Genotyping and traceability analysis of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau by MLVA / 中华地方病学杂志

Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.

CRISPR genotyping of Yersinia pestis in the plague natural foci of Qinghai-Tibet Plateau / 中华地方病学杂志

Objective:To investigate the genotypes of clustered regularly interspaced short palindromic repeats (CRISPR) of Yersinia pestis ( Y. pestis) and it's regional distribution in the plague natural foci of Qinghai-Tibet Plateau. Methods:In this study, 1 004 strains of Y. pestis isolated from different hosts and vectors in different regions from 1954 to 2011 were selected as experimental objects, which preserved in Qinghai Institute for Endemic Disease Prevention and Control, and DNA of Y. pestis was extracted by traditional phenol-chloroform method. Three CRISPR loci (YPa, YPb and YPc) were amplified by PCR and sequenced, respectively, and the CRISPR sequences were compared with the CRISPRDictionary database recently reported in the literature to identify CRISPR spacer. For the spacer discovered for the first time at each CRISPR locus, Blast sequence alignment was performed in the National Center for Biotechnology Information (NCBI) database to speculate the source of gene sequence. Genotyping of Y. pestis from Qinghai-Tibet Plateau was performed based on the polymorphism of CRISPR spacer array. Results:A total of 53 spacers were found in 1 004 strains of Y. pestis, of which 6 spacers were newly discovered, namely a105, a106, a107, b51, b52 and c14. The 1 004 strains of Y. pestis were divided into 10 groups with 44 different CRISPR genotypes, and 15 new genotypes were found. The dominant genotypes of Y. pestis were G26-a1', G7, G22, G24-a1', G22-a1', G9 and G26-a1'a60 in Himalayan Marmot plague natural foci, and G37-a6' in Qinghai Microtinae plague natural foci. Conclusion:The CRISPR genotypes of Y. pestis in the plague natural foci of Qinghai-Tibet Plateau are high diverse and have significant regional distribution characteristics.

Research progress on identification and monitoring of the appropriate habitat of Himalayan marmot / 中华地方病学杂志

Qinghai-Tibet Plateau is one of the major plague foci in China. Himalayan marmot (hereinafter referred to as the marmot) is the main host animals of the foci. Effective identification and monitoring of its habitat in Qinghai-Tibet Plateau is important for active prevention and effective control of the marmot plague epidemic. Spatial information technology is an important means for rapid identification and effective monitoring of marmot. This review analyzed the factors that affect the distribution patterns of marmot habitat, including topographical features, soil properties, vegetation types and cover, climatic conditions and human activities, and reviewed and summarized the progress and shortcomings of relevant research on identifying and monitoring marmot habitat based on spatial information technologies such as remote sensing and geographic information system. Meanwhile, combined with the emerging low-altitude remote sensing, crowdsource data acquisition, machine learning and other big data mining methods, from the perspective of multi-source data, integration of emerging technologies and comprehensive evaluation system, the new ideas of using spatial information technology to quickly identify and effectively monitor the habitat of marmot, was discussed. It is expected to provide technical support for prediction and effective control of marmot plague in Qinghai-Tibet Plateau under the background of changing global climate and increasing human activities.

Epidemic status of alveolar echinococcosis in Tibetan children in south Qing-hai Province / 中国血吸虫病防治杂志

Objective To understand the epidemic status of alveolar echinococcosis in Tibetan children in south Qinghai Province. Methods The imageology and serology methods were applied to investigate the epidemic status of alveolar echinococ?cosis of elementary school students in Yushu and Guoluo prefectures. Results Totally 19 629 children were investigated by the portable B ultrasound and 221 children were diagnosed as alveolar echinococcosis with the morbidity of 1.13%. Totally 9 888 were investigated by the indirect ELISA,and the positive rate of serum tests was 12.59%(190/9 888). The total morbidity of al?veolar echinococcosis of children in Yushu Prefecture was 0.42%(31/7 454)and it was 1.57%(190/12 175)in Guoluo Prefec?ture and there was a significant difference between them(χ2=53.42,P0.05). The total morbidity of alveolar echinococcosis of children in different age groups was from 1.00%to 1.13%,and there was no significant difference among the age groups(χ2=2.73,P>0.05). The P1a type,P1b type and P2 type accounted for 67.76%(145/214),10.75%(23/214)and 16.36%(35/214)respectively among the alveolar echinococcosis patients. The diameters of nidus of 87.38%(187/214)patients were less than 5 cm,and 2.80%(6/214)were more than 10 cm. Conclusions The epidemic status of alveolar echinococcosis of Tibetan children in south Qinghai Province is serious,but most of the patients were at early stage of the disease. The screening work of al?veolar echinococcosis should be strengthened in Tibetan children,so as to perform the early diagnosis and treatment.

Epidemiological characteristics of canine Echinococcus infection in Qinghai⁃ Tibet Plateau of China / 中国血吸虫病防治杂志

@#The Qinghai⁃Tibet Plateau is known as one of the highest endemic areas of echinococcosis However，the dog infection rates of Echinococcus granulosus in the plateau regions were similar to other non⁃Tibetan areas with the high endemic，and most of the rates were below 40%. The infected dogs with E. multilocularis were ubiquitous in Ganzi Prefecture of Sichuan Province and Qinghai Province where many survey data were done and available，which was much different from those in non⁃Ti⁃betan areas where the geographical distribution of dogs infected with E. multilocularis was sporadic. The total infection rates of Echinococcus in dogs kept stable from 1983 to 2009 in Ganzi Prefecture of Sichuan Province and did not show much variation from 2000 to 2014 in Qinghai Province as well. Since 2006，the national comprehensive prevention and control strategy and mea⁃ sures against echinococcosis have been launched in China，and significant progress has been made. In the endemic Tibetan area of Sichuan，the general Echinococcus infection rates in dogs were 28.10%，15.87%，19.22%，3.28% and 1.11% from 2009 to 2013，respectively，and the Echinococcus coproantigen⁃positive rate in Gannan Prefecture of Gansu Province and parts of Qing⁃ hai Province also decreased. This paper reviews the literature on the characteristics of dog infections in the Qinghai⁃Tibetan Plateau，so as to provide useful information to support echinococcosis control and prevention there.

Endangered situation and conservation strategy of Tibetan medicine in Qinghai-Tibet Plateau / 中国中药杂志

With the rapid development of Tibetan medicine industry, the study on plateau medicinal plants' endangered status is not enough, measures to protect is weak and the plateau ecological environment' inherent vulnerability, resulted in the shortage of Tibetan medicinal resources and affect the sustainable development . According to the existing endangered information of Tibetan medicine resources, how to formulate feasible protection plan, is an urgent problem of the rational development and utilization of Tibetan medicine resources to be solved. To find out the endangered Tibetan medicines in Qinghai Tibet Plateau, the Grade division method of Chinese Rare and Endangered Plants was applied, the endangered species were sorted out, which divided into class one (threatened) eleven species, class two (rare) twenty-one species, and class three (fading) forty-two species,a total of seventy-four species.In addition to national protection list in "Chinese rare and endangered plants". It's proposed to increase the endangered Tibetan medicinal species. Finally, according to the endangered status of the resources,from the survey of endangered Tibetan medicinal species regularly, the germplasm repository establishment of endangered Tibetan medicine, in situ conservation, artificial cultivation research and renew the idea, reasonable development and utilization, a total of 5 aspects to discussed the protection strategy, to provide a scientific basis for the protection and sustainable utilization of Tibetan medicine resources in Qinghai-Tibet Plateau.