This study was aimed at understanding the Babesia species makeup and distribution in small mammals in Jinsha River Basin of Yunnan Province,and the Babesia carriage status in small mammals in this area,to provide a scientific basis for the preven-tion and control of Babesia disease.A total of 1 493 small mammals belonging to 5 orders,10 families,25 genera,and 54 species were captured from 10 counties(cities)in the Jinsha River Basin of Yunnan Province in various agricultural and forest environments.DNA was extracted from liver and tick tissues,and 150 bp fragments of Babesia 18S rRNA were detected through molecular biological methods.The positive samples showed amplification of a 1 600 bp target fragment of 18S rRNA.Species characteristics were assessed through sequence comparison and phylogenetic analysis.A total of 14 small mammals infected with Babesia were detected in six coun-ties(cities)of Jinsha River Basin,Yunnan Province,with a positivity rate of 0.93%(14/1 493).The Otsu and Kobe types of Babesia voles were analyzed,and their sequences were compared with the sequences from human Babesia cases with high similarity and close evolutionary relationships.The positivity rates were 2.34%(3/128)in Qiaojia County,2.06%(2/97)in Yongshan County,1.88%(4/213)in Yuanmou County,1.03%(3/291)in Deqin County,0.95%(1/105)in Shangri-La City,and 0.78%(1/128)in Shuifu County.The positive small mammals belonged to one order,two families,six genera,and the following eight species:P.leucurus 5.56%(1/18),R.brunneusculus 3.36%(4/119),M.minutus 3.33%(1/30),E.custos 2.94%(1/34),N.confucianus 2.65%(3/113),N.fulvescens 2.35%(2/85),A.latronum 1.16%(1/86),and A.draco 0.98%(1/102).The detection of Babesia in M.minutus was re-poorted first time.Small animals infected with Babesia were detected in all three habitats and altitudes,and higher infection rates were observed in forest regions between 1 500 and 2 500 meters and high-altitude residential areas.Babesia infection was found in many small mammals in several counties(cities)along Jinsha River in Yunnan Province,and the epidemic status of Babesia in these areas warrants attention.

OBJECTIVE To establish the HPLC fingerprint of Xintong granules and the quantitative analysis of multi-components by single-marker method(QAMS)to determine the contents of 7 components,so as to provide a scientific basis for their quality control.METHODS HPLC method was used to establish the fingerprints for 10 batches of Xintong granules(No.S1-S10),and similarity evaluation,cluster analysis(CA)and partial least squares-discriminant analysis(PLS-DA)were performed.At the same time,the contents of seven components,including puerarin,daidzin,calycosin-7-O-β-D-glucoside,stilbene glycoside,naringin,icariin and tanshinone ⅡA,were determined by QAMS method,and were compared with the results of external standard method.RESULTS A total of 18 common peaks were marked and 7 peaks were identified in the HPLC fingerprints for 10 batches of Xintong granules,namely puerarin(peak 4),daidzin(peak 7),calycosin-7-O-β-D-glucoside(peak 9),stilbene glycoside(peak 10),naringin(peak 12),icariin(peak 17),and tanshinone ⅡA(peak 18);the similarities among them were more than 0.990,and CA and PLS-DA results showed that S4-S5,S8-S10,S1-S3 and S6-S7 were clustered into three categories,respectively.Using naringin as the internal standard,the contents of puerarin,daidzin,calycosin-7-O-β-D-glucoside,stilbene glycoside,icariin and tanshinone ⅡA were determined to be 7.868 1-10.181 2,1.709 2-2.374 1,0.285 2-0.326 3,1.024 1-1.523 9,0.140 2-0.290 4,and 0.077 1-0.219 4 mg/g,respectively,by the QAMS.These results showed no significant differences compared to those obtained by the external standard method.CONCLUSIONS Established HPLC fingerprint and QAMS method are convenient,stable and accurate,which can provide a basis for the quality evaluation of Xintong granules.

This study was aimed at understanding the Babesia species makeup and distribution in small mammals in Jinsha River Basin of Yunnan Province,and the Babesia carriage status in small mammals in this area,to provide a scientific basis for the preven-tion and control of Babesia disease.A total of 1 493 small mammals belonging to 5 orders,10 families,25 genera,and 54 species were captured from 10 counties(cities)in the Jinsha River Basin of Yunnan Province in various agricultural and forest environments.DNA was extracted from liver and tick tissues,and 150 bp fragments of Babesia 18S rRNA were detected through molecular biological methods.The positive samples showed amplification of a 1 600 bp target fragment of 18S rRNA.Species characteristics were assessed through sequence comparison and phylogenetic analysis.A total of 14 small mammals infected with Babesia were detected in six coun-ties(cities)of Jinsha River Basin,Yunnan Province,with a positivity rate of 0.93%(14/1 493).The Otsu and Kobe types of Babesia voles were analyzed,and their sequences were compared with the sequences from human Babesia cases with high similarity and close evolutionary relationships.The positivity rates were 2.34%(3/128)in Qiaojia County,2.06%(2/97)in Yongshan County,1.88%(4/213)in Yuanmou County,1.03%(3/291)in Deqin County,0.95%(1/105)in Shangri-La City,and 0.78%(1/128)in Shuifu County.The positive small mammals belonged to one order,two families,six genera,and the following eight species:P.leucurus 5.56%(1/18),R.brunneusculus 3.36%(4/119),M.minutus 3.33%(1/30),E.custos 2.94%(1/34),N.confucianus 2.65%(3/113),N.fulvescens 2.35%(2/85),A.latronum 1.16%(1/86),and A.draco 0.98%(1/102).The detection of Babesia in M.minutus was re-poorted first time.Small animals infected with Babesia were detected in all three habitats and altitudes,and higher infection rates were observed in forest regions between 1 500 and 2 500 meters and high-altitude residential areas.Babesia infection was found in many small mammals in several counties(cities)along Jinsha River in Yunnan Province,and the epidemic status of Babesia in these areas warrants attention.