Development and field evaluation of a novel sugar bait device for controlling residential vector mosquitoes in Zhejiang Province, China.

Background: Mosquito-borne diseases pose serious public health threats in Zhejiang Province, China, and vector control is believed to be the primary method for reducing transmission. Due to severe resistance problems, effective and sustainable methods without chemical insecticides are urgently required to control mosquito vectors. Attractive toxic sugar baits (ATSB) are newly developed methods to control mosquitoes in recent decades with the core element sugar bait, which was invented according to the sugar-feeding behavior of mosquitoes. In this study, we developed a Novel Sugar Bait Device (NSBD) trap by combining sugar bait and physical adhesive capture technology. The study aimed to evaluate the effect of the NSBD trap on controlling mosquitoes in residential environments and to identify the optimal sugar solution concentration in the sugar bait of the NSBD for real use. Methods: Four residential villages in Ningbo City with similar geographic environments and mosquito densities were selected for field trials in 2022. One village (site 1) was designated as the control group, and three villages (sites 2-4) served as the test groups to assess the effectiveness of NSBD traps with different sugar solution concentrations (6, 8, and 10%) in the sugar bait. Larval and adult mosquito densities were monitored monthly before and semi-monthly after the trials using the CDC light trap and larval pipette method. Results: Before the trials, we monitored mosquito density for 3 months to confirm the baseline mosquito density among the four sites, and no statistical differences in adult and larval mosquitoes were found (adult, F = 3.047, p > 0.05; larvae, F = 0.436, p > 0.05). After the trials, all NCBD traps effectively controlled larval and adult mosquito densities, with the highest standard decrease rates of larval and adult mosquito densities at 57.80 and 86.31%, respectively, observed in site 4. The most suitable sugar solution concentration in the sugar bait was 10%. Conclusion: NSBD traps effectively controlled mosquitoes in residential environments during field trials. Without the use of insecticides, this may be a promising choice for mosquito vector control to prevent mosquito-borne diseases.

The ecological and etiological investigation of ticks and rodents in China: results from an ongoing surveillance study in Zhejiang Province.

Objectives: This study aimed to analyze the population density of vector ticks and reservoir hosts rodents, and to investigate the relevant pathogen infection in Zhejiang Province, China. Methods: In this surveillance study, the data of ticks density were collected with the tick picking method on animal body surface and the drag-flag method, while the rodent density with the night trapping method. The samples of ticks were examined for the severe fever with thrombocytopenia syndrome virus (SFTSV), and blood serum and organs from rodents were subjected for SFTSV, hantavirus, Leptospira, Orientia tsutsugamushi (O. tsutsugamushi) and Yersinia pestis (Y. pestis) screening in the laboratory. Results: From 2017 to 2022 in Zhejiang Province, 16,230 parasitic ticks were found in 1848 positive animals, with the density of parasitic ticks of 1.29 ticks per host animal, and a total of 5,201 questing ticks were captured from 1,140,910 meters of vegetation distance with the questing tick density of 0.46 ticks/flag·100 m. Haemaphysalis longicornis (H. longicornis) was the major species. A total of 2,187,739 mousetraps were distributed and 12,705 rodents were trapped, with the density of 0.58 per 100 trap-nights. Rattus norvegicus was the major species. For SFTSV screening, two groups nymphal ticks of H. longicornis were tested to be positive. For the rodents samples, the Leptospira had a positive rate of 12.28% (197/1604), the hantavirus was 1.00% (16/1604), and the O. tsutsugamushi was 0.15% (2/1332). No positive results were found with SFTSV and Y. pestis in the rodents samples. Conclusion: Findings from this study indicated that the ticks and rodents were widely distributed in Zhejiang Province. Particularly, the positive detection of SFTSV, Leptospira, hantavirus and O. tsutsugamushi in ticks or rodents from this area suggested that more attention should be paid to the possibilities of relevant vector-borne diseases occurrence.

A Review of Pathogens Transmitted by the Container-Inhabiting Mosquitoes, Aedes Albopictus, A Global Public Health Threat.

Dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus (ZIKV) are highly pathogenic human arboviruses transmitted by the Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) or Ae. Albopictus mosquito. These arboviruses are responsible for causing fever, hemorrhagic conditions, and neurological diseases in humans post-bite from an infected Aedes mosquito. Over the past 80 years, the Ae. albopictus has infested every habitable continent, bar Antarctica, thereby escalating the probability of global insect-borne infectious disease outbreaks. This research follows the global transmission pattern of Ae. albopictus and provides a summary of disease prevention and control strategies for mosquito-borne infections, as implemented by the World Health Organization (WHO) and both Asian and European countries. Consequently, this study can aid in the prevention and control of mosquito-borne diseases while acting as a basis for international collaboration on effectively managing arbovirus infection issues in public health.

Evaluation of the effectiveness of the human-baited double net traps and BG traps compared with the human landing catches for collecting outdoor Aedes albopictus in China.

Dengue fever is one of the biggest threats to public health in China, causing huge disease burden and economic loss. Aedes-mosquito surveillance could be a cornerstone for predicting the risk of Aedes-borne diseases and evaluating the effect of vector management during diseases outbreaks. The human landing catch (HLC) method is regarded as the "gold standard" for catching Aedes mosquitoes, but it potentially exposes field professionals to vectors of known or unknown pathogens. Human-baited double net (HDN) was recommended to replace HLC for emergency monitoring in China when Aedes-borne diseases break out, but it had been reported with low efficiency for capturing Aedes mosquitoes. In this study, we compared HLC with HDN and BG traps for field Aedes albopictus monitoring, with the aim of evaluating the effectiveness of HDN replacing HLC and finding an effective and safe alternative to the HLC for monitoring Aedes albopictus. Six sites in Hangzhou, Shaoxing, and Yiwu, Zhejiang Province, China, were chosen to conduct outdoor HLC, HDN, and BG trap catches from June to October 2021. The tests were performed 3 h apart: 8:30-9:30 AM, 16:30-17:30 PM, and 17:30-18:30 PM. A total of 2330 adult mosquitoes were collected, and Aedes albopictus was the most abundant species in all three catches with 848(98.95%), 559(97.39%) and 867 (96.44%) caught in HLC, HDN and BG traps respectively. Compared to HLC, HDN collected significantly less Ae. albopictus and Ae. albopictus females per trapping period (P < 0.001, P < 0.001), whereas no statistical differences were found between the HLC and BG trap (P = 0.970, P > 0.05). Statistically significant positive spatial correlations for Ae. albopictus sampling was found between HLC and HDN traps (r = 0.543, P < 0.001) and HLC and BG traps (r = 0.658, P < 0.001). In conclusion, both the BG trap and HDN have a significant positive spatial correlation with HLC, making them safer alternatives to HLC for Ae. albopictus monitoring in China. However, with better a sampling efficiency, being less labor intensive, and no human-baited attraction bias, the BG trap could be a better choice than the HDN trap.

A Review of the Surveillance Techniques for Aedes albopictus.

Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) transmits a variety of arboviruses (arthropod-borne viruses) and acts as one of the most dangerous mosquito species in the world. Mosquito surveillance is the main means of evaluating vector density, vector-borne disease risk, and the efficacy of vector-control operations. The larval density of Ae. albopictus can be reflected by means of Breteau index and Route index, and egg density can be monitored by ovitrap and mosq-ovitrap, whereas mosquito surveillance methods mainly include human landing catch, human-baited double net trap, BG-Sentinel trap, autocidal gravid ovitrap, gravid Aedes trap, and mosquito magnet. This article describes different methods of Ae. albopictus surveillance and offers suggestions to improve surveillance.

Practice of integrated vector surveillance of arthropod vectors, pathogens and reservoir hosts to monitor the occurrence of tropical vector-borne diseases in 2020 in Zhejiang Province, China.

Background: Vector-borne diseases have become one of the most serious local public health threats. Monitoring and controlling vectors are important means of controlling vector-borne diseases. However, traditional vector surveillance systems in China mainly monitor vector density, making its early-warning effect on vector-borne diseases weak. In this study, we applied an integrated surveillance system of multiple arthropod vectors and reservoir host containing ecology, etiology, and drug resistance monitoring to obtain better knowledge on vector populations and provide early warning of suspicious vector-borne infectious disease occurrence. Methods: An ecology surveillance of mosquitoes, rodents, ticks, and chigger mites, a pathogen infection survey on mosquitoes and rodents, and a drug resistance survey on Aedes albopictus were conducted in 12 cities in Zhejiang Province in 2020. Results: A total of 15,645 adult mosquitoes were collected at a density of 19.8 mosquitoes per Centers for Disease Control and Prevention light trap. Culex tritaeniorhynchus (72.76%) was the most abundant species. The Breteau index of Ae. albopictus was 13.11. The rodent density was 0.91 rodents per hundred traps; the most abundant species was Rattus norvegicus (33.73%). The densities of dissociate and ectoparasitic ticks were 0.79 ticks per hundred meters and 0.97 ticks per animal, respectively. The most abundant tick species was Haemaphysalis longicornis (56.38%). The density of chigger mites was 14.11 per rodent; two species were identified, with the most abundant species being Walchia spp. mite (68.35%). No flavivirus or alphavirus was found in mosquito etiology monitoring, whereas the positivity rates of hantavirus, the pathogenic bacteria Leptospira spp., Orientia tsutsugamushi, and Bartonella spp. detected in rodent etiology monitoring were 1.86, 7.36, 0.35 and 7.05%, respectively. Field populations of Ae. albopictus in Zhejiang Province were widely resistant to pyrethroids but sensitive to most insecticides tested, including organophosphorus and carbamate insecticides. Conclusion: Integrated surveillance systems on multiple arthropod vectors (mosquitoes, ticks, mites) and animal reservoirs (rodents) can provide important information for the prevention and control of epidemic emergencies.

Fine-scale genetic structure and wolbachia infection of aedes albopictus (Diptera: Culicidae) in Nanjing city, China.

Background: Aedes albopictus is an indigenous primary vector of dengue and Zika viruses in China. Wolbachia is a gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Compared with research on the dispersion of Ae. albopictus at the macrospatial level (mainly at the country or continent level), little is known about its variation and Wolbachia infection at the microspatial level, which is essential for its management. Meanwhile, no local cases of dengue fever have been recorded in the history of Nanjing, which implies that few adulticides have been applied in the city. Thus, the present study examines how the Ae. albopictus population varies and the Wolbachia infection status of each population among microspatial regions of Nanjing City. Methods: The genetic structure of 17 Aedes albopictus populations collected from urban, urban fringe, and rural regions of Nanjing City was investigated based on 9 microsatellite loci and the mitochondrial coxI gene. The Wolbachia infection status of each population was also assessed with Wolbachia A- and Wolbachia B-specific primers. Results: Nine out of 58 tested pairs of microsatellite markers were highly polymorphic, with a mean PIC value of 0.560, and these markers were therefore chosen for microsatellite genotyping analysis. The Na value of each Ae. albopictus population was very high, and the urban area populations (7.353 ± 4.975) showed a lower mean value than the urban fringe region populations (7.866 ± 5.010). A total of 19 coxI haplotypes were observed among 329 Ae. albopictus individuals via haplotype genotyping, with the highest diversity observed among the urban fringe Ae. albopictus populations (Hd = 0.456) and the lowest among the urban populations (Hd = 0.277). Each Ae. albopictus population showed significant departure from HWE, and significant population expansion was observed in only three populations from the urban (ZSL), urban fringe (HAJY), and rural areas (HSZY) (p < 0.05). Combined with DAPC analysis, all the Ae. albopictus populations were adequately allocated to two clades with significant genetic differences according to population structure analysis, and the best K value was equal to two. AMOVA results showed that most (96.18%) of the genetic variation detected in Ae. albopictus occurred within individuals (FIT = 0.22238, p < 0.0001), while no significant positive correlation was observed via isolation by distance (IBD) analysis (R 2 = 0.03262, p = 0.584). The TCS network of all haplotypes showed that haplotype 1 (H1) and haplotype 4 (H4) were the most frequent haplotypes among all populations, and the haplotype frequency significantly increased from urban regions (36.84%) to rural regions (68.42%). Frequent migration was observed among Ae. albopictus populations from rural to urban regions via the urban fringe region, with four direct migration routes between rural and urban regions. Furthermore, Wolbachia genotyping results showed that most of the individuals of each population were coinfected with Wolbachia A and Wolbachia B. The independent infection rate of Wolbachia A was slightly higher than that of Wolbachia B, and no significant differences were observed among different regions. Conclusion: In the microspatial environment of Nanjing City, the urban fringe region is an important region for the dispersion of Ae. albopictus populations between rural and urban areas, and Wolbachia A and Wolbachia B coinfection is the most common Wolbachia infection status in all Ae. albopictus populations among different regions.

Screening and analysis of immune-related genes of Aedes aegypti infected with DENV2.

Dengue virus type Ⅱ (DENV2) is a primary serotype responsible for the dengue fever epidemic, and Aedes aegypti is the main DENV2 vector. Understanding the Aedes aegypti immune mechanism against DENV2 is the basis for research on immune blockade in mosquitoes. Some preliminary studies lack validation in the literature, so this study was performed to further study and validate the potential target genes to provide a further basis for screening key target genes. We screened 51 genes possibly related to Aedes aegypti infection and immunity from the literature for further verification. First, bioinformatic methods such as GO, KEGG and PPI analysis were used, and then RT-qPCR was used to detect the changes in mRNA expression in the midguts and salivary glands of Aedes aegypti infected with DENV2.Bioinformatic analysis showed that mostly genes of the glucose metabolism pathway and myoprotein were influenced. In salivary glands, the Gst (xa) and Toll (xb) expression levels were significantly correlated with DENV2 load (y, lg[DENV2 RNA copies]), y = -3436xa+0.2287xb+3.8194 (adjusted R2 = 0.5563, F = 9.148, PF = 0.0045). In midguts, DENV2 load was significantly correlated with the relative Fba(R2 = 0.4381, t = 2.497, p < 0.05, df = 8), UcCr(R2 = 0.4072, t = 2.344, p < 0.05, df = 8) and Gbps1(R2 = 0.4678, t = 2.652, p < 0.05, df = 8) expression levels, but multiple regression did not yield significant results. This study shows that genes related to glucose metabolism and muscle proteins contribute to the interaction between Aedes aegypti and dengue virus. It was confirmed that SAAG-4, histone H4, endoplasmin, catalase and other genes are involved in the regulation of DENV2 infection in Aedes aegypti. It was revealed that GST and Toll in salivary glands may have antagonistic effects on the regulation of DENV2 load. Fba, UcCr and Gbps1 in the midgut may increase DENV2 load. These study results further condensed the potential target gene range of the Aedes aegypti immune mechanism against DENV2 infection and provided basic information for research on the Aedes aegypti in vivo blockade strategy against DENV2.

Entomological Investigation and Detection of Dengue Virus Type 1 in Aedes (Stegomyia) albopictus (Skuse) During the 2018-2020 Outbreak in Zhejiang Province, China.

Mosquito-borne diseases are still threats to public health in the Zhejiang province of China. Surveillance of mosquitoes and the mosquito-borne pathogen is a vital approach for early warning, prevention, and control of the infectious disease. In this study, from 2018 to 2020, a total of 141607 female mosquitoes were caught by means of the light trap method. The main species were Culex pipiens quinquefasciatus/pallens (41.32%), Culex tritaeniorhynchus (47.6%), Aedes albopictus (2.5%), Anopheles sinensis (5.87%), Armigeres subalbatus (2.64%) and other mosquito species (0.07%). Cx. pipiens s.l. were the dominant species in two urban habitats and rural residential areas while Cx. tritaeniorhynchus was the main dominant species in the rural livestock sheds. In terms of seasonal fluctuation, Cx. pipiens s.l fluctuated at a high level from May to October. The peaks of Cx. tritaeniorhynchus, An. sinensis and Ar. subalbatus were in July. In addition, a total of 693 Ae. albopictus were collected with Biogents Mosquitaire CO2 traps in emergency surveillance of dengue fever (DF) and screened for dengue virus infection. There were three circumstances of collection: The first: the sampling time before mosquito control during the local outbreak of DF in Lucheng of Wenzhou, 2019; The second circumstance: the sampling time after mosquito control during the local outbreak of DF of other cities in 2018-2019; The third circumstance: past DF epidemic areas the sampling time before mosquito control during the local outbreak of DF in Lucheng, Wenzhou, Zhejiang, 2019. The pools formed by mosquitoes collected in these three circumstances were 3 (6.1%), 35 (71.5%), and 11 (22.4%) respectively. Of the 49 pools tested, only one in the first circumstance was positive. The full-length dengue virus sequence of ZJWZ/2019 was obtained by sequencing and uploaded to the NCBI as number OK448162. Full-length nucleotide and amino acid homology analyses showed that ZJWZ2019 and Wenzhou DF serum isolates ZJWZ-62/2019 (MW582816) and ZJWZ-18/2019 (MW582815) had the highest homology. The analysis of full genome and E gene phylogenetic trees showed that ZJWZ2019 belonged to serotype 1, genotype I, lineage II, which was evolutionarily related to OK159963/Cambodia/2019. It implies that ZJWZ2019 originated in Cambodia. This study showed the species composition, seasonal dynamics of mosquitoes in different habitats in Zhejiang province and confirmed the role of Ae. albopictus in the transmission cycle of in outbreak of DF in the Lucheng district of Wenzhou in 2019, suggesting the importance of monitoring of vector Aedes infected dengue virus in the prevention and control of DF.

From SARS to the Omicron variant of COVID-19: China's policy adjustments and changes to prevent and control infectious diseases.

The COVID-19 pandemic has been the biggest public health crisis in a century. Since it was initially reported in 2019, the duration and intensity of its impacts are still in serious question around the world, and it is about to enter its third year. The first public health revolution failed to achieve its ultimate targets, as previously contained infectious diseases seem to have returned, and new infectious diseases continue to emerge. The prevention and control of infectious diseases is still a public health priority worldwide. After SARS, China adjusted a series of its infectious disease policies. In order to ensure the effectiveness and implementation of prevention and control interventions, the government should integrate the concept of public health. Perhaps we need a global public health system at the government level to fight the potential threat of infectious disease. This system could include multifaceted strategies, not just specific prevention and control interventions, and it could also be a comprehensive system to ensure unimpeded communication and cooperation as well as sustainable development.

Vectors, Hosts, and the Possible Risk Factors Associated with Severe Fever with Thrombocytopenia Syndrome.

Severe fever with thrombocytopenia syndrome (SFTS) is a disease caused by infection with the SFTS virus (SFTSV). SFTS has become a crucial public health concern because of the heavy burden, lack of vaccines, effective therapies, and high-fatality rate. Evidence suggests that SFTSV circulates between ticks and animals in nature and is transmitted to humans by tick bites. In particular, ticks have been implicated as vectors of SFTSV, where domestic or wild animals may play as the amplifying hosts. Many studies have identified antigens and antibodies against SFTSV in various animals such as sheep, goats, cattle, and rodents. Besides, person-to-person transmission through contact with blood or mucous of an infected person has also been reported. In this study, we reviewed the literature and summarized the vectors and hosts associated with SFTS and the possible risk factors.

Knockdown Resistance (kdr) Mutations I1532T and F1534S Were Identified in Aedes albopictus Field Populations in Zhejiang Province, Central China.

Aedes albopictus is the only vector that can transmit the dengue virus in Zhejiang Province, central China, and it can develop insecticide resistance due to long-term exposure to pyrethroids. The presence of knockdown resistance (kdr) mutations is one of the mechanisms responsible for pyrethroid resistance, and has been reported in some Ae. albopictus populations in southern China. However, little is known about the DNA diversity of the voltage-gated sodium channel (VGSC) gene in Ae. albopictus populations in central China. Four Ae. albopictus field populations were collected, in Yiwu (YW), Quzhou (QZ), Wenzhou (WZ), and Jiaxing (JX) from Zhejiang Province, central China. The susceptibility of Ae. albopictus adults to three pyrethroids (beta-cypermethrin, deltamethrin, and permethrin) was tested using the WHO tube assay, and Kdr mutations were identified via PCR and sequencing. The relationship between kdr mutations and pyrethroid phenotypes was also analyzed. Of the four populations, none was sensitive to any pyrethroid tested, and the YW population showed the strongest pyrethroid resistance. Non-synonymous kdr mutations were detected in codons 1532 and 1534, domain III. At codon 1534, one mutant allele, TCC(S), was detected in the four populations with a frequency of 42.08%, while at codon 1532, one mutant allele, ACC(T), was detected in the JX and QZ populations, with frequencies of 4.22 and 3.03%, respectively. The F1534S mutant allele was positively correlated with both beta-cypermethrin and deltamethrin resistance phenotypes (OR > 1, P < 0.05), whereas the I1532T mutant allele was possibly negatively correlated with beta-cypermethrin, deltamethrin, and permethrin resistance phenotypes (OR < 1, P > 0.05). In conclusion, resistance and resistance mutations regarding to three pyrethroids are already present in the Ae. Albopictus populations from Zhejiang, central China, which prompts the need to use non-insecticide-based methods of insect control.

Update on the use of sildenafil in neonatal pulmonary hypertension: a narrative review of the history, current administration, and future directions.

Pulmonary hypertension (PH) is a life-threatening syndrome in neonates and has multiple and varied etiologies. However, few clinical studies have systematically evaluated the treatment regimens for this population. Phosphodiesterase (PDE) inhibitors, such as milrinone, tadalafil, dipyridamole, and sildenafil, are the most important regulators of vascular relaxation in the normal pulmonary vascular transition after birth, and these agents are widely used in the treatment of PH. Sildenafil, a representative PDE-5 inhibitor, has an important role as a single mode of therapy. However, the lack of evidence from pharmacokinetic and clinical trials has limited the emergence of standardized treatment regimens for sildenafil. There are also differing opinions among researchers regarding the best route of sildenafil administration. Due to the interindividual variability in the neonatal population, it is worth selecting the most suitable route of sildenafil administration according to the specific conditions of the neonatal population. These may be evaluated using the oxygenation index (OI), pulmonary artery pressure, mean blood pressure, and the serological index. This article reviews the clinical data on the use of sildenafil, focusing on the current and promising alternative routes of administration, which may affect subsequent clinical research in term and preterm neonates.

Field evaluation of two mosquito traps in Zhejiang Province, China.

Mosquito-borne Diseases are a common but severe public health threat. However, there is a lack of consensus on the effect of different mosquito trapping devices in China. This study firstly compared the BGM trap with the CDC light trap, commonly used in Chinese mosquito surveillance. Field trials of traps' efficiency were conducted in Yiwu city, China, from May 21st, 2018 to November 31st, 2018. Sixty-five comparisons were completed in five different biotopes (an urban residential area, a rural residential area, a park, a hospital, and a pig shelter). Concerning the number of mosquitoes per trap, the BGM trap outperformed three out of five biotopes. In contrast, the CDC light trap only showed better performance in the pig shelter. For specific species, the BGM trap outperformed in capturing Ae. albopictus, while the CDC light trap caught significantly more Cx. tritaeniorhynchus. Regarding Ae. albopictus and Cx. pipiens s.l. surveillance, the BGM trap is more suitable. The BGM trap shows significantly higher or similar efficiency than the CDC light trap in trapping common mosquito species in China, except in the pig shelter. Therefore, we recommend that Chinese researchers and public health practitioners use the BGM trap in future mosquito surveillance.

Effect of different carbon dioxide (CO2) flows on trapping Aedes albopictus with BG traps in the field in Zhejiang Province, China.

Carbon dioxide (CO2) attracts host-seeking adult mosquitoes; this fact is exploited for mosquito monitoring, which is important for evaluating the effects of mosquito-control operations. A field experiment was designed to explore the relationship between the CO2 flow rate and the trapping effect of BG traps. The aim was to select an appropriate flow rate for monitoring Aedes albopictus. Six sampling sites were selected for field experiments in Hangzhou city, Zhejiang Province, China. A total of six CO2 flow rates (0.00 L/min, 0.075 L/min, 0.15 L/min, 0.30 L/min, 0.60 L/min and 1.20 L/min) were tested to compare their effects on mosquito trapping. The catches were performed in six trapping periods between 15:30 and 18:30, and each catch period lasted 0.5 h. A total of 3068 adult mosquitoes were captured at six sampling sites in six days using BG traps (with BG-Sweetscent), among which 86.96% were Ae. albopictus. The total number of Ae. albopictus (males and females) captured at a flow rate of 0.00 L/min was significantly lower than the numbers captured at 0.075 L/min, 0.15 L/min, 0.30 L/min, 0.60 L/min and 1.20 L/min (P<0.001, P<0.001, P<0.001, P<0.001, and P<0.001 respectively). The total number of Ae. albopictus captured and the number of Ae. albopictus females captured increased with increasing CO2 flow and peaked at 0.3 L/min, above which these capture numbers did not increase significantly. In conclusion, the appropriate CO2 flow rate for monitoring Ae. albopictus with BG traps was 0.3 L/min.

Susceptibility of Armigeres subalbatus Coquillett (Diptera: Culicidae) to Zika virus through oral and urine infection.

BACKGROUND: Zika virus (ZIKV) disease outbreaks have been occurring in South America since 2015, and has spread to North America. Because birth defects and cases of Guillain Barré have been associated with infection with ZIKV, this has drawn global attention. ZIKV is generally considered an Aedes-transmitted pathogen. The transmission of ZIKV through blood by Aedes mosquito bites has been recognized as the major transmission route. However, it is not clear whether there are other transmission routes that can cause viral infection in mosquitos. The aim of the present study is to describe the susceptibility of Armigeres subalbatus, which often develop in human waste lagoons, to ZIKV, through oral infection in adult mosquitoes and urine infection in larvae. METHODOLOGY/PRINCIPAL FINDINGS: Five-day-old female Ar. subalbatus ingested infectious blood meals containing ZIKV. After 4, 7, and 10 days of ingesting infectious blood meals, ZIKV could be detected in the midguts, salivary glands, ovaries, and collected saliva of mosquitoes. The ZIKV infection rate (IR) on day 10 reached 40% in salivary glands and 13% in saliva, indicating that these mosquitoes were able to transmit ZIKV. In addition, ZIKV infection was also discovered in mosquito ovaries, suggesting the possibility of vertical transmission of virus. Moreover, Ar. subalbatus transmitted ZIKV to infant mice bitten by infectious mosquitoes. In a second experiment, 1st-instar larvae of Ar. subalbatus were reared in water containing ZIKV and human urine. After pupation, pupae were placed in clean water and transferred to a mosquito cage for emergence. Although ZIKV RNA was detected in all of the larvae tested, ZIKV was not detected in the saliva of any adult Ar. subalbatus. Considering that there are more uncontrollable factors in nature than in the laboratory environment, the possibility that the virus is transmitted to adult mosquitoes via larvae is very small period. CONCLUSIONS/SIGNIFICANCE: Adult Ar. subalbatus could be infected with ZIKV and transmit ZIKV through mosquito bites. Therefore, in many rural areas in China and in undeveloped areas of other Asian countries, the management of human waste lagoons in the prevention and control of Zika disease should be considered. Corresponding adjustments and modifications should also be made in prevention and control strategies against ZIKV.

Insecticide Resistance of Aedes albopictus in Zhejiang Province, China.

From 2003 until 2018, a total of 12 outbreaks with 1,654 confirmed dengue cases have been reported in Zhejiang Province. The emergence of insecticide resistance in mosquitoes will affect the control of dengue. Our study aims to investigate the current situation of insecticide resistance of Ae. albopictus in Zhejiang Province and compares it with the situation in 2016. Ae. albopictus were collected from 12 Zhejiang Province cities in 2019. Resistance to three major categories of insecticides, including 8 commonly used insecticides, was evaluated according to the tube test protocol recommended by China CDC. Ae. albopictus in all cities, except Hangzhou, Wenzhou, Lishui and Shaoxing, showed decreased susceptibility to beta-cypermethrin, deltamethrin and permethrin. For malathion, 3 cities Ae. albopictus have developed resistance, 3 cities Ae. albopictus have decreased susceptibility. For propoxur, in 3 cities Ae. albopictus showed decreased susceptibility with mortality ranging from 94.24% to 96.67%. The resistance to alpha-cypermethrin, lambda-cyhalothrin and fenitrothion is rare in Ae. albopictus in that only Zhoushan's mosquitoes showed decreased susceptibility to alpha-cypermethrin. The resistance to beta-cypermethrin, deltamethrin and permethrin was significantly correlated with each other. Compared to the situation in 2016, the insecticide resistance of Ae. albopictus in Zhejiang Province has become more common in 2019. In the emergency preparedness for future mosquito-borne diseases, two things should be done: 1) the selection of insecticides should be made based on information from insecticide resistance surveillance 2) the use of insecticide should follow scientific guidance.

Relationships between traditional larval indices and meteorological factors with the adult density of Aedes albopictus captured by BG-mosquito trap.

OBJECTIVES: Larval indices have been used for Ae. albopictus surveillance for many years, while there is limited use in assessing dengue transmission risk and adult mosquito emergence. This study is aimed to explore the relationships between larval indices and the Ae. albopictus density captured by BG-mosquito trap (BG-trap) method, with considering the meteorological factors. METHODS: Data on larval density, adult mosquito density and meteorology factors were collected in an entomological survey carried out in Quzhou City, Zhejiang Province of China in 2018. The Spearman's rank correlation and Pearson correlation were used for the analysis on the correlation of density indices. Generalized additive models were established to analyze the influencing factors of mosquito density. RESULTS: Breteau index (BI), House index (HI) and Container index (CI) were highly correlated with each other (r>0.7, p<0.05). The Ae. albopictus density was significantly correlated with CI (rs = 0.260, p<0.05), CI pre one week (rs = 0.259, p<0.05), and CI pre three weeks (rs = 0.329, p<0.05). BI was correlated with female Ae. albopictus density pre 4 weeks (r = -0.299, p<0.05). Female Ae. albopictus density was correlated with CI pre 3 weeks (rs = 0.303, p<0.05). The influencing factors of BI were average wind speed pre 1 week, average temperature and female Ae. albopictus density pre 4 weeks. The influencing factors of CI were average humidity pre 3 weeks and average temperature. The influencing factors of HI were average temperature and precipitation pre 4 weeks. The influencing factor of Ae. albopictus density and female Ae. albopictus density was temperature. CONCLUSIONS: The adult Ae. albopictus density had low correlation with certain larval indices. Some of the meteorology factors played significant roles in the density of adult Ae. albopictus and larva with or without a time lag.

Vector Competence and Vertical Transmission of Zika Virus in Aedes albopictus (Diptera: Culicidae).

Zika virus (ZIKV) is an emerging mosquito-borne pathogen belonging to the genus Flavivirus of the family Flaviviridae. Aedes albopictus is widely distributed in China. However, little is known about the vector competence of Ae. albopictus in China. The present study presents the oral susceptibility and vector competence of Ae. albopictus Guangzhou strain to ZIKV. Additionally, vertical transmission of ZIKV is described. The results demonstrated the susceptibility of local Ae. albopictus mosquitoes to ZIKV with an extrinsic incubation period of 6 days. Disseminated infection was observed in Ae. albopictus starting on day 2 postinfection (PI). Starting on day 6 PI, the saliva of Ae. albopictus exhibited ZIKV infection, and the transmission rate was 36.4%. Vertical transmission was observed during the first gonotrophic cycle. The minimum infection rate was observed in third-to-fourth instar larvae.

RNA interference of odorant receptor CquiOR114/117 affects blood-feeding behavior in Culex quinquefasciatus.

The odorant receptors (ORs) play a critical role for mosquitoes in the identification of blood-feeding hosts and other physiological processes. The OR8 subfamily in mosquitoes has been shown to be strongly involved in the detection the mammalian host associated odor, 1-octen-3-ol. CquiOR114/117 has been shown to be an orthologous OR8 in Culex quinquefasciatus Say. In this study, the expression of CquiOR114/117 in the different developmental stages of Cx. quinquefasciatus was detected by the amplification of CquiOR114/117 with real-time fluorescence quantitative polymerase chain reaction (PCR). RNA interference (RNAi) technology was used to interfere with the expression of CquiOR114/117 in females to observe the blood-feeding behavior change. The results showed that the expression level of CquiOR114/117 in the egg-to-pupa stage was significantly lower than that in the adult stage and that the expression level of the female mosquitoes peaked on the third day after emergence. The expression of CquiOR114/117 was significantly decreased in the 2-6 days after the injection of dsRNA compared with the control groups. The analysis of the blood-feeding behavior showed a significant positive correlation between CquiOR114/117 expression and the engorgement rate of the mosquitoes. CquiOR114/117 is speculated to have an effect on the blood-feeding behavior of Cx. quinquefasciatus.