Objective To identify rodent residues using DNA barcoding technology in plague areas of Shaanxi and to analyze the feasibility of DNA barcoding method.Methods DNA extraction,PCR,electrophoresis and sequence determination and alignment were used to determine the cytochrome C oxidase Ⅰ (COI) gene sequence from 62 residues of 7 species identified by morphology in 8 different parts.COI gene sequence was analyzed using BLAST software of American National Center for Biotechnology Information (NCBI) for sequence homology comparison and the phylogenetic tree was constructed by using the neighbor joining(NJ)method of COI gene sequence.Results In addition to the hair,CO I genes of the feet,the tail,the fur,the muscle,the ribs,the ear,and the eye were amplified,respectively,and the size of amplified fragment was similar to the size of the product with an expected fragment(700 bp),and the fragment was a single band.COI genes of 59 residue specimen were obtained by DNA sequencing and there were high degree of homologies between CO I gene sequences of Meriones unguiculatus,Cricetulus barabensis,Meriones meridianus,Dipus sagitta,Phodopus roborovskii,Rattus norvegicus and Allactaga sibirica and their corresponding host genes in NCBI(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ method showed that 7 NJ phylogenetic trees were constructed with COI sequence of 59 species residues.The CO I sequences of same rodent with different residues were clustered into one group.Condusion DNA barcoding technology can identify host animal residues in plague areas,and the identification results are reliable.

We studied the genetic characteristics of Spermophilus in Shaanxi Province,China.The COI,Cyt-b gene were sequenced and the results were compared with those of dauricus from Inner Mongolia Keyouzhong Banner and Zhengxiangbai Banner,and S.alaschanicus from Haiyuan County of Ningxia.And genetic distance was analyzed and Neighbor-Joining tree was built.Results showed that the genetic distance of COI gene sequences between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤0.5％,and the genetic distance was ranged from 7.9％ to 9.3％ with Citellus dauricus from Inner Mongolia.The genetic distance of Cyt-b gene between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤2.2％,and ranged from 8.9％ to 11.2％ with Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI,Cyt-b gene showed two major clusters.One of them were clustered by Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia,and another one was Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI gene showed that all samples from Shaanxi Province clustered in a group.In conclusion,the Spermophilus in Shaanxi Province were S.alaschanicus.

Objective To apply DNA barcoding technology for exploring its taxonomic status and differences in the molecular biology of Cricetulus barabensis in Shaanxi Province.Methods Sixty-five samples of Cricetulus barabensis were collected from Dingbian,Jingbian Counties in northern of Shaanxi and Dali County in Guanzhong plain (Dingbian 58 samples,Jingbian 2 samples,and Dali 5 samples).According to the mitochondrial cytochrome C oxidase subunit I gene (CO I) sequence,the genetic distance was calculated and Neighbor-Joining tree was constructed.Results The genetic distance between two samples (13.16,13.21) and other 56 samples of Dingbian was 9.2％-10.0％.The genetic distance between the 56 samples of Dingbian and Jingbian was less than 1％ and Dali was 7.2％-8.3％;the average intraspecific genetic distance of Jingbian and Dali was less than 1％.The Neighbor-Joining tree showed that all the Cricetulus barabensis samples from the three counties were separated into two large branches.The samples of 13.16,13.21 from Dingbian together were classified into a class and the rest of the samples into another separate branch.At the same time,other samples from Dingbian except 13.16,13.21 and Jingbian were distributed in a small branch,and Dali samples were occupied another small branch.Conclusion Using the DNA barcoding technology,we have determined three subspecies of Cricetulus barabensis in Shaanxi Province,Dingbian has two kinds and Dali has a different subspecies.

Objective:To analyze the epidemiological characteristics of public health emergencies caused by human brucellosis in Shaanxi Province in 2020, and to provide scientific basis for prevention and control of human brucellosis in Shaanxi Province.Methods:Descriptive epidemiological analysis of public health emergencies caused by human brucellosis in Shaanxi Province in 2020 was carried out, and the handling of the epidemic was studied.Results:A total of 13 public health emergencies caused by human brucellosis were reported in Shaanxi Province in 2020, 3 in northern Shaanxi, 10 in Guanzhong area, involving 65 confirmed cases of human brucellosis. Gender distribution: 47 males and 18 females; occupational distribution: 35 farmers, 13 breeders, 8 students, 6 scattered children, 2 workers, and 1 child in kindergarten; age distribution: the age of onset was mainly 50 to 69 years old (30 cases); time distribution: 13 public health emergencies occurred in April to August, with the peak in May; route of infection: 11 cases were infected by occupational exposure, 1 case was infected by drinking raw goat milk, and 1 case was infected by contacting the dogs infected by sick sheep. And 13 public health emergencies were handled in a timely and effective manner, including incident reporting, epidemiological investigation, high-risk population screening, pathogen cultivation, animal epidemic disposal, disinfection, publicity and education, etc.Conclusion:The brucellosis epidemic is active in the old epidemic areas in northern Shaanxi and the new epidemic areas in Guanzhong area, Shaanxi Province, and targeted prevention and control measures should be formulated according to the epidemiological characteristics of the epidemic to ensure accurate prevention and control of human brucellosis.

Objective:To construct a follow-up evaluation index system for "Diagnosis of Brucellosis" (WS 269-2019), and provide a reference basis for the next revision and improvement of the standard.Methods:The evaluation index system for "Diagnosis of Brucellosis" (WS 269-2019) was preliminarily established by consulting relevant references and materials. The experts in the field of diagnosis, treatment, prevention and control of brucellosis were selected, and two rounds of expert consultation were carried out in the form of questionnaires using the Delphi method. The necessity and availability of evaluation indicators were scored, and suggestions for modifying and adding indicators were put forward. Based on this, a standard follow-up evaluation index system was established. At the same time, a judgment matrix was constructed combined with the Saaty scale, and the analytic hierarchy process was used to calculate the weight of each index in the system.Results:After 2 rounds of expert ( n = 10) consultation, a standard follow-up evaluation index system for "Diagnosis of Brucellosis" (WS 269-2019) was constructed with 3 first-level indexes, 8 second-level indexes and 21 third-level indexes. The positive coefficients of experts in 2 rounds of questionnaires were both 100%; the coefficient of authority of experts was 0.82; the Kendall's coefficients of concordance of first-level, second-level and third-level indexes were 0.722, 0.260, and 0.181, respectively, with P < 0.05. Among the first-level indexes, the weight of standard quality evaluation was the highest (0.364), and the weight of standard implementation status was the lowest (0.278); among the second-level indexes, the combined weight of social benefits was the highest (0.186), and the combined weight of advanced nature was the lowest (0.043); among the third-level indexes, the combined weight of timely diagnosis rate was the highest (0.096), and the combined weight of consistency with technical data was the lowest (0.009). Conclusions:The constructed follow-up evaluation index system for "Diagnosis of Brucellosis" (WS 269-2019) is scientific and reliable, which evaluated qualitatively and quantitatively, reduces the defects of a single evaluation, and provides a basis for subsequent revision and improvement of the standard.

Objective:To analyze the epidemiological characteristics and related indicators of plague in Yulin City, Shaanxi Province, and to evaluate the risk of plague epidemic in the future and formulate scientific and reasonable prevention and control measures.Methods:The plague surveillance data of national (Dingbian County) and provincial (Yuyang District, Jingbian County, Hengshan District, Shenmu City, Fugu County) plague monitoring sites in Yulin City from 2011 to 2021 were collected through the China Disease Prevention and Control Information System Pestis Prevention and Control Management Information System, and the plague epidemic situation among humans and animals, the main host animals and their flea infection were analyzed by descriptive epidemiological methods. The expert consultation method and plague risk assessment tool V1.0 were used to assess the epidemic risk of the plague in Yulin City.Results:There was no human and animal plague epidemic in Yulin City from 2011 to 2021. In national monitoring site, the density of main host animal was 5.79/hm 2, ranging from 3.02/hm 2 to 9.08/hm 2,the dominant species was Mongolian gerbil, accounting for 98.21% (3 402/3 464); the capture rate of wild nocturnal rodents was 1.27% (350/27 600), ranging from 0.21% to 3.83%, the dominant species was Cricetulus barabensis, accounting for 44.86% (157/350); the flea infection rate of the rodent body was 16.91% (768/4 541), with a flea index of 0.40, the dominant flea species was the Nosopsyllus laeviceps kuzenkov, accounting for 66.54% (1 203/1 808). In provincial monitoring site, the density of main host animals was 0.49/hm 2, ranging from 0.31/hm 2 to 0.67/hm 2, the dominant species was Alashan ground squirrel, accounting for 63.61% (194/305); the capture rate of nocturnal rodents in the wild was 1.76% (560/31 795), ranging from 0.89% to 3.93%, the dominant species was Roborovski dwarf hamster, accounting for 26.61% (149/560); the capture rate of domestic rats was 2.37% (397/16 750), ranging from 1.48% to 3.10%, the rodents included Rattus norvegicus (47.36%, 188/397) and house mouse (52.64%, 209/397); the flea infection rate of the rat body was 13.26% (182/1 373), with a flea index of 0.40; the dominant flea species was Ophthalmopsylla jettmari, accounting for 45.23% (251/555). The 5 281 pathogenic culture samples and 2 110 serological test samples were all negative. The Pearson correlation analysis results showed that there was no correlation between rodent density and flea infection rate or flea index at national and provincial monitoring sites ( r = - 0.26, - 0.48, 0.09, 0.12, P > 0.05), while flea infection rate and flea index were positively correlated ( r = 0.67, 0.81, P < 0.05). In 2022, Yulin City, Shaanxi Province was not ruled out the possibility of human plague epidemic, and the risk of plague epidemic among animals was high. Conclusions:From 2011 to 2021, the density of the main host animals at the national and provincial monitoring sites in Yulin City has remained at a low level, and the flea index has increased. There is a risk of plague epidemic in Yulin City, so the monitoring work should be further strengthened, and emergency supplies and capacity reserves should be well prepared.

Objective To apply the DNA barcoding technology for identification on host animal and to establish the host animal DNA bar code database on natural foci of plague in Shaanxi.Methods 139 host animals belonging to 3 orders,6 families and 12 genera and 62 residues belonging to 7 species from 8 different parts of the province,were detected.DNA barcoding technology was used to analyze the DNA CO Ⅰ gene sequence on the natural foci of plague in Dingbian county.Results The intra-specific genetic distance was less than 2％ while the inter-specific distance ranged from 8.9％ to 15.1％.Fourteen major clusters were apparently showed on a Neighbor-Joining tree.Residue samples could be detected regarding the objective gene.Alashan ground squirrel was previously noticed to carry 14 major clusters,which were previously mistakenly named as Citellus dauricus in Dingbian county.Conclusion DNA barcoding technology could overcome the shortcomings caused by the morphological identification so could be used to identify the host animal and residues in the natural focus of plague.

Objective To investigate the ability in diagnosing and reporting brucellosis of primary medical personnel in Suide County of Shaanxi Province,and provide scientific basis for making prevention and control strategies.Methods Convenience sampling method was adopted in October 2016.A questionnaire survey was conducted among the county hospitals,township hospitals,village clinics and individual clinics in Suide County to collect information on the ability in diagnosing and reporting brucellosis of primary medical personnel.The specific content includes the basic information,the awareness of brucellosis diagnosis and reporting capabilities,the situation of brucellosis training,and the views about the diagnosis of brucellosis.SPSS 18.0 software was used to carry out descriptive statistical analysis.Results A total of 77 primary healthcare workers were surveyed.Their average age was (38 ± 9) years old,57.1％ (44/77) workers were undergraduates,and 53.2％ (41/77) were working in county hospitals.9.1％ (7/77) were completely aware of the major infectious diseases,29.9％ (23/77) were fully aware of the transmission mute,28.6％ (22/77) and 66.2％ (51/77) known the high risk population and the epidemiological history of brucellosis.Twenty-six percent (20/77) of the medical staff were thoroughly aware of the main clinical manifestations,68.8％ (53/77) were clearly aware of the clinical stage of brucellosis;24.7％ (19/77) could understand the specificity of the laboratory tests of brucellosis;44.2％ (34/77) had a strong ability in reporting brucellosis.71.4％ (55/77) of the medical staff received the training about the knowledge of brucellosis;49.4％ (38/77) thought that the clinical manifestation was the most difficult to grasp in the criteria about diagnosis of brucellosis.Conclusions The ability of diagnosis and reporting of brucellosis is weak in primary healthcare workers in Suide County,the epidemiology and clinical knowledge of brucellosis are not comprehensive.It is suggested to strengthen the understanding of brucellosis among medical staff,especially in primary healthcare workers.We should carry out the training of disease epidemiology and clinical knowledge of brucellosis and standardize patient management.

Objective To learn the diagnosis and treatment of brucellosis patients in Suide County of Shaanxi Province in 2015,and to provide a scientific basis for making accurate prevention and control measures.Methods All the cases diagnosed as brucellosis in Suide County from January 1 to December 31 in 2015 and lived in this county were retrospectively investigated by case questionnaire survey,including basic information,medical procedures,and epidemiological contacts.Descriptive statistical analysis was performed using SPSS 18.0 software.Results In 2015,75 cases of brucellosis were diagnosed in Suide County,75 questionnaires were distributed,with 53 completed questionnaires returned.The average age of respondents was (49 ± 14) years old,of which 84.9％ (45/53) were male and 94.3％ (50/53) were farmers,except one case was actively monitored by Suide County Centre for Disease Control and Prevention then treated,the rest of the patients were treated after the onset of the disease.Of the 52 patients,one patient was diagnosed after one visit,accounting for 25.0％ (13/52);one patient was diagnosed after at least 4 visits,accounting for 3.8％ (2/52).The cumulative diagnosis rate at provincial-level hospitals was 1/3 and that at disease prevention and control institutions was 92.7％ (51/55),there were no confirmed cases in municipal hospitals,county hospitals,township hospitals,village clinics and individual clinics.For the first reason to see a doctor,65.4％ (34/52) of the patients were fever or accompanied by sweating,fatigue,arthralgia and waist and leg pain.The medians from onset to treatment between the first diagnosis,and 2,3,4 visits were 15,18,27,45 days,respectively;the median from onset to diagnosis was 21 days,ranging from 2 to 182 days.Totally 84.9％ (45/53) patients had a history of exposure to animals,of which 97.8％ (44/45) had contacted the sheep.Conclusions The cases in Suide County are mainly adult male farmers,and the diagnosis rate at hospitals below municipal level is low.It is recommended to strengthen the health intervention of high-risk groups and improve the level of diagnosis and treatment of primary medical staff.

Objective To introduce the concept of health promotion to brucellosis comprehensive prevention and control and then the effect was evaluated.We expect this article could provide scientific basis for prevention and control of brucellosis.Methods The study began in March 2014,a high-incidence district of brucellosis in Dali County was selected.Our study populations were divided to primary target groups and secondary target groups by the extent of exposure to sheep.We implemented general health campaigns,targeted health education,intervention and inducement,promotion of establish leading groups,and released relevant policies and rules for target population.According to the information from the questionnaire survey designed by us,before and after the intervention,the difference between the awareness rate of knowledge on brucellosis control,the health behavior formation rate and the effects of different intervention measures in primary target groups,were compared.The correct diagnostic rate in time (in 3 months),the eligible rate of trained health works,the change of the incidence of brucellosis in human cases,the perfection of the relevant policies on brucellosis control were analyzed,and the effectiveness of comprehensive prevention and control and health promotion strategies were evaluated.Results The disease awareness rate of target audience was 62.49％ (12 695/20 315) after the intervention which was 20.23％ (3 746/18 517) before the intervention.The awareness rate of primary target groups and secondary target groups was increased by 39.69％ [before intervention:33.75％ (1 624/4 812),after intervention:73.44％ (3 569/4 860)] and 43.57％ [before intervention:15.48％ (2 122/13 705),after intervention:59.05％ (9 126/15 455)],respectively.South of Luohe and north of Luohe increased by 37.70％ [before intervention:36.81％ (731/1 986),after intervention:74.51％ (1 453/1 950)] and 35.86％ [before intervention:35.06％ (711/2 028),after intervention:70.92％ (1 366/1 926)] after the intervention.The success rate of prevention and control skill on brucellosis in medical staff was 88.52％ (594/671);the correct diagnostic rate of new cases was 93.73％ (370/383).The proportion of new cases in the selected county in 2014 was 18.26％ (269/1 473) of the whole Shaanxi Province,the ratio decreased compared to that of the previous year.The proportion of new cases in 2015 was 6.14％ (75/1 221).The proportion of new cases in 2016 was 3.48％ (33/948).Conclusions Through the implementation of health promotion,health education and health promotion comprehensive measures,the disease awareness rate in target population has increased substantially.These measures are also able to lead focused population to develop health behavior and to decrease the incidence of brucellosis among people,and to realize the sustainable control of human brucellosis.