Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a food-borne bacterial pathogen that causes various diseases in both humans and animals such as hemorrhagic colitis and hemolytic uremic syndrome. Because cattle are the main reservoir of this microorganism, undercooked meat and meat byproducts contaminated with EHEC O157:H7 are most commonly associated with epidemic disease outbreaks. As an enteric pathogen, EHEC O157:H7 enters the body via a fecal-oral route and must survive passage through the gastric stomach at pH 1.5 to 3.5 to establish an infection within the gastrointestinal tracts. Therefore, the ability to resist such acidic environments is important to the pathogenesis of EHEC O157:H7 during a host infection. In this review, we will discuss on the acid resistance (AR) mechanisms induced by EHEC O157:H7 when E. coli encounters acidic environments.

Poultry play an important role in meeting the protein demand through the supply of chicken meat and eggs, and antimicrobial-resistant bacteria can be transmitted from poultry to humans through the food chain. In this study, 716 E. coli isolates were collected from poultry industry in Korea during the period from 2016-2018. Among the cephalosporin antimicrobial, the rate of resistance to first-cephalosporins (cefazolin and cephalexin) was more than 18.0% but secondcephalosporins (cefoxitin and cefuroxime) and four-cephalosporins (cefepime) was less than 10.0%, without any differences based on the poultry. In quinolone antimicrobial, the rate of resistance to nalidixic acid was more than 46.0% in all poultry but ciprofloxacin was more than 44.0% in broiler farm and chicken meat and less than 18.0% in layer farm and egg. In addition, the rate of resistance to ampicillin and tetracycline was more than 30.0% without any differences in poultry, but the rate of resistance to amoxicillin - clavulanate, trimethoprim - sulfamethoxazole, imipenem and gentamicin was higher in broiler farm and chicken meat than in layer farm and egg. MDR was detected in 120 (60.6%), 113 (79.6%), 80 (37.0%), and 75 (46.9%) isolates from broiler farm, chicken meat, layer farm, and egg, respectively, with an overall prevalence of 54.2% (388/716).

노거수는 사람으로 따지면 노년기에 해당하여 다른 나무에 비해 기후변화 등 환경변화에 취약하며, 강한 바람과 눈 피해로 가지 부러짐, 쓰러짐 등 큰 피해를 받을 수 있다. 이러한 위험요인으로부터 노거수를 건강하게 보존하기 위해서는 체계적인 진단과 정기적인 모니터링이 이루어져야 한다. 다행히 천연기념물로 지정한 노거수를 대상으로는 ｢문화 재보호법｣에 따라 5년마다 정기조사를 실시하고 ｢천연기념물(식물) 상시관리 지침｣에 따라 예방적 모니터링을 하고 있다. 모니터링 항목으로는 나무 생육상태 진단을 위해 수세, 수형, 신초, 잎 크기, 가지고사, 병해충, 토양 건습도 등을 조사하고 있으나 항목은 정성적으로 평가되며, 육안조사 위주의 주관적 판단에 따라 생육진단이 이루어지고 있어 조사자에 따라 평가에 차이가 발생할 수 있다. 객관적인 생육상태 평가를 위해 생장추, 천공저항, 사이고미터(Shigometer), X-ray 등의 방법이 사용되고 있으나 생장추와 천공은 나무에 구멍을 내기 때문에 나무에 물리적 영향을 미치거나 직접적인 부후 확산의 가능성이 있고, 사이고미터는 조사자와 측정 환경에 따른 값의 변화폭이 크고 수목에 따라 정확도에 차이가 있을 수 있다. 천연기념물 노거수나 보호수의 경우 물리적으로 구멍을 뚫는 천공 방식을 적용하는데 한계가 있기 때문에 비파괴적으로 나무 내부를 진단하고 정량적으로 생육상태를 평가할 수 있는 단층촬영방법을 적용할 수 있다. 국내에 관련 연구는 거의 없는 실정으로 본 연구는 내부 단층촬영법을 이용해 노거수 생육상태 평가 연구의 기초자료 구축을 목적으로 한다. 조사는 2018년 3월 ~ 2018년 7월 동안 천연기념물 노거수 및 보호수로 지정된 느티나무, 매실나무, 소나무 10주를 대상으로 PiCUS 장비를 이용해 음파 단층촬영(sonic tomography) 및 전기저항 단층촬영(electric resistance tomography)을 실시하였다. 두 단층촬영 결과를 종합하여 나무 내부의 생육상태를 진단하고 대상 나무의 부후(decay)와 공동(cavity) 유무, 위치, 부후수준, 훼손된 목재 면적(damaged areas) 등을 도출하였다. 음파 및 전기저항 단층촬영법은 노거수 생육현황 조사시 육안조사를 보완하여 생육진단의 정확성을 높이고 측정 결과를 정량화하며 생육상태 모니터링 DB 구축에 도움이 될 수 있을 것이다. 나무 내부단층촬영법은 2000대 초부터 국외를 중심으로 연구가 이루어졌으나 국내 자생 수종을 대상으로 진행된 바가 거의 없으며 노거수 등 직경 1m 이상의 나무를 대상으로 한 연구도 미흡한 실정이다. 본 연구는 나무 내부단층촬영법을 시범 적용한 연구로 후속적으로는 주요 자생 수종에 본 방법을 적용하여 장비의 정확성을 검증하는 기초 연구를 실시하고, 다음 단계로 이를 활용해 노거수 생육상태를 객관적으로 진단할 수 있는 평가기준을 마련하여 천연기념물 노거수와 보호수를 효율적으로 관리해야 할 것이다.

Molecular diagnostic markers are necessary for establishing highthroughput screening systems to support insecticide-resistant population management. Here, we identified single amino acid substitution mutations related to carbamate resistance in Laodelphax striatellus Fallén type-1 acetylcholinesterase (Lsace1) using carbofuran-selected strains. The phenotypic resistance profiles of the final selection strain (SEL9) compared to the susceptible strain revealed a 14-fold higher resistance ratio based on topical application, 1.2-fold higher general esterase activity, and 4.3- fold higher acetylcholinesterase insensitivity based on the 50% inhibitory concentration (I50), suggesting that insensitivity of the target site could occur as a resistance factor. Comparison of the nucleotide sequences of Lsace1 of five strains (SUS, SEL0, SEL3, SEL6, and SEL9) revealed two amino acid substitutions (F330Y and F331H). To understand the roles of these mutations, we determined the allele frequency of both point mutations in the selected strains using quantitative sequencing methods. In addition, several quantitative genotypic traits (e.g., gene copy numbers and transcript levels of Lsace1, Lsace2, and LS.CarE1) were assessed. A correlation analysis of genotypic and phenotypic traits revealed strong correlations between resistance level and I50 with F331H allele frequency. Interestingly, the F331H mutation was negatively correlated with transcript levels of Lsace1, suggesting that selection pressure might result in a reduction of the target gene. Overall, the F331H mutation and reduced mRNA are important factors in the development of carbamate resistance. Furthermore, the point mutation can be used to monitor rapid carbofuran resistance in conjunction with molecular diagnostic methods such as quantitative sequencing.

The eggs of Asian tiger mosquito, Aedes albopictus, possess high desiccation resistance, which contribute the rapid spread of this mosquito across the world. Melanization of eggshell appear to play a role in the resistance to desiccation. Dopachrome-conversion enzyme (DCE, Yellow) significantly accelerates the melanization of the eggshell. In this study, we demonstrated functional importance of two yellow genes, AalY-g and AalY-g2, in the chorion formation. Both genes were highly induced in the ovary at 48 h after blood meal. Injection of dsRNA for AalY-g or AalY-g2 into adult females had no effect on fecundity. However, the outermost colorless exochorion of the eggs obtained from both dsRNA-treated females was fragile and peeled off in places, and melanization of the endochorion was obviously delayed by several hours. In addition, unlike eggs from control females which acquired high desiccation resistance between 18 and 24 h after oviposition (HAO), 60-70% 24 HAO eggs from either AalY-g- or AalY-g2-deficient females were collapsed when they were moved to an air-dry condition, and the desiccation resistance was not increased in later stages of embryonic development analyzed. TEM analysis revealed that abnormal morphology and ultrastructure of the endochorion, particularly outer-endochorion, in the 24 HAO and older eggs from either AalY-g-and AalY-g2-deficient females. These results indicate that AalY-g and AalY-g2 are required for morphology and formation of the endochorion (outer-endochorion), a structure that appears to be critical for desiccation resistance of the Ae. albopictus eggs. This work was supported by NRFs (NRF-2015R1A6A3A04060323 and NRF-2018R1A2B6005106)

Mature silkworm of Bombyx mori is known to contain various functional materials. However, it is too hard to chew or digest for humans when it is cooked or lyophilized. In Korea, the Rural Development Administration recently developed and patented a method for making mature silkworms edible. In this study, therefore, we examined the effects of a larval powder of steamed mature silkworm (SMSP) on skin pigmentation and melanogenesis. For elucidating the depigmenting activity, lightness of a designated site on the murine dorsal skin was measured in vivo. During the experiment, hyperpigmentation was induced on the skin by cumulative exposure to ultraviolet B (UVB) irradiation. At the end of the experiment, melanin production on the skin was visualized by Fontana-Masson staining. Orally administered SMSP of pistachio cocoon strain for at least a month or longer significantly and reduced abnormal pigmentation caused by UVB on the murine dorsal skin. SMSP also showed a potential anti-melanogenic efficacy in modulating UVB-induced hyperpigmentation. Taken together, SMSP was identified as a potential candidate for a novel anti-melanogenic agent, which showed depigmenting efficacy against UVB-induced hyperpigmentation in vivo when administered orally.

Recently, phosphine resistance of Sitophilus oryzae has been reported from China, India, Brazil and Australia. In this study, susceptibility of three fumigants were assesses on phosphine resistant and susceptible S. oryzae to investigate domestic phosphine resistance level and to use base data for resistance control. On susceptible insects, LCT50 of phosphine was 0.440mg h/L for egg, 0.602mg h/L for early larvae, 3.901mg h/L for late larvae, 6.171mg h/L for pupae and 0.295mg h/L for adult stage, respectively. LCT50 of methyl bromide was 9.997mg h/ L for egg, 12.113mg h/L for early larvae, 18.952mg h/L for late larvae, 21.104mg h/L for pupae and 17.824mg h/L for adult stage, respectively. LCT50 of ethyl formate was 75.795mg h/L for egg, 60.110mg h/L for early larvae, 160.491mg h/L for late larvae, 255.797mg h/L for pupae and 77.711mg h/L for adult stage, respectively. On resistant insects, LCT50 of phosphine was 6.959mg h/L for egg, 28.456mg h/L for early larvae, 48.170mg h/L for late larvae, 29.106mg h/L for pupae and 16.550mg h/L for adult stage, respectively. LCT50 product of methyl bromide was 17.842mg h/L for egg, 14.900mg h/L for early larvae, 25.840mg h/L for late larvae, 43.520mg h/L for pupae and 16.397mg h/ L for adult stage, respectively. LCT50 of ethyl formate was 60.034mg h/L for egg, 64.450mg h/L for early larvae, 149.028mg h/L for late larvae, 140.408mg h/L for pupae and 66.043mg h/L for adult stage, respectively. Domestic resistant S. oryzae showed 4 to 56 times higher resistance rate than susceptible insects.

The two-spotted spider mite, Tetranychus urticae, is a worldwide agricultural pest that invades a wide range of host crops and rapidly develops resistance to pesticides. T. urticae can be resistant to one acaricide or exhibit multiple resistances or cross resistance to various other acaricides. Acequinocyl inhibits respiration in mitochondria at the ubiquinol oxidation site (Q0) of Complex III of the electron transfer chain. Pyridaben is a METI acaricide that inhibits mitochondrial electron transport at complex I. In this study, we investigated the cross resistance to seven acaricides in acequinocyl- and pyridaben-resistant strain. Furthermore, the frequencies of the I256V and N321S mutations in mitochondrial cytochrome b (cytb) of pyridaben-resistant and fieldcollected strain was analyzed.

Entomopathogenic fungi have been known as promising candidates for biological control of insect pests. Recently, researchers consider the fungal thermotolerance in formulations and field applications. In this study, we investigated the production of thermotolerant Isaria javanica and I.fumosorosea conidia through grain-based solid cultures and exposure to light stress. As results, of the ten grain substrates, Italian millet, rice, perilla seed and sesame, rice, sorghum produced highly thermotolerant conidia in the strains. The two strains were exposed to a light stress and showed enhanced thermal stability compared to control, when exposed to 45°C for 2 hours. This work suggests that heatresistant entomopathogenic fungal conidia can be produced by grainbased solid cultures and exposure to light stress.

Residual contact vial (RCV) method was used to monitor insecticide resistance in field populations of the melon thrips, Thrips palmi. Median lethal doses (LD50) at 8 h post-treatment of six insecticides (chlorfenapyr, cyantraniliprole, cypermethrin, dinotefuran, emamectin benzoate and spinosad), which are commonly used for T. palmi control, were determined at 8 h post-treatment using a susceptible RDA strain. The diagnostic doses for on-site resistance monitoring of the six insecticides, which were determined as two-fold higher doses of LD90 for the RDA strain, were in the range of 0.299 to 164,25 μg-1cm2. To test the applicability of RCV for T. palmi, insecticide resistance levels in three field populations (Gyeonggi; GG_AS, Chungbuk; CB_CJ, Jeonbuk; JB_KJ) were evaluated. Field populations showed reduced mortality (0-50% mortality) to spinosad, cypermethrin, cyantraniliprole and emamectin benzoate, that they have different degree of resistances to these insecticides. In particular, all test field populations exhibited 0% mortality to spinosad, suggesting wide spread of spinosad resistance in the field. Moreover, no detectable mortality to emamectin benzoate was observed in JB_KJ strain, suggesting uneven distribution of emamectin benzoate resistant population of T. palmi. To provide more precise information on resistance profiles and distribution in T. palmi populations, it would be necessary to conduct a large scale resistance mapping for broad geographical regions.

Laodelphax striatellus is an important pest of rice due to not only sucking rice seedlings, but also transmitting serious plant viruses. Among various kinds of insecticide groups (carbamates, organophosphorus, neonicotinoids, etc.), carbofuran, a systemic carbamate insecticide, has been most extensively used to control rice pests including L. striatellus, resulting in widespread carbamate resistance in Korea and other Northeast Asia countries. To identify the genes associated with carbofuran resistance, we obtained a 14-fold higher resistant strain (SEL9) from the mixed-field population (SEL0) by consecutive selection. A transcriptome-based analysis was conducted and differentially expressed genes (DEG) were compared between the SEL9 and SEL0 strains. A total of 96,185,150 reads were analyzed, of which 62,860,430 reads were mapped. From these reads, 15,356 transcripts were annotated. A total of 327 up-regulated and 275 down-regulated genes were identified in the resistant SEL9 strain compared to SEL0 strain by DEG analysis. Gene ontology (GO) analysis was performed using DEGs showing statistical significance (P < 0.05). The GO analysis identified 1,320 genes in biological process group, 1,100 genes in cellular component group and 428 genes in molecular_function group.

Phosphine (PH3) resistance in the stored-products insect pests has been reported throughout the world in various insect species, including Rhyzopertha dominica, Tribolium castaneum, and Cryptolestes ferrugineus, leading farmers and fumigators to identify new fumigation tools to control PH3-resistant insect pests in storage facilities. Understanding PH3-resistance mechanisms in insects might contribute to providing clues for the development of new chemicals, including fumigants, to control various PH3-resistant insects. A proteomic study has shown 15 decreased proteins in the PH3-resistant R. dominica (CRD343 strain) in comparison to the PH3-susceptible R. dominica, and among those 15 proteins, dihydrolipoamide dehydrogenase (DLD), a protein involved in the Krebs cycle, was identified (Park et al., 2008). The DLD polymorphisms responsible for genetic resistance have disulfide active sites for PH3 binding and are highly sensitive to arsenic exposure after mutagenesis in insects (R. dominica and T. castaneum) and Caenorhabditis elegans (Schlipalius et al., 2012). Here, two PH3- resistant S. oryzae strains were used to understand the development of PH3 resistance in these insects. Acute toxicity test by PH3 on the two PH3-resistant strains was undertaken followed by ethyl formate inhibition study on cytochrome c oxidase activity. The Lineweaver-Burk plots after inhibition studies showed there were significantly difference in inhibition mode between the resistant strains and the control. The RT-qPCR analysis and the next-generation sequencing of the mitochondrial DNA revealed significant changes in metabolism and energy production. Taken together, the PH3 resistance in S. oryzae was definitely acquired by the overall transformation of biochemical reactions to overcome PH3 toxicity.

4,4’-dichlorodiphenyltrichloroethane (DDT) has been re-recommended by the World Health Organization for malaria mosquito control in Africa. Previous DDT use has resulted in predisposition of resistance, and with continued use resistance will increase further in terms of level and extent. Drosophila melanogaster is a model dipteran that has many available genetic tools, has been widely used for elucidating insecticide resistance mechanisms, and is related to malaria mosquitoes allowing for extrapolation. The 91-R strain of D. melanogaster is highly resistant to DDT (>1500-fold); however, there is no mechanistic scheme that accounts for this level of resistance. Recently, reduced penetration, increased detoxification, and direct excretion have been identified as resistance mechanisms in the 91-R strain. Their interactions, however, remain unclear. Use of Gal4/UAS-RNAi transgenic lines of D. melanogaster allowed for the targeted knockdown of genes putatively involved in DDT resistance and has identified the role of several cuticular proteins (Cyp4g1 and Lcp1), cytochrome P450 monooxygenases (Cyp6g1 and Cyp12d1), and ATP binding cassette transporters (mdr50, mdr65, and mrp1) in increased sensitivity to DDT. These findings have been further validated in 91-R flies using a nanoparticle-enhanced RNAi strategy, directly implication these genes in DDT resistance in 91-R flies.

The cotton bollworm, Helicoverpa armigera is one of the most polyphagous and cosmopolitan pest species in world wide including Korea. It also showed high level of resistance against various types of insecticide. To set the integrated resistant management program, first, we tried to identify the insecticide resistant mechanism via transcriptome analysis of coding and non-coding RNAs using 4 strains (Australian susceptible and resistant strains, Korean and Brazilian resistant strains, additional a sub-lethal dose insecticide treatment in Korean resistant strain). From the illumina based RNAseq data sets with genome information, some resistant involved detoxification genes and long non-coding RNAs were discovered. Second, following molecular markers were developed from resistant strain specific amino acid substitution from those candidate genes. Third, functionally identified genes’ markers were applied in field populations using some molecular diagnostic tools such as LAMP (loop mediated isothermal amplification). Now additional functional analysis is going on for these un-characterized candidate genes and long non-coding RNAs.