Objective: To screen, classify, and analyze the tissue expression in different tissues and function of gustatory receptor (Gr) genes in Haemaphysalis verticalis, so as to provide a theoretical basis for the future regulation of its behavior using molecular biology techniques. Methods: H. verticalis ticks collected from the body surface of Spermophilus dauricus were starved for one month, after which their transcriptome was sequenced using Illumina HiSeq high-throughput technology. Key Gr genes were screened from the transcriptome using bioinformatics methods, followed by phylogenetic analysis and tissue expression profiling in adult ticks. The three-dimensional structures of these Gr proteins were predicted using homology modeling, and the rationality of the predicted models was evaluated. The primary pheromones on the body surface of S. dauricus (the host of H. verticalis) were identified using gas chromatography-mass spectrometry. Molecular docking between the Grs and pheromone ligands was performed using AutoDock 1.5.6 software. Results: A total of two Gr genes, Haemaphysalis verticalis Gr1 (HverGr1) and Haemaphysalis verticalis Gr2 (HverGr2), were identified in the transcriptome of H. verticalis. Phylogenetic analysis revealed that these Gr genes were relatively conserved. Tissue expression profiling in adult ticks revealed that both HverGr1 and HverGr2 were significantly highly expressed in the chemosensory organs compared to other tissues (all P < 0.05), showing a high expression pattern. Molecular docking demonstrated that stable intermolecular forces, primarily hydrophobic interactions, could form between the Grs of H. verticalis and the body surface pheromones of S. dauricus. Conclusions: Two putative Gr genes with significantly high expression in the chemosensory organs of H. verticalis were identified. Molecular docking suggested that these receptors can interact with the body surface pheromones of the host S. dauricus. These findings provide key targets for elucidating the gustatory recognition mechanisms of H. verticalis and establish a theoretical foundation for developing tick control strategies based on host pheromones.

Objective: To investigate the effects of different doses of exogenous 20-hydroxyecdysone (20E) on the growth and development of Aedes albopictus pupae, clarify its dose-effect relationship, and provide a theoretical basis for the development of mosquito control strategies. Methods: Pupae were collected at 8 h after pupation, and divided into groups receiving 0 (control group), 0.01, 0.05, 0.1, and 0.5 μg of 20E. Microinjection was used to administer 20E to mosquito pupae. The total numbers of survivors, eclosed individuals, and malformed individuals were recorded every 12 h. Survival rate, eclosion rate, malformation rate, and pupal survival time were calculated. The relative expression levels of key genes in the 20E signaling pathway in mosquito pupae were detected by reverse transcription quantitative real-time polymerase chain reaction. Differences in overall survival rate and overall eclosion rate were analyzed using the log-rank test. Pupal survival time was analyzed using one-way analysis of variance. Malformation rates were analyzed using the Pearson Chi-square test. Gene expression differences were analyzed using the independent samples t-test. Results: The 180-hour survival rates of Ae. albopictus in the 0.05, 0.1, and 0.5 μg 20E groups were (66.67±6.67)%, (15.55±3.85)%, and (0.00±0.00)%, respectively, all lower than that of the control group (95.55±3.85)%. The differences in overall survival rate between these groups and the control group were statistically significant (χ²=22.870, 111.400, 180.900, all P < 0.001). The pupal survival time in the 0.1 μg and 0.5 μg groups was 81.80 h and 125.20 h, respectively. The differences between these two groups and the control group (65.30 h) were statistically significant (q=5.327, P=0.024; q=19.350, P < 0.001). The eclosion rates of Ae. albopictus in the 0.05, 0.1, and 0.5 μg 20E groups were (66.67±6.67)%, (15.55±3.85)%, and (0.00±0.00)%, respectively, all lower than that of the control group (95.55±3.85)%. The differences in overall eclosion rates between these groups and the control group were statistically significant (χ²=71.680, 131.000, 157.700, all P < 0.001). The malformation rates of Ae. albopictus in the 0.05 and 0.1 μg 20E groups were (22.22±5.09)% and (52.22±8.39)%, respectively, both higher than that of the control group (0.00±0.00)%. The differences in malformation rates between these groups and the control group were statistically significant (χ²=22.500, 63.610, all P < 0.001). At 48 h after treatment, the body color of the 0.5 μg 20E group was significantly lighter than that of the control group. At 1 h after treatment with 0.1 μg 20E, the expression levels of AaEcR, AaBR-C, AaE75, AaHR3, and AaFTZ-F1 were upregulated by 1.89, 2.37, 2.96, 2.29, and 2.16 times, respectively (t=9.030, 11.960, 17.800, 34.260, 16.450, all P < 0.001). At 6 h after treatment, the expression levels of AaBR-C, AaE75, AaHR3, and AaFTZ-F1 were upregulated by 1.42, 1.26, 1.88, and 1.48 times, respectively (t=5.375, P=0.006; t=6.662, P=0.003; t=6.891, P=0.002; t=7.419, P=0.002), while the expression level of AaEcR was downregulated by 13.45% (t=5.531, P=0.005). At 12 h after treatment, the expression levels of AaEcR, AaBR-C, AaE75, AaHR3, and AaFTZ-F1 were upregulated by 1.59, 1.85, 1.48, 1.79, and 1.90 times, respectively (t=10.550, 30.360, 12.010, 18.280, 9.297, all P < 0.001). At 24 h after treatment, the expression levels of AaE75 and AaHR3 were upregulated by 2.18 and 2.79 times, respectively (t=5.745, P=0.005; t=19.660, P < 0.001), while the expression levels of AaEcR and AaFTZ-F1 were downregulated by 16.56% and 26.76%, respectively (t=3.540, P=0.024; t=2.787, P=0.050). Conclusions: Exogenous 20E can reduce survival rate and eclosion rate of Ae. albopictus pupae, prolong pupal development duration, and affect pupal body color, with doses of 0.05 and 0.1 μg showing significant teratogenic effects. A dose of 0.1 μg can regulate the expression of key genes in the mosquito pupal ecdysone signaling pathway, which may be an important molecular mechanism underlying developmental abnormalities.

Objective: To investigate the effects of rainfall washout on the survival of Aedes albopictus larvae and examine the role of container characteristics in modulating these effects, so as to provide scientific evidence for mosquito density surveillance, early warning, and the optimization of control strategies. Methods: Field simulation experiments were conducted in Shenzhen in August 2024. Containers with different capacities (100-2 000 ml), contents (no added material, longan leaves, soil), and types (beakers and enamel basins) were set up in experimental and control groups. Thirty Ae. albopictus larvae were placed into each container. Experiments were carried out during heavy rainfall periods. The numbers of washed-away and dead larvae were recorded. Meteorological data were collected, including rainfall amount, rainfall intensity, wind speed, atmospheric pressure, air temperature, and humidity. Data analysis was performed using R 4.5.3 software. The paired t-test and Wilcoxon signed-rank test were used to analyze the differences in the numbers of lost and dead larvae under conditions with and without rainfall washout. A generalized linear mixed model was employed to examine the effects of container capacity, contents, container types, and meteorological factors on larva loss. Results: Significant differences were observed between the experimental and control groups in the number of lost larvae (t=11.180, P < 0.001) and dead larvae (W=1 019.500, P < 0.001). Container capacity was significantly negatively correlated with both the number of lost larvae (β=-0.264, P < 0.001) and the number of washed-away larvae (β=-0.500, P < 0.001). The presence of added contents in containers reduced larva loss, among which the protective effect of adding soil was significant (β=-0.528, P=0.005), while that of adding longan leaves was not significant (β=-0.296, P=0.088). Under the condition of the same capacity, the difference in the number of lost larvae between enamel basins and beakers was not significant (β=-0.286, P=0.264). Under the condition of the same height, the larva loss in enamel basins was significantly lower than that in beakers (β=-0.763, P < 0.001). Among the meteorological factors, cumulative rainfall had a significant positive effect on larva loss (β=0.069, P < 0.001), while wind speed had a significant negative effect (β=-1.157, P < 0.001). Conclusions: Rainfall washout under natural conditions exerts significant adverse effects on the survival of Ae. albopictus larvae in small containers through direct washout and increased mortality risk. These effects are modulated by container capacity, types, and contents. It is recommended to incorporate rainfall washout and container characteristics into mosquito vector density assessment and disease risk prediction models, in order to improve the accuracy of early warning and the effectiveness of targeted control measures.

Objective: To systematically analyze national fly surveillance data from 2020 to 2025, clarify the species composition, spatiotemporal density distribution, and regional differences of flies in China, so as to provide a scientific basis for precise vector control. Methods: Data were collected from 92 nationwide fly surveillance sites from 2020 to 2025. Most sites in southern China conducted year-round surveillance, whereas those in northern China mainly carried out seasonal surveillance, usually from April or May to September or October. One-way analysis of variance or the Kruskal-Wallis H test was used to compare differences in fly density among different habitats, months, and geographical regions. Linear regression was used to analyze the annual trend of fly density. Results: During the surveillance period, a total of 127 871 cages were deployed, and 558 816 flies were captured, with an average density of 4.37 flies/cage. The top four taxa in terms of proportion were Musca domestica (37.34%), Lucilia sericata (15.26%), Sarcophagidae (14.29%), and Muscina stabulans (8.53%). Chrysomya megacephala ranked six, with a proportion of 4.61%. Fly density differed significantly among habitats (F=7.891, P=0.001), with the highest density in farmers' markets (5.38 flies/cage). Seasonal fluctuation showed a single-peak pattern, with density reaching the maximum in July (9.29 flies/cage). National fly density showed a significant downward trend (β=-0.344, P=0.002). Regional analysis revealed there was a significant difference in fly density among different provinces (H=48.720, P=0.028), with relatively high densities in northwest China (Xinjiang Uygur Autonomous Region and Qinghai Province) and parts of southwest China (Guizhou Province), and the lowest density in south China (Guangdong Province). Conclusions: From 2020 to 2025, the composition of dominant fly species in China has changed substantially, with synanthropic flies becoming dominant. Fly density shows strong spatiotemporal heterogeneity and an overall downward trend. Future fly control should take regional differences into account and implement region-specific and category-based precise management strategies.

Objective: To investigate the changes in mosquito population density and the current status of resistance of Aedes albopictus to commonly used insecticides in Fangshan District, Beijing, and to provide a scientific basis for regional mosquito-borne disease prevention and control as well as the use of insecticides in chemical control. Methods: Mosquito population density and insecticide resistance surveillance data from 2021 to 2024 in Fangshan District were collected. R version 4.5.2 was used to analyze the seasonality and long-term trend of density using a seasonal-trend decomposition model based on locally weighted regression (STL). Kruskal-Wallis H rank sum test was used to compare mosquito density differences among different habitats. Bonferroni-corrected Mann-Whitney U test was used for multiple comparisons. For insecticide resistance surveillance, adult mosquitoes were tested using the WHO tube method to determine the 24 h mortality rate against 0.7% beta-cypermethrin, 0.5% malathion, and 0.05% propoxur. Larvae were tested using the larval immersion method to determine the median lethal concentrations and resistance ratios (RRs) for beta-cypermethrin, propoxur, and temephos. The data were fitted to the Probit model using the drc package in R 4.5.2 to obtain regression equations and 95% confidence intervals. Results: A total of 1 440 (lamp·nights) were set from 2021 to 2024. A total of 871 mosquitoes belonging to 2 genera and 2 species were captured, with an average density of 0.30 individuals/(lamp·hour). Significant differences in mosquito density were found among habitats (H=27.583, P < 0.001). Adult resistance surveillance showed that in 2021 and 2023, adult Ae. albopictus exhibited resistance to 0.7% beta-cypermethrin, 0.5% malathion, and 0.05% propoxur (24 h mortality rate: 47.78%~76.67%). Larvae showed low resistance to beta-cypermethrin (RR=4.05-9.60), moderate resistance to propoxur (RR=13.55-14.36), and a shift from susceptibility to low resistance for temephos, with the RR increasing from 1.71 to 3.59. Conclusions: The main mosquito populations in Fangshan District, Beijing are Culex pipiens pallens and Ae. albopictus. Among them, Ae. albopictus has developed widespread resistance to commonly used insecticides. It is necessary to adhere to the strategy of prioritizing environmental management supplemented by chemical control, optimize insecticide rotation, avoid excessive dosage and off-label application, and continuously advance the balance between mosquito vector control and insecticide resistance management.

Objective: To analyze the density seasonal variation, habitat distribution, and insecticide resistance level of Aedes albopictus, the dengue vector, in key surveillance areas of Xi'an City, China from 2020 to 2024, so as to provide a scientific basis for formulating targeted prevention and control measures for mosquito-borne diseases. Methods: Ae. albopictus mosquitoes were monitored by the Breteau index (BI) method (for larvae) and the double-layered mosquito net method (for adults) in Chang'an District, which is adjacent to the main urban area of Xi'an City. The insecticide resistance of Ae. albopictus was monitored by the larval immersion method and the adult's WHO tube method. The data were statistically described and analyzed using SPSS 16.0. The comparison of measurement data was conducted using analysis of variance (data conformed to a normal distribution with homogeneity of variance) or Kruskal-Wallis H rank sum test, and the count data were analyzed using the Chi-square test. Results: In 2020-2024, the mean net trap index of Ae. albopictus was 5.92 mosquitoes/(net·h) in Chang'an District, Xi'an City; the density of the adults reached its peak in July and August; there was no statistically significant difference in the net trap index among different habitats (F=2.854, P=0.070). The mean BI was 5.46. Larval-positive water was primarily observed in tires and waste tires (42/105, 40.00%), idle containers (121/362, 33.43%), and small-scale standing water in green belts (33/118, 27.97%). The BI reached its peak from May to August. Adult Ae. albopictus showed resistance to 0.03% deltamethrin (24 h mortality rate: 48.33%), 0.08% beta-cypermethrin (24 h mortality rate: 72.88%), and 0.4% permethrin (24 h mortality rate: 55.56%), and sensitivity to 0.05% propoxur (24 h mortality rate: 100.00%) and 0.5% malathion (24 h mortality rate: 100.00%). Larvae of Ae. albopictus showed high resistance to deltamethrin (resistance ratio [RR]=377.40), beta-cypermethrin (RR=95.15), and permethrin (RR=279.12), low resistance to propoxur (RR=8.56) and temephos (RR=7.58), and sensitivity to dichlorvos (RR=1.51). Conclusions: The density of Ae. albopictus in key surveillance areas of Xi'an City from 2020 to 2024 was generally high. Aedes vector should be closely monitored in July and August in Xi'an City, and the focus of prevention and control should be adjusted promptly in accordance with the surveillance data. The cleanup and remediation of mosquito breeding sites should be conducted proactively in March, with particular attention to idle containers and standing water within the community environment. The efficacy of mosquito-killing insecticides such as pyrethroids should be evaluated in control practice, and low-resistance or sensitive insecticides should be alternated to enhance the mosquito-killing effect and prevent or reduce the transmission risk of dengue fever.

Objective: To investigate the baseline status of vectors at the China-Vietnam border ports in Guangxi Zhuang Autonomous Region (Guangxi ports), analyze their species composition, density levels, and variation characteristics across different port types, assess the risks of cross-border transmission of vector-borne diseases, and provide a scientific basis for precise vector control at ports. Methods: Long-term surveillance was conducted at six waterway, eight highway, three aviation, and one railway ports in Guangxi Zhuang Autonomous Region from 2016 to 2024. Rodent snap trapping, cage trapping, CO₂ light trapping, fly cage trapping, sticky trapping, and the dragging-flagging methods were employed to monitor rodents, mosquitoes, flies, cockroaches, and questing ticks. Data were analyzed using SPSS 25.0 software. One-way ANOVA (with Tukey's honestly significant difference post-hoc test) or the Kruskal-Wallis H test (with Dunn's post-hoc test using Bonferroni correction) was used for inter-group comparisons of vector density. Differences in vector species composition among different types of ports were analyzed using the Pearson Chi-square test. Results: A total of 71 vector species of rodents, mosquitoes, flies, cockroaches, and questing ticks were captured (excluding on-host ectoparasites on rodents). Among 11 rodent species, Rattus norvegicus was the dominant species in Guangxi ports (667/1 206, 55.31%) and also the dominant species at highway (318/465, 68.39%), waterway (241/412, 58.50%), and railway (57/89, 64.04%) ports, while the dominant species at aviation ports was R. tanezumi (163/240, 67.92%). The average rodent density differed significantly across different port types (H=7.824, P=0.020), with significantly higher rodent density at highway ports than at waterway ports (Z=3.860, P=0.018). Four species of fleas, two species of ticks, and one species of mite were collected as ectoparasites on rodents. Among 26 mosquito species captured, Culex tritaeniorhynchus was not only the dominant species in Guangxi ports (61 160/132 854, 46.04%) but also the dominant species at aviation ports (37 126/49 055, 75.68%) and highway (22 082/68 302, 32.33%), while the dominant species was Aedes albopictus (3 972/9 235, 43.01%) at waterway ports and Cx. pipiens quinquefasciatus (2 199/6 262, 35.12%) at railway ports. The average mosquito density differed significantly across different port types (H=9.642, P=0.008), and the density at aviation ports was significantly higher than at waterway ports (Z=3.270, P=0.039). The average container index for immature mosquitoes was 2.02%. Among 25 fly species collected, Chrysomya megacephala was the dominant species in Guangxi ports (5 378/22 120, 24.31%), and fly density was relatively high at railway ports. Among 5 cockroach species collected, Blattella germanica was the dominant species in Guangxi ports (2 756/4 147, 66.46%). A total of 22 questing ticks from four species were collected, with Rhipicephalus sanguineus being the dominant species in Guangxi ports (11 individuals). The overall difference in the distribution of vector categories among different port types was statistically significant (χ²=5 428.763, P < 0.001). The distribution of mosquitoes differed significantly among port types (χ²=8 962.357, P < 0.001), with mosquitoes dominating at both highway ports (68 302/76 517, 89.26%, rank 1) and aviation ports (49 055/54 829, 89.47%, rank 1). The distribution of flies also differed significantly among port types (χ²=12 345.678, P < 0.001), showing a uniquely high proportion at waterway ports (10 569/20 900, 50.57%, rank 1). The distribution of cockroaches and ticks differed significantly among port types (χ²=234.567, P < 0.001), with the highest proportion at aviation ports (1 954/54 829, 3.57%, rank 3). The distribution of rodents differed significantly among port types (χ²=156.789, P < 0.001), with the highest proportion at waterway ports (412/20 900, 1.97%, rank 4). Conclusions: The vector community structure in Guangxi ports is shaped by both species-specific ecological habits and port habitat suitability. There are potential risks of cross-border transmission of multiple vector-borne diseases. The foundational database established in this study provides a scientific basis for precise vector control at ports and the construction of national health barriers.

Objective: To investigate the population density, seasonal fluctuation, breeding habits, and Dengue virus carriage and its characteristics in dengue vector Aedes mosquitoes in the China-Myanmar border area of Gengma Dai and Wa Autonomous County, Lincang City, Yunnan Province, China, so as to provide a basis for formulating effective local dengue prevention and control measures. Methods: From April to December 2024, the density of Aedes larvae was investigated using the Breteau index (BI) method, adult mosquitoes were captured using a backpack aspirator, and Dengue virus in mosquitoes were detected by reverse transcription quantitative PCR. Positive PCR products were subjected to Sanger sequencing of the viral E gene. Data were organized using Excel 2010 software and statistical analysis was performed using SPSS 27.0 software. The Chi-square test or Fisher's exact probability test was used to compare the positive rates of water containers and Aedes breeding status. Results: A total of 1 680 households were surveyed, of which 268 were positive vector Aedes, with a house index of 15.95. A total of 4 626 water containers were found and 329 positive containers were identified, with a BI of 19.58 and a container index of 7.11. Buckets were the predominant type of positive containers, and the positive rates of vector Aedes differed significantly among container types (χ²=38.720, P < 0.001). Larval breeding was found from April to December, and larval population density peaked in June (BI=38.50). All collected Aedes larvae were identified as Ae. aegypti or Ae. albopictus, with a ratio of 14.63∶1. A total of 385 adult Ae. aegypti were captured, adults were active from April to December, and the density peak occurred in October (124/385, 32.21%). A total of 335 adult Ae. albopictus were captured, adults were active from June to December, with a peak in June (241/335, 71.94%). The Dengue virus type 4 (DENV-4) was detected in Ae. aegypti, with a carrying rate (pool positive rate) of 5.00%, Sequencing of the E gene revealed that it belonged to DENV-4 genotype Ⅰ and was highly similar to the 2017 Myanmar strain (MW788995.1). No Dengue virus was detected in Ae. albopictus. Conclusions: The population densities of Ae. aegypti and Ae. albopictus are relatively high in Gengma County, and adult Ae. aegypti carries DENV-4. These findings suggest that local authorities should strengthen routine monitoring and precise control of vector Aedes mosquitoes, enhance surveillance for cross-border importation of dengue, and implement joint prevention and control measures.

Objective: To analyze the epidemiological characteristics and spatiotemporal distribution of hemorrhagic fever with renal syndrome (HFRS) in Qianjiang City, Hubei Province, from 2014 to 2023, and provide references for identifying HFRS high-risk areas and formulating targeted scientific prevention and control measures. Methods: Based on the surveillance data from the China Information System for Disease Control and Prevention, descriptive epidemiological methods were used for statistical analysis. The Chi-square test was applied for rate comparisons, and Spearman correlation analysis was used to examine the associations of incidence with cultivated land area and rodent density. Spatiotemporal scan analysis was performed using SaTScan 9.4 software, and maps were generated using ArcGIS 10.8 software. Host animals were captured using the night trapping method. Rodent density was calculated and rodent species were identified. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect Hantavirus in rodent lungs and determine the virus genotype. Results: From 2014 to 2023, a total of 628 HFRS cases were reported in Qianjiang City, with 8 deaths, resulting in a case fatality rate of 1.27% and an average annual reported incidence of 6.23/100 000. The incidence increased and then decreased from 2014 to 2023, with the highest incidence of 21.22/100 000 in 2018. Two incidence peaks were observed annually: one in late spring and early summer (May-June) and another in winter (December-January of the following year). The male-to-female ratio of the cases was 2.19∶1. Cases were mainly concentrated in the population aged 40 to < 70 years, accounting for 73.25% (460/628). Farmers constituted the majority of cases, accounting for 76.75% (482/628). Spatiotemporal scan identified three clusters: the primary cluster was concentrated in the western and southwestern areas of Qianjiang City, including 8 towns (streets) such as Haokou Town (relative risk [RR]=5.47, log-likelihood ratio [LLR]=121.13, P < 0.001); the secondary cluster was concentrated in Zongkou Management District and its four adjacent towns (streets) in the southeastern area of Qianjiang City (RR=5.47, LLR=53.36, P < 0.001); the tertiary cluster was located in Zhouji Street (RR=6.67, LLR=18.68, P < 0.001). From 2014 to 2023, a total of 8 840 effective rodent traps were deployed in Qianjiang City, with 568 small mammals captured, resulting in an average density of 6.43%. In indoor habitats, Rattus norvegicus was the dominant species, accounting for 62.40% (161/258); in outdoor habitats, Apodemus agrarius predominated, accounting for 82.58% (256/310). The highest Hantavirus positive rate in small mammals was observed in 2018 at 17.14% (6/35). Conclusions: From 2014 to 2023, HFRS mainly occurred in middle-aged and elderly male farmers in Qianjiang City, with distinct seasonality (high in late spring, early summer and winter) and regional clustering (high in western and southern areas). Targeted interventions and control measures should be implemented before the epidemic seasons, focusing on high-risk areas and populations.

Objective: To understand the epidemic status of animal plague and the prevalence of Yersinia pestis infection in the human population in the natural foci of plague in Hebei Province, China in 2024, and to assess the risk of plague transmission. Methods: Animal plague surveillance was carried out in the natural foci in Hebei Province. The density of wild rodents was surveyed using the 24-hour bow trap method, and the capture rate of wild nocturnal rodents was surveyed using the five-meter trap line method. Differences in wild rodent density and nocturnal rodent capture rate across different habitats were analyzed. Rodent body fleas and nest fleas were collected, and differences in flea infestation rates and flea indices of the two types of fleas across different months were compared. Liver and spleen samples from all host animals were collected aseptically, imprinted, and cultured on self-made Plongeur medium. Identified fleas were homogenized and spot-inoculated onto the same medium and the growth of Y. pestis was observed after 24-48 hours of cultivation. Serum from live rodents or heart extracted from dead rodents was collected. The indirect hemagglutination assay (IHA) (microplate) was used for initial screening of Y. pestis F1 antibody, with suspicious positive sera subjected to duplicate testing for confirmation. Permanent residents of the plague focus were selected for serological surveillance of plague. The F1 antibody of Y. pestis was initially screened by IHA and confirmed by Western blot assay. Data were organized using Excel 2010 software and statistical analysis was performed with SPSS 26.0 software. The nonparametric Kruskal-Wallis H rank-rum test was used for intergroup comparisons of measurement data, while the Chi-square test or Fisher's exact test was used for enumeration data. Results: In 2024, the average density of wild rodents in the natural plague foci of Hebei Province was 0.17 inds/hm². Meriones unguiculatus was the dominant species, accounting for 61.67% (37/60) of all captured wild rodents. Wild rodent density differed significantly across habitats (H=16.975, P=0.002). The hillside habitat had the highest rodent density (0.41 inds/hm²), with Spermophilus dauricus as the dominant species (14/15). Cultivated land had the next highest density, with M. unguiculatus as the sole species captured (17 individuals). The capture rate of nocturnal rodents was 1.47% (100/6 800), with Cricetulus barabensis as the dominant species (74/100, 74.00%). The annual flea infestation rate on rodent bodies was 33.55% (210/626), with an average flea index of 1.41 (880/626). The flea infestation rate in rodent nests was 39.47% (15/38), with an average flea index of 1.55 (59/38). The flea indices for body fleas peaked in July (H=11.430, P=0.043). Bacteriological and serological tests on host animals and fleas were all negative. For human serological surveillance, 1 265 serum samples were collected and tested. Seventeen samples were initially positive by IHA, and 12 were confirmed positive by Western blot assay, yielding a positive rate of 0.95%. The antibody titers ranged from 1:8 to 1:128, and all the positive individuals were over 60 years old. Conclusions: In 2024, animal plague in the natural plague foci in Hebei Province was in a quiescent state. However, a potential risk of Y. pestis infection exists in the human population, necessitating continued strengthening of surveillance, prevention, and control.

Objective: To investigate the spatiotemporal distribution of questing ticks in urban parks in Shanghai City and determine the correlations between questing tick density and key meteorological factors, so as to provide a scientific basis for the prevention and control of ticks and tick-borne diseases in urban parks. Methods: From March to December 2024, a survey of questing tick infestation was conducted in urban parks across Shanghai using stratified sampling and the dragging-flagging method. Additionally, a systematic survey was carried out in five representative urban parks. Spearman rank correlation analysis was employed to examine the correlations between questing tick density and meteorological factors, and negative binomial regression models were used to analyze the effects of meteorological factors on tick population density. Results: A total of 135 urban parks were surveyed, with an overall questing tick infestation rate of 48.15% (65/135). Haemaphysalis flava (686/1 188, 57.74%) and H. longicornis (492/1 188, 41.41%) were the dominant species. The density of questing ticks exhibited significant seasonal fluctuation, appearing in March, peaking in September [46.35 ticks/(flag·100 m)], and virtually disappearing by December. Negative binomial regression analysis indicated a significant inverted U-shaped nonlinear relationship between monthly mean temperature and questing tick density (P＜0.05), revealing developmental stage-specific thermal niche differentiation: the optimal activity temperature range for nymphs (19.8-21.3 ℃) was significantly lower than that for larvae (24.1 ℃) and the total tick population (24.5-27.0 ℃). Regarding covariates, relative humidity had a significant positive effect on larval density (β=0.343, P=0.003), while sunshine duration had a significant negative effect on total tick density (β=-0.452, P=0.031). Conclusions: Questing ticks were detected in nearly half of the surveyed urban parks in Shanghai City, with H. longicornis and H. flava identified as the dominant species. Questing ticks exhibited distinct seasonality, and temperature was the core driver of their seasonal population dynamics. High humidity and low-light conditions favored their population proliferation. It is recommended to strengthen tick surveillance and control in urban parks to mitigate the risk of tick-borne diseases among urban residents.

Objective: To investigate the impact of meteorological factors on the density of Aedes albopictus and their lag effects in Changsha City, and to provide a basis for optimizing mosquito vector control strategies in local dengue fever prevention and control. Methods: The Ae. albopictus density data monitored by mosquito trap lamps every half month from April to November during 2017-2024 in Changsha City and corresponding meteorological data were collected. Spearman rank correlation test was used to analyze their correlations. The distributed lag nonlinear model combined with generalized linear model and natural spline function was applied to analyze the nonlinear and lagged relationships between meteorological factors and Ae. albopictus density. Results: The distributed lag nonlinear model showed that half-month mean temperature had significant effects on Ae. albopictus density at lags of 0-30 days, with the maximum risk effect observed at half-month mean temperature of 33.01℃ and a lag of 0 days (relative risk [RR]=1.81, 95% confidence interval [CI]: 1.13-2.89). Half-month cumulative precipitation had significant effects on Ae. albopictus density at lags of 0-19.5 days, with the highest RR at a lag of 0 days and a half-month cumulative precipitation of 234.00 mm (RR=1.31, 95%CI: 1.00-1.72). Half-month mean wind speed showed significant effects on Ae. albopictus density at lags of 9-30 days. When the half-month mean wind speed ranged from 1.60 to 2.50 m/s, the RR values were 1.00, peaking at a lag of 30 days and a half-month mean wind speed of 1.60 m/s (RR=1.89, 95%CI: 1.28-2.78). Conclusions: The effects of half-month mean temperature, half-month cumulative precipitation, and half-month mean wind speed on Ae. albopictus density exhibit nonlinear and lag characteristics. In the prevention and control of Ae. albopictus, dynamic risk assessment based on meteorological factors and their lag effects on Ae. albopictus density is recommended to identify critical intervention windows.

Objective: To establish a meteorology-driven vector Aedes density forecasting model based on the daily meteorological factors and concurrent Breteau index (BI) data in Ruili City from 2017 to 2020 and from May to October 2023, so as to provide a quantitative basis for the early warning of dengue fever and other Aedes-borne infectious diseases. Methods: Pearson correlation analysis was used to determine the association between meteorological factors lagged by 0-30 days and BI. A BI forecasting equation was constructed using multiple linear stepwise regression. The equation was subjected to back calculation using data in 2019 and 2020, and independently validated with data in 2023. Results: From 2017 to 2020, BI in Ruili City exhibited a typical rainy season peak pattern, with the highest values in July and August. BI was significantly positively correlated with monthly mean precipitation (r=0.865, P < 0.001) and monthly mean temperature (r=0.774, P < 0.001), and showed the significant negative correlation with the monthly mean diurnal temperature range (Td) (r=-0.960, P < 0.001). Lagged correlation analysis revealed that the mean BI in the next 7 days (BI₇) was significantly positively correlated with the mean precipitation in the past 7 days (R₇) (r=0.622, P < 0.001), and negatively correlated with the daily Td (r=-0.748, P < 0.001). The forecasting model constructed by the multiple stepwise regression method was: BI₇=12.958 6+1.500 4R₇-0.679 8Td+ϵ. Back calculation showed that the correlation coefficients (r) for 2019 and 2020 were 0.851 and 0.875, respectively, and the independent validation for 2023 showed r=0.514 (all P < 0.05). The trends of the forecasted values were consistent with those of the measured values. Conclusions: This study establishes the first quantitative meteorology-based BI forecasting model for Ruili City. The daily mean precipitation and Td in the past 7 days are the core meteorological driving factors for Aedes density changes in Ruili City. This model can be used for early warning of dengue fever and other mosquito-borne infectious diseases, precise mosquito control, and public health decision-making in the context of climate warming.

Objective: To clarify the species composition characteristics and pathogen-carrying status of small mammals in Ningbo City, so as to provide a scientific basis for the precise prevention and control, risk assessment, and strategy formulation of rodent-borne zoonoses in the region. Methods: From March to November 2025, small mammals were monitored using the night cage-trapping method across multiple habitat types in 10 counties (cities and districts) of Ningbo City. After classification and identification of the captured individuals, liver, spleen, lung, and kidney tissues were collected. Twelve common zoonotic pathogens were detected using (RT-)qPCR. The species composition of small mammals was analyzed. The differences in pathogen carriage among different species, habitats, and regions were compared using the Chi-square test or Fisher's exact probability test. Results: A total of 538 small mammals were captured, covering 4 families and 11 species, with Muridae as the dominant family. Rattus norvegicus (225/538, 41.82%), Apodemus agrarius (109/538, 20.26%), and R. tanezumi (54/538, 10.04%) were the most abundant species, accounting for 72.12% (388/538) of the total captures. Ten of the 12 target pathogens were detected, with a total positive rate of 34.39% (185/538), a single pathogen infection rate of 25.09% (135/538), and a mixed infection rate of 9.29% (50/538). A single animal could simultaneously carry up to 4 pathogens. Bartonella had the highest detection rate (116/538, 21.56%), followed by pathogenic Leptospira (60/538, 11.15%), while Dabie bandavirus and Orientia tsutsugamushi were not detected. Statistical analysis showed significant differences in the total positive rates of pathogens among different species, habitats, and regions (all P < 0.001). The positive rates in Niviventer lotipes (33/45, 73.33%), rural residential areas (23/40, 57.50%), and Beilun District (29/48, 60.42%) were the highest in their respective categories. Conclusions: Small mammals in Ningbo City have high species diversity and generally carry multiple pathogenic zoonotic agents, posing a potential public health risk for human infection. The regional pathogen epidemic risk was closely related to the dominant species composition, species-specific pathogen-carrying spectrum, and habitat types. It is necessary to strengthen the precise density control of rodents and other small mammals, occupational protection of key populations, and routine pathogen monitoring in multiple habitats, as well as to establish a stratified and classified prevention and control system to build a solid defense line for regional zoonosis prevention and control.

Objective: To investigate the infection status of rodent-borne pathogens in rodents from the Marmota himalayana plague natural foci and Neodon fuscus plague natural foci in Yushu Tibetan Autonomous Prefecture, Qinghai Province, in the Sanjiangyuan area, so as to provide a reference for the prevention and control of rodent-borne diseases in the region. Methods: Rodents were captured in Batang Township of Yushu City and Zhenqin Town of Chengduo County in Yushu Tibetan Autonomous Prefecture using noose trapping and trap-line methods. After dissection, liver, spleen, kidney, and lung tissues were collected. DNA and RNA were extracted using the Chelex 100 method or the column membrane method. TaqMan probe-based qPCR or RT-qPCR was used to detect eight pathogens: Hantavirus, Dabie bandavirus (DBV), Leptospira interrogans, Rickettsia typhi, Orientia tsutsugamushi, Bartonella spp., Anaplasma phagocytophilum, and Francisella tularensis. Data were processed using Excel 2016 and SPSS 22.0 softwares. Intergroup comparisons were performed using the Chi-square test or Fisher's exact test. Results: A total of 200 rodent samples were captured, including 70 Ochotona curzoniae (35.00%), 90 N. fuscus (45.00%), and 40 M. himalayana (20.00%). The detection rate was 1.50% for Hantaan virus, 14.00% for the DBV L segment, 4.50% for the DBV M segment, 13.00% for R. typhi, 9.00% for O. tsutsugamushi, 10.00% for Bartonella spp., 1.00% for A. phagocytophilum, and 25.50% for F. tularensis. Seoul virus, the DBV S segment, and L. interrogans were not detected. The detection rates of pathogens varied greatly among rodent species. Only the DBV L segment was detected in O. curzoniae, with a detection rate of 10.00% (7/70). L. fuscus had the highest detection rate of F. tularensis, reaching 44.44% (40/90). No significant differences were found in the detection rates of different pathogens between males and females among the three rodent species (all P > 0.05). The detection rate of F. tularensis in Zhenqin Town of Chengduo County (24/50, 48.00%) was significantly higher than that in Batang Township of Yushu City (27/150, 18.00%) (χ²=17.765, P < 0.001). A total of 39 rodents showed co-infection with pathogens, accounting for 19.50% of the total. M. himalayana had the highest co-infection rate (24/40, 60.00%), and co-infection with four pathogens was observed. Conclusions: Rodents in the plague natural foci of the Sanjiangyuan area in Qinghai Province are infected with rodent-borne pathogens such as Bartonella spp., DBV, and F. tularensis. Surveillance of rodent-borne pathogens carried by local rodents should be strengthened, and prevention and control measures targeting rodents such as M. himalayana and N. fuscus should be actively implemented to reduce the risk of human infection with plague and other rodent-borne diseases.

Objective: To investigate the populations of small mammals and their carriage of pathogens in the eastern region of Tiantai County, Zhejiang Province, and to provide a basis for the prevention and control of local rodent-borne diseases. Methods: From March 2024 to November 2025, small mammals were captured using the night-trapping method (snap or cage traps) in Shiliang Town, Sanhe Town, and Tantou Town in eastern Tiantai County. Following the Taizhou Municipal Rodent-borne Pathogen Surveillance Protocol, organs including liver, spleen, kidney, lung, and rectum were collected for nucleic acid extraction. qPCR or RT-qPCR was used to detect the presence of the following 10 pathogens in the samples: Dabie bandavirus, Hantavirus (Hantaan virus and Seoul virus), Bartonella, Rickettsia typhi, Orientia tsutsugamushi, Leptospira, spotted fever group rickettsiae, Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Borrelia burgdorferi. The Chi-square test or Fisher's exact test was used to compare the differences in pathogen positive rates among different towns and small mammal species. Results: A total of 362 small mammals were captured, belonging to 9 species, 8 genera, 3 families, and 2 orders. Among them, rodents accounted for 99.45% (360/362), and eulipotyphlans accounted for 0.55% (2/362). The Apodemus agrarius (176/362, 48.62%) and the Rattus norvegicus (113/362, 31.22%) were the dominant species, jointly accounting for 79.84% (289/362) of the total captures. Among the 362 small mammals, 69 were positive for pathogens, with an overall positivity rate of 19.06%. Specifically, 44 were positive for Leptospira (12.15%), 20 for Bartonella (5.52%), 6 for Seoul virus (1.66%), and 1 for B. burgdorferi (0.28%). Two A. agrarius individuals were co-infected with Leptospira and Bartonella. The pathogen positivity rates in Sanhe Town, Tantou Town, and Shiliang Town were 40.63% (39/96), 13.46% (28/208), and 3.45% (2/58), respectively. The difference in pathogen positivity rate among different towns was statistically significant (χ²=47.082, P<0.001). The difference in pathogen positivity rates among different small mammal species was statistically significant (P<0.001) and A. agrarius had the highest pathogent positivity rate of 27.27% (48/176). Conclusions: The small mammals in Tiantai County are dominated by A. agrarius and R. norvegicus, which carry multiple zoonotic pathogens. Pathogen positivity rates vary among different towns and small mammal species. It is necessary to strengthen surveillance and control measures targeting the dominant reservoir hosts and high-risk areas to reduce the risk of disease transmission.

Objective: To investigate the resistance levels of Culex pipiens quinquefasciatus adults and larvae to commonly used public health insecticides in the main urban areas of Kunming City, Yunnan Province, so as to provide a theoretical basis for government departments to use public health insecticides scientifically and rationally. Methods: During the peak mosquito activity period from August to October 2024 in Kunming City, field populations of Cx. pipiens quinquefasciatus were collected from five main urban districts (Wuhua, Guandu, Panlong, Xishan, and Chenggong) using methods such as dipping method and pipette method. The mosquitoes were reared in the laboratory to the F₁ generation. The resistance levels of adults and larvae to six commonly used public health insecticides were determined using the WHO tube test and the larval immersion method, respectively. The Probit method in the regression analysis of SPSS 19.0 statistical software package was used to calculate the median lethal concentration (LC₅₀) and their 95% confidence intervals for the tested populations. Results: The larvae of Cx. pipiens quinquefasciatus in the main urban areas of Kunming City showed high resistance to deltamethrin and permethrin, with resistance ratios (RR) of 89.47 and 40.50, respectively; moderate resistance to beta-cypermethrin (RR=15.19); and low resistance to temephos (RR=4.69). However, they remained sensitive to dichlorvos and propoxur, with RR of 0.35 and 1.31, respectively. Adult mosquitoes showed resistance to 0.025% beta-cypermethrin, 0.25% permethrin, and 0.025% deltamethrin, with mortality rates of 12.63%, 9.00%, and 2.14%, respectively. Suspected resistance was observed to propoxur and malathion, with mortality rates of 93.68% and 96.67%, respectively. Conclusions: The resistance profile of Cx. pipiens quinquefasciatus in Kunming City is consistent with the trends of resistance observed in other major vector mosquito species in Yunnan Province. The long-term and frequent use of pyrethroid insecticides may be the main factor leading to the development of resistance. Against the dual background of current vector-borne disease control and the patriotic public health campaign, there is an urgent need to implement a comprehensive strategy based on environmental management, insecticide rotation, compound formulations deployment, and continuous resistance surveillance, in order to delay the development of resistance and ensure the effectiveness of mosquito control measures.

Objective: To investigate the distribution, density, and pathogen carriage of Culex tritaeniorhynchus in Nanjing City in 2025, and to provide a scientific basis for assessing the risk of mosquito-borne disease transmission in Nanjing City. Methods: From March to November 2025, the density of Cx. tritaeniorhynchus was monitored using light traps in five types of habitats (residential areas, parks, hospitals, rural households, and livestock sheds) across 12 districts of Nanjing City. From June to September, Cx. tritaeniorhynchus samples were collected from two habitat types (rural households and livestock sheds) in Liuhe District and Lishui District. Japanese encephalitis virus (JEV) was detected using quantitative real-time PCR. SPSS 25.0 software was employed for statistical analysis. Kruskal-Wallis H test was used to compare mosquito density across different districts and habitats, Fisher's exact test was used to compare JEV positivity rates across different districts, and the Chi-square test was used to compare JEV positivity rates across different habitats. Results: The highest density of Cx. tritaeniorhynchus observed in Jiangning District, at 0.17 mosquitoes/(light·h), with a statistically significant difference observed among different districts (H=61.960, P < 0.001). Livestock sheds had the highest density of 0.12 mosquitoes/(light·h), but the difference in Cx. tritaeniorhynchus density among different habitats was not significant (H=8.272, P=0.082). The density exhibited a seasonal fluctuation characterized by an overall trend of initial increase followed by a decrease, peaking in July at 0.13 mosquitoes/(light·h). Pathogen detection showed that the JEV pool positivity rate of Cx. tritaeniorhynchus was 12.86% (18/140). The difference in JEV pool positivity rate between Liuhe District and Lishui District was significant (P=0.024), while the difference in JEV pool positivity rate between rural households and livestock sheds was not significant (χ²=1.800, P=0.180). Conclusions: The distribution of Cx. tritaeniorhynchus in Nanjing City is characterized by high density in suburban areas and low density in urban areas. JEV was detected in samples from both livestock sheds and rural households, indicating a risk of Japanese encephalitis transmission. Subsequent surveillance and control measures should be strengthened.

Objective: To evaluate the training scores for plague prevention and control knowledge among medical and health personnel at municipal and county levels in Hebei Province, analyze the key influencing factors, and provide a reference for further optimizing the plague prevention and control training program and enhancing training quality in the province. Methods: The training was delivered through a combination of theoretical lectures and practical operation sessions from 2024 to 2025. A comprehensive assessment of the training outcomes covering both theoretical knowledge and practical skills was conducted upon the completion of training, Excel 2021 and SPSS 27.0 software were employed for data compilation and statistical analysis of the training assessment results, binary logistic regression was adopted to analyze influencing factors. Results: A total of 342 medical and health personnel from 202 institutions across 77 counties in 9 cities of Hebei Province were trained. A total of 340 (99.42%) trainees were very satisfied with the overall quality of the training, and their average assessment score was (96.05±2.93) points. There were statistically significant differences in training assessment scores among trainees stratified by region, gender, institutional administrative level, professional title, and job position (all P < 0.05). Trainees from Shijiazhuang (96.87±1.91), Langfang(96.76±2.39), and Chengde (96.74±2.74) achieved higher scores; female (96.26±2.82) trainees scored higher than male (95.81±3.03) trainees; trainees from county-level (96.20±2.86) institutions scored higher than those from municipal-level (95.24±3.16) institutions; trainees with intermediate professional titles (96.68±2.19) achieved the highest scores; and doctors (95.93±2.89) achieved the lowest scores. Sex, institutional administrative level, professional title, and job position were important influencing factors for training scores (all P < 0.001). Conclusions: There are differences in training assessment scores among medical and health personnel across different regions, genders, professional titles, and job positions. Plague prevention and control training has improved the plague prevention and control capacity of medical and health personnel to a certain extent. However, in terms of curriculum design and assessment methods, targeted training and evaluation should be implemented based on job responsibilities and institutional administrative levels, so as to better enhance the overall level of plague prevention and control in Hebei Province.

Ticks are a group of blood-sucking ectoparasites that infest the body surface of livestock, wild animals, and humans. They not only directly endanger host health, but also transmit a variety of zoonotic diseases, posing serious threats to the livestock industry and human health. Tick specimens carry information on tick morphology, genetics, and pathogen carriage, and are key materials for studies on tick classification, phylogeny, biological characteristics, biogeography, and tick-borne diseases. However, ticks are small in size, most have a body surface covered with chitinous scuta, and exhibit stage-specific blood-feeding habits. Therefore, appropriate methods should be adopted for specimen processing according to different research objectives. This article integrates reported techniques for tick specimen collection, preparation, and preservation. Based on the authors' long-term research experience, this review systematically describes the relevant operational procedures to provide technical support for research on ticks and tick-borne diseases.