Objective To establish a high-throughput semi-automatic bacterial whole-genome sequencing system to meet the needs of bacterial pathogen surveillance based on the genome sequence. Methods An automated liquid handler, disposable mechanical pipette tips, a 96-well nucleic acid extraction kit, an automated sequencing library construction system, and a sequencer were used to realize the semi-automation of bacterial genome sequencing. Results The distance between the mechanical pipette tip and the bottom of the sample well, the mixing speed of liquid, and the volume ratio of the mixed liquid to the sample each time were closely related to the cross-contamination of the samples. When the distance between the mechanical pipette tip and the bottom of the sample well was 2 mm, the mixing speed of liquid was 15% of the full speed, and the volume ratio of the mixed liquid to the sample was 50% each time, the contamination rate was 0. Compared with the manual procedure, this high-throughput semi-automatic bacterial whole-genome sequencing system improved the work efficiency by 4-fold. Conclusion A high-throughput semi-automatic bacterial whole-genome sequencing system is established, which can meet the needs of bacterial pathogen surveillance based on the genome sequence.

Bed bugs ( Cimex lectularius and C. hemipterus) feed on the blood of humans and animals and severely affect people's lives and health. After the World War II, bed bugs were almost eradicated in most parts of the world with the application of insecticides. However, since the late 1990 s, bed bugs have resurged all over the world (except Antarctica). Due to their cryptic behavior, nocturnality, and increasingly serious insecticide resistance, bed bugs have become one of the most difficult urban pests to control. During the investigation of bed bug infestations and control conducted in some areas of Guangdong province, China, from 2017 to 2019, it was found that bed bug infestations were increasing, and there were many challenges in the current bed bug control. Thus, how to effectively suppress the resurgence of bed bugs and adopt effective control measures has become an urgent concern. This paper reviewed literature on bed bug prevention and control, proposed corresponding countermeasures to the problems existing in the current bed bug control in China, and bed bug control strategies in China.

Objective To investigate the epidemiological features and etiology of imported chikungunya fever and related prevention and control measures, and to provide experience for future prevention and control of this disease. Methods An epidemiological investigation was performed for two patients with chikungunya fever imported from Bangladesh in August 2017 in Dongyang. Blood samples were collected and RT-PCR was used for nucleic acid detection of chikungunya virus. Results The blood samples of these two patients tested positive for nucleic acid of chikungunya virus. They both travelled to Bangladesh before the onset of this disease, and thus they were confirmed as imported cases from Bangladesh. Both patients were cured after symptomatic/supportive treatment. Conclusion Integrated prevention and control measures including isolation, mosquito control at epidemic spots, and clearance of breeding sites can effectively prevent local spread of this disease.

Objective To investigate the infection status and genotypes of Wolbachia in Nycteribiidae and Streblidae in Yunnan province, China. Methods From 2012 to 2017, Nycteribiidae and Streblidae were collected from bats in 9 spots in Yunnan province, i.e., Mangshi, Tengchong, Wanding, Menglian, Mengla, Xiangyun, Mojiang, Yongde county, and Baoshan city. The 16S rRNA and wsp genes of Wolbachia were amplified and sequenced by PCR. Phylogenetic analyses were performed on all the 16S rRNA and wsp genes. Results From 2012 to 2017, a total of 747 (48 batches) Nycteribiidae flies and 19 (6 batches) Streblidae flies were collected from the fur of bats in the 9 collection spots in Yunnan province. A total of 24 pairs of 16S rRNA and wsp genes of Wolbachia were identified in the 54 batches of samples. For the Nycteribiidae from the fur of bats in Yunnan province, Wolbachia infection was positive in 2 out of 6 batches of Phthiridium and 19 out of 37 batches of Eucampsipoda, but negative in any batch of Nycteribia (0/4) and Penicillidia (0/1). All 6 batches of Streblidae belonged to Brachytarsina, 3 out of which were positive for Wolbachia infection. Phylogenetic analysis of the 16S rRNA gene showed that the 24 Wolbachia strains shared the same evolutionary branch with one from termites (DQ837204), with a 98.80%-100% homology in nucleotide sequence. Phylogenetic analysis of the wsp gene revealed that the 24 Wolbachia strains belonged to the supergroups B and F. Conclusion Wolbachia has been widely distributed in Nycteribiidae and Streblidae in Yunnan province for a long time. Phylogenetic analysis of the wsp sequences suggests that all identified Wolbachia strains belong to the supergroups B and F.

This paper reports 2 genera, 16 species of hematophagous midges from Lushui, Fugong, Gongshan, 3 counties of Yunnan province in China-Burma border, 2017, among which 15 species in Culicoides and 1 species in Lasiohelea. A new species C. ( Culicoides) lushuiensis Liu et Feng, sp.nov. is described. Three species: C. cylindratus, C. newsteadi, La. hainana are recorded for the first time in Yunnan province. Culicoides ( Culicoides) lushuiensis Liu et Feng, sp.nov. is closely allied to C. dubius Arnaud, 1956, but is distinctly different from distal portion anal cell of wing with a ouvm shap pale spot, PR 3.00 of female; and ninth sternum with wide "V" caudomedion excavationninth, tergum apically with fine break out, Basistyle with rectangulate ventra root, aedeagus distal portion short, stout, aedeagus basal arch extending nearly to half of total length of male of C. dubius Arnaud, 1956. The type specimens are deposited in the Shenyang Command Center for Disease Control and Prevention (Shenyang 110034).

Objective To investigate the natural infection with Dengue virus(DENV) and Japanese encephalitis virus (JEV) in mosquitoes in Jinghong city, Xishuangbanna prefecture, Yunnan province, China, and provide the scientific basis for its prevention and control. Methods Mosquito samples were collected from urban area of Jinghong in August and September 2015. Mosquito samples were tested for the nucleic acid of DENV and JEV by RT-PCR assay, and positive specimens were sequenced for C/PrM gene for homology and phylogenetic analyses. Results A total of 2 009 mosquitoes were collected in urban area of Jinghong. Aedes albopictus, Ae. aegypti, Culex(species not identified) and Ae. vexans were 896, 477, 634, and 2 of the total, respectively. The JEV RNA were detected from one pool of Culex mosquitoes and one pool of Ae. albopictus mosquitoes. The sequences of C/PrM genes of the 2 JEV strains were obtained. Homology and phylogenetic analyses showed that the 2 JEV strains are consistent with the genotypeⅠ(G-Ⅰ) and its clade of the same G-Ⅰ strains of JEV from GenBank, and the JEV strains from Jinghong were closely related to the JEV G-Ⅰ strains from Honghe prefecture of Yunnan in 2009, Dehong prefecture of Yunnan in 2010 and Gansu province, China in 2008. Mosquito samples tested negative for DENV in the study. Conclusion Existence of JEV G-Ⅰ in Jinghong city of Xishuangbanna prefecture was firstly confirmed from mosquitoes in 2015. It should be strengthened to monitor and control Japanese encephalitis epidemics in this area.

Objective Application of aqueous suspension of Bacillus thuringiensis subsp. israelensis ( Bti)on the control of blackfly in the lower reaches of Irtysh River in Xinjiang Uygur Autonomous Region. Methods The pesticidal activity of Bti on blackfly larvae was tested through bioassay and field experiments, respectively. The LC ₅₀ and LC ₉₀ were computed by the Probit regression analysis method in the SPSS 19.0 software. Results The Bti aqueous suspension is an effective pesticide for blackfly control. The bioassay result suggested the LC ₅₀ was 0.034×10 ^-6 g/ml and the LC ₉₀ was 0.118×10 ^-6 g/ml. In field tests, the control efficacy of blackfly larvae was more than 83.21% in 12 h and more than 97.07% in 24 h. From 2013 to 2015, the adult population density of blackfly in the continuous application area decreased by 89.43%. Conclusion Bti aqueous suspension is an effective pesticide to control blackfly larvae, which is efficient for the prevention and control of adult blackfly in the lower reaches of the Irtysh River in Xinjiang Uygur Autonomous Region.

Objective Through intensive surveillance of three years in a row, Yersinia pestis was found within the focus, which highly supports the inner preservation theory of plague. Methods From 2013 to 2015 in four plague foci where the outbreak took place, Xingyi county in Guizhou province, Xilin and Longlin counties in Guangxi Zhuang Autonomous Region and Luoping county in Yunnan province, samples of rodents were examined for Y. pestis, serum detection of rodent hosts and indicator animals were tested for F1 antibody; samples of rodents, dogs and pigs were isolated for Y. enterocolitica and Y. pseudotuberculosis. Pig and dog samples from plague free area in Guangxi and Guizhou provinces were also examined for Y. enterocolitica and Y. pseudotuberculosis. Diarrhea children ≤5 years old were surveyed for these two enteropathogens in plague free area of Guangxi. Results Rodents and indicator animals were found carrying the F1 antibody in natural plague foci during rest period. The isolation rate of Y. enterocolitica in animals of natural plague foci was 0.80%, 1.55% and 3.59% for rodents, dogs and pigs, the rate in plague free region was 3.86% and 12.71% for dogs and pigs. For pathogenic strains, it was 0.39% and 3.19% for dogs and pigs in natural plague foci, 1.19% and 7.43% in plague free regions. Yersinia pseudotuberculosis was only isolated in plague free areas. The infection rate in children (≤5 years old) with diarrhea in plague free area of Guangxi was 0.64%. Conclusion Yersinia pestis can still be found in natural plague foci during test period, proven by F1 antibody positive rodents and indicator animals, which highly supports the inner preservation theory of plague. The isolation rate of pathogenic Y. enterocolitica was higher in plague free area than plague foci, which is an evidence of cross immunity theory.

Objective To observe the biological characteristics of Rhipicephalus sanguineus under different laboratory conditions. Methods Rhipicephalus sanguineus were cultured at different temperature in the laboratory. The growth, development and oviposition were recorded. Results Rhipicephalus sanguineus can be preserved for long term under proper conditions, and complete a generation for about 87 d at 25℃. It cannot be preserved in the laboratory when the temperature is less than 15℃. According to the prediction of the binomial model, the developmental zero temperature is 12.7℃ and the developmental peak temperature is 28.5℃. Conclusion The biological characteristics of Rh. sanguineus have been obtained under laboratory conditions. It will provide reference for colonization and scientific experiments of Rh. sanguineus.

Objective To study species distribution, community composition and bloodsucking rate of hematophagous midges in 3 cities (countries) of China-Laos border. Methods Hematophagous midges were captured with light trap to generate data on the species distribution in China-Laos border in June to August, 2015. Results A total of 78 336 host-seeking midges belonging to 54 species and 3 genera were collected; genus Leptoconops with 4 species, genus Lasiohelea with 7 species, genus Culicoedes with 43 species. Seven species, C. gentiloides, C. huffi, C. kibabaluensis, C. lansangensis, C. parahumeralis, C. variatus, La. adita are recorded for the first time in Yunnan province. The species distribution of hematophagous midges was recorded 48 species in Mengla, 22 species in Jiangcheng, 16 species in Jinghong. The predominant species included C. jacobsoni, C. parahumeralis and C. palpifer, and their composition were 25.93%, 19.25%, and 17.13% respectively. The community composition and the bloodsucking rate of midges varied with different counties and habitats. The bloodsucking rate was 32.26% in residence district, and 28.24% in cowshed. Conclusion The investigation provides the reference for species distribution, community composition and studies of infectious disease by hematophagous Culicoides.

Objective To understand the distribution of rodents, dynamics of rodent density and composition in hydro-fluctuation area of Three Gorges Reservoir after experimental storage, and provide evidence for the risk prediction and control of rodent-borne diseases. Methods Four surveillance sites were set in upper, middle and down reaches of the Three Gorges Reservoir and surveillance were carried out twice after water drainage and before water storage every year. The small mammal density and the species composition in different regions, altitude and periods were analyzed. Results During 2010-2014, a total of 116 small mammals were captured and the average capture rate was 1.10% and on the decline overall. The captured small mammals were classified into six species. Apodemus agrarius was the predominant species accounting for 46.55% and Anourosorex squamipes was next with 25.86%. The average capture rate after water drainage (1.23%) was higher than that before water storage (0.90%). All captured small mammals distributed in all three altitude and the composition decreased from high to low latitude. The predominant species was different in three altitude. The predominant species in 166-175 m altitude was A. squamipes accounting for 44.45%, followed by Rattus tanezumi (18.52% ); In 145-155 m and 156-165 m altitude, Ap. agrarius was predominant species accounting for 84.00% and 72.97%, respectively. Conclusion The average capture rate was low and there was various species of rodent in hydro-fluctuation area. The density and species composition of small mammals varies with different regions, altitude and monitoring time. The host animals of hemorrhagic fever of renal syndrome, Leptospinosis and commensal rodent plague may be of risks in hydro-fluctuation area of Three Gorges Reservoir. It should be strengthened to monitor the host animals and take the corresponding control measures in the area based on the risk assessment.

Objective To study the changes in ecological habits of important malaria vectors in the epidemic-prone regions of malaria in Yunnan province, China. Methods Light traps were used to collect mosquitoes in households and cowsheds all night, and human landing collection was used to study the pattern of mosquito activities at night in Jinghong city, Yuanjiang county, Suijiang county, and Longling county. The host source of mosquito blood meal was identified by multiplex PCR analysis. Results A total of 44 635 mosquitoes from 18 species were collected, of which Anopheles sinensis, An. minimus, An. lesteri, An. kunmingensis, and other species accounted for 59.30% (26 468), 5.44% (2428), 0.22% (100), 3.63% (1620), and 31.41% (14 019), respectively. The peaks of night activity for different Anopheles mosquito species were as follows: between 23:00 and 00:00 for An. minimus in Jinghong, between 20:00 and 21:00 for An. sinensis in Jinghong and Yuanjiang, between 02:00 and 03:00 for An. sinensis in Suijiang, and before 22:00 for An. kunmingensis. The expected lifespans of An. sinensis, An. minimus, and An. kunmingensis were relatively long, between 0.90 and 1.00. The human blood indices for An. sinensis, An. minimus, and An. kunmingensis were 0.30, 0.50, and 0.30, respectively. Conclusion The ecological habits of An. minimus, An. sinensis, and An. kunmingensis have changed, and whether these changes lead to alterations in the endemic characteristics of malaria in those regions awaits further investigation.

Objective To investigate the species composition and spatial distribution of small mammals in the mountainous region of southern Pu'er, Yunnan province, China. Methods During 2011 to 2012, a field survey was carried out at 19 villages and towns in four counties (districts) in southern Pu'er, i.e., Simao, Lancang, Menglian, and Jiangcheng. Small mammals were captured with small beast traps in an altitude range of 700-2000 m, covering six altitude gradient belts (Ⅰ-Ⅵ) and five major habitats (A-E). The richness of animal species, genera, and families was analyzed using the Shannon-Wiener diversity index. Results A total of 787 individual small mammals were captured and classified into 21 species of 11 genera, which belonged to 5 families of 3 orders, i.e., Rodentia, Insectivora, and Scandentia. Of these, the predominant species were Rattus tanezumi (30.24%), R. brunneusculus sladeni (19.95%), and Crocidura dracula dracula (10.04%). Geologically, the richness of small mammal genera and species presented a gradually increasing trend from east to west. In different vertical zones, there were 11 species (5 genera, 3 families) of small mammals in belt Ⅰ (700 m-), 14 species (10 genera, 4 families) in belt Ⅱ (900 m-), 17 species (9 genera, 5 families) in belt Ⅲ (1100 m-), 7 species (4 genera, 4 families) in belt Ⅳ (1300 m-), 13 species (7 genera, 5 families) in belt Ⅴ (1500 m-), and 12 species (7 genera, 4 families) in belt Ⅵ ( 1700 m-). By comparison, the richness of small mammal species, genera, and families was higher in belts Ⅱ and Ⅲ (around 1000 m) than in the other belts. In the major habitats, there were 13 species (8 genera, 4 families) of small mammals in type A broadleaf forest, 19 species (11 genera, 5 families) in type B farmland in broadleaf forest, 17 species (10 genera, 4 families) in type C mountainous farming area, 5 species (4 genera, 4 families) in type D shrubbery, and 9 species (6 genera, 3 families) in type E forest tea-growing area. Among different habitats, the richness of animal genera and species was the highest in the farmland in broadleaf forest. Conclusion The richness of small mammal genera and species presents a gradually increasing trend from east to west in the mountainous region of southern Pu'er, Yunnan. Among the 21 species of small mammals, R. tanezumi occurs as the predominant species with strong spatial distribution ability. There exist certain differences between different small mammal species in their selection and orientation of the environment as well as the ability of space resource utilization.

Objective To investigate the species composition and spatial distribution of small mammals in the mountainous region of southern Pu’er, Yunnan province, China. Methods During 2011 to 2012, a field survey was carried out at 19 villages and towns in four counties (districts) in southern Pu’er, i.e., Simao, Lancang, Menglian, and Jiangcheng. Small mammals were captured with small beast traps in an altitude range of 700-2000 m, covering six altitude gradient belts (Ⅰ-Ⅵ) and five major habitats (A-E). The richness of animal species, genera, and families was analyzed using the Shannon?Wiener diversity index. Results A total of 787 individual small mammals were captured and classified into 21 species of 11 genera, which belonged to 5 families of 3 orders, i.e., Rodentia, Insectivora, and Scandentia. Of these, the predominant species were Rattus tanezumi (30.24%), R. brunneusculus sladeni (19.95%), and Crocidura dracula dracula (10.04%). Geologically, the richness of small mammal genera and species presented a gradually increasing trend from east to west. In different vertical zones, there were 11 species (5 genera, 3 families) of small mammals in belt Ⅰ (700 m-), 14 species (10 genera, 4 families) in belt Ⅱ (900 m-), 17 species (9 genera, 5 families) in belt Ⅲ (1100 m-), 7 species (4 genera, 4 families) in belt Ⅳ (1300 m-), 13 species (7 genera, 5 families) in belt Ⅴ (1500 m-), and 12 species (7 genera, 4 families) in belt Ⅵ ( 1700 m-). By comparison, the richness of small mammal species, genera, and families was higher in belts Ⅱ and Ⅲ (around 1000 m) than in the other belts. In the major habitats, there were 13 species (8 genera, 4 families) of small mammals in type A broadleaf forest, 19 species (11 genera, 5 families) in type B farmland in broadleaf forest, 17 species (10 genera, 4 families) in type C mountainous farming area, 5 species (4 genera, 4 families) in type D shrubbery, and 9 species (6 genera, 3 families) in type E forest tea-growing area. Among different habitats, the richness of animal genera and species was the highest in the farmland in broadleaf forest. Conclusion The richness of small mammal genera and species presents a gradually increasing trend from east to west in the mountainous region of southern Pu’er, Yunnan. Among the 21 species of small mammals, R. tanezumi occurs as the predominant species with strong spatial distribution ability. There exist certain differences between different small mammal species in their selection and orientation of the environment as well as the ability of space resource utilization.

Objective To investigate the species composition and population density of mosquitoes before impoundment and after recession in the fluctuating zones of the Three Gorges Reservoir region and to provide a basis for surveillance of mosquito vector and mosquito-borne diseases. Methods Four surveillance sites were established in the typical fluctuating zones at upper, middle, and lower streams of the Three Gorges Reservoir region. For each surveillance site, 3 lamp trapping cages were placed at each of the three different altitudes (10 m altitude difference). The surveillance was carried out before impoundment and after recession in 2010. Results In 2010, a total of 101 mosquitoes were captured with 90 lamp-times, which belonged to 4 species: Culex tritaeniorhynchus, which accounted for 21.78% (n＝22); Anopheles sinensis, which accounted for 16.83% (n＝17); Cx. pipiens, which accounted for 16.83% (n＝17); Armigeres subalbatus, which accounted for 6.93% (n＝7); other species, which accounted for 37.63% (n＝38). Conclusion The fluctuating zones of the Three Gorges Reservoir region provide breeding sites for mosquitoes. Mosquito density is low in these areas, and shows a decreasing trend with increasing altitude. Vector of Japanese encephalitis, Cx. tritaeniorhynchus, and malaria vector, An. sinensis, exist in these areas.

Objective To investigate the population dynamics of mosquitoes in Beitun of Xinjiang Uygur Autonomous Region,China, and to provide countermeasures for the control of mosquitoes in Beitun. Methods The population dynamics of mosquitoes were monitored continuously from April to September during 2011-2013. The number of immature mosquitoes in different aquatic habitats was monitored every two days. The water bodies positive for immature mosquitoes were sprayed with biological agents (Bacillus thuringiensissubsp. israelensis please specify the product used) and continuously monitored for 48 hours. The number of adult mosquitoes was surveyed by light trap capture from 10p.m. to 12 p.m. every day at the city square of Beitun. Results Immature mosquitoes appeared from May to July, with a peak density from late May to early June. The density of immature mosquitoes in breeding sources was 18 240 mosquitoes/m ². The control efficacy of biological agent (B.t.i) reached up to 96%. Adult mosquitoes usually appeared from April to September, with a peak activity period from June to July. The highest density of adult mosquitoes was 85 mosquitoes per day (determined by light trap). Mosquito density showed decline curve by two times of aerial spraying insecticides including biological pesticides B.t.i(Titer 480 ITU/mg) and 2.5% lambda-cyhalothrin EC,10% acetamiprid EC and silicone additives to control mosquito. Conclusion Mosquito density used to be high in Beitun, but was decreased by 75% through integrated control for three years.

Objective To investigateMusca domesticaresistance to commonly used insecticides in Tianjin in 2013, and to study the trend of insecticide resistance by comparison with historical data. Methods Biological assay was applied to determine the resistance levels of M. domesticato deltamethrin, DDVP, and propoxur. The results were compared with the historical data in Tianjin. Results Musca domesticain Tianjin showed different levels of resistance to DDVP, deltamethrin, and propoxur. Statistics revealed that the resistance to DDVP (19.33 fold) was the highest, and that the resistance to propoxur was the lowest (2.34 fold). The resistance to propoxur was significantly different from the resistance to the other two insecticides. In comparison with historical data, it was found that the insecticide resistance ofM. domesticawas decreased in Tianjin in the past 10 years. Conclusion Routine surveillance of the insecticide resistance ofM. domesticaprovides evidence for the screening and rational use of chemical pesticides, and therefore effectively prevents and delays the insecticide resistance ofM. domestica.

Objective To survey the knowledge and behaviors about vector control among residents and their impact factors in Tianjin, China and to provide a basis for effective intervention strategy. Methods A questionnaire survey was conducted in 630 residents who were selected from 18 districts or counties of Tianjin by simple random sampling. Results Of all residents surveyed, 62.1% said that their residence had been infested with vectors; only 44.6% had the knowledge about vector control, and there were relatively few residents with the knowledge about control of mosquitoes and cockroaches. Age, career, and education level were significant impact factors for the knowledge about vector control in Tianjin residents (P＜0.05); the proportion of participants with knowledge about vector control increased as the age (OR＝2.417) and education level (OR＝1.764) increased, and it was higher in officials than in farmers (OR＝2.374). The residents knew little about vector control strategy; only 31.5% of the participants thought integrated vector management based on breeding source management was the key to preventing vector infestation in urban areas. Only 5.3% of the participants were willing to hire PCO professionals for pest control after finding vector infestation. The main approach to gaining knowledge about vector control was radio or television (37.7%), and few residents gained the knowledge through health education in the community (19.7%). Conclusion The knowledge about vector control among the residents in Tianjin is insufficient, and their behaviors conducive to vector breeding source creation need to be adjusted. The health education about vector?borne diseases and training on vector control need to be implemented among residents, especially farmers, retired people, young genetrations and those with an education level of junior high school or lower.

Objective To establish the evaluation index system for vector control by Delphi method. Methods Literature method was used to establish a preliminary evaluation index system. Based on this, the suitable indices were selected and their weights were determined according to the average scores of importance and coefficients of variation. Results After a two-round expert consultation, the evaluation index system was established; it consisted of 6 first-level indices, 27 second?level indices, and 33 third?level indices. The response rate of each round was more than 90%; the expert authority coefficient was 0.82; the coordination coefficients were 0.23 (χ2＝126.07, P＜0.05) and 0.41 (χ2＝212.26, P＜0.05). Conclusion The evaluation index system for vector control has been preliminarily established, which provides a quantitative basis for vector control capability evaluation.

Objective To address the current issues of pest control operation (PCO) and pest control workers in Tianjin, China and to promote the specialized and standardized development of PCO companies in Tianjin. Methods A questionnaire survey was conducted in 165 pest control workers who were trained in 2010-2011 to address their general concerns and to determine their awareness of pest control knowledge. One pest control worker was randomly selected from each PCO company. Results The pest control workers in Tianjin had a young age structure; there were still some workers with low educational levels; the workers had short lengths of service and low wage levels. The PCO companies in Tianjin are small in size, have small market coverage and distinct regional characteristics, severe price competition in the low-end market, and low degree of specialization. Some companies sought short-term profits and had no operation standardization. The pest control workers in Tianjin had a relatively high awareness of fly and rodent control knowledge but a relatively low awareness of mosquito and cockroach control knowledge, and their awareness of integrated control of insect-borne diseases was particularly low. Conclusion The Pest Management Association must regulate and provide professional training of pest control workers, and establish the principle that chemical control is only part of the integrated pest management. It is necessary to promote category-specific regulations for pesticides, adaptation of relevant regulations and standards, quality control of third-part service, establishment of liability insurance, etc., so that the PCO companies are transformed into pest management professional companies and are more environment-friendly, specialized, and institutionalized.

Objective To assess the risk of vector-borne diseases in Tianjin, China by risk recognition and assessment methods and to prevent the incidence and prevalence of vector-borne diseases. Methods The risk of vector-borne diseases was graded by comprehensively analyzing the probability and consequence of risk events. Results In Tianjin, the density of rodents decreased year by year from 2008 to 2012 (0.64%-0.19%); the captured rodents had a negative indirect hemagglutination test for plague F1 antibody; hantavirus was detected in 0.85% of rodents in 2011 but not in 2012. The mean densities of mosquitoes were 14.11, 15.04, 16.64, 18.82, and 11.09 mosquitoes/lamp·h; no pathogens were detected in mosquitoes in recent years. In Tianjin, the prevalence of mosquito-borne diseases, such as malaria, dengue fever, and Japanese encephalitis, and rodent-borne diseases, such as plague and hemorrhagic fever with renal syndrome, were at low risk. Conclusion Certain risk of vector-borne diseases does exist in Tianjin. In order to prevent and control the incidence and prevalence of vector-borne diseases, it is necessary to strengthen the surveillance of vector density and pathogens carried by main vectors, enhance the knowledge among residents, and make good preparation for the potential introduction and disposal of vector-borne pathogens.

Objective To investigate the degree and characteristics of Blattella germanica infestation in the restaurants of Tianjin, China and the influential factors, and to provide a scientific basis for the development of cockroach control measures. Methods The infestation rate, population density, and density index of B. germanica were monitored using sticky traps in 540 restaurants in 18 counties or districts of Tianjin. Results The mean infestation rate of B. germanica in the restaurants of Tianjin was 43.9%, and the mean population density and density index were 3.7 individuals/trap and 20.6 individuals/room, respectively. The population density of B. germanica was relatively high in the operating room, cold meat room, pastry room, and food warehouse, where the refrigerator radiator, dressing storage cabinet, and repository of pastry materials and sweets were the main places infested. The infestation rates of B. germanica in urban areas and villages/towns were 43.0% and 44.8%, respectively, without significant difference between them (P>0.05). The infestation rates of B. germanica in small and medium-sized restaurants and large restaurants were 43.4% and 45.1%, respectively, without significant difference between them (P>0.05). The infestation rates of B. germanica in restaurants with an operating period less than 5 years, restaurants with an operating period of 5-10 years, and restaurants with an operating period greater than 10 years were 33.9%, 57.6%, and 56.9%, respectively, with significant differences between them (P<0.01). The univariate logistic regression analysis showed that the operating period was an influential factor for the infestation rate of B. germanica in restaurants (OR=1.833, P<0.05). Conclusion The restaurants in Tianjin are heavily infested with B. germanica, so surveillance and early control should be enhanced, particularly at the refrigerator radiator, dressing storage cabinet, and repository of pastry materials and sweets in operating room, cold meat room, pastry room, and food warehouse.

Objective To investigate the anti-tick skills and their influential factors among the residents in Tianjin, China, and to provide a basis for effective control measures and health education. Methods A questionnaire survey was conducted in 360 residents aged above 15 years who were randomly selected from 18 districts or counties of Tianjin. Results Of all respondent residents, 61.7% (221/358) mastered anti-tick skills, including anti-tick methods (81.8%), knowledge about the habits and characteristics of tick (59.2%), self-protection methods (72.1%) like wearing light-colored clothes (35.8%) and applying repellent (57.8%), and post-biting treatment methods (76.3%). Among the residents aged above 50 years, 50.0% (38/76) mastered anti-tick skills, compared with 64.9% (183/282) in the rest of the residents ( P ＜ 0.05). Among the residents with college diplomas, 73.6% (156/212) mastered anti-tick skills, compared with 50.0% (36/72) in those with only high-school diplomas and 39.2% (29/74) in the rest of the residents ( P ＜ 0.01). Among the residents with a monthly income of less than 2000 yuan, 38.5% (42/109) mastered anti-tick skills, compared with 71.4% (147/206) in those with a monthly income of 2000-4000 yuan and 74.4% (32/43) in those with a monthly income more than 4000 yuan ( P ＜ 0.01). The logistic regression analysis showed that career and monthly income were the significant factors affecting the anti-tick skills of residents in Tianjin ( P ＜ 0.05). Conclusion Career and monthly income are the main influential factors for the anti-tick skills of the residents in Tianjin. Health education should be enhanced among the workers in high-risk industry and low-income residents to improve their anti-tick skills.

Objective To compare the mosquito-trapping effects of lamp trapping method and labor hour method, and to provide a reference for scientific and standard investigation and surveillance of mosquitoes. Methods Three sites were selected for study based on their different distances from the Three Gorge Dam in the Three Gorge Reservoir area. The mosquito species composition was investigated by lamp trapping method and labor hour method in the residential area and its surrounding area at each of the three sites. Results Both lamp trapping method and labor hour method could be used for successfully trapping the main species of mosquitoes in the Three Gorge Reservoir area, but with difference in the species composition of captured mosquitoes. More Culex tritaeniorhynchus was captured with lamp trapping method than with labor hour method, while much more Armigeres subalbatus was captured with labor hour method than with lamp trapping method. The application scope of lamp trapping method was wider than that of labor hour method. The lamp trapping method could be applied in the places without space limitation, such as forest and rice field. Conclusion Each mosquito trapping method has its own advantages and disadvantages. Lamp trapping method is recommended in the long-time investigation and surveillance of mosquitoes.

Objective To establish a method for identifying Haemaphysalis flava and H. campanulata with molecular markers, and to learn the phylogenetic relationship of these two species of ticks. Methods Ticks were collected from animals in Hubei province, and identified by their morphological characteristics. The 12S rDNA, 16S rDNA and mitochondrial cytochrome oxidase subunitⅠ gene (COⅠ) from the genomes of the two kinds of ticks were amplified by PCR and sequenced for homology analysis. PAUP 4.0 software was used to construct the individual phylogenetic trees of the three gene sequences for phylogenetic analysis. Results The identities between the two species of ticks were 90.8% for 12S rDNA, 90.4% for 16S rDNA, and 86.8% for COⅠ, with the homology of the three gene fragments between collected H. flava and known H. flava being 100%, 99.5% and 99.7%, respectively. On the phylogenetic trees based on the nucleotide sequences of the 12S rDNA, 16S rDNA and COⅠ gene, H. flava was clustered together with their respective class. H. flava and the H. campanulata formed a distinct branch. However, the phylogenetic relationships between different species of Haemaphysalis were different on the trees based on these three genes. Conclusion As far as identification of the ticks with similar morphological features is concerned, molecular markers in combination with the conventional morphological classification can make it more accurate, contributing to better understanding of their evolutionary relationships. Further phylogenetic analysis of gene sequences also provides a basis for the prevention and control of tick-borne diseases.

Objective To Evaluate the resistance of Rattus norvegicus to warfarin and bromadiolone in Chengdu, providing information for the control of rodents in the area. Methods The resistance of R. norvegicus to warfarin and bromadiolone was tested with the BCR method. Results The 50% effective dose (ED50) of warfarin for R. norvigicus was 0.873 mg/kg for male and 1.439 mg/kg for female respectively, with that of bromadiolone being 1.091 mg/kg for male and 1.296 mg/kg for female. According to the baseline of susceptible R. norvegicus approved by ARRPC,the resistance level of the population was between 0.578 and 0.676 fold for warfarin and between 2.125 and 2.321 for bromadiolone. Conclusion R. norvigicus populations have developed resistance to bromadiolone in Chengdu, but not to warfarin, which can be used as an alternative rodenticide applicable to the control of rodents in Chengdu.

Objective To develop a highly specific and sensitive Real-time PCR assay to detect Orientia tsutsugamushi. Methods A pair of primers and a TaqMan-MGB probe were designed according to the 56 kD outer membrane gene sequence. Results A linear relationship between the threshold cycle (Ct) of the quantitative Real-time PCR and the DNA copy number was demonstrated (r=0.99). The standard curve showed that 26 copies target genes per reaction could be detected by this method. The lowest detection limit of this method was 2 copies per μl. The method showed high species specificity and good reproducibility. Conclusion These results suggested that the Real-time PCR with TaqMan-MGB assay is highly specific and sensitive for the detection of O. tsutsugamushi, which might be applied for the diagnosis of this infection.