Objective:To study the comparative genomic characteristics of Marmota himalayana-derived (referred to as marmota-derived) Brucella abortus (B.ab). Methods:The species and types of one strain of marmota-derived Brucella and one strain of human-derived Brucella isolated from the brucellosis epidemic area in Qinghai Province in the same year were identified. Meanwhile, DNA was extracted for whole genome sequencing and comparative genomics analysis (including phylogenetic tree construction, gene family clustering analysis, common/specific gene analysis, and genomic structural variation analysis, etc.). Results:Two Brucella strains from different hosts were identified as B.ab. By constructing a phylogenetic tree, the marmota-derived B.ab strain was grouped with strains from Heilongjiang Province and showed genetic correlation with strains from Russia. Human-derived B.ab strain was classified as a strain in Inner Mongolia Autonomous Region, Hebei Province, Beijing City, and Gansu Province. The multilocus sequence typing (MLST) of the two strains belonged to the ST2 type. Multiple locus variable-number tandem repeat analysis (MLVA) belonged to two new MLVA-8 and MLVA-11 genotypes, which were clustered in two subclusters of the same cluster and clustered with the strains from Inner Mongolia Autonomous Region, Xinjiang Uygur Autonomous Region, and Hebei Province. The pan-genome numbers of the marmota-derived B.ab and human-derived B.ab were 283 and 8, respectively; the number of core genes (common genes) was 68 and 2, respectively; and the number of unique genes was 3 and 4, respectively. The unique gene encoded proteins were inconsistent. In marmota-derived B.ab, the main ones were the ABC transporter ATP-binding protein, N-terminal acetyltransferase, and glucose/galactose transporter. The number of homologous genes of the marmota-derived B.ab and human-derived B.ab was 16 and 20, respectively; the number of translocation and inversion genes was 13 and 8, respectively; the number of deletion mutation genes was 11 and 14, respectively. Pathogenicity analysis showed that both strains had the mprF resistance gene, and the marmota-derived B.ab strain also carried bacitracin and macrolide resistance genes. Conclusions:Brucella exhibits cross-species genetic diversity. The proteins encoded by the unique genes of the marmota-derived B.ab mainly play a role in metabolic and epigenetic regulation. The strains cluster with B.ab strains from northern China, providing a reference for molecular epidemiology and pathogen tracing of B.ab infection.

Objective:To study the comparative genomic characteristics of Marmota himalayana-derived (referred to as marmota-derived) Brucella abortus (B.ab). Methods:The species and types of one strain of marmota-derived Brucella and one strain of human-derived Brucella isolated from the brucellosis epidemic area in Qinghai Province in the same year were identified. Meanwhile, DNA was extracted for whole genome sequencing and comparative genomics analysis (including phylogenetic tree construction, gene family clustering analysis, common/specific gene analysis, and genomic structural variation analysis, etc.). Results:Two Brucella strains from different hosts were identified as B.ab. By constructing a phylogenetic tree, the marmota-derived B.ab strain was grouped with strains from Heilongjiang Province and showed genetic correlation with strains from Russia. Human-derived B.ab strain was classified as a strain in Inner Mongolia Autonomous Region, Hebei Province, Beijing City, and Gansu Province. The multilocus sequence typing (MLST) of the two strains belonged to the ST2 type. Multiple locus variable-number tandem repeat analysis (MLVA) belonged to two new MLVA-8 and MLVA-11 genotypes, which were clustered in two subclusters of the same cluster and clustered with the strains from Inner Mongolia Autonomous Region, Xinjiang Uygur Autonomous Region, and Hebei Province. The pan-genome numbers of the marmota-derived B.ab and human-derived B.ab were 283 and 8, respectively; the number of core genes (common genes) was 68 and 2, respectively; and the number of unique genes was 3 and 4, respectively. The unique gene encoded proteins were inconsistent. In marmota-derived B.ab, the main ones were the ABC transporter ATP-binding protein, N-terminal acetyltransferase, and glucose/galactose transporter. The number of homologous genes of the marmota-derived B.ab and human-derived B.ab was 16 and 20, respectively; the number of translocation and inversion genes was 13 and 8, respectively; the number of deletion mutation genes was 11 and 14, respectively. Pathogenicity analysis showed that both strains had the mprF resistance gene, and the marmota-derived B.ab strain also carried bacitracin and macrolide resistance genes. Conclusions:Brucella exhibits cross-species genetic diversity. The proteins encoded by the unique genes of the marmota-derived B.ab mainly play a role in metabolic and epigenetic regulation. The strains cluster with B.ab strains from northern China, providing a reference for molecular epidemiology and pathogen tracing of B.ab infection.

Objective:To analyze the surveillance results of human brucellosis in Qinghai Province, to study the epidemic characteristics of brucellosis, and to provide scientific basis for formulating brucellosis prevention and control strategies and measures.Methods:By using descriptive epidemiological method, surveillance data from 4 national and 26 provincial brucellosis surveillance sites in Qinghai Province in 2022 and brucellosis related information from the "Disease Surveillance Information Reporting and Management System" in 2022 were collected and summarized. The population, time, regional distribution and epidemic characteristics of brucellosis in Qinghai Province were analyzed.Results:In 2022, a total of 12 483 people were monitored at 4 national and 26 provincial brucellosis surveillance sites. In Rose Bengal plate agglutination test (RBPT), 714 patients were positive, and the positive rate was 5.72% (714/12 483). In tube agglutination test (SAT), 508 individuals tested positive, and the positive rate was 4.07% (508/12 483). There were 1 156 reported cases in 2022, including 910 males and 246 females, with a gender ratio of 3.70 ∶ 1.00. The age range was mainly between 30 and 59 years old, accounting for 77.68% (898/1 156). Most of them were farmers engaged in breeding and transportation, accounting for 81.40% (941/1 156). The onset time was concentrated from June to August, accounting for 50.26% (581/1 156). The distribution area was mainly in the northeast of Qinghai Province (Haibei Tibetan Autonomous Prefecture, Xining City, Haidong City), accounting for 86.33% (998/1 156).Conclusions:The epidemic of human brucellosis in Qinghai Province is still relatively serious, especially in the northeastern agricultural area. Comprehensive measures should be taken to further curb the outbreak of human brucellosis.

Objective:To analyze the surveillance results of human brucellosis in Qinghai Province, to study the epidemic characteristics of brucellosis, and to provide scientific basis for formulating brucellosis prevention and control strategies and measures.Methods:By using descriptive epidemiological method, surveillance data from 4 national and 26 provincial brucellosis surveillance sites in Qinghai Province in 2022 and brucellosis related information from the "Disease Surveillance Information Reporting and Management System" in 2022 were collected and summarized. The population, time, regional distribution and epidemic characteristics of brucellosis in Qinghai Province were analyzed.Results:In 2022, a total of 12 483 people were monitored at 4 national and 26 provincial brucellosis surveillance sites. In Rose Bengal plate agglutination test (RBPT), 714 patients were positive, and the positive rate was 5.72% (714/12 483). In tube agglutination test (SAT), 508 individuals tested positive, and the positive rate was 4.07% (508/12 483). There were 1 156 reported cases in 2022, including 910 males and 246 females, with a gender ratio of 3.70 ∶ 1.00. The age range was mainly between 30 and 59 years old, accounting for 77.68% (898/1 156). Most of them were farmers engaged in breeding and transportation, accounting for 81.40% (941/1 156). The onset time was concentrated from June to August, accounting for 50.26% (581/1 156). The distribution area was mainly in the northeast of Qinghai Province (Haibei Tibetan Autonomous Prefecture, Xining City, Haidong City), accounting for 86.33% (998/1 156).Conclusions:The epidemic of human brucellosis in Qinghai Province is still relatively serious, especially in the northeastern agricultural area. Comprehensive measures should be taken to further curb the outbreak of human brucellosis.

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.

Objective:To learn about the epidemic situation and trend of human brucellosis in Menyuan Hui Autonomous County (referred to as Menyuan County) of Qinghai Province, and to provide reference for formulating brucellosis prevention and control measures in Menyuan County.Methods:Data on human brucellosis in Menyuan County reported by the Infectious Disease Reporting Information Management System of China Disease Control and Prevention Information System from 2013 to 2020 were collected and analyzed by descriptive epidemiological analysis (three distribution).Results:A total of 186 cases of brucellosis were confirmed from 2013 to 2020, with an average annual incidence of 14.553/100 000. The annual incidence rate was increasing year by year(χ 2trend = 22.08, P = 0.002). The cases were distributed in 67 villages of 12 towns; cases were more common in the age group of 15-< 65 years old (96.24%, 179/186), and there were more men than women cases (sex ratio was 3.89∶1.00, 148/38). Conclusions:The incidence rate of brucellosis cases in Menyuan County is increasing year by year, and the scope of its impact is constantly expanding. A multi-sectoral joint prevention and control mechanism should be established to strengthen management, increase the prevention and control of brucellosis, and stop the spread of the epidemic to surrounding counties and cities.

Objective:To master the epidemic trend of human brucellosis in Qinghai Province, so as to provide basis for scientific prevention and control of the disease.Methods:In 2019 and 2020, at the national and provincial brucellosis monitoring sites in Qinghai Province, a total of 18 counties (cities and districts, hereinafter referred to as counties), no less than 400 serum samples were sampled every year for brucellosis Rose-Bengal plate agglutination test (RBPT) and serum tube agglutination test (SAT), which would be tested and judged according to the criteria of "Diagnosis for Brucellosis" (WS 269-2019).Results:In 2019, a total of 1 612 people were monitored in national brucellosis monitoring sites, 93 were RBPT positive, 54 were SAT positive, 54 were diagnosed, and the prevalence rate was 3.35% (54/1 612). In 2020, 1 677 people were monitored in national brucellosis monitoring sites, 151 were RBPT positive, 80 were SAT positive, 80 were diagnosed, and the prevalence rate was 4.77% (80/1 677). There were significant differences in RBPT positive rate, SAT positive rate and prevalence rate among national monitoring sites between the two years (χ 2 = 12.52, 4.24, 4.24, P < 0.05). In 2019, a total of 6 043 people were monitored in provincial brucellosis monitoring sites, 128 were RBPT positive, 91 were SAT positive, 87 were diagnosed, and the prevalence rate was 1.44% (87/6 043). In 2020, 5 664 people were monitored, 108 were RBPT positive, 59 were SAT positive, 52 were diagnosed, and the prevalence rate was 0.92% (52/5 664). There was no significant difference in RBPT positive rate among provincial monitoring sites between the two years (χ 2 = 0.66, P = 0.416), and the differences in SAT positive rate and prevalence rate were statistically significant among provincial monitoring sites between the two years (χ 2 = 4.98, 14.57, P < 0.05). Conclusion:In 2019 and 2020, there are human brucellosis in national and provincial brucellosis monitoring sites in Qinghai Province.

Objective:To analyze the causes and epidemiological characteristics of an outbreak of brucellosis in Haidong City Qinghai Province, and provide a scientific basis for prevention and control of brucellosis.Methods:Using the retrospective research method, the epidemic data on brucellosis, field case investigations, serological test results in Haidong City, Qinghai Province in 2019 were collected from the "China Disease Prevention and Control Information System", and the above data were analyzed by descriptive epidemiological analysis.Results:There were 3 confirmed brucellosis patients, who were breeders of the farm; 1 case of latent infection was found in key population, who was villager from the same village as the confirmed patients; 4 people had a history of close contact with infected sheep. A total of 110 sheep serum and 7 bovine serum samples were collected in the animal breeding area, of which 26 were positive, all were sheep serum; 568 sheep serum and 17 bovine serum samples in the same village were negative.Conclusions:The key occupational groups contacted with infected sheep without protection is the main cause of this epidemic. Health education on brucellosis prevention and control should be vigorously carried out to improve the awareness of the people to prevent the disease; the monitoring of animal brucellosis by relevant departments should be strengthened to effectively control the spread of animal brucellosis, and prevent the occurrence of human brucellosis.

Objective:Combined with the actual work requirements of brucellosis in Qinghai Province, to design a management information system for brucellosis patients, in order to realize the information of diagnosis and treatment of brucellosis patients.Methods:Based on the theory of management information system, according to the functional requirements of the actual work and performance parameters, the general framework of management information system was designed, the establishment of the management information system was gradually completed, and the system was improved through the system testing.Results:The system was easy to use and easy to operate, and patient's detailed information, diagnosis and treatment results and treatment plan could be obtained quickly. At the same time, follow-up, trace investigation, and brucellosis prevention and treatment propaganda could be carried out.Conclusions:The established brucellosis management information system can realize the electronic management of patient information, master the disease development and diagnosis and treatment results of brucellosis patients. Through follow-up investigation, the system can increase active screening and early warning of suspected cases, and provide health education to realize early diagnosis and early treatment of the patients.

Objective:To detect cartilage oligomeric matrix protein (COMP) and C-terminal telopeptide of collagen type Ⅱ (CTX-Ⅱ) among brucellosis patients at early period, and provide a better understanding of the osteoarticular changes in this disease at early stage.Methods:Using case-control research method, the early brucellosis patients who visited the Department for Brucellosis Disease Prevention and Control, Qinghai Institute for Endemic Disease Prevention and Control from 2013 to 2016 were selected as the case group; through the principle of random matching, the healthy physical examination population was set as the control group. Serum COMP and CTX-Ⅱ were detected by enzyme-linked immunosorbent assay.Results:A total of 128 patients were investigated, including 61 patients (41 males and 20 females) with early brucellosis, aged (39.41 ± 10.51) years old, and 67 controls (44 males and 23 females), aged (40.31 ± 12.52) years old. The median of serum COMP in the case group was 1.70 μg/L, which was significantly higher than that in the control group (0.79 μg/L). The difference between the two groups was statistically significant ( Z = 3.614, P < 0.05); the median of serum CTX-Ⅱ in the case group was 116.44 pg/ml, and the median of serum CTX-Ⅱ in the control group was 128.48 pg/ml, and the difference was not statistically significant ( Z = 0.044, P > 0.05). Conclusion:The serum COMP levels of brucellosis patients at early period are increased, indicating osteoarticular changes of human brucellosis at early stage.