Frankliniella occidentalis is a major pest in agriculture. Following overuse of insecticides, high resistance has developed due to its high reproduction rate and short generation time. To control the resistant strains of the thrips, the ingestion RNAi- based control was established. A total of 67 genes were selected, and their double-stranded RNAs (dsRNA) were delivered to thrips via the leaf disc-feeding method. Among the genes screened, the dsRNA of Toll-like receptor 6 (TLR6) and coatomer protein subunit epsilon (COPE) resulted in the highest mortality (3.8- and 2.8-fold faster LT50 compared to control, respectively) when ingested by thrips. The dsRNA-fed thrips showed 53% and 83% reduced transcription levels of TLR6 and COPE, respectively. This result demonstrates that the observed mortality of thrips following dsRNA ingestion was due to RNAi, and this lethal genes can be employed as a practical tool to control thrips in the field.

Mosquitoes are primary medical insect pests due to their diseases transmission as vectors. In Korea, the insecticide-resistant populations of disease vector mosquito species, such as Anopheles sinensis, Culex pipiens and Culex tritaeniorhynchus, have constantly increased. Thus, management of insecticide resistance to major insecticides including pyrethroids and organophosphates is required for more efficient control of resistant populations. In this study, the quantitative sequencing (QS) protocols were established to detect the frequencies of three mutations (the L1014F on voltage sensitive sodium channel and the G119S and F331W on acetylcholinesterase 1) that are associated with either pyrethroids or organophosphates. Based on the QS protocol using newly designed non-polymorphic primers, resistance allele frequencies (RAFs) were estimated in field populations of An. sinensis, Cx. pipiens and Cx tritaeniorhynchus collected from an identical site in Korea. The dynamics of each resistance allele frequency over time in the same populations were also evaluated.

A residual contact vial plus water (RCVpW) bioassay method was established to monitor insectiside resistance in field populations of the melon thrips, Thrips palmi. Resistance level against six major insecticides were evalutated in five regions to test applicability of RCVpW as an on-site resistance monitoring tool. Reduced mortality in response to six test insecticides were exhibited compared to the RDA susceptable strain showing 100 % mortality, indicating different degree of resistance. An apparently reduced mortality to emamectin benzoate and chlofenapyr was observed in some field populations, suggesting uneven distribution of resistance to these insecticides in field populations. In addition, spinosad resistance was high and widely distributed in the test regions. Synergistic bioassay revealed that cytochrome P450-mediated metabolic factor is involved in spinosad resistance in the Korean population.

One of major mechanisms of insecticide resistance is the reduced susceptibility caused by mutations on the main target sites, such as sodium channel and acetylcholinesterase. Bioassay is a useful method to diagnose insecticide resistance of mosquitoes; however, it is hard to establish regional/annual resistance database based on bioassay. Recently, various markers of mutation and copy number variation have been identified through insecticide mechanism studies. Thus, molecular detection of resistance based on the resistance marker is now feasible, which can be readily implemented as a novel resistance monitoring tool to complement or replace the conventional bioassay method. In Korea, the density of vector mosquitoes native to the subtropical areas has increased due to climate change. Therefore, it is required to establish an efficient resistance monitoring system based on molecular markers to facilitate the construction of a nation-wide resistance map for more effective control of mosquitos. In addition, alternative insecticides should be introduced to the areas where mosquitoes develop high levels of resistance to maximize control efficacy against resistant populations

To be better fit for highways, pavement systems are required to provide comfortable and safe driving and be structurally durable. Composite pavements can be an effective option as they are more durable by placing a high functional asphalt overlay on a rigid concrete base layer. In order to apply a composite pavement system to the field, it is necessary not only to develop technologies that prevent reflecting crack and deterioration of the base layer, but also to improve bonding performance of materials and ensure structural performance as a pavement system against traffic loading. In advanced countries like Japan, USA and Europe, high-functional composite pavement systems are being put into practice across new highway networks. In this study, we evaluated structural performance (rutting, reflecting crack, and deflection) by applying traffic loads of actual highways through an accelerated pavement tester (APT) of a composite pavement section made up of a quiet porous surface laid over a water-proofing layer, a continuously reinforced concrete base, and a lean concrete sub-base layer, which was developed with new pavement methods used for each layer prior to field application. The APT specimen was constructed with paving materials and equipment actually used on site in the same dimensions (W3.5m*L14m*H2m) as actual highway sections in Korea, and 3-axle double-wheel heavy load (45ton) cart type KALES(Korean Accelerated Loading and Environmental Simulator) traveling on the specimen in both directions was used to simulate traffic loading. After applying around 8,574,000 ESALs of traffic loads, no reflecting crack occurred on the asphalt surface of the composite pavement, without surface distress except for rutting. In order to examine what causes rutting of pavements, we surveyed thickness of pavements by layer and measured asphalt density.

The common bed bug, Cimex lectularius, possesses a cholinesterase expressed exclusively in the salivary gland (ClSChE). In this paper, we investigated the molecular structure, tissue distribution patterns, and biochemical properties of ClSChE and showed that ClSChE exists as a soluble monomeric form or a soluble dimeric form connected by a disulfide bridge. Immunohistochemical analysis confirmed that ClSChE was expressed in the epithelial cells of both the salivary gland and the duct. In addition, the secretion of monomeric ClSChE through the proboscis during feeding was detected by western blotting using a ClSChE-specific antibody. To predict the role of ClSChE injected into the tissue of an animal host, we analyzed the extent of sequestration and hydrolysis of acetylcholine (ACh)/choline (Ch) by ClSChE by ultra-performance liquid chromatography-tandem mass spectrometry. Kinetic analysis revealed that ClSChE possesses extremely low Km (high affinity to ACh) and Vmax values. These findings suggest that ClSChE functions as a sequestering enzyme specific to ACh (not to Ch) by having a very strong affinity to ACh but an extremely long turnover time.

Perturbation of normal behaviors (e.g., nursing and foraging) in honey bee colonies by any external factors would immediately reduce the colony’s capacity for brood rearing, which can eventually lead to collapse of entire colony. To investigate the effects of brood rearing suppression in the biology of honey bee workers (nurse and forager), the gene-set enrichment analysis (GSEA) was performed for the transcriptomes of worker bees with or without their brood rearing being suppressed, from which functional profiles of pathways under influences by each condition were identified. Blocking of normal labor (i.e., nursing or foraging) induced the over-representation of pathways related with reshaping of worker bee physiology, suggesting that transition of labor is physiologically reversible. In addition, brood rearing suppression appeared to result in the reduction of neuronal excitability and aggressiveness in both forager and nurse, which would be necessary to manage the in-hive stress under unfavorable conditions

Strawberry jelly as a universal design food was developed using strawberry juice (SJ), sugar, xanthan gum (XG), and locust bean gum (LBG). Experimental variables included SJ concentration (30-40% (w/w)), sugar concentration (7.5-10.0% (w/w)), and the ratio of XG/LBG (0.3-4.0% (w/w)), and response variables were textural (hardness, gumminess, chewiness) and color properties. The formulation of strawberry jelly was optimized against hardness and the interactions among variables were predicted using the response surface methodology. Controlled storage test at 5 or 15°C was conducted to determine the values of the jelly at different temperatures. The optimal SJ and sugar concentrations and the ratio of XG/LBG against hardness were 40, 10, and 1.5%, respectively. The color did not change significantly during storage at 5 and 15°C (p>0.05). However, the textural characteristics during storage increased significantly at 5°C (p<0.05) and the hardness was appropriate to be used as a criterion for determining the shelf life of the jelly. The shelf life at 5°C generated from a zero-order kinetics (R²=0.96) was 40 d according to a criterial value, 1.8 N, of hardness. The Q10 value was calculated as 0.6, which allowed prediction of the shelf life values at different temperatures. The results from this study suggested a formulation of strawberry jelly as a universal design food and allowed determination of the shelf life of the food product.

Human body and head lice are obligatory human ectoparasites. Although both body and head lice belong to a single species, Pediculus humanus, only body lice are known to be a vector of several bacterial diseases. The higher vector competence of body lice is assumed to be due to their weaker immune response than that of head lice. To test this hypothesis, immune reactions were compared between body and head lice following infections by two model bacteria, Staphylococcus aureus and Escherichia coli, and a human pathogen, Bartonella quintana. Following dermal or oral challenge, the number of these bacteria increased both in hemocoel and alimentary tract of body lice but not in head lice and the viability of the B. quintana was significantly higher in body louse feces, the major route of infection to human. In addition, body lice showed the lower basal/induced transcription level of major immune genes, cytotoxic reactive oxygen species and phagocytosis activity compared with head lice. These findings suggest that a reduced immune response may be responsible, in part, for the increased proliferation and excretion of viable bacteria which are associated with the high level of human infectivity seen in body versus head lice.

Tropilaelaps mercedesae is an ectoparasite of immature honey bees belonging to the genus Tropilaelaps (Acari: Laelapidae). T. mercedesae has become a major threat to the Western honey bee Apis mellifera in Asia, including Korea, and is expanding its geographical range to northern regions due to global warming. To establish gene resources of T. mercedesae, the whole transcriptome was analyzed by RNA sequencing. An mRNA-focused library was generated from total RNA extracted from the mixed stages using the TruSeq RNA Library Preparation kit and sequenced using the HiSeq 2000 platform. A total of 6.0 Gb reads were obtained with 85% Q30 value. Trimmed sequence data were de novo assembled using the CLC Assembly Cell v 4.2. A total of 64,868 non-duplicate contigs were finally obtained and annotated by the Blast2GO using the NCBI nr database. The most abundant species in the resulting 14,336 Blast hits (22.1%) was Metaseiulus occidentalis, a predatory mite, followed by Ixodes scapularis and Tribolium castaneum, suggesting that the T. mercedesae transcriptome matches well with closely related other arthropod species, including mites and ticks. In order to provide basic information for efficient control and monitoring of potential resistance in T. mercedesae, acaricide target genes were annotated and characterized. One voltage-sensitive sodium channel gene encoding the molecular target of fluvalinate, a pyrethroid acaricide most widely used for the control of T. mercedesae, was identified and its molecular properties were investigated. In addition, other acaricide target genes, including acetylcholinesterase and glutamate (or GABA)-gated chloride channel, were identified and characterized.

The acetylcholinesterase 1 (AmAChE1) of the honey bee is known to be abundantly expressed both in the central and peripheral nervous systems. AmAChE1 exists mostly in the soluble form with little catalytic activity and has non-neuronal functions. Our preliminary observation showed that AmAChE1 expression fluctuated between the forages and nurses. A more systematic expression profiling of AmAChE1 over a year cycle on a monthly basis revealed that AmAChE1 was predominantly expressed during the winter months with being moderately expressed during the rainy summer time. However, no significant difference in AmAChE1 expression was noticed between the nurse and forager workers. Interestingly, AmAChE1 expression was inhibited when bees were allowed for brooding by placing overwintering bee hives in strawberry green houses with the supplement of pollen diets whereas it was resumed when the bee hives were removed from the green houses, thereby suppressed brooding. To confirm whether brooding status is a main determining factor for the suppression of AmAChE1 expression, active bee hives were placed in a screen tent, thereby hindering foraging, until brooding was completely suppressed, and then allowed to restore brooding by removing the screen. The AmAChE1 expression in the head was up-regulated when brooding was suppressed whereas its expression was down-regulated when brooding was resumed. These finding demonstrates that AmAChE1 expression in the central nervous system (i.e., head) is related with brooding status of honey bee. To understand the connection between the AmAChE1 expression and other pathways related with brooding, currently in progress are the analyses of head transcriptomes of honey bee workers with or without their brooding suppressed.

Leptotrombidium pallidum is the major vector mites for Orientia tsutsugamushi, the causative agent of scrub typhus. To understand the molecular mechanism of L. pallidum, we sequenced the whole genome using Illumina sequencing technology. Totally four genomic libraries with different insert sizes ranging from 280 bp to 8 kb were used to generate 45.1 Gb of genome in the combination of paired-end and mate-pairs sequencing reads. Quality filtering and correction of paired-end reads for very small and/or bad-quality sequences yielded 26.9 Gb of high-quality sequences, which are used to estimate the genome size as 175 Mbusing kmer methods and assembled into a 193.7 Mb genomic sequence scaffolds with N50 length of 92,945 bp. Furthermore, 94% of CEGMA completeness score were obtained from genome scaffold assembly. To facilitate gene annotation, we used a combination of de novo and homology based tools to predict gene models in the chigger mite genome. A combination of evidence-based and de novo approaches predicted 15,842 high-confidence protein-coding genes with an average transcript length of 1,511 bp and 2.4 exons per gene which corresponds to about 12.4% total gene length. Bacterial endosymbiosis are very common in mite species and can range from mutualistic to pathogenic associations. Henceforth, the endosymbionts in L. pallidum were predicted using the NCBI microbial draft genomes and mitochondrial genome. Besides, this L. pallidum draft genome can be used as a significant reference for comparative genomic studies across mite species.

Leptotrombidium pallidum is the major vector mite for Orientia tsutsugamushi, the causative agent of scrub typhus, in Asian countries, including Korea. The genome size of L. pallidum was previously estimated to be 191 ± 7 Mb (Kim et al., 2014). Genomic DNA (gDNA) was extracted from a single female from a 9-generation inbred L. pallidum colony and used for whole genome amplification (WGA). The resulting amplified gDNA was used for the construction of paired-end and mate-pair libraries and sequenced using Illumina platforms (HiSeq2000 and MiSeq). An unamplified gDNA sample extracted from 20 female mites was also used for sequencing in parallel. More than 45Gb sequence reads from both paired-end and mate-pair libraries of the WGA gDNA were trimmed and then de novo assembled using the CLC Asembly Cell v.4.0 for contig assembly and SSPACE for scaffolding. The assembly generated approximately 6,545 scaffolds with N50 value of 92,945 and total size of ~193Mb, which was in a good agreement with our previous estimation. Repeat analysis showed that about 30% of genome (~58Mb) was masked as repeats, most of which were unclassified novel elements. For gene predictions, generated were the PASA models based on genomic alignments of RNA-seq reads from 4 different chigger mite samples (i.e. male, female, larva, and protonymph) and the GeneWise models based on genomic alignments of protein sequences from 4 closely related species with chigger mite. Independently, ab initio gene predictions were performed with AUGUSTUS and FgeneSH with custom trained matrices optimized for L. pallidum and GENEID with pre-trained matrix for Acyrthopsiphon pisum. By combining all together, 15,842 genes were predicted finally. Manual curation is in progress for various groups of genes, including chemosensory receptor genes, immune-related genes, acaricide target genes, etc.