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Plague is a highly virulent fleaborne zoonotic disease caused by the bacterium Yersinia pestis (1,2). Humans can acquire plague through contact with infectious animal tissues or through inhalation. Human contact [...]

We analyzed epidemiologic characteristics and distribution of 1,067 human plague cases and 5,958 Yersinia pestis isolates collected from humans, host animals, and insect vectors during 1950-2019 in 4 Marmota plague foci in China. The case-fatality rate for plague in humans was 68.88%; the overall trend slowly decreased over time but fluctuated greatly. Most human cases (98.31%) and isolates (82.06%) identified from any source were from the Marmota himalayana plague focus. The tendency among human cases could be divided into 3 stages: 1950-1969, 1970-2003, and 2004-2019. The Marmota sibirica plague focus has not had identified human cases nor isolates since 1926. However, in the other 3 foci, Y. pestis continues to circulate among animal hosts; ecologic factors might affect local Y. pestis activity. Marmota plague foci are active in China, and the epidemic boundary is constantly expanding, posing a potential threat to domestic and global public health. [ABSTRACT FROM AUTHOR]

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.

Objective: To identify the epidemiology and etiology characteristics of Tibetan sheep plague in Qinghai plateau., Methods: The background materials of Qinghai Tibetan sheep plague found during 1975 to 2009 were summarized, the regional, time and interpersonal distribution, infection routes, ecological factors for the spread were used to analyze; followed by choosing 14 Yersinia pestis strains isolated from such sheep for biochemical test, toxicity test, virulence factors identification, plasmid analysis, and DFR genotype., Results: From 1975 to 2009, 14 Yersinia pestis strains were isolated from Tibetan sheep in Qinghai province. Tibetan sheep, as the infection source, had caused 10 cases of human plague, 25 plague patients, and 13 cases of death. All of the initial cases were infected due to eating Tibetan sheep died of plague; followed by cases due to contact of plague patients, while all the initial cases were bubonic plague. Cases of bubonic plague developed into secondary pneumonic plague and septicemia plague were most popular and with high mortality. Most of the Tibetan sheep plague and human plague occurred in Gannan ecological zone in southern Gansu province, which was closely related to its unique ecological and geographical landscape. Tibetan sheep plague coincided with human plague caused by Tibetan sheep, especially noteworthy was that November (a time for marmots to start their dormancy) witnesses the number of Yersinia pestis strains isolated from Tibetan sheep and human plague cases caused by Tibetan sheep. This constituted the underlying cause that the epidemic time of Tibetan sheep plague lags obviously behind that of the Marmot plague. It was confirmed in the study that all the 14 strains were of Qinghai-Tibet Plateau ecotype, with virulence factors evaluation and toxicity test demonstrating strains as velogenic. As found in the (Different Region) DFR genotyping, the strains isolated from Yushu county and Zhiduo county were genomovar 5, the two strain isolated from Nangqian county were genomovar 5 and genomovar 7, while those isolated Delingha region were genomovar 8., Conclusion: Tibetan sheep were vulnerable to plague infection, hence causing human plague as the infectious source. The Yersinia pestis strains isolated from Tibetan sheep plague carried pathogen characteristics of Qinghai-Tibet plateau plague, developing many new characteristics of such plague.

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.

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.

Objective: To analyze the results of etiology and serology of plague among human and infected animals in Qinghai province from 2001 to 2010., Methods: Thirty-seven cases of human infected with plague, 53 541 different animal samples, 5 685 sets of vector insects flea and 49 039 different animal serum samples were obtained between 2001 and 2010. A total of 7 811 samples of serum from healthy farmers and herdsmen in 14 counties in Qinghai from 2005 to 2007 were collected. Yersinia pestis (Y. pestis) were detected in visceral and secretions from human, infected animals and vector insects, respectively. Plague antigen was detected by reverse indirect hemagglutination assay (RIHA) in those samples. Indirect hemagglutination assay (IHA) was used to test plague FI antibody in serum of human and infected animals., Results: 37 human plague cases were confirmed, 21 strains of plague Y. pestis were isolated from human cases and 14 positive were detected out. 133 of 7 811 samples of human serum were IHA positive, with the positive rate at 1.7%. A total of 146 strains of plague were isolated from infected animals and vector insects, 99 out of which were from infected animals, with a ratio of Marmota himalayan at 72.7% (72/99) and the other 47 were from vector insects, with a ratio of callopsylla solaris at 68.1% (32/47). The number of IHA and PIHA positive were 300 and 10, respectively. A total of 3 animals and 3 insects species were identified as new epidemic hosts for plague. The natural plague focus of Microtus fuscus was discovered and confirmed and coexisted with natural focus of Marmota himalayan in Chengduo county, Yushu prefecture. The epidemic situation of plague is distributed mainly in Haixi, Yushu and Hainan prefectures., Conclusion: From 2001 to 2010, animal infected with plague was detected in successive years and human plague was very common in Qinghai. New infected animals and vector insects species and new epidemic areas were confirmed, hence the trend of plague prevalence for humans and animals is very active in Qinghai province.

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.