Genetic Differentiation of the Bloodsucking Midge Forcipomyia taiwana (Diptera: Ceratopogonidae): Implication of the Geographic Isolation by the Central Mountain Ranges in Taiwan.

Forcipomyia (Lasiohelea) taiwana, a small bloodsucking midge, thrives in moderately moist habitats and is commonly found in grassy and bushy areas at an elevation below 250 m. This species exhibits a diurnal biting pattern and shows a marked preference for human blood. Although not known to transmit arthropod-borne diseases, the bites of F. taiwana can induce severe allergic reactions in some individuals. As a significant nuisance in Taiwan, affecting both daily life and the tourism industry, comprehensive studies on its population genetics across different geographical regions remain scarce. The central mountain ranges in Taiwan, comprising more than two hundred peaks above 3000 m in elevation, extend from the north to the south of the island, creating distinct eastern and western geographical divisions. This study utilizes microsatellite markers to explore the genetic differentiation of F. taiwana populations located in the eastern and western regions of the mountain ranges. Our findings reveal substantial genetic differentiation among populations inhabiting Taiwan's western region compared to those in the eastern region. This indicates that the topographical barriers presented by the mountain ranges significantly restrict gene flow, particularly given the species' limited active flight ability and habitat preferences. Although passive dispersal mechanisms, like wind or human activity, could contribute, this study concludes that the gene flow of F. taiwana between the western and eastern regions is primarily influenced by topographical constraints.

Nationwide survey of ticks on domesticated animals in Taiwan: Revealing the hidden threat to animal and public health.

Ticks are not only bloodsucking ectoparasites but also important vectors of tick-borne diseases (TBDs), posing significant threats to public and animal health. Domesticated animals serve as critical hosts for numerous ticks, highlighting the importance of understanding tick infestations in Taiwan. To address this knowledge gap, we conducted a nationwide survey to identify ticks on domesticated animals and associated environments in 2018 and 2019. A total of 6,205 ticks were collected from 1,337 host animals, revealing the presence of seven tick species, with Rhipicephalus microplus, and Rhipicephalus sanguineus being the dominant species. High infestation rates and widespread distribution of ticks were observed on domesticated animals, especially on dogs and cattle (yellow cattle and angus cattle), and the neighbouring grassland of yellow cattle. While this study has certain limitations, it provides valuable insights into the distribution and prevalence of ticks on domesticated animals in Taiwan and their implications for controlling TBDs. Further research is needed to comprehensively understand the complex interactions among ticks, hosts and pathogens.

Occurrence and surveillance of Taiwanese bovine arboviruses using hematophagous insects in dairy farms during 2012-2019.

Culicoides-borne viruses are an important arbovirus group causing bovine diseases. During 2012-2019, 2,525 pools consisting of 108,937 specimens of vectors were subjected to PCR detection of bovine arbovirus belonging to Orthobunyavirus, Orbivirus, and Ephemerovirus. Twelve virus RNAs, of which 6, that is, Shuni virus, Shamonda virus, and Sathuperi virus in Orthobunyavirus and Sathuvachari virus and epizootic hemorrhagic disease virus serotypes 4 and 7 in Orbivirus were detected for the first time in the area. Potential vector species were evaluated by the minimum infection rate, and the population abundance of Culicoides oxystoma, Culex tritaeniorhynchus, and Anopheles sinensis indicated that they were the main potential vector species in dairy farms in Taiwan.

Joint influence of architectural and spatiotemporal factors on the presence of Aedes aegypti in urban environments.

BACKGROUND: Urbanization has led to the proliferation of high-rise buildings, which have substantially influenced the distribution of dengue vectors, such as Aedes aegypti (L.). However, knowledge gaps exist regarding the individual and combined effects of architectural and spatiotemporal factors on dengue vector. This study investigated the interrelationship between Ae. aegypti presence, building architectural features, and spatiotemporal factors in urban environments. RESULTS: The mosquito Ae. aegypti presence varied by location and seasons, being higher in outdoor environments than in indoor environments. Lingya (Kaohsiung City, Taiwan) had the highest mosquito numbers, particularly in basement and first floor areas. Ae. aegypti was found on multiple floors within buildings, and their presence was greater in summer and autumn. The XGBoost model revealed that height within a building, temperature, humidity, resident density, and rainfall were key factors influencing mosquito presence, whereas openness had a relatively minor impact. CONCLUSION: To effectively address the problems caused by urbanization, the three-dimensional distribution of Ae. aegypti, including their spatial distribution across heights and areas within the urban environment, must be considered. By incorporating these multiple factors, this approach provides valuable insights for those responsible for urban planning and disease management strategies. Understanding the interplay between architectural features, environmental conditions, and the presence of Ae. aegypti is essential for developing targeted interventions and mitigating the adverse impacts of urbanization on public health. © 2023 Society of Chemical Industry.

Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3-11 Amphipathic Helices to Favor Membrane Interaction.

We investigated the antimicrobial activity and membrane disruption modes of the antimicrobial peptide mastoparan-AF against hemolytic Escherichia coli O157:H7. Based on the physicochemical properties, mastoparan-AF may potentially adopt a 3-11 amphipathic helix-type structure, with five to seven nonpolar or hydrophobic amino acid residues forming the hydrophobic face. E. coli O157:H7 and two diarrheagenic E. coli veterinary clinical isolates, which are highly resistant to multiple antibiotics, are sensitive to mastoparan-AF, with minimum inhibitory and bactericidal concentrations (MIC and MBC) ranging from 16 to 32 µg mL-1 for E. coli O157:H7 and four to eight µg mL-1 for the latter two isolates. Mastoparan-AF treatment, which correlates proportionally with membrane permeabilization of the bacteria, may lead to abnormal dents, large perforations or full opening at apical ends (hollow tubes), vesicle budding, and membrane corrugation and invagination forming irregular pits or pores on E. coli O157:H7 surface. In addition, mRNAs of prepromastoparan-AF and prepromastoparan-B share a 5'-poly(A) leader sequence at the 5'-UTR known for the advantage in cap-independent translation. This is the first report about the 3-11 amphipathic helix structure of mastoparans to facilitate membrane interaction. Mastoparan-AF could potentially be employed to combat multiple antibiotic-resistant hemolytic E. coli O157:H7 and other pathogenic E. coli.

The First Nationwide Surveillance of Severe Fever with Thrombocytopenia Syndrome in Ruminants and Wildlife in Taiwan.

Since the first discovery of severe fever with thrombocytopenia syndrome virus (SFTSV) in China in 2009, SFTSV has rapidly spread through other Asian countries, including Japan, Korea, Vietnam and Pakistan, in chronological order. Taiwan reported its first discovery of SFTSV in sheep and humans in 2020. However, the prevalence of SFTSV in domestic and wildlife animals and the geographic distribution of the virus within the island remain unknown. A total of 1324 animal samples, including 803 domestic ruminants, 521 wildlife animals and 47 tick pools, were collected from March 2021 to December 2022 from 12 counties and one terrestrial island. The viral RNA was detected by a one-step real-time reverse transcription polymerase chain reaction (RT-PCR). Overall, 29.9% (240/803) of ruminants showed positive SFTSV RNA. Sheep had the highest viral RNA prevalence of 60% (30/50), followed by beef cattle at 28.4% (44/155), goats at 28.3% (47/166), and dairy cows at 27.5% (119/432). The bovine as a total of dairy cow and beef cattle was 27.8% (163/587). The viral RNA prevalence in ticks (predominantly Rhipicephalus microplus) was similar to those of ruminants at 27.7% (13/47), but wild animals exhibited a much lower prevalence at 1.3% (7/521). Geographically the distribution of positivity was quite even, being 33%, 29.1%, 27.5% and 37.5% for northern, central, southern and eastern Taiwan, respectively. Statistically, the positive rate of beef cattle in the central region (55.6%) and dairy cattle in the eastern region (40.6%) were significantly higher than the other regions; and the prevalence in Autumn (September-November) was significantly higher than in the other seasons (p < 0.001). The nationwide study herein revealed for the first time the wide distribution and high prevalence of SFTSV in both domestic animals and ticks in Taiwan. Considering the high mortality rate in humans, surveillance of other animal species, particularly those in close contact with humans, and instigation of protective measures for farmers, veterinarians, and especially older populations visiting or living near farms or rural areas should be prioritized.

Global population genetic structure and lineage differentiation of the stable fly, Stomoxys calcitrans.

The bloodsucking fly, Stomoxys calcitrans (Diptera: Muscidae), is a cosmopolitan pest that transmits potential pathogens mechanically. We conduct phylogeographic analyses of S. calcitrans to resolve its global population genetic structure for establishing baseline of molecular studies. Results from mitochondrial gene suggested that the major divergence of S. calcitrans predominantly occurred 0.32-0.47 million years ago (Mya) and the subsequent diversifications took place during 0.13-0.27 Mya. The Ethiopian region was deduced as the most likely origin of S. calcitrans and the Nearctic lineages were considered to have originated from Oriental or Palaearctic regions. Our results further revealed that each biogeographic region of S. calcitrans likely maintains its genetic specialty, and yet, those non-monophyletic relationships were possibly caused by ancestral retention, dispersal with mammals, long-distance migration, and the international livestock industries. Moreover, the three highly diverged Ethiopian lineages may be putative cryptic species that require clarification of their veterinary importance. Unravelling the genetic structure of stable fly and preventing gene flow among biogeographic regions through anthropogenic activities are thus pivotal in livestock industry administration, particularly genetic exchange among differentiated lineages that might lead to the consequence of ecological trait alterations.

Impact of temperature on infection with Japanese encephalitis virus of three potential urban vectors in Taiwan; Aedes albopictus, Armigeres subalbatus, and Culex quinquefasciatus.

Japanese encephalitis (JE) is an important mosquito-borne infectious disease in rural areas of Asia that is caused by Japanese encephalitis virus (JEV). Culex tritaeniorhynchus is the major vector of JEV, nevertheless there are other mosquitoes that may be able to transmit JEV. This study confirms that the midgut, head tissue, salivary glands, and reproductive tissue of Aedes albopictus, Armigeres subalbatus, and Culex quinquefasciatus are all able to be infected with JEV after a virus-containing blood meal was ingested by female mosquitoes. Even though the susceptibility to JEV of the different tissues varies, the virus-positive rate increased with the number of days after JEV infection. Moreover, once JEV escapes the midgut barrier, the oral transmission rates of JEV were 16%, 2%, and 21% for Ae. albopictus, Ar. subalbatus, and Cx. quinquefasciatus at 14 days after infection at 30 °C, respectively. There is no supporting evidence to suggest vertical transmission of JEV by the tested mosquitoes. Collectively, raising the temperature enhances JEV replication in the salivary gland of the three mosquito species, suggesting that global warming will enhance mosquito vector competence and that this is likely to lead to an increase in the probability of JEV transmission.

Evaluation of the Potential Entomopathogenic Fungi Purpureocillium lilacinum and Fusarium verticillioides for Biological Control of Forcipomyia taiwana (Shiraki).

Forcipomyia taiwana (Diptera: Ceratopogonidae) is a nuisance blood-sucking pest to humans in Taiwan. An F. taiwana bite causes itching and redness and usually causes serious harassment to human outdoor activity. In terms of F. taiwana control, chemical pesticides are ineffective. Therefore, other efforts are needed. Fungal mycosis in the larvae, pupae, and emerging F. taiwana adults was found during the rearing of F. taiwana. In this study, six fungal isolates were isolated from infected cadavers and subjected to molecular identification. In addition, their biocontrol potential was evaluated against different life stages of F. taiwana. Based on the pathogenicity screening, two fungal isolates, NCHU-NPUST-175 and -178, which caused higher mortality on the fourth instar larvae of F. taiwana, were selected for virulence tests against different life stages of F. taiwana larvae. The results of the phylogenetic analysis indicated that the NCHU-NPUST-175 and -178 belonged to Purpureocillium lilacinum and Fusarium verticillioides, respectively. Bioassay against different life stages of F. taiwana with different spore concentrations (5 × 105 to 5 × 107 conidia/mL) revealed a dose-dependent effect on larvae for both fungal isolates, while only 38% and 50% mortality was found in highest concentration (5 × 107 conidia/mL) at fourth instar larvae by Pl-NCHU-NPUST-175 and Fv-NCHU-NPUST-178, respectively. Moreover, reductions in egg-hatching rate and adult emergence rate were found, when the last stage of F. taiwana was inoculated with both fungal isolates, indicating the ovicidal potential and the impact of entomopathogenic fungi on the development of F. taiwana. In conclusion, Pl-NCHU-NPUST-175 and Fv-NCHU-NPUST-178 showed larvicidal activity, ovicidal activity, and impact on adult emergence on F. taiwana.

Molecular epidemiology of Akabane virus in Taiwan.

BACKGROUND: Akabane virus (AKAV) is a teratogenic and neuropathogenic arbovirus that infects livestock and wild animals. AKAVs are endemic arboviruses from dairy farms in Taiwan in 1989, and the first sequence was detected in cattle with nonsuppurative encephalitis in 1992. OBJECTIVES: This study aims to understand the epidemiological relationships of the akabane viruses between Taiwan and nearby places. METHODS: In this study, 17 specimens were identified or isolated from vector insects, and ruminant fetuses collected from 1992 to 2015 were sequenced and analysed. RESULTS: Sequence analyses revealed all Taiwanese AKAVs belonged to genogroup Ia but diverged into two clusters in the phylogenetic trees, implying that at least two invasive events of AKAV may have occurred in Taiwan. CONCLUSIONS: The two clusters of AKAVs could still be identified in Taiwan in 2015, and a reassortment event was observed, indicating that the two clusters of AKAVs are already endemic in Taiwan.

The cecropin-prophenoloxidase regulatory mechanism is a cross-species physiological function in mosquitoes.

This study's aim was to investigate whether the cecropin-prophenoloxidase regulatory mechanism is a cross-species physiological function among mosquitoes. BLAST and phylogenetic analysis revealed that three mosquito cecropin Bs, namely Aedes albopictus cecropin B (Aalcec B), Armigeres subalbatus cecropin B2 (Ascec B2), and Culex quinquefasciatus cecropin B1 (Cqcec B1), play crucial roles in cuticle formation during pupal development via the regulation of prophenoloxidase 3 (PPO 3). The effects of cecropin B knockdown were rescued in a cross-species manner by injecting synthetic cecropin B peptide into pupae. Further investigations showed that these three cecropin B peptides bind to TTGG(A/C)A motifs within each of the PPO 3 DNA fragments obtained from these three mosquitoes. These results suggest that Aalcec B, Ascec B2, and Cqcec B1 each play an important role as a transcription factor in cuticle formation and that similar cecropin-prophenoloxidase regulatory mechanisms exist in multiple mosquito species.

Insecticide Resistance Profiles and Synergism of Field Aedes aegypti from Indonesia.

Information on the insecticide resistance profiles of Aedes aegypti in Indonesia is fragmentary because of the lack of wide-area insecticide resistance surveillance. We collected Ae. aegypti from 32 districts and regencies in 27 Indonesian provinces and used WHO bioassays to evaluate their resistance to deltamethrin, permethrin, bendiocarb, and pirimiphos-methyl. To determine the possible resistance mechanisms of Ae. aegypti, synergism tests were conducted using piperonyl butoxide (PBO) and S,S,S-tributylphosphorotrithioates (DEF). The Ae. aegypti from all locations exhibited various levels of resistance to pyrethroids. Their resistance ratio (RR50) to permethrin and deltamethrin ranged from 4.08× to 127× and from 4.37× to 72.20×, respectively. In contrast with the findings of other studies, most strains from the highly urbanized cities on the island of Java (i.e., Banten, Jakarta, Bandung, Semarang, Yogyakarta, and Surabaya) exhibited low to moderate resistance to pyrethroids. By contrast, the strains collected from the less populated Kalimantan region exhibited very high resistance to pyrethroids. The possible reasons are discussed herein. Low levels of resistance to bendiocarb (RR50, 1.24-6.46×) and pirimiphos-methyl (RR50, 1.01-2.70×) were observed in all tested strains, regardless of locality. PBO and DEF synergists significantly increased the susceptibility of Ae. aegypti to permethrin and deltamethrin and reduced their resistance ratio to less than 16×. The synergism tests suggested the major involvement of cytochrome P450 monooxygenases and esterases in conferring pyrethroid resistance. On the basis of our results, we proposed a 6-month rotation of insecticides (deltamethrin + synergists ➝ bendiocarb ➝ permethrin + synergists ➝ pirimiphos-methyl) and the use of an insecticide mixture containing pyrethroid and pyrimiphos-methyl to control Ae. aegypti populations and overcome the challenge of widespread Ae. aegypti resistance to pyrethroid in Indonesia.

Assessment of packing density and transportation effect on sterilized pupae and adult Aedes aegypti (Diptera: Culicidae) in non-chilled conditions.

Long-distance transportation from a radiation facility to a target site potentially affects the quantity and quality of sterile male mosquitoes. This study tested the effects of multi-hour land transportation on the survival, longevity, and mating performance of gamma-rays sterilized adult and pupal male mosquitoes at different densities in non-chilled condition. The results demonstrated that mortality rate, longevity, induced sterility (IS) level, and mating competitiveness (C index) were significantly affected by life stage, transportation treatment, and density. Transportation was detrimental to the survival and longevity of the adults, and transporting pupae was restricted by the overcrowding effect; particularly, those packing density of 200 pupae. The longevity of transported mosquitoes were 1-5 days shorter than that of non-transported mosquitoes regardless of packing density. The irradiated transported adult males exhibited an equal IS and C index to their non-transported counterparts. Although there was no evidence suggested an association between low mating competitiveness and packing density in the transported adults, the mating competitiveness of adult mosquitoes decreased with increased packing density. Additionally, the effects of transportation and packing density on the mating ability of transported pupal males were also notable. The results indicate the factors of packing density and life stages in transporting sterile males under non-chilled conditions should be taken into account in formulating the procedure in SIT operation.

Ovitrap surveillance of dengue vector mosquitoes in Bandung City, West Java Province, Indonesia.

Larval surveillance is the central approach for monitoring dengue vector populations in Indonesia. However, traditional larval indices are ineffective for measuring mosquito population dynamics and predicting the dengue transmission risk. We conducted a 14-month ovitrap surveillance. Eggs and immature mosquitoes were collected on a weekly basis from an urban village of Bandung, namely Sekejati. Ovitrap-related indices, namely positive house index (PHI), ovitrap index (OI), and ovitrap density index (ODI), were generated and correlated with environmental variables, housing type (terraced or high-density housing), ovitrap placement location (indoor or outdoor; household or public place), and local dengue cases. Our results demonstrated that Aedes aegypti was significantly predominant compared with Aedes albopictus at each housing type and ovitrap placement location. Ovitrap placement locations and rainfall were the major factors contributing to variations in PHI, OI, and ODI, whereas the influences of housing type and temperature were subtle. Indoor site values were significantly positively correlated to outdoor sites' values for both OI and ODI. OI and ODI values from households were best predicted with those from public places at 1- and 0-week lags, respectively. Weekly rainfall values at 4- and 3-week lags were the best predictors of OI and ODI for households and public places, respectively. Monthly mean PHI, OI, and ODI were significantly associated with local dengue cases. In conclusion, ovitrap may be an effective tool for monitoring the population dynamics of Aedes mosquitoes, predicting dengue outbreaks, and serving as an early indicator to initiate environmental clean-up. Ovitrap surveillance is easy for surveyors if they are tasked with a certain number of ovitraps at a designated area, unlike the existing larval surveillance methodology, which entails identifying potential breeding sites largely at the surveyors' discretion. Ovitrap surveillance may reduce the influence of individual effort in larval surveillance that likely causes inconsistency in results.

Interpopulation variations in life history traits and reproductive tactics in Aedes aegypti: A test on populations 50 km apart.

The interpopulation variation in life history traits of a species reflects evolutionary adaptation in response to a local environment regime. We examined the life history traits of Aedes aegypti populations from 2 cities in southern Taiwan separated by 50 km. Results revealed a high level of trait differentiation in immature developmental time and survival of Ae. aegypti between the 2 cities. The Kaohsiung populations exhibited total pupation of 40%-60% on day 8; this was significantly lower than that of the Tainan populations and laboratory-reared KHsm mosquitos, which exhibited a pupation rate of 70%-90%. The slow immature development of the Kaohsiung populations was reflected in the low percentage of adult emergence (22%-26%) on day 10. The prolonged immature development did not select larger adults with longer life spans because the Kaohsiung populations had a shorter life span (≈37 d) than that of the Tainan populations (≈42 d). By contrast, immature development and longevity did not differ between populations within each region, indicating weak local differentiation. Three field populations exhibited male-bias sex ratio because of differential mortality of female immatures. The effect of female size on adult life history was nonsignificant. Two reproduction tactics were detected, representing the balanced-mortality hypothesis and the bet-hedging hypothesis. Despite their differential life history strategies and reproductive tactics, these mosquito populations apparently counterbalanced any shortcomings in traits to produce similar population growth. Maintaining optimal population density is essential for Aedes mosquitos to increase the probability of encountering mates and reduce the Allee effect.

The Invasion and Encapsulation of the Entomopathogenic Nematode, Steinernema abbasi, in Aedes albopictus (Diptera: Culicidae) Larvae.

The Asian tiger mosquito, Aedes albopictus, is of crucial concern to the public and veterinary health because of its vector role in transmission of several mosquito-borne diseases. Over the past decades, entomopathogenic nematodes (EPNs) have been used to control important agricultural insect pests and are considered to be effective against mosquitoes as well. The objectives of this study were to investigate the mosquitocidal effects of Steinernema abbasi to Ae. albopictus and the encapsulation processes of invading nematodes in the mosquito host. In this study, we found that S. abbasi was pathogenic to 3rd and 4th instar larvae of Ae. albopictus by entering the hemocoel of the 3rd and 4th instar larvae mainly through mouth and gastric caecum or by penetrating pupae through the intersegmental membrane or trumpet. The mosquito larvae infected with a single nematode caused a high mortality. Although EPNs in the hemocoel of mosquitoes were melanized and encapsulated, most Ae. albopictus larvae failed to survive after infection with S. abbasi. Overall, we demonstrated that S. abbasi is pathogenic to Ae. albopictus larvae, suggesting that this S. abbasi isolate has potential as a biocontrol agent for managing this vector mosquito.

A simplified method for blood feeding, oral infection, and saliva collection of the dengue vector mosquitoes.

A simple device using folded Parafilm-M as an artificial blood feeder was designed for studying two important dengue vector mosquitoes, Aedes aegypti and Aedes albopictus. The efficiency of the artificial blood feeder was investigated by comparing the numbers of engorged mosquitoes that fed on the artificial blood feeder versus mice as a live blood source. Significantly more engorged females Aedes aegypti fed on the artificial blood feeder than on mice. In addition, the artificial feeder could serve as a useful apparatus for oral infection via artificial blood meals, and for saliva collection in mosquitoes. Our method enabled us to collect saliva from multiple mosquitoes at once, providing sufficient infected saliva for determination of the virus titer by plaque assay analysis. Our artificial feeder has the advantage that it is simple, inexpensive, and efficient.

Oogenesis of Hematophagous Midge Forcipomyia taiwana (Diptera: Ceratopogonidae) and Nuage Localization of Vasa in Germline Cells.

Forcipomyia taiwana is an irritating hematophagous midge that preferentially attacks humans and affects leisure industries in Taiwan. Understanding the female reproductive biology of such pests would facilitate the development of pest control strategies. However, knowledge about oogenesis in the genus Forcipomyia is unavailable. Accordingly, we examined the ovariole structure and features of oogenesis in terms of the oocyte and the nurse cell. After being blood-fed, we observed a high degree of gonotrophic harmony-the synchronization of developing follicles. The follicle of the F. taiwana has only one nurse cell connected to the oocyte, which is distinct among hematophagous midges. In the nurse cell, we identified the perinuclear localization of the germline marker, Vasa. The Vasa localization is reminiscent of the nuclear envelope-associated nuage observed by electron microscopy. To determine whether F. taiwana Vasa (FtVasa) is an authentic nuage component, we produced transgenic flies expressing FtVasa in the female germline and proved that FtVasa was able to be localized to Drosophila nuage. By characterizing the oogenesis and Vasa expression in the germline cells of F. taiwana, this study extends knowledge about the female reproductive biology of hematophagous midges.

The first discovery of severe fever with thrombocytopenia syndrome virus in Taiwan.

Severe fever with thrombocytopenia syndrome (SFTS), an emerging tick-borne zoonosis, has been rapidly spread in many Asian counties since 2010, which raises the great concern in East Asia. Nevertheless, the infection status of SFTS in Taiwan remains unclear. To investigate the existence of SFTSV in Taiwan, a total of 151 serum samples collected from 31 sheep, 63 bovine and 57 dogs were enrolled this study. Furthermore, 360 adult female Rhipicephalus microplus were also included. One-step RT-nested PCR and IgG ELISA were conducted to test SFTSV specific RNA and antibodies, respectively. The result provided the first evidence of the existence of SFTSV RNA and antibodies in ruminants and ticks in Taiwan.

Effects of larval diets and temperature regimes on life history traits, energy reserves and temperature tolerance of male Aedes aegypti (Diptera: Culicidae): optimizing rearing techniques for the sterile insect programmes.

BACKGROUND: Producing high quality sterile males is vital in Aedes aegypti rear-and-release birth control strategies. Larval diets, rearing temperatures, and their interactions determine the accumulation rates of essential nutrients in larvae, but these factors have been understudied in relation to mass-rearing techniques for producing eminent males. METHODS: We compared the effects of two larval diets, a cereal-legume-based diet (Khan's diet) and a standard larval diet developed in the FAO/IAEA Insect Pest Control Laboratory (IAEA 2 diet). Diets were tested at selected temperatures for both larval and male adult life history traits, adult extreme temperature tolerance, and mating capacity relative to energy reserves of reared male adult Ae. aegypti. RESULTS: Khan's diet resulted in shorter immature development time at each test temperature (except for 25 °C) than an IAEA 2 diet. Larvae reared at 28 °C and 32 °C with Khan's diet demonstrated low pupation rates (c.80%). We accounted for these phenomena as secondary sex ratio manipulation, because a higher proportion of male adults emerged at 28 °C and 32 °C than that for the IAEA 2 diet. In general, the pupal development time shortened as temperature increased, resulting in higher teneral energy reserves in male mosquitoes. High energy reserves allowed male mosquitoes reared with Khan's diet to have higher adult longevity (5-6 days longer when sugar-fed and 2-3 days longer when water-fed) and tolerance of heat stress than those fed on the IAEA 2 diet. The IAEA 2 diet produced larger male mosquitoes than Khan's diet did: mosquitoes fed on Khan's diet were 1.03-1.05 times smaller than those fed on the IAEA 2 diet at 28 °C and 32 °C. No evidence indicated reduced mating capacity for small mosquitoes fed on Khan's diet. CONCLUSIONS: Larvae reared at 28 °C and 32 °C with Khan's diet were characterized by shorter immature development time compared with those fed on the IAEA 2 diet. Adult mosquitoes produced from that larval rearing condition exhibited a significant male bias, long lifespan, and better endurance against extreme temperatures relative to energy reserves. Thus, the larval diet at rearing temperature of 28 °C and 32 °C optimized rearing techniques for the sterile insect programmes. However, mating competitiveness and flight performance of adult males require further investigation.