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Objective:To analyze the plague monitoring results of Marmota sibirica plague natural focus in Hulunbuir Plateau, and to study the characteristics of development and variation of the plague focus. Methods:From 2009 to 2020, monitoring was conducted in Manzhouli City, Prairie Chenbarhu Banner, Xin Barag Left Banner, and Xin Barag Right Banner from May to September each year. The one-day bow-clip method was used to investigate the density of the Spermophilus dauricus, and the 5-meter clamp method was used to investigate other small rodents. The classification and identification of ectoparasite fleas were done by using low power microscope. According to "Diagnostic Criteria for Plague" (WS 279-2008), the detectable materials of rats and fleas were tested, and Yersinia pestis was isolated and cultured. The serums of collected rats were detected by indirect hemagglutination test. Results:From 2009 to 2020, the average density of dauricus in Marmota sibirica plague natural focus of Hulunbuir Plateau was 0.45/hm 2, and the capture rate of other small rodents was 1.25% (787/63 091). A total of 9 species, 8 genera and 4 families of rodents were captured. The total flea infection rate of Spermophilus dauricus was 28.37% (1 945/6 856) and the fleas index was 1.04. A total of 3 species, 3 genera and 2 families of fleas were collected. A total of 6 674 Spermophilus dauricus were dissected and 6 787 fleas were cultured in 2 537 groups. The Yersinia pestis test was negative. A total of 6 795 serum samples from Spermophilus dauricus were detected, of which 11 were positive with a positive detection rate of 0.16%. The highest titer of positive serum was 1∶1 280. Indirect hemagglutination test positive materials were found for 5 consecutive years from 2010 to 2014; the positive serums were found in Manzhouli City and Xin Barag Right Banner. Conclusions:The Yersinia pestis antibady positive serum of Spermophilus dauricus has been detected in Marmota sibirica plague focus of Hulunbuir Plateau for many years, which is worthy of high vigilance. In the future, continuous surveillance should be carried out, bacterial detection should be strengthened, possible plague outbreaks should be detected in time, and various plague prevention and control should be done well.
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@#Abstract: Objective To detect and analyze the antiserum of Yersinia pestis phage in Marmota himalayana blood from the natural plague foci of Qinghai-Tibet Plateau by micro-bolus technique, to provide a theoretical basis for interaction between phages and mammalian immunology, phage therapy and interaction between bacteriophage and ecology in future. Methods Using diagnostic Yersinia pestis phage and 3 wild plague phages from Qinghai-Tibet Plateau Natural Plague Foci as antigens, 847 serums of Marmota Himalayana blood, from Tongde, Guinan, Gonghe, Xinghai, Tianjun foci counties in Qinghai Plateau, were collected from July to September in 2020, 2021 and determined on antiserum of Yersinia pestis phage by microplate method and double agar plate method. Results The neutralization reaction experiment lasted for 24 hours between 4 phage and 847 serums by microplate method independently. These mixtures were tested by double agar plate method. All results were negative on antiserum of Yersinia pestis bacteriophage. Conclusions The positive antiserum of Yersinia pestis phage in Marmota himalayana were not found the natural plague foci of Qinghai-Tibet Plateau, which agreed with plague epidemiology in 5 foci counties in Qinghai plateau from 2020-2021, that was a characteristic of the resting period. In other words, it was in the absence of plague pathogen. It also showed indirectly that the absence or weak presence of Yersinia pestis bacteriophage in the plague foci. It showed a lower frequency on host animals coming into contact with phages naturally. The antiserum of Yersinia pestis phage may be related to the form of plague infection and the intensity of the disease.
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Antiviral nucleoside drugs represented by zidovudine, lamivudine, telbivudine, ribavirin, etc. have become the first choice drugs for clinical treatment of viral diseases such as AIDS, hepatitis B, herpes. However, the above drugs not only can inhibit virus replication, they can also interfere with the host mitochondrial mtDNA replication through similar mechanisms, leading to mitochondrial toxicity-related adverse reactions. Therefore, both the routine safety evaluation and mitochondrial toxicity evaluation are required for the development of nucleoside drugs. The research progress of mitochondrial toxicity evaluation models of nucleoside drugs are summarized for reference.
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Objective To investigate the correlation of meteorological factors with the numbers of marmots and parasitic fleas in the plague foci of Himalayan marmot in Subei County and Sunan County of Gansu Province.Methods The surveillance data (from Gansu Provincial Center for Disease Control and Prevention) of natural plague foci of Subei,Sunan of Himalayan marmot from 1982 to 2016 and the meteorological factors data (from China Meteorological Data Sharing Service Network) such as annual average temperature,rainfall and relative humidity during the same period were collected.The correlation between marmot density,body flea index and meteorological factors were analyzed by SAS 9.3 software.Based on the meteorological data as an independent variable in current year and previous 1,2,and 3 years (relative to previous 1,2,and 3 years as the reference),the marmot density as the dependent variable,the time series semiparametric generalized additive model (GAM) was established to quantitatively study the nonlinear effect of meteorological factors on the marmot density by R2.3.2 software.Results From 1982 to 2016,the marmot density in Subei County of Gansu Province was 0.29-6.63 per hectare,and the body flea index range was 0.04-4.56;the marmot density in Sunan County of Gansu Province was 1.22-7.92 per hectare,and the body flea index range was 0.06-0.42.In Subei County,the marmot density was correlated with the previous 1 year rainfall and relative humidity,respectively (r =0.43,0.36,P＜ 0.05),and negatively correlated with the average temperature of the year and the previous 1,3 years (r =-0.58,-0.55,-0.41,P ＜ 0.05).In Sunan County,the marmot density was correlated with the relative humidity of the year and the previous 1 year rainfall and relative humidity,respectively (r =0.36,0.57,0.43,P ＜ 0.05).In Subei County,the body flea index was negatively correlated with the average temperature of the year and the previous 1,2,3 years,respectively (r =-0.57,-0.43,-0.31,-0.37,P ＜ 0.05).In Sunan County,there was a negative correlation between the body flea index and the average temperature of the year and the previous 1,2,3 years (r =-0.71,-0.46,-0.38,-0.40,P ＜ 0.05).After controlling other influencing factors,the results of the GAM showed that for every 1 mm increase in rainfall of the year in Subei County,the marmot density increased 0.93％;for every 1％ increase in relative humidity of the previous 3 year in Subei County,the marmot density increased 25.34％;for every 1 mm increase in rainfall of the previous 3 year in Sunan County,the marmot density increased 1.69％.Conclusions The numbers of marmots in the plague foci of Himalayan marmot in Subei County and Sunan County of Gansu Province are closely related to rainfall and relative humidity,and the influence has different hysteresis effects.Surface temperature is a important factor affecting the population of parasitic fleas.
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Objective@#To investigate the correlation of meteorological factors with the numbers of marmots and parasitic fleas in the plague foci of Himalayan marmot in Subei County and Sunan County of Gansu Province.@*Methods@#The surveillance data(from Gansu Provincial Center for Disease Control and Prevention) of natural plague foci of Subei, Sunan of Himalayan marmot from 1982 to 2016 and the meteorological factors data (from China Meteorological Data Sharing Service Network) such as annual average temperature, rainfall and relative humidity during the same period were collected. The correlation between marmot density, body flea index and meteorological factors were analyzed by SAS 9.3 software. Based on the meteorological data as an independent variable in current year and previous 1, 2, and 3 years (relative to previous 1, 2, and 3 years as the reference), the marmot density as the dependent variable, the time series semiparametric generalized additive model (GAM) was established to quantitatively study the nonlinear effect of meteorological factors on the marmot density by R2.3.2 software.@*Results@#From 1982 to 2016, the marmot density in Subei County of Gansu Province was 0.29-6.63 per hectare, and the body flea index range was 0.04-4.56; the marmot density in Sunan County of Gansu Province was 1.22-7.92 per hectare, and the body flea index range was 0.06-0.42. In Subei County, the marmot density was correlated with the previous 1 year rainfall and relative humidity, respectively (r= 0.43, 0.36, P < 0.05), and negatively correlated with the average temperature of the year and the previous 1, 3 years (r=-0.58,-0.55,-0.41, P < 0.05). In Sunan County, the marmot density was correlated with the relative humidity of the year and the previous 1 year rainfall and relative humidity, respectively (r= 0.36, 0.57, 0.43, P < 0.05). In Subei County, the body flea index was negatively correlated with the average temperature of the year and the previous 1, 2, 3 years, respectively(r=-0.57,-0.43,-0.31,-0.37, P < 0.05). In Sunan County, there was a negative correlation between the body flea index and the average temperature of the year and the previous 1, 2, 3 years (r=-0.71,-0.46,-0.38,-0.40, P < 0.05). After controlling other influencing factors, the results of the GAM showed that for every 1 mm increase in rainfall of the year in Subei County, the marmot density increased 0.93%; for every 1% increase in relative humidity of the previous 3 year in Subei County, the marmot density increased 25.34%; for every 1 mm increase in rainfall of the previous 3 year in Sunan County, the marmot density increased 1.69%.@*Conclusions@#The numbers of marmots in the plague foci of Himalayan marmot in Subei County and Sunan County of Gansu Province are closely related to rainfall and relative humidity, and the influence has different hysteresis effects. Surface temperature is a important factor affecting the population of parasitic fleas.
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Objective To analyze the relationship between meteorological variables and the prevalence of marmot plague in the Himalayan marmot foci of Subei County,Gansu Province.Methods The surveillance data (from Gansu Provincial Center for Disease Control and Prevention) on human plague,the meteorological factors (average rainfall,temperature,relative humidity) data (from China Meteorological Data Sharing Service Network) of the current and preceding (relative to current year as a reference) first,second and third years and meteorological factors data of the current and preceding first,second and third months in Subei of Himalayan marmot foci from 1973 to 2014 were collected.The association of positive rate of plague bacteriological detection and meteorological factors was evaluated by using SAS 9.3 software.The influence of meteorological factors on the epidemic of marmot plague was analyzed by the logistic regression analysis.Results From 1973 to 2014,6 human plague cases with 5 dead cases occurred in Subei County.The positive rate of plague bacteriological detection was positively correlated with the average relative humidity of current year and the average temperature of preceding first year (r =0.366,0.385,P ＜ 0.05),and it was negatively correlated with the average relative humidity of preceding third year (r =-0.304,P ＜ 0.05);the positive rate of plague bacteriological detection was positively correlated with the average rainfall of preceding first month (r =0.212,P ＜ 0.05),and it was negatively correlated with the average relative humidity of preceding second and third months (r =-0.196,-0.201,P ＜ 0.05).The risk factor of animal plague epidemic in marmot foci in Subei County was average temperature of current month [odds ratio (OR) =1.304,95％ confidence intervals (CI):1.195-1.422],and the protective factor was average relative humidity of preceding third month (OR =0.960,95％CI:0.932-0.988).Conclusions The prevalence of marmot plague is closely associated with meteorological factors in the natural plague foci of Subei County of Gansu Province.The temperature not only plays a role in promoting the prevalence of plague,but also has a lagging effect.Therefore,the meteorological factors can be considered as a reference of the plague prevalence in Himalayan marmot plague foci.
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Objective To understand the relationship between habitat choice of Himalaya marmot and vegetation elements such as vegetation coverage,grassland type,vegetation type and so on by using geographic information technology in Qinghai.Methods Himalayan marmot field survey data from Qinghai Provincial Institute for Endemic Disease Control and Prevention and vegetation coverage,grassland type and other remote sensing and geographic information data was processed for spatial overlay using ArcGIS tool.The vegetation characteristic information of the location of Himalayan marmot was searched and analyzed statistically,and then the initial quantitative analysis of Himalayan marmot on the selection of habitat vegetation conditions was made.Results After screening,84 samples of Himalayan marmot were retained.The mean of vegetation coverage was 0.708 5,the range was 0.313 3-1.000 0.Totally 54.76％ (46/84) of the Himalayan marmot samples was in the vegetation coverage of 0.70 to 1.00.Among the nine grassland types,the alpine meadows accounted for 39.78％ of total area of Qinghai,with 59 Himalayan marmot sample points distributed,which accounting for 70.24％ (59/84).Himalayan marmot mainly feed on some of the roots of grassland plants,including Poa pratensis,Alpine Kobresia,Potentilla,Polygonum viviparum,Kobresia cordata,Oxytropis,Aster and other miscellaneous grass.Conclusions Vegetation conditions is one of the important factors of Himalayan marmot in habitat selection.Quantitative analysis of the relationship between Himalayan marmot spatial distribution and vegetation factors by using geographic information technology,can deepen the understanding of marmot habitat selection,and provide a basis for grassland plague surveillance and control.
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We studied Yersinia infection and carriage in shepherd dog on the natural foci of Marmota himalayana plague and investigated the substitution or antagonism relationship among three pathogenic Yersinia in nature,providing a novel approach for prevention and control of plague.Blood sample and rectal swab specimens from shepherd dogs were collected.Rectal swab specimens were detected and isolated for Y.enterocolitican and Y.pseudotuberculosis.IHA was used to measure the positive rate of F1 antibody.Results showed that among 88,94,70 and 64 serum specimens respectively from Akesai,Subei,Sunan,and Tianzhu,IHA positive rates were 31.82％,32.98％,2.86％ and 1.56％ separately.A total of 236 anal swab specimens of shepherd dog were collected,among which one Y.kristensenii strain and 2 non-pathogenic Y.enterocolitica strain were recovered from Akesai and Subei respectively,where plague was violently prevalent in animals.All the results indicate that shepherd dogs in natural foci of M.himalayana plague where plague in animals are prevalent with non-pathogenic Y.enterocolitica,is an evidence of antagonism relationship in three pathogenic Yersinia.
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Objective@#To establish mouse intestinal tissue and its application in the proliferation of Marmota Himalayana hepatovirus (MHHAV).@*Methods@#The intestinal crypts of Balb/C mice were digested with EDTA at a high concentration, cultured in Matrigel and passaged into frozen intestinal enteroids. The monolayers were inoculated on 96-well plates and infected with MHHAV. Real-time qPCR virus concentrations at different time points were compared with those of a number of cell lines routinely used to develop hepatitis A virus.@*Results@#The mouse intestinal tissue culture system was successfully established. MHHAV did not proliferate in the cell lines commonly used for the culture of hepatitis A virus, but proliferated well in the intestinal tissues of mice. The virus multiplied about 3 700 fold in 4 days.@*Conclusions@#Mice intestinal enteroids can be used for the isolation and culture of MHHAV.
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Objective To predict the spatial distribution of Himalayan marmot using geographic information system (GIS) and Genetic Algorithm for Rule-set Production (GARP) model based on the monitoring data of Himalayan marmot in Qinghai Province.Methods Based on the data of routine monitoring and field survey by Global Position System (GPS) of Himalayan marmot,the position data was processed by spatial mapping using ArcGIS software.Ecological environment variables related to Himalayan marmot including terrain,slope,temperature,precipitation,vegetation,land use and other related variables were extracted and analyzed.The prediction model of Himalayan marmot distribution was constructed based on GARP model and ArcGIS software,and the spatial distribution mapping and analysis were carried out.Results Totally 198 points of Himalayan marmot were obtained by combining the recovery position with GPS information.The average model error of omission was 1.998 through the GARP modeling,while the optimal 100 model were highly statistically significant (all x2 ＞ 163.03,all P ＜ 0.01).The spatial distribution of predicted probability was divided into three grades including less than ＜ 40％,40％-＜ 80％ and 80％-100％ using ArcGIS,and the area with the prediction probability of 80％-100％ was the most suitable distribution area of Himalayan marmot.Conclusions The spatial distribution of plague host animal is predicted successfully using GIS and GARP ecological niche model.The result is more accurate compared to the statistic area by administrative region,which can provide important reference for plague prevention and control.
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Objective To investigate the internal parasites infection in albino Marmota himalayan,and to provide a basis for seting up the quality standard of Marmota himalayana.Methods 21 wild albinism and 30 wild nomal Marmota himalayana from the same origin were detected by method as intestinal parasites in stool examination and serological testing, toxoplasma gondii and echinococcosis parasite infections of each animal, then detected the internal parasites and eggs under platoscope and microscope.Results The natural infectious rate of the roundworm in wild albinism and nomal Marmota Himalayana were respectively 71.4% (15 /21) and 66.7% (20 /30), using the exact probability method of inspection, there was no significant difference in the roundworm infection rate between two groups marmota; antibody test results of Serum toxoplasma gondii and Echinoocosis were negative.Application of albendazole and Ivermectin injection drug expelling and purification, effect to be trusted.Conclusions Monitoring results and the cure method can be used as an albino Himalayan marmot displacement experiment of animal parasite quality monitoring index and purification method.
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Objective To investigate the histological changes of the right ventricular remodeling in Xinjiang Mar-mota baibacina which are adapted to high-altitude hypoxia .Methods Hematoxylin and eosin and Masson staining was used to examine the pathological changes and degree of fibrosis , and immunohistochemical technique was used to detect connexin 43 ( CX43) expression in the right ventricular myocardium .Results In the right ventricular myocardium , cardio-myocyte hypertrophy and increased amount of collagen fibers were observed .The right ventricular hypertrophy index and weight index of Marmota baibacina were significantly higher , and expression of CX43 was significantly reduced than that of normal rats ( P<0.05 for both ) .Conclusions High-altitude hypoxia induces right ventricular remodeling in Xinjiang Marmota baibacina.It can be used as an ideal animal model for studies on the mechanisms of hypoxia adaptation on high -altitude plateau .
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Objective To master of the characteristics of development and variation in plague natural foci of Marmota sibirica, and to analyze the plague monitoring results from 2004 to 2008. Methods From May to September every year, we monitored plague in Manzhouli, Chenbaerhuqi, Xinbaerhuzuoqi, Xinbaerhuyouqi,Ewenkeqi and Yakeshi city. The monitored area was 20 000 - 40 000 hm2 in every county. The density of Marmota,ground squirrel and moonlighting rats was surveyed by path method, one-day bow-clip method and a clamp was placed every five meters, respectively. The classification and identification of ectoparasite fleas were done by using low power lens. The fleas in the ground squirrel hole were collected by flannel rubber stick, then classified and identified. According to National Standard of Plague Diagnostic Criteria(GB 15991-1995), the collected rats and fleas were detected by isolation and cultured Yersinia pestis, the serums of collected rats were tested by indirect hemagglutination test. Results In five years, the density of Marmota and ground squirrel was 0.010/hm2 and 0.602/hm2, respectively. The capture rate of moonlighting rats was 2.69％ (258/9600). The flea infection rate of Marmota was 17.54％(10/57) and the fleas index was 2.54. The fleas infection rate of ground squirrel was 28.40％(213/750) and the fleas index was 1.01. Flea infection rat of ground squirrel hole was 5.60％(46/822) and the fleas index was 0.17. The total number of various hosts of Yersinia pestis detected was 1351, the groups number of the variety of cultured fleas was 127, and the pathogen test results were negative. The number of serum tested was 1064, positive number was 43, and the detection rate was 4.04％(43/1064). The highest positive titer was 1 : 1280. Other than 2006, the remaining four years were found positive for blood clotting material; positive serum was found in a total of three regions, they were Manzhouli, Xinbaerhuzuoqi, and Xinbaerhuyouqi; and 30, 12 and 1 copies of hemagglutination-positive sera were detected. Conclusions The epidemic of plague natural foci of Marmota sibirica is in a active state, and gradually expands the scope. We must continue to strengthen the inspection of the bacteria, bearing in mind the replacement of the region's main host, make every effort to prevent and control of human plague.
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Objective Throush identify biochemical characteristics and virulence factors of 2 strains suspected Yersinia pestis(Y.pestis)isolated from the dead Marmota himalayana(M.himalayana)to confirm the nature epidemic focus in Dege County,Sichuan Province.Methods Y.pestis was analyzed by specific staining and shape,culturing characteristics,splitting-test by bacteriophage,test of biochemical characteristics and glycolysis ability,virulence factors,virulence,nutritional requirement,plasmid,genetic test and genetic type. Results The tested strains were Gram staining bacilus.The main biochemical characteristics were Arabinose(+)、 Rhamnose(-),Maltose(+),Melibiose(-),Glycerol(+),Denitrification(+).The virulence factors with FI+.VW+, Pgm+,Pst I+;and with the common 6.0×106,45.0×106,65.0×106 plasmids,also with the virulence-relative plasmid gene.Both their absolutely lethal dose(LD100)in mice were 50 bacteria.The nutritional requirement appeared which were depended on Phenylalanine and Methionine.With the Genomovar 5 genotype characteristics of M.himalayana plague foci of Qinghai-Tibet plateau.The difference between tested strains and Yersinia pseudotubercuosis on the 3 different culture medium was obvious.The tested strains had a Y.pestis' specific 3a fragment,Pst I and FI-Ag,at 22 ℃,the strains could be split by bacteriophage completely.Conclusions According to the diagnostic criteria of plague in China,the 2 suspected strains isolated from Dege County,Sichuan Province ale confirmed as Y.pestis.both with powerful virulenceand with the characteristics of the Y.pestis of M.himahtyana in Qinghai-Tibet plateau plague natural focus.
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Objective To explore the existence of natural loci on Marmota himalayana plague in Sichuan province and to provide basis for prevention and control of the disease. Methods Both epidemiological investigation and laboratory tests were used to provide the host animal and fleas of the vectors with Yersinia pestis carriers. Results 30 species of animals were found to belong to 10 orders. Ochotona curzoniae and M.himalayana were the most common ones while 7 species of the fleas belonged to 7 genera and 3 families. M.himalayana was the main reservoirs while Callopsylla dolabris and Oropsylla silantiewi served as vectors. The 13 Y.pestis were identified from 43 Marmota samples. 8 samples were identified under IHA, with the highest titer of herding-dogs serum as 1 : 10 240. 19 samples were F1 antigen positive using RIHA and the highest titer of M.himalayana serum was 1:409 600. The major foci was 4545 km2, distributed at Dege county in Sichuan province. Conclusion We have confirmed the existence of natural foci on M. Himalayana plague in Sichuan province.
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The objective of this study is to express the carbohydrate recognition domain (CRD) of the asialoglycoprotein receptor (ASGPR) H1 and H2 subunits of Marmota himalayan in vitro, and develop polyclonal antibodies against the recombinant proteins. RT-PCR was used to amplify ASGPR CRDH1 and CRDH2 from the liver tissue of Marmota himalayan. The products of amplification were subcloned into prokaryotic expression vector pRSET-B, and expressed in E. coli BL21(DE3)plysS. The recombinant proteins were purified using Ni-NTA spin column. The purified proteins were inoculated into BALB/c mice to develop polyclonal antibodies. The sensitivity and specificity of antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohistochemical staining (IHC). The polyclonal antibodies showed high sensitivity and specificity against both denaturated and native ASGPR proteins. We successfully amplified and expressed the ASGPR CRDs of Marmota himalayan. The nucleic sequences of ASGPR CRDH1 and CRDH2 of Marmota himalayan have been submitted to Genbank and the sequence ID are DQ 845465 and DQ845466, respectively. The proteins and antibodies prepared can be used for targeting gene therapy in a new animal model-Marmota himalayan-for the research of infectious diseases of hepatitis viruses and liver cancer treatment.