Objective To establish the DNA barcoding identification techniques for 49 common fly species in Calliphoridae, and provide references data for the identification of these species with DNA barcodes, as to solve the difficulties in the accurate identification of the females, individuals with lost morphological features, and larvae of these species. Methods Totally 166 individuals in Calliphoridae were identified with the morphological characters which were collected from different areas in China during 2012 to 2015, DNA barcodes of the males and females were obtained, an N-J tree was constructed in combination with published data. Consistency of the identification results by morphology and DNA barcodes were tested as well. Results All the 166 sequences of these 49 species were submitted to the public data base, DNA barcodes sequences of 28 species were obtained for the first time. The differences of the intra-species was less than 1%, and the inter-species DNA barcode differences increasing with the morphology differences, and the identification results based on DNA barcodes were consistent with that based on morphology. Conclusion The application of DNA barcodes were strong supplemental to the morphology identification of the species, especially to the incomplete, non-adult and female individuals, and can increase accuracy of identification, and it is very important for the rapid and accurate species identification of the medical vectors at the ports.

Objective To identify the flesh fly from an entry ship rapidly and accurately with‘Barcoding and Morphology’ techniques. Methods A flesh fly specimen was collected by Zhongshan CIQ on an entry ship from Australia with grapes. Morphological identification was conducted, and genomic DNA were extracted from a hind-leg of the fly. Target fragments were amplified with animal DNA barcoding universal primers (LCO1490 and HCO2198), then purified, cloned, sequenced and blasted in BOLD, a NJ tree was built based on the original data. Results From the morphological identification, it is a female Sarcophaga sp. Because of the lack of the references for female flesh flies, we could only identify it to a female Sarcophaga sp. Sequence of the DNA barcode of it was 100% identical to the published sequence of Sarcophaga australis in BOLD.‘Science and technology novelty search’result showed that it is a non-recorded species in China. Conclusion Based on the results, it could be concluded that the fly was a female S. australis. It is intercepted for the first time at China port.‘B & M’techniques is a good tool to identify the intercepted non-recorded fly species.

Objective To apply DNA barcoding in species identification of exotic vectors. Methods Morphology and DNA barcoding based identification was performed on two blowflies from Australia that were intercepted at the Shenwan port (Zhongshan, Guangdong) in September 2013. The identification results were confirmed with specialists in Australia and compared to data of a similar species, Calliphora augur. Results The two blowflies were identified as C. dubia, which represents exotic species in China based on sci-tech novelty search. Conclusion DNA barcoding is a strong supplementary tool for conventional taxonomy, which plays an important role in frontier health and quarantine inspection by timely identifying vector insects, reducing the spread of vector insects and the carried pathogens, and mitigating threats to public health.

Objective To apply DNA barcoding in species identification of exotic vectors. Methods Morphology and DNA barcoding based identification was performed on two blowflies from Australia that were intercepted at the Shenwan port (Zhongshan, Guangdong) in September 2013. The identification results were confirmed with specialists in Australia and compared to data of a similar species, Calliphora augur. Results The two blowflies were identified as C. dubia, which represents exotic species in China based on sci?tech novelty search. Conclusion DNA barcoding is a strong supplementary tool for conventional taxonomy, which plays an important role in frontier health and quarantine inspection by timely identifying vector insects, reducing the spread of vector insects and the carried pathogens, and mitigating threats to public health.

Objective To ensure the purity of template in amplification of barcode DNA from the microtissues of medical vectors and to minimize the morphological damage to specimens, a method of direct PCR without DNA extraction was established.Methods Individuals of mites, fleas, and ticks, as well as the first tarsus of metapodium from flies, were used as samples in this study. The concentration of lysis buffer and the ratio of lysisvs. stop buffer were optimized to determine the reaction conditions.KOD FX DNA polymerase was used instead of Taq DNA polymerase to directly amplify barcode DNA. The PCR product was sequenced and aligned with GenBank sequences using Blast to test whether the sequences were contaminated. Results The optimized lysis buffer was 50 mmol/L NaOH. The optimized ratio of lysisvs. stopbufferwas180μl∶20μl. The optimized reaction system(50μl) was determined as follows: 2×KOD FX DNA polymerase buffer (containing Mg²⁺) 25μl, 2mmol/LdNTP 10μl , KOD FX DNA polymerase (1U/μl)1 μl, forward primerLCO1490(20μmol/L)1μl, andreverseprimer HCO2198(20μmol/L)1μl. The reaction conditions were optimized as follows: 95℃3 min for pre-heating; 98℃10 s, 50℃30 s, and 68℃1 min for35 cycles, followed by extension 7 min at 68℃. No contamination was found by Blast alignment of amplified sequences.Conclusion The method established in this study is easy to operate, and omission of DNA extraction will save time and expenses. This method is suitable for direct amplification of barcode DNA from mites, fleas, ticks, and even the first tarsus of fly metapodium.

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.

【Abstract】 Objective To investigate control effect of flies in temporary shelters of post-earthquake disaster areas and to provide the science evidence for its control. Methods Manage environmental sanitation and spray periodically insecticide to control flies in temporary shelters. Investigate fly population by cage-trap method, monitor flies density by eye balling method in washrooms, trash piles(cans), inside and outside of tents, and survey the management of flies breeding sites and prevention and control measure of civilians by field observation. Results Houseflies was the dominant species （96.85%） in the temporary shelters. The average fly density was less than one fly/m2 around temporary shelters after the earthquake for 4-8 weeks. At the fifth week post-earthquake, all the shelters were equipped with washroom. About 76.92% trash containers were equipped, and 69.23% of domestic garbage were cleaned up and transported away without delay. 94.01% of food-leavings was covered, and 93.00% of tableware was deposited in the cupboard. Conclusion It was effective to take the integrated pest management measures to control flies in the temporary shelters. The control of breeding sites should be strengthened in the future.