First detection of a Vssc allele V1016G conferring a high level of insecticide resistance in Aedes albopictus collected from Europe (Italy) and Asia (Vietnam), 2016: a new emerging threat to controlling arboviral diseases.

IntroductionAedes albopictus (Skuse) is an important vector of arboviral diseases, including dengue, chikungunya and Zika virus disease. Monitoring insecticide resistance and mechanisms by which the mosquito develops resistance is crucial to minimise disease transmission.AimTo determine insecticide resistance status and mechanisms in Ae. albopictus from different geographical regions.MethodsWe sampled 33 populations of Ae. albopictus from Asia, Europe and South America, and tested these for susceptibility to permethrin, a pyrethroid insecticide. In resistant populations, the target site for pyrethroids, a voltage-sensitive sodium channel (Vssc) was genotyped. Three resistant sub-strains, each harbouring a resistance allele homozygously, were established and susceptibilities to three different pyrethroids (with and without a cytochrome P450 inhibitor) were assayed.ResultsMost populations of Ae. albopictus tested were highly susceptible to permethrin but a few from Italy and Vietnam (4/33), exhibited high-level resistance. Genotyping studies detected a knockdown resistance (kdr) allele V1016G in Vssc for the first time in Ae. albopictus. Two previously reported kdr alleles, F1534C and F1534S, were also detected. The bioassays indicated that the strain homozygous for the V1016G allele showed much greater levels of pyrethroid resistance than other strains harbouring F1534C or F1534S.ConclusionThe V1016G allele was detected in bothAsian and Italian Ae. albopictus populations, thus a spread of this allele beyond Italy in Europe cannot be ruled out. This study emphasises the necessity to frequently and regularly monitor the V1016G allele in Ae. albopictus, particularly where this mosquito species is the main vector of arboviruses.

Role of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) in local dengue epidemics in Taiwan.

BACKGROUND: Aedes mosquitoes in Taiwan mainly comprise Aedes albopictus and Ae. aegypti. However, the species contributing to autochthonous dengue spread and the extent at which it occurs remain unclear. Thus, in this study, we spatially analyzed real data to determine spatial features related to local dengue incidence and mosquito density, particularly that of Ae. albopictus and Ae. aegypti. METHODS: We used bivariate Moran's I statistic and geographically weighted regression (GWR) spatial methods to analyze the globally spatial dependence and locally regressed relationship between (1) imported dengue incidences and Breteau indices (BIs) of Ae. albopictus, (2) imported dengue incidences and BI of Ae. aegypti, (3) autochthonous dengue incidences and BI of Ae. albopictus, (4) autochthonous dengue incidences and BI of Ae. aegypti, (5) all dengue incidences and BI of Ae. albopictus, (6) all dengue incidences and BI of Ae. aegypti, (7) BI of Ae. albopictus and human population density, and (8) BI of Ae. aegypti and human population density in 348 townships in Taiwan. RESULTS: In the GWR models, regression coefficients of spatially regressed relationships between the incidence of autochthonous dengue and vector density of Ae. aegypti were significant and positive in most townships in Taiwan. However, Ae. albopictus had significant but negative regression coefficients in clusters of dengue epidemics. In the global bivariate Moran's index, spatial dependence between the incidence of autochthonous dengue and vector density of Ae. aegypti was significant and exhibited positive correlation in Taiwan (bivariate Moran's index = 0.51). However, Ae. albopictus exhibited positively significant but low correlation (bivariate Moran's index = 0.06). Similar results were observed in the two spatial methods between all dengue incidences and Aedes mosquitoes (Ae. aegypti and Ae. albopictus). The regression coefficients of spatially regressed relationships between imported dengue cases and Aedes mosquitoes (Ae. aegypti and Ae. albopictus) were significant in 348 townships in Taiwan. The results indicated that local Aedes mosquitoes do not contribute to the dengue incidence of imported cases. The density of Ae. aegypti positively correlated with the density of human population. By contrast, the density of Ae. albopictus negatively correlated with the density of human population in the areas of southern Taiwan. The results indicated that Ae. aegypti has more opportunities for human-mosquito contact in dengue endemic areas in southern Taiwan. CONCLUSIONS: Ae. aegypti, but not Ae. albopictus, and human population density in southern Taiwan are closely associated with an increased risk of autochthonous dengue incidence.

Susceptibility of Aedes albopictus and Aedes aegypti to three imported Chikungunya virus strains, including the E1/226V variant in Taiwan.

BACKGROUND/PURPOSE: An E1/226V variant Chikungunya virus (CHIKV) efficiently transmitted by Aedes albopictus to humans poses a significant threat to public health for those areas with the presence of Aedes albopictus, including Taiwan. METHODS: We infected three imported CHIKV isolates including the E1/226V variant with Ae. albopictus and Aedes aegypti in the laboratory to understand the disease risk. Viral RNA was measured by real time reverse transcription polymerase chain reaction. RESULTS: The viral susceptibility varied by virus strain and mosquito species and strain. The Asian virus strain started to replicate at 5-6 days post infection (dpi) with the maximum virus yield, ranging from 10(3.63) to 10(3.87) at 5-10 dpi in both species. The variant CHIKV Central/East/South African (CESA) virus genotype replicated earlier at 1 dpi with the maximum virus yield ranging from 10(5.63) to 10(6.52) at 3-6 dpi in Ae. albopictus females while the nonvariant virus strain replicated at 1-2 dpi with the maximum virus yield ranging from 10(5.51) to 10(6.27) at 6-12 dpi. In Ae. aegypti, these viruses replicated at 1-2 dpi, with maximum yields at 4-5 dpi (range from 10(5.38) to 10(5.62)). CONCLUSION: We concluded that the risk of CHIKV in Taiwan is high in all distribution areas of Ae. aegypti and Ae. albopictus for the CESA genotype and that the E1/226V variant virus strain presents an even higher risk.

Molecular Identification of Species Belonging to Culex vishnui Subgroup (Diptera: Culicidae), Vectors of Japanese Encephalitis Virus, in Taiwan.

Classification of mosquitoes with overlapping features remains problematic when using traditional morphological identification alone. In this study, we used molecular methods to elucidate the taxonomic status of Culex tritaeniorhynchus, Culex annulus, and Culex pseudovishnui species as vectors of the Japanese encephalitis virus belonging to the Culex vishnui subgroup and gene flow among them. In this study, 76, 59, and 3 samples of Cx. annulus, Cx. tritaeniorhynchus, and Cx. pseudovishnui, respectively, were collected around Taiwan. Phylogenetic analysis and genetic divergence were based on genomic sequence variations in ribosomal DNA and the internal transcribed spacer (rDNA) and cytochrome c oxidase subunit I (COI). Our results revealed that Cx. annulus and Cx. vishnui are genetically similar and share a gene pool among the species from Taiwan and other Asian countries. However, two hidden taxa of Cx. tritaeniorhynchus, which clustered together according to the rDNA sequences, were discovered based on the COI sequences. In addition, Cx. pseudovishnui has different gene pools from those of the strains from other countries, implying that the population from Taiwan is probably either a unique strain or a sibling species. This study provides molecular information on the taxonomic status of the species in the Cx. vishnui subgroup in Taiwan and gene flow between these species, providing valuable information for vector control operations and the delineation of the evolutionary process.

First molecular detection of a novel Babesia species from Haemaphysalis hystricis in Taiwan.

Newly recorded ticks and emerging tick-borne pathogens have recently been reported in subtropical and tropical East Asia. In this study, a total of 1,615 ticks (259 Haemaphysalis hystricis, 1334 Rhipicephalus microplus, 19 H. flava, and 3 R. haemaphysaloides) were collected by flagging from vegetation in Taiwan during 2019-2021. All 1,615 captured tick samples tested negative for SFTSV and Borrelia, but 12 of 356 tick samples tested positive for PCR amplification of a fragment of the 18S rRNA gene of Babesia spp., with an infection rate of 3.37 % (12/356) and a minimum infection rate of 0.74 % (12/1,615). Among the 12 detected Babesia spp., 11 were identified as Babesia bigemina in R. microplus, and the other one, detected in H. hystricis, was classified as an unnamed novel Babesia sp. Interestingly, the 18S rRNA sequence from the isolate detected in H. hystricis shared 98.79 % to 99.50 % identity with those of recent isolates from Japan, China and Nigeria. The exact origin of the Babesia species is not known, but the findings highlight the importance of international cooperation and the exchange of information on ticks and tick-borne pathogens. This represents a rare report of a Babesia sp. identified in H. hystricis, a tick species that has been proposed as a novel vector for some Babesia spp. This study supports H. hystricis as a possible vector of Babesia spp.

A dengue vector surveillance by human population-stratified ovitrap survey for Aedes (Diptera: Culicidae) adult and egg collections in high dengue-risk areas of Taiwan.

Aedes aegypti L. is the primary dengue vector in southern Taiwan. This article is the first report on a large-scale surveillance program to study the spatial-temporal distribution of the local Ae. aegytpi population using ovitraps stratified according to the human population in high dengue-risk areas. The sampling program was conducted for 1 yr and was based on weekly collections of eggs and adults in Kaohsiung City. In total, 10,380 ovitraps were placed in 5,190 households. Paired ovitraps, one indoors and one outdoors were used per 400 people. Three treatments in these ovitraps (paddle-shaped wooden sticks, sticky plastic, or both) were assigned by stratified random sampling to two areas (i.e., metropolitan or rural, respectively). We found that the sticky plastic alone had a higher sensitivity for detecting the occurrence of indigenous dengue cases than other treatments with time lags of up to 14 wk. The wooden paddle alone detected the oviposition of Ae. aegypti throughout the year in this study area. Furthermore, significantly more Ae. aegypti females were collected indoors than outdoors. Therefore, our survey identified the whole year oviposition activity, spatial-temporal distribution of the local Ae. aegypti population and a 14 wk lag correlation with dengue incidence to plan an effectively proactive control.

The role of voltage-gated sodium channel genotypes in pyrethroid resistance in Aedes aegypti in Taiwan.

BACKGROUND: Aedes aegypti is the major vector of dengue that threatens public health in tropical and subtropical regions. Pyrethroid-based control strategies effectively control this vector, but the repeated usage of the same insecticides leads to resistance and hampers control efforts. Therefore, efficient and prompt monitoring of insecticide resistance in local mosquito populations is critical for dengue control. METHODOLOGY/PRINCIPAL FINDING: We collected Ae. aegypti in southern Taiwan in March and October 2016. We analyzed the voltage-gated sodium channel (vgsc) genotypes of parentals (G0) and G1 adults after cypermethrin insecticide bioassay. Our results showed that four VGSC mutations (S989P, V1016G, F1534C, and D1763Y) associated with resistance were commonly detected in field-collected Ae. aegypti. The frequencies of these four mutations in the local mosquito population were significantly higher in October (0.29, 0.4, 0.27 and 0.11) than in March (0.09, 0.16, 0.18 and 0.03). Specific vgsc combined genotypes composed of the one to four such mutations (SGFY/SGFY, SVCD/SVCD, SGFY/PGFD, SVCD/SGFY, PGFD/PGFD, and SVCD/PGFD) shifted towards higher frequencies in October, implying their resistance role. In addition, the cypermethrin exposure bioassay data supported the field observations. Moreover, our study observed an association between the resistance level and the proportion of resistance genotypes in the population. CONCLUSIONS/SIGNIFICANCE: This is the first study to demonstrate the role of four-locus vgsc genotypes in resistance evaluation in a local Ae. aegypti population in Taiwan. This alternative method using resistance-associated genotypes as an indicator of practically insecticide resistance monitoring is a useful tool for providing precise and real-time information for decision makers.

The National Laboratory Response to the COVID-19 Pandemic in Taiwan.

From April 23 to November 2021, a wave of COVID-19 infections caused by a new Alpha variant swept across Taiwan, resulting in 14,458 positive cases and 830 deaths among over 3.8 million people tested. To cope with the sudden increase in sample volume, as of December 14, 2021, a network of 249 laboratories with a total diagnostic capacity of 158,492 real-time reverse transcription polymerase chain reaction tests per day was established in 22 administrative regions. As of April 2022, over 9.5 million specimens were tested. Fully automated high-throughput and point-of-care nucleic acid testing, and rapid antigen testing, were simultaneously implemented to expand the country's daily diagnostic capacity. Saliva testing and sample pooling were also introduced to increase screening efficiency in certain situations. Antibody testing and genomic sequencing were also adopted for more precise epidemic investigation. Other challenges encountered and overcome include a lack of resources and interfacing of laboratory information management systems for case reporting, limited specimen allocation and delivery, and limited staff for diagnostic processing.

Delayed correlation between the incidence rate of indigenous murine typhus in humans and the seropositive rate of Rickettsia typhi infection in small mammals in Taiwan from 2007-2019.

Murine typhus is a flea-borne zoonotic disease with acute febrile illness caused by Rickettsia typhi and is distributed widely throughout the world, particularly in port cities and coastal regions. We observed that murine typhus was an endemic disease (number of annual indigenous cases = 29.23±8.76) with a low incidence rate (0.13±2.03*10-4 per 100,000 person-years) in Taiwan from 2007-2019. Most (45.79%, 174/380) indigenous infections were reported in May, June, and July. The incidence rates in both May and June were statistically higher than those in other months (p<0.05). Correspondingly, sera collected from small mammals (rodents and shrews) trapped in airports and harbors demonstrated anti-R. typhi antibody responses (seropositive rate = 8.24±0.33%). Interestingly, the ports with the highest seropositivity rates in small mammals are all inside/near the areas with the highest incidence rates of indigenous murine typhus. In addition, incidence rates in humans were positively correlated with the 1-month and 2-month prior seropositive rates in small mammals (R = 0.31 and 0.37, respectively). As early treatment with appropriate antibiotics for murine typhus could effectively shorten the duration of illness and reduce the risk of hospitalization and fatality, flea-related exposure experience should be considered in clinics during peak seasons and the months after a rise in seropositivity rates in small mammals. Surveillance in small mammals might be helpful for the development of real-time reporting or even early reminders for physicians of sporadic murine typhus cases based on the delayed correlation observed in this study.

Discovery of super-insecticide-resistant dengue mosquitoes in Asia: Threats of concomitant knockdown resistance mutations.

Aedes aegypti (Linnaeus, 1762) is the main mosquito vector for dengue and other arboviral infectious diseases. Control of this important vector highly relies on the use of insecticides, especially pyrethroids. The high frequency (>78%) of the L982W substitution was detected at the target site of the pyrethroid insecticide, the voltage-gated sodium channel (Vgsc) of A. aegypti collected from Vietnam and Cambodia. Alleles having concomitant mutations L982W + F1534C and V1016G + F1534C were also confirmed in both countries, and their frequency was high (>90%) in Phnom Penh, Cambodia. Strains having these alleles exhibited substantially higher levels of pyrethroid resistance than any other field population ever reported. The L982W substitution has never been detected in any country of the Indochina Peninsula except Vietnam and Cambodia, but it may be spreading to other areas of Asia, which can cause an unprecedentedly serious threat to the control of dengue fever as well as other Aedes-borne infectious diseases.

Commensal Rodent Habitat Expansion Enhances Arthropod Disease Vectors on a Tropical Volcanic Island.

On volcanic islands, the release of animals from predators and competitors can lead to increased body size and population density as well as the expanded habitat use of introduced animals relative to their mainland counterparts. Such alterations might facilitate the spread of diseases on islands when these exotic animals also carry pathogenic agents; however, this has rarely been investigated. The commensal Asian house rat (Rattus tanezumi) is confined to human residential surroundings in mainland Taiwan but can be observed in the forests of nearby Orchid Island, which is a tropical volcanic island. Orchid Island is also a hot spot for scrub typhus, a lethal febrile disease transmitted by larval trombiculid mites (chiggers) that are infected primarily with the rickettsia Orientia tsutsugamushi (OT). We predicted an increase in chigger abundance when rodents (the primary host of chiggers) invade forests from human settlements since soils are largely absent in the latter habitat but necessary for the survival of nymphal and adult mites. A trimonthly rodent survey at 10 sites in three habitats (human residential, grassland, and forest) found only R. tanezumi and showed more R. tanezumi and chiggers in forests than in human residential sites. There was a positive association between rodent and chigger abundance, as well as between rodent body weight and chigger load. Lastly, >95% of chiggers were Leptotrombidium deliense and their OT infection rates were similar among all habitats. Our study demonstrated potentially elevated risks of scrub typhus when this commensal rat species is allowed to invade natural habitats on islands. Additionally, while the success of invasive species can be ascribed to their parasites being left behind, island invaders might instead obtain more parasites if the parasite requires only a single host (e.g., trombiculid mite), is a host generalist (e.g., L. deliense), and is transferred from unsuitable to suitable habitats (i.e., human settlements on the mainland to forests on an island).

Building the National SARS-CoV-2 Laboratory Diagnostic Capacity in Taiwan.

Through early and proactive laboratory testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes novel coronavirus 2019, Taiwan has demonstrated an efficient and rapid control response to contain the outbreak. Two days after the World Health Organization announced the complete viral genome sequence, the national laboratory of the Taiwan Centers for Disease Control developed a specific real-time reverse transcription polymerase chain reaction (real-time RT-PCR) test for SARS-CoV-2. The national laboratory network was further strengthened through the recruitment of medical centers and regional hospitals distributed throughout most geographical regions of the country. Ultimately, a network of 60 laboratories with a capacity of 7,342 real-time RT-PCR tests per day was established. Between January 14 and August 5, 2020, a total of 158,772 tests were conducted, corresponding to 120,487 cases. Test results were obtained within 24 hours, enabling an efficient and rapid control response.

Voltage-gated sodium channel intron polymorphism and four mutations comprise six haplotypes in an Aedes aegypti population in Taiwan.

BACKGROUND: Knockdown resistance (kdr) to dichlorodiphenyltrichloroethane (DDT) and pyrethroids is known to link amino acid substitutions in the voltage-gated sodium channel (VGSC) in Aedes aegypti. Dengue fever primarily transmitted by Ae. aegypti is an annual public health issue in Taiwan. Accordingly, pyrethroid insecticides have been heavily used for decades to control mosquito populations in the summer and autumn. In Taiwan, an Ae. aegypti population with two VGSC mutations, V1016G and D1763Y, was described previously. METHODOLOGY/PRINCIPAL FINDING: Aedes aegypti (G0) were collected in Tainan and Kaohsiung in southern Taiwan. The VGSC gene polymorphisms of the kdr mutations and the intron flanked by exons 20 and 21 were verified. The first generation offspring (G1) were used to measure the resistance level to cypermethrin, a pyrethroid insecticide currently used in Taiwan. In addition to V1016G and D1763Y, we describe two new mutations, S989P and F1534C, which have not been reported in Taiwan. Moreover, we also identify two types (groups A and B) of introns between exons 20 and 21. Intriguingly, the kdr mutations S989P, V1016G and D1763Y are strictly located on the haplotype harboring the group A intron, whereas F1534C links to the group B intron. When those data were taken together, we proposed the following six haplotypes for VGSC genes in Taiwan today: (i)S989-intron A-V1016-F1534-D1763, (ii)S989-intron A-V1016G-F1534-D1763, (iii)S989P-intron A-V1016G-F1534-D1763, (iv)S989-intron A-V1016G-F1534-D1763Y, (v)S989-intron B-V1016-F1534-D1763 and (vi)S989-intron B-V1016-F1534C-D1763. Triple heterozygous mutations of either S989P/V1016G/F1534C or V1016G/F1534C/D1763Y can be found in one single Ae. aegypti mosquito. The proportions of the VGSC mutations were relevant to cypermethrin resistance. Notably, the presence of S989P and V1016G in the population could be a helpful reference to predict the resistance level to cypermethrin. This is the first study to demonstrate the coexistence of four kdr mutations in a population of Ae. aegypti. CONCLUSIONS/SIGNIFICANCE: Four kdr mutations (S989P, V1016G, F1534C and D1763Y) and two intron forms (Group A and B) were commonly found in local Ae. aegypti populations in Taiwan.

The resting sites and blood-meal sources of Anopheles minimus in Taiwan.

BACKGROUND: The WHO declared Taiwan free from malaria in 1965, but in 2003 the reporting of two introduced cases in a rural area suggested a possible local transmission of this disease. Therefore, understanding the resting sites and the blood sources of Anopheles minimus is crucial in order to provide information for implementing vector control strategies. METHODS: During a two-year survey, mosquitoes were collected in houses and their surrounding areas and at the bank of larval habitats by backpack aspirators in 17 villages in rural areas of southern and eastern Taiwan for 1 hr. On the same day, blacklight traps were hung downward overnight. Blood-fed mosquito samples were analysed by PCR. RESULTS: Of the 195 total households surveyed by backpack aspirators, no Anopheles adults were collected inside the houses, while a single Anopheles minimus and a single Anopheles maculatus were collected outside of the houses. On the same day, 23 An. minimus, two An. maculatus, two Anopheles ludlowae, two Anopheles sinensis, and one Anopheles tessellatus were collected along the bank of larval habitats. In blacklight traps hung outside of the houses in the villages, 69 An. minimus, 62 An. ludlowae, 31 An. sinensis, and 19 An. maculatus were collected. In larval habitats, 98 An. ludlowae, 64 An. minimus, 49 An. sinensis, and 14 An. maculatus were collected. Of a total of 10 blood-fed samples, An. minimus fed on four animals including bovine (60%), dogs (20%), pig (10%), and non-chicken avian (10%). CONCLUSION: Anopheles minimus, an opportunist feeder in Taiwan, was not collected inside the houses, but was found outside of the houses in villages and surrounding larval habitats. Therefore, an outdoor transmission of malaria is likely to occur and, thus, the bed nets, which are favoured for controlling the late biting of An. minimus, should be a very efficient and effective method for those local residents who sleep outdoors. Additionally, space spray of insecticides for Anopheles at night, as well as residual spray inside animal huts and selective larval habitats, are also helpful to control female adults.

Critical low temperature for the survival of Aedes aegypti in Taiwan.

BACKGROUND: Taiwan is geographically located in a region that spans both tropical and subtropical climates (22-25°N and 120-122°E). The Taiwan Centers for Disease Control have found that the ecological habitat of Aedes aegypti appears only south of 23.5°N. Low temperatures may contribute to this particular habitat distribution of Ae. aegypti under the influence of the East Asian winter monsoon. However, the threshold condition related to critically low temperatures remains unclear because of the lack of large-scale spatial studies. This topic warrants further study, particularly through national entomological surveillance and satellite-derived land surface temperature (LST) data. METHODS: We hypothesized that the distribution of Ae. aegypti is highly correlated with the threshold nighttime LST and that a critical low LST limits the survival of Ae. aegypti. A mosquito dataset collected from the Taiwan Centers for Disease Control was utilized in conjunction with image data obtained from the moderate resolution imaging spectroradiometer (MODIS) during 2009-2011. Spatial interpolation and phi coefficient methods were used to analyze the correlation between the distributions of immature forms of Ae. aegypti and threshold LST, which was predicted from MODIS calculations for 348 townships in Taiwan. RESULTS: According to the evaluation of the correlation between estimated nighttime temperatures and the occurrence of Ae. aegypti, winter had the highest peak phi coefficient, and the corresponding estimated threshold temperatures ranged from 13.7 to 14 °C in the ordinary kriging model, which was the optimal interpolation model in terms of the root mean square error. The mean threshold temperature was determined to be 13.8 °C, which is a critical temperature to limit the occurrence of Ae. aegypti. CONCLUSIONS: An LST of 13.8 °C was found to be the critical temperature for Ae. aegypti larvae, which results in the near disappearance of Ae. aegypti during winter in the subtropical regions of Taiwan under the influence of the prevailing East Asian winter monsoon.

Identification of Three Distinct Groups of Anopheles lindesayi in Japan by Morphological and Genetic Analyses.

Anopheles (Anopheles) lindesayi Giles consists of 5 subspecies. In Japan, only one subspecies, An. l. japonicus Yamada, has been reported. Its geographical populations are morphologically diverse; however, they are regarded as a single subspecies. In this study, we re-evaluated the taxonomic status of An. l. japonicus in Japan, and that of another subspecies, An. l. pleccau, distributed in Taiwan, by comparative morphological and molecular analyses based on the gene sequences of mitochondrial DNA cytochrome c oxidase I (COI) and ribosomal DNA internal transcribed spacer 2 (ITS2). Nucleotide sequence divergence was calculated using the Kimura-two-parameter (K2P) distance model. Phylogenetic trees based on COI and ITS2 sequences showed 3 distinct clades: Eastern Japan, Western Japan, and the Ryukyus. The sequences of the Ryukyu specimens were located within the same clade as that of the sequences of the Taiwanese specimens. Regarding the COI sequences, the 3 geographical groups in Japan were genetically distinct. The following morphological characteristics distinguished the groups: larval seta 1-S, pupal setae 5 through segments IV-VII, and pupal setae 6 on segments IV-VII. Based on these results, it was revealed that An. l. japonicus included 3 genetically and morphologically distinct groups: 2 groups of An. l. japonicus and a group in the Ryukyus, which was a synonym of An. l. pleccau.

Differential survival of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) larvae exposed to low temperatures in Taiwan.

Aedes aegypti (L.) and Aedes albopictus (Skuse) differ in their distribution in Taiwan. The former species is distributed in the south of Taiwan, whereas the latter is found throughout the island. One possible explanation proposes that low temperatures in the winter prevent the expansion of Ae. aegypti. Hence, the impact of low temperatures on immatures of both species was studied in the laboratory and in the field. Our study showed that, under most conditions, Ae. aegypti were more sensitive to low temperatures than Ae. albopictus both in the laboratory and in the field. The survival rates at 10 degrees C for the first and fourth instars of Ae. albopictus were significantly better than those of Ae. aegypti. At 2.5 and 5.0 degrees C, the first instars of Ae. albopictus survived better than the same stadium of Ae. aegypti, but the fourth instars of Ae. aegypti survived better. Short exposures to low temperatures did not affect the acclimatization of Ae. aegypti immatures but longer exposures did increase the physiological adaptation to low temperatures. For Ae. albopictus, exposure to low temperatures increases the acclimatization of this species. In field experiments, Ae. aegypti larvae had a significantly higher mortality than Ae. albopictus during exposures to cold fronts in the 2004 winter. We conclude that low temperatures in northern and central Taiwan have a negative impact on the distribution of Ae. aegypti, but this factor alone is not sufficient to prevent this species from occupying the rest of Taiwan.

Emergency vector control in a DENV-2 outbreak in 2002 in Pingtung City, Pingtung County, Taiwan.

This paper reports the strategy and effectiveness of an emergency control program conducted in Pingtung City, Taiwan in response to dengue outbreaks. In our control strategy, we carried out 3 insecticide space sprays with an interval of 6-7 days and 2 source reductions to cover the entire duration of dengue virus exposure in humans and mosquito vectors. The control effect was demonstrated by a significant reduction in the Breteau (51.1%) and larval (80.0%) indices, but no such effect was demonstrated by alterations in the adult index (54.9%), house index (45.0%), container index (33.8%), or by indoor (15.8%), outdoor (31.2%), or total water-filled containers (22.7%) per 100 premises examined. The contribution made by the reduction in the number of positive containers was primarily in the outdoor (77.2%), and not the indoor containers (-6.0%). This reduction attributed to an overall reduction of 96.0% Aedes albopictus larvae and 71.0% Aedes aegypti. Therefore, 4 weeks after this extensive emergency control measure, the number of dengue cases dropped to one. Finally, due to both the decrease in temperature resulting from the upcoming winter, and to the sustained effort toward source reduction, the transmission cycle of DENV-2 in Pingtung City was interrupted at the beginning of 2003, and no additional cases were identified in late 2003.

Taiwan's Public Health National Laboratory System: Success in Influenza Diagnosis and Surveillance.

Taiwan's National Laboratory System is one of the action packages of the Global Health Security Agenda, which was launched by the World Health Organization (WHO) to promote health security as an international priority and to encourage progress toward full implementation of the WHO International Health Regulations (IHR) 2005. The mission of each national laboratory system is to conduct real-time biosurveillance and effective laboratory-based diagnostics, as measured by a nationwide laboratory system able to reliably conduct diagnoses on specimens transported properly to designated laboratories from at least 80% of the regions in the country. In Taiwan, the national laboratory system for public health is well-established and coordinated by the Taiwan Centers for Disease Control (CDC), which is the government authority in charge of infectious disease prevention and intervention. Through the national laboratory system, Taiwan CDC effectively detects and characterizes pathogens that cause communicable diseases across the entire country, including both known and novel threats, and also conducts epidemiologic analyses of infectious diseases. In this article, we describe the national laboratory system for public health in Taiwan. We provide additional information on the national influenza laboratory surveillance network to demonstrate how our national laboratory systems work in practice, including descriptions of long-term seasonal influenza characterization and successful experiences identifying novel H7N9 and H6N1 influenza viruses.