of the Qinghai-Tibet Plateau belonged to genomovar 14. CONCLUSION: The distribution of genomovars of Y. pestis in the Qinghai-Tibet plateau had remarkable characteristics geographically. Based on the distribution of genomovars of Y. pestis, the routes of transmission and microevolution of Y. pestis were proposed.