Abstract:This study aimed to determine the positivity rate and main pathogen species of bovine piroplasmosis in different areas of Xinjiang,providing a reference for establishing a scientific monitoring and prevention system.A total of 121 bovine blood samples were randomly collected from Turpan and Ili for DNA polymerase chain reaction(PCR)testing.Positive samples were subjected to DNA sequencing,and a phylogenetic tree of the pathogens was constructed.The results showed that the overall positivity rate for Theileria annulata in the 121 collected blood samples was 27.27%(33/121).Specifically,the positivity rate in Turpan was 7.14%(3/42),while in Gongliu County,Ili,it was 37.97%(30/79).Additionally,nine positive cases for Anaplasma were detected in the 42 blood samples from Turpan,with a positivity rate of 21.43%.No Babesia was detected in any of the samples.Through sequencing analysis,the strains isolated from Turpan and Ili in this study were identified as Theileria annulata.Phylogenetic evidence suggests that they may be derived from Indian strains and local Xinjiang strains.The study indicates significant regional differences in the prevalence of piroplasmosis between Turpan(7.14%)and Gongliu County,Ili(37.97%),with Theileria annulata being the dominant pathogen for piroplasm infection in these areas,and no Babesia infection was detected.

Abstract:Porcine circovirus(PCV)is a significant pathogen that severely jeopardizes the global swine industry.Among its types,porcine circovirus type 2(PCV2)and porcine circovirus type 3(PCV3)have become the focus of current research due to their highly similar clinical features and frequent co-infections.This article reviews the latest research progress in the dual fluorescent PCR detection technology for PCV2 and PCV3.It summarizes the advantages of dual fluorescent PCR in improving detection efficiency,reducing costs,and achieving accurate differential diagnosis.The practical application value of this technology in epidemiological investigations,vaccine efficacy evaluation,and the diagnosis of co-infections is discussed.Furthermore,future development directions are proposed to provide a reference for the prevention and control of porcine circovirus diseases.

Abstract:The experiment aimed to investigate the growth and reproductive performance of hybrids between Silver King pigeons and Gray King pigeons,and to analyze the effects of incubation and brooding on the reproductive energy consumption of breeding pigeons.A total of 350 pairs each of Silver King pigeons(Y),Gray King pigeons(H),and Silver King(♂)×Gray King(♀)hybrids(YH)were randomly selected for rearing observation over a one-year experimental period.The results showed that the number of laying nests and total egg production for Group Y were significantly higher than those for Groups H and YH(P<0.05).The number of hatched chicks in Group Y was significantly higher than in Group YH(P<0.05)and extremely higher than in group H(P<0.01).The number of dead chicks in group YH was extremely lower than in groups Y and H(P<0.01),and the survival rate at 28 days of age was extremely higher than in group H(P<0.01).Heterosis was highest for the number of dead chicks,at-58.14%;followed by the number of infertile eggs and dead embryo eggs,at-1.97%and-1.57%,respectively;the heterosis rates for fertilization rate and squab survival rate at 28 days were 0.39%and 0.52%,respectively.There were certain differences in the growth performance among Groups H,Y,and YH,with egg weight exceeding 22 g for all groups.The correlation between both egg production number and laying interval,and the number of infertile eggs and return-to-lay time,exceeded 0.5.The laying interval showed a highly significant negative correlation with the number of infertile eggs and dead embryo eggs.The laying interval was highest for breeding pigeons that completed incubation of two eggs.The laying interval for pigeons that incubated eggs for only 5 days was extremely lower than that of the other groups(P<0.01).The laying interval for pigeons that completed incubation of one egg was extremely higher than for those that did not complete incubation of two eggs(P<0.01).The return-to-lay time for pigeons incubating one egg was extremely lower than for those incubating two eggs(P<0.01).The laying interval and return-to-lay time for pigeons brooding two squabs were approximately 3 days longer compared to those brooding one squab(P<0.01).The study indicates that the reproductive performance of group Y is superior to that of group H,and the 28-day survival rate of YH squabs exhibits heterosis.The laying interval and return-to-lay time are highly correlated with the number of eggs produced.The number and duration of incubated eggs,as well as the number of brooded squabs,significantly influence the laying interval and return-to-lay time.

Abstract:The study aims to understand the prevalence and drug resistance of Salmonella in a large-scale broiler farm in the central Yunnan region.Chicken samples suspected of Salmonella infection in the farm were collected for bacterial isolation,and the isolates were identified by biochemical identification,multiplex PCR and serological methods.The drug sensitivity test of the isolates was conducted by the disk AGAR diffusion method,and the drug resistance genes were detected by PCR technology simultaneously.The results showed that eight strains of Salmonella were isolated from the suspected diseased chicken samples,among which seven were Salmonella Gallinarum and one was Salmonella Newport.The resistance rates of eight Salmonella strains to penicillin,tetracycline and norfloxacin were 100.0%,the resistance rates to nine drugs such as oxacillin and cefazolin ranged from 50.0%～87.5%,and the resistance rate to gentamicin was only 37.5%.All eight Salmonella strains were multi-drug resistant,and they were resistant to a minimum of six and a maximum of 13 drugs.Among the 17 drug resistance genes detected,the detection rates of blaCTX-M,blaOXA,blaTEM-1b,tetA,sul1,sul2,sul3,aadA1,aadA2,aacC4,and cmlA in eight Salmonella strains were all greater than 87.5%.Studies have shown that the prevalence of Salmonella in this field is mainly Salmonella Gallinarum,and the phenomenon of multi-drug resistance in the isolates is serious.Based on the results of the drug sensitivity test,it is recommended to use kanamycin or gentamicin for treatment.

Abstract:The research aims to summarize the diagnosis and treatment methods of sheep mastitis,with the goal of reducing the incidence and minimizing the economic losses of livestock farms.The study took all the ewes in the pens of a sheep farm in Xinjiang as the experimental subjects.Based on the clinical symptoms and the test results of the diagnostic reagent for mastitis,milk samples of the sick sheep were collected.Through enrichment,isolation and selective culture,etiological research was conducted,and drug sensitivity tests and molecular biological identification were carried out.The results showed that during the seasonal change in the farm,a large number of sheep mastitis infections and outbreaks occurred.Among the 174 ewes examined,102 were diagnosed with mastitis,with an incidence rate as high as 58.62%.Among them,the incidence rate of clinical mastitis was 13.22%(23/174),and that of latent mastitis was 45.40%(79/174).After isolation and culture,eight isolates suspected to be Staphylococcus aureus were obtained.The isolates showed sensitivity to penicillin,streptomycin,norfloxacin and compound sulfamethoxazole,and the homology with Staphylococcus aureus all exceeded 96.00%.Based on the results of the drug sensitivity test,the treatment plan was designed.Except for a few ewes and some lambs that died,most ewes improved significantly after treatment,and the cure rate was 95.10%(97/102).Studies have shown that mastitis in lactating ewes in this farm is mainly caused by Staphylococcus aureus.The combination of drug treatment and strengthened feeding management is effective in both the treatment and prevention of mastitis.

Abstract:This study aimed to analyze the bacterial carriage and drug resistance of yaks in the Nagqu region of Xizang,providing a reference for disease prevention and control in high-altitude yak farming.Bacteriological methods were used to isolate and purify bacteria from yak fecal samples collected in Nagqu.Identification was performed using microbial mass spectrometry,and drug resistance was determined by the micro-broth dilution method to preliminarily understand the characteristics of intestinal flora in yaks from high-altitude regions.The results showed that 10 strains of Escherichia coli,38 strains of Arthrobacter gandavensis,four strains of Acinetobacter calcoaceticus,one strain of Arthrobacter koreensis,two strains of Bacillus silvestris,and two strains of Enterococcus casseliflavus were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS).Drug susceptibility testing revealed that Escherichia coli exhibited high resistance to sulfonamide antibiotics,while it was sensitive to β-lactam antibiotics,aminoglycoside antibiotics,chloramphenicol antibiotics,tetracycline antibiotics,and polypeptide antibiotics,with a resistance rate of 0.Yak-derived Acinetobacter calcoaceticus showed high resistance to β-lactam antibiotics,chloramphenicol antibiotics(florfenicol),sulfonamide antibiotics,and polypeptide antibiotics(colistin).However,it was sensitive to aminoglycoside antibiotics,chloramphenicol antibiotics,and tetracycline antibiotics,with a resistance rate of 0.Arthrobacter strains exhibited low resistance rates.Gram-positive bacteria demonstrated severe antibiotic resistance,with resistance rates of 100%to penicillin,erythromycin,clindamycin,ceftiofur,cefoxitin,sulfisoxazole,vancomycin,tiamulin,tilmicosin,and linezolid.The resistance rate to amoxicillin/clavulanic acid and trimethoprim/sulfamethoxazole was 50%.These bacteria were sensitive to enrofloxacin,ofloxacin,oxacillin,tetracycline,florfenicol,and gentamicin,with a resistance rate of 0.The study indicates that yak-derived microorganisms are diverse,and the drug resistance data suggest an increased resistance of yak-derived Escherichia coli to sulfonamide antibiotics,which warrants attention.

Abstract:This study focused on the effects of dietary supplementation with Gynostemma pentaphyllum powder on the body weight,serum biochemical parameters,and immune organ development of broilers.A total of 120 one-day-old AA broilers(half male and half female)with similar body weights were randomly divided into four groups,with five replicates per group and six broilers per replicate.The control group was fed a basal diet,while the experimental groups received the basal diet supplemented with 0.5%,1.0%,and 2.0%Gynostemma pentaphyllum powder,respectively.The trial lasted for 42 days.The results showed that the body weight of broilers at 42 days of age in the 1.0%group was significantly higher than that in the 2.0%group(P<0.05),and was higher than that in the control and 0.5%groups,although these differences were not statistically significant(P>0.05).Compared with the control group,there were no significant differences in the contents of serum uric acid,total protein and albumin,as well as the activities of alanine aminotransferase and aspartate aminotransferase in broilers of each Gynostemma pentaphyllum powder addition group(P>0.05).The serum globulin content of broilers in the 1.0%group was significantly higher than that in the other groups(P<0.05).There was no significant difference in the spleen index and bursa of Fabricius index among the groups of broilers(P>0.05),and the 1.0%group was slightly higher than the other groups.The thymus index of broilers in the 1.0%group was significantly higher than that in the control group(P<0.05),but there was no significant difference compared with the 0.5%group and the 2.0%group(P>0.05).Morphological observation of the thymus revealed clearer structure,distinct demarcation between the cortex and medulla,tightly arranged cells,and a tendency for medullary area expansion in the 1.0%group.The study indicates that dietary supplementation with 1.0%Gynostemma pentaphyllum powder can promote broiler growth and thymus development,thereby contributing to improved broiler health.

Abstract:This research employed network pharmacology and molecular docking techniques to investigate the inhibitory mechanism of Scutellaria baicalensis against Aeromonas salmonicida.The susceptibility of Aeromonas salmonicida JMAV14 to the drug was screened through in vitro antibacterial assays of traditional Chinese medicine,and the minimum inhibitory concentration(MIC)was determined.Active components of Scutellaria baicalensis were collected from the TCMSP database,and their targets were obtained via the PubChem,Swiss Target Prediction,and Swiss ADME databases.Targets related to'Aeromonas salmonicida'were retrieved from the Genecards,OMIM,and TTD databases.Intersecting targets were identified using the Microbioinformatics Venny platform.Protein-protein interaction(PPI)analysis of the intersecting targets was performed on the STRING platform,while GO functional enrichment and KEGG pathway enrichment analyses were conducted on the DAVID platform.Molecular docking was carried out using AutoDockTools software.The results showed that 36 active components and 445 potential targets were screened from Scutellaria baicalensis,with 13 targets associated with inhibiting Aeromonas salmonicida,yielding two intersecting targets.PPI analysis revealed that Bcl2l1 and Mcl1 were core targets.GO and KEGG enrichment analyses identified 16 functional terms and three pathways related to the inhibition of Aeromonas salmonicida,with the apoptosis pathway being the most significantly affected.Both wogonin and acacetin were able to bind to the core targets MCL1 and BCL2L1.The study demonstrates that active components in Scutellaria baicalensis,such as wogonin and acacetin,act on the targets Bcl2l1 and Mcl1,inhibiting Aeromonas salmonicida by regulating the apoptosis pathway,JAK-STAT signaling pathway,and PI3K-Akt signaling pathway.

Abstract:This study aimed to breed new high-quality meat sheep varieties by crossing Suffolk(♂)and Kazakh(♀)sheep,analyzing the growth performance,slaughter traits,and the correlation between body measurements and body weight in the F1 generation,thereby providing a reference for meat sheep breeding selection.Thirty-two 110-day-old hybrid male lambs were selected,fattened for 50 days,and then slaughtered for measurement.Multidimensional analysis was conducted using SPSS 27.0 software:phenotypic statistics and stepwise regression analysis of body weight and body measurement indicators(body length,height at withers,chest circumference,cannon bone circumference);correlation analysis between body weight and feed efficiency traits;phenotypic statistics of slaughter traits and organ indicators;and establishing the optimal regression equation between body weight and body measurements through correlation coefficient decomposition.The results showed that the correlation coefficients between body weight and body length,height at withers,chest circumference,and cannon bone circumference in Suffolk×Kazakh F1 male lambs were 0.737,0.720,0.663,and 0.458,respectively.The correlation coefficients between mid-term body weight and initial weight,and final weight reached 0.977 and 0.978,respectively.The slaughter rate was 49.74%,net meat percentage 68.47%,carcass weight 23.27 kg,loin eye area 24.55 cm²,meat-to-bone ratio 3.51%,and tail fat weight 0.90 kg.The established optimal regression equation was:Y=-64.715+0.603X₁(body length)+0.348X₂(height at withers)+0.239X₃(chest circumference)+1.731X₄(cannon bone circumference)(R²=0.865,P<0.05).The research indicates that the Suffolk×Kazakh F1 generation exhibits good growth performance,high feed utilization efficiency,and outstanding slaughter traits,providing important references for the selection and breeding of new meat sheep varieties.

Abstract:The study aims to investigate the effects of two different dietary structures on weight gain and oral disease prevention in captive white kangaroos,in order to identify a more suitable dietary structure for them.The experiment selected 10 healthy adult white kangaroos,which were randomly divided into two groups,with five kangaroos in each group.Group Ⅰ was fed a diet with a higher proportion of roughage,while Group Ⅱ was fed a diet with a lower proportion of roughage.The trial lasted for 21 days.The results showed that the apparent digestibility of crude protein in Group Ⅰ was significantly higher than that in Group Ⅱ(P<0.05).The dietary structure with an increased proportion of roughage(GroupⅠ)significantly increased the body weight of the white kangaroos,and the roughage intake remained at a high level without symptoms related to lumpy jaw disease.In contrast,the dietary structure with a reduced proportion of roughage(Group Ⅱ)led to a decrease in body weight,significant fluctuations in roughage intake,and one white kangaroo showed obvious swelling on the left cheek.Additionally,compared with the same period,the incidence of lumpy jaw disease in white kangaroos decreased during the trial.The study indicates that a dietary structure with a higher proportion of roughage is more suitable for captive white kangaroos,helping to improve their health and prevent the occurrence of oral diseases.

Abstract:A male Exotic Shorthair cat developed symptoms of anorexia,severe depression,and abnormal urination following environmental changes after long-distance transportation.Based on clinical examination,laboratory tests,and imaging findings,the cat was diagnosed with feline lower urinary tract disease.Conservative management was implemented according to clinical symptoms and test results,including medication to correct hydration status,address electrolyte imbalances,and reduce hyperkalemia.A follow-up examination two weeks after discharge indicated a good prognosis.

Abstract:Poria cocos polysaccharide is one of the main active components of Poria cocos,possessing various pharmacological effects such as hepatoprotective,anti-inflammatory,antioxidant,anti-tumor,and immunomodulatory activities.As a feed additive,it can promote animal growth and development,enhance antioxidant capacity and immune function,and improve intestinal barrier function.This article reviews and summarizes the research progress on the main pharmacological effects of Poria cocos polysaccharide and its application in livestock and poultry farming,aiming to provide references for its further development and application.

Abstract:This study aimed to screen microbial strains with high efficiency in degrading sulfites in corn starch processing by-products.Strains were isolated from corn by-products,feed raw materials,and other sources.All isolated and purified strains were identified by MALDI-TOF mass spectrometry and preserved.Strains listed in the Feed Additive Catalog were selected for preliminary screening of sulfite degradation capability.Secondary screening was conducted through solid-state fermentation of corn germ meal,followed by acid tolerance evaluation of the top six strains with the highest degradation rates.Results showed that 194 strains-including bacilli,yeasts,and lactic acid bacteria-were selected for preliminary screening.The yeast strain JM06 exhibited the highest degradation rate(72.46%),significantly outperforming lactic acid bacteria(the best-performing lactic acid bacterium was Lactiplantibacillus plantarum ZW21,with a degradation rate of 34.44%).In secondary screening,yeast and lactic acid bacteria showed comparable degradation rates for sulfites in corn germ meal,with JM06(63.36%)and ZW21(62.26%)being the top performers.Based on these results,JM06,ZW21,JM02,SS01,JM05,and SS03 were selected for acid tolerance evaluation,revealing significantly superior acid resistance in yeasts versus lactic acid bacteria.This study demonstrates that yeasts outperform lactic acid bacteria in both sulfite degradation capability and acid tolerance,identifying JM06 as the optimal strain for sulfite degradation.

Abstract:This experiment aimed to investigate the therapeutic effect of Yiqi Jianwei Powder on piglets with diarrhea due to spleen deficiency.Eighty piglets diagnosed by veterinarians with diarrhea due to spleen deficiency were randomly divided into two groups,with 40 piglets in each group.Group A was treated with 0.04 g/kg colistin sulfate soluble powder mixed with feed,and Group B was treated with 1 g/kg Yiqi Jianwei Powder mixed with feed.Both were administered three times a day for 7 days.The results showed that after the treatment,the body weight of piglets in both groups increased compared to that before the treatment,with the final weight and average daily feed intake in Group B being significantly higher than those in Group A(P<0.05).Additionally,the symptom scores of piglets in both groups improved compared to pre-treatment levels,and all scores in Group B were significantly lower than those in Group A(P<0.05).The proportions of piglets in Group B that achieved recovery,marked effectiveness,and effectiveness were 42.50%,25.00%,and 15.00%,respectively,all higher than those in Group A(50.00%,30.00%,and 17.50%,respectively).The inefficiency rate of treatment in Group B was 2.50%,lower than that in Group A(17.50%).The study indicates that using Yiqi Jianwei powder to treat piglets with diarrhea due to spleen deficiency can alleviate symptoms,promote weight gain,and facilitate recovery.

Abstract:The experiment aimed to understand the antibody levels of swine fever virus and porcine pseudorabies virus in some large-scale pig farms in Bazhou.Blood samples of 1 078 pigs from seven large-scale pig farms in Bazhou were collected,and serological tests(ELISA)were conducted to detect the antibodies of swine fever virus and porcine pseudorabies virus.The results showed that the total positive rate of swine fever virus antibodies in the seven farms was 73.19%,meeting the qualified standard.Among them,the positive rate of 107-day-old fattening pigs was 66.67%,that of 114-day-old fattening pigs was 60.00%,that of 17-day-old piglets was 50.00%,that of 30-day-old piglets was 62.30%,that of 50-day-old piglets was 16.00%,and that of 57-day-old piglets was 43.33%.The positive rate of 60-day-old piglets was 58.89%,and that of nursery pigs was 50.00%.The antibody titers of pigs in the above eight stages were relatively low.The total positive rate of gB antibody against porcine pseudorabies virus was 98.61%,83.33%for 107-day-old fattening pigs,98.36%for 30-day-old piglets,90.00%for 57-day-old piglets,94.44%for 60-day-old piglets,and 98.33%for nursery pigs.The positive rates of antibodies in the remaining samples were all 100.00%.The total positive rate of gE antibody against porcine pseudorabies virus was 0,which proved that the sampled pig farm was not infected by wild porcine pseudorabies virus.Research shows that the positive rate of antibodies against swine fever virus in the seven large-scale pig farms in Bazhou Prefecture is generally at a passing level,while the positive rate of antibodies against porcine pseudorabies virus is at an excellent level.In the next step,on the basis of maintaining the excellent level of the positive rate of antibodies against porcine pseudorabies virus,the level of antibodies against swine fever virus should be improved and optimized.

Abstract:The research aims to establish a comprehensive prevention and control system for epidemic diarrhea in piglets.High-titer immune serum against porcine epidemic diarrhea virus(PEDV)was prepared by optimizing the refeeding procedure(two immune stimulations with a 72-hour interval).After the quality was tested,the prevention and treatment trials of epidemic diarrhea in newborn piglets were carried out.The results showed that the average titer of serum specific immunoglobulin G(IgG)against PEDV was 1.82,and the pathogen screening met the safety standards for biological products(the detection limit of multiplex PCR was<10³/μL).The incidence rate of suckling piglets in the serum intervention group was 9.09%(2/22),significantly lower than 38.46%(10/26)in the control group,and the protection rate was 76.37%.The clinical cure rate of the serum intervention group reached 82.61%(19/23),significantly higher than that of the drug control group,which was 16.67%(4/24).The mortality rate of the serum intervention group was 17.39%(4/23),while that of the control group was as high as 83.33%(20/24).Studies have shown that the preparation of high-immunity serum against PEDV is simple to operate,safe in quality and has a good preventive and therapeutic effect on epidemic diarrhea in piglets.

Abstract:Canine corneal ulcer is a common and serious ophthalmic disease that widely affects the visual health of dogs.Its causes are complex and diverse,mainly including bacterial infection,mechanical injury and immune-related factors,etc.This article reviews the epidemiological characteristics,main pathogenic bacteria and their drug resistance changes,pathological mechanisms,and clinical diagnostic methods of canine corneal ulcers.It also compares traditional and emerging treatment strategies and explores future research directions,with the aim of providing references for the clinical diagnosis and treatment of canine corneal ulcers,promoting the standardized management of this disease,and improving the therapeutic effect.

Abstract:The study aims to investigate the antibiotic resistance of Staphylococcus aureus in milk samples of dairy cows with mastitis in Ningxia region,and to provide a reference for the scientific prevention and treatment of Staphylococcus aureus.The biofilm formation ability of 234 strains of Staphylococcus aureus was determined by semi-quantitative crystal violet method,and the formation process of the biofilm was observed by scanning electron microscopy.The resistance of Staphylococcus aureus to 15 kinds of antibiotics was detected by broth dilution method,and the carrying status of genes related to the biofilm of Staphylococcus aureus was detected by polymerase chain reaction(PCR).The results showed that the positive rate of biofilm formation among 234 strains of Staphylococcus aureus was as high as 79.06％(185/234).Under the scanning electron microscope,it can be seen that the bacteria are closely connected into a network structure,forming a biofilm.As time goes by,the biofilm structure becomes increasingly compact.The resistance rate of Staphylococcus aureus to penicillin was the highest(94.87％,222/234),followed by ampicillin(85.90％,201/234).The carrying rates of biofilm-related genes such as icaA in Staphylococcus aureus biofilm-positive bacteria were all higher than those in negative strains.Studies have shown that there is a risk of multi-drug resistance to Staphylococcus aureus in the raw milk of cows with mastitis.The drug resistance of Staphylococcus aureus is related to the formation of biofilms.It is necessary to strengthen the monitoring of drug resistance in cows with mastitis,and pay attention to rational drug use and scientific prevention and treatment.

Abstract:Protein feed resources are tight in China.High-protein corn has a high protein content and can be used as a substitute for protein feed.It has the advantages of reducing costs and improving the growth performance of livestock and poultry.Conducting research on high-protein corn can effectively alleviate the shortage of protein feed resources in our country.Although significant breakthroughs have been made in the study of high-protein genes in corn,the breeding of high-protein corn varieties and their wide application in livestock and poultry production still require more data support.This article reviews the current breeding status,nutritional characteristics,application in livestock and poultry breeding,and the problems existing in the promotion of high-protein corn,with the aim of providing a reference for the application research of high-protein corn as livestock and poultry feed.

Abstract:The research aims to prepare terbinafine hydrochloride(TNH)spray and conduct quality control on it.The orthogonal experiment was adopted to optimize the formula of TNH spray,and the content of TNH spray was determined by high performance liquid chromatography(HPLC).The therapeutic effect of this spray on skin tinea was also investigated.The results show that the optimal formula of TNH spray was:1.5 g/100 mL of propylene glycol(PG),8 g/100 mL of polyethylene glycol(PEG),and 0.2 g/100 mL of anhydrous sodium acetate(SA).The light transmittance(T value)of the three batches of TNH sprays prepared according to the optimal formula was 98.8%,98.2%,and 98.6%respectively.The pH values were all qualified,and the stability was all grade 3.The HPLC determination range was within 0.058 8～0.196 0 g/L.The linear regression equation was Y=26 557 391 X-31 461(r2=0.999 4).The relative standard deviation(RSD value)of the precision test was 1.18%,and the solution was stable within 24 hours after preparation.The average recovery rate was 104.82%,the RSD value of the recovery rate was 1.56%,and the content determination values of the TNH spray in three consecutive batches were 100.2%,102.0%,and 101.9%respectively.The average cured area of TNH spray on SD rats with skin tinea reached 1.4 cm2,and the cure rate was 74%.Research shows that the TNH spray formula optimized by orthogonal experiments meets its quality requirements.The HPLC method has good precision,high accuracy and repeatability,and can effectively control its quality.The TNH spray has a relatively good effect in treating skin tinea.