비브리오패혈증균은 세계에서 치사율이 50%에 달하는 가장 치명적인 수인성식품매개 병원균으로 해수에서 흔히 있으며, 특히 따뜻한 계절에 발생한다. 본 연구는 제주도 의 해수, 유통 수산물, 수족관물에서 분리한 비브리오패혈 증균에 대해서 RT-PCR을 이용한 독소 유전자, Vitek을 이 용한 항생제 내성, PFGE를 이용한 유전적 특성을 조사하 였다. 총 487개의 시료를 조사한 결과 비브리오패혈증균 46주(중복 균주 포함)가 해수에서 44주, 유통수산물에서 1 주, 수족관물에서 1주 분리되었다. rtxA, viu와 같은 독소 유전자는 각각 8주(17.4%), 9주(19.6%) 검출되었고, vvhA 와 같은 독소 유전자는 모든 균주에서 검출되었다. 항생 제 내성 실험결과 cefoxitin 항상제에 대해서 100% 내성 이 나타났다. 비브리오패혈증균 46주에 대한 PFGE 분석결과 총 6유형이 100% 상동성을 보였고, 유사도는 81.3- 98.0%로 나타났다. 수산물과 수족관물에서 분리된 비브리 오패혈증균은 해수와의 상동성 결과 유사도는 불일치로 나타났고 지역과 시료 사이에는 유사성이 없었다. 독소 유 전자를 가진 비브리오패혈증균에 의한 식중독 환자가 제 주도에서 발생한 점을 고려해볼 때, 해수, 유통 수산물, 수 족관물에서 분리한 비브리오패혈증균에 대한 모니터링이 지속되어야 할 것으로 보인다.

The purpose of this study is to investigate the distribution of toxin genes and antimicrobial resistance of Vibrio parahaemolyticus isolated from seafood in Gwangju. A total of 335 seafood, including 163 shellfish, 97 fish, and 36 mollusk, were tested in this study. As a result, V. parahaemolyticus was detected in 123 (36.7%) of 335 seafood. The tdh gene was not detected in all strains, while the trh gene was detected in 3 strains (2.4%). According to antimicrobial susceptibility test, 116 strains (94.3%) represent resistance to ampicillin, and 1 strain (0.8%) represents resistance to trimethoprim/sulfametoxazole. However, all strains were sensitive to 9 antimicrobial agents, including amikacin, chloramphenicol, tetracycline, and more. Therefore, the risk of V. parahaemolyticus isolated from seafood in Gwangju is considered low, but continuous monitoring of V. parahaemolyticus in seafood is required.

본 연구에서는 기 선발한 Enterobacter asburiae ObRS-5 균주를 1×108 cfu mL-1 농도로 고추에 관주 처리했을 때 Phytophthora capsici에 의한 고추역병을 74.6% 방제하는 효과가 있었다. E. asburiae ObRS-5 균주에 의한 고추역병 방제 메커니즘을 확인하기 위해 고추의 PR1, PR4 및 PR10 유전자를 특이적으로 증폭하는 프라이머를 이용하여 quantitative PCR을 수행하였다. 그 결과 E. asburiae ObRS- 5 균주를 처리한 고추에서 대조구와 비교하여 상기 세 가지 유전자의 발현이 모두 높은 수준을 유지하였다. 또한 E. asburiae ObRS-5 균주는 고추의 생육을 억제하지 않으면서 ISR 반응을 유도하는 것으로 나타났다. 이러한 결과를 통하여 P. capsici이 침입할 때 E. asburiae ObRS-5 균주가 매개 하는 ISR 메커니즘을 통해 Phytophthora 역병의 제어가 가능한 것으로 사료된다.

본 연구에서는 비브리오 패혈증 예방과 치료에 유용한 항생제를 제시하기 위해 V. vulnificus의 독소유전자 분포와 항생제 내성을 분석하였다. 2015년부터 2017년까지 3년간 전남지역에서 발생한 비브리오 패혈증 환자로부터 분리되어 보관된 18균주와 전남지역에서 채취된 어패류 및 횟집 수족관수에서 분리된 5균주, 총 23균주를 대상으로 하였다. 실험에 사용된 V. vulnificus 23균주 모두 V. vulnificus로 재확인되었다. V. vulnificus 균주의 독소유전자를 분석한 결과, 23균주 중 19균주(82.6%)에서 RtxA 독소 유전자가 확인되었고, 23균주 모두에서 viuB와 vvhA 독소 유전자가 검출되었다. 이러한 결과는 독소유전자의 검출율이 기존 보고에 비해 높은 것이며, 실험에 사용한 모든 V. vulnificus 균주가 1개 이상의 독소유전자를 보유한 것으로 생선회 섭취와 상처를 통한 비브리오 패혈증 감염의 위험성이 상존하고 있었다. 따라서 횟집 종사자 등에 대한 비브리오 패혈증 예방 교육이 필요한 것으로 판단되었다. V. vulnificus에 대한 항생제 내성 실험결과 cefoxitin 항생제에 94.4%가 내성을 나타내었고, chloramphenicol과 tetracycline 등 14종의 항생제에 감수성을 나타내었다. 비브리오 패혈증 치료에 chloramphenicol과 tetracycline 항생제를 사용하는 현 치료법이 유용한 것으로 판단되었다.

Staphylococcus pseudintermedius는 개에서 기회감염을 유발하는 병원체이며, 공중보건학적으로도 주요한 인수공통 병원체이다. 개에서 분리된 S. pseudintermedius 균주들은 주로 항생제 내성 및 개에서 피부 감염을 유발하는 주요 원인균으로 연구되어 왔지만, 가축에서 분리된 S. pseudintermedius 균주들의 항생제 내성 및 장내 독소 생성에 대한 정보는 매우 제한적이다. 본 연구에서는 개, 돼지, 육우에서 분리된 S. pseudintermedius 균주들에서 18가지의 장내 독소 (staphylococcal enterotoxin; SE) 유전자와 toxic shock syndrome toxin 유전자(tst-1)의 분포양상을 조사 하였다. 또한, S. pseudintermedius 균주들의 항생제 내성 양상과 더불어 mecA 유전자 및 SCCmec type 또한 확인하였다. 육우에서 분리한 하나의 균주를 제외한 모든 개와 돼지 분리주들이 4개 이상의 항생제에 내성을 보였으며, 개에서 분리된 6개의 균주 중 4개의 S. pseudintermedius 균주 들이 메티실린 내성과 더불어 SCCmec V를 가진 것으로 확인 되었다. 총 11개의 SE 유전자들 (seb, sec, see, seg, sei, sej, sel, seo, sep, seq, seu) 및 tst-1가 개, 돼지 및 육 우로부터 분리된 S. pseudintermedius 균주들에서 확인 되었으며, 대부분의 분리주들 (83%)에서 2개 이상의 SE 유전자들이 확인 되었고, 그 중 sel (42%) 및 sep (42%)가 가장 빈번하게 검출 되었다. 본 연구를 통하여 반려견에서 뿐만 아니라 주요 가축에서 존재하는 S. pseudintermedius 균주들에서 높은 항생제 내성 양상을 확인 하였으며 , 항생제 내성과 더불어 여러 staphylococcal enterotoxin 및 tst-1유전자들을 전파 할 가능성을 확인 하였다 .

본 연구는 옥수수 재배 시 환경에 영향을 미치는 노균병 저항성과 관련된 유전자 후보군을 탐색해서 노균병으로 인한 토양오염과 옥수수 생산량 감소를 해결하기 위하여 노균병 저항성 품종을 효율적으로 발굴하기 위한 연구이다. 옥수수의 6번 염색체의 152,892,333과 154,335,437 사이에 있는 노균병 저항성 유전자를 탐색하였으며 이 부분에 존재할 것으로 예상되는 전사체에서 38개의 프라이머 세트를 디자인하여 이 중 16개의 예측 전사체를 가려 내었다. 또한 RT-PCR을 수행하여 감염된 Ki11의 발현이 높은 7개의 전사체로 5개의 품종에 대하여 건강한 샘플과 감염된 샘플을 검정하였고 최종 5개의 후보 유전자군[알려지지 않은 미확인 유전자 2개, OFP transcription factor, bZIP transcription factor, pentatricopeptide repeat (Ppr)]이 발견 되었다. 본 연구의 결과로 추가적인 실험 설계를 통해 5개의 후보 유전자군에 대한 재검정을 통하여 확실한 노균병 저항성 유전자를 발굴하고 이를 노균병 저항성 품종 개발 및 방재에 이용할 수 있을 것으로 사료된다.

Many β-lactam antimicrobials, including cephalosporins, have been used in both veterinary and human medicine in the treatment of zoonotic and infectious diseases. Especially, third-generation cephalosporins such as ceftiofur have been approved for systemic use in food-producing animals, which has resulted in the emergence of β-lactamase genes. This study aimed to investigate the occurrence of β-lactamase-producing E. coli isolated from commercial layers and characterized their antimicrobial resistance and virulence genes. Among the 85 cefotaxime (CTX)-resistant E. coli, all isolates showed resistance to at least one antimicrobial, and the rates of resistance to nalidixic acid, cephalothin, ampicillin, and cefazolin were more than 50.0%. In particular, 28 isolates were identified as containing b-lactamase genes. The extended-spectrum β-lactamase (ESBL) and plasmid-mediated AmpC genes blaCTX-M-1, blaCTX-M-14, blaCTX-M-15, and blaCMY-2 were detected in 1, 6, 5, and 4 isolates, respectively. The non-ESBL/pAmpC gene blaTEM-1 was detected in 12 isolates. The distribution of antimicrobial resistance genes in 28 β-lactamase-producing E. coli was as follows: aac(3)-II (64.3%), sul2 (32.1%), tetA (28.6%), sul1 (25.0%), cmlA gene (25.0%), and tetB (14.3%). In total, 6 virulence genes (astA, eaeA, escV, fimH, iucC, and papC) were also identified and the rates in virulence gene were as below: fimH (92.9%), iucC (25.0%), astA (21.4%), papC (10.7%), eaeA (7.1%) and escV (7.1%). Our findings suggest that antimicrobials used in commercial layer must be regulated in Korea, and comprehensive surveillance is necessary to prevent the dissemination of resistant isolates.

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.

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 ability of antimicrobial resistant Salmonella strains to cause invasive disease can be attributed to various virulence genes. In this study, the virulence genes located in SPI-1, SPI-2, SPI-5, SPI-11 were found in all antimicrobial-resistant Salmonella isolates. This suggests that these genes play important roles in Salmonella invasion, growth, or survival in the host. The association between the presence of virulence genes and antimicrobial resistance was assessed using the Spearman’s correlation coefficient, and there is a positive association between the gatC, tcfA, hylE, spiA, pagC, msgA, invA, sipB, prgH, spaN, orgA, tolC, iroN, sitC, lpfC, and sopB genes, and resistance to CF, NA and S. This suggests that the association between antimicrobial agents and virulence genes has been shown to vary with the types of antibiotics that are commonly used in different countries. These different associations can be explained by the mechanisms underlying pathogenicity and the acquisition of resistance genes by Salmonella.

본 연구는 서울시내에서 시판중인 식육에서 E. faecalis 를 분리하고 이 균들의 항생제 내성 패턴, 항생제 유출 펌 프 유전자 및 병독성 유전자의 분포를 분석하였다. 총 277 개의 식육시료에서 93균주의 E. faecalis 를 분리하였다. 이 균주들의 항생제 내성비율은 ampicillin에는 35.5%, chloramphenicol에 6.4%, ciprofloxacin에 4.3%, eryhtromycin 에 18.3%, quinupristin-dalfopristin에 76.3%, tetracycline에 45.2%의 내성이었으며 levofloxacin, teiconplanin 및 vancomycin에는 모든 균이 감수성이었다. 약물 유출펌프인 MFS 타입의 eme(A)와 ABC 타입의 efr(A)유전자는 모든 균주 (100%)에서 확인되었으며 efr(B)는 98.9%, lsa는 91.4%의 균주에서 확인되었다. 병독성 인자인 gel(E)는 68.8%, ace 는 90.3%, asa1는 47.3%, efaA는 91.4%, esp는 12.9%의 균주에서 확인되었다. 본 연구는 시판 식육에서 분리한 지 표 미생물의 하나인 E. faecalis의 항생제 내성, 약물유출 펌프 및 병독서 유전자의 분포를 분석한 연구로 지속적인 모니터링을 하여야 할 것으로 사료된다.

Insect resistance to crop protecting chemicals is developing very rapidly, which is a major problem in the production of crops. Recently, the mechanism of resistance to various pseticides s has been revealed in Helicoverpa armigera. However, to date, no technology has been developed to quickly diagnose resistance yet. In this study, we present the results of developing method of rapid diagnosis of resistance

In order to establish symbiotic host-bacterial relationships, symbionts in insects evolved a mechanism to overcome host immune responses. Here we provide the resistance of symbiotic bacteria on the insect immune system. As a result, through the transposon mutagenesis, we found a salivary gland (SG) susceptible mutant. The disrupted gene was identified as nlpB involved in lipoprotein synthesis. The nlpB, bla double deletion mutant was sensitive to SG like nlpB-Tn5 inserted mutant. This mutant increases outer membrane permeability. It provides an explanation for SG susceptibility, because the antimicrobial peptide in SG would be able to translocate across the outer membrane more easily than in the wild type. These results indicate that nlpB and bla are likely to be important factors in terms of determining resistance against SG of Riptortus that is connected with the successful colonization of the Riptortus midgut.

Polydnaviruses (PDVs) are a group of insect viruses and symbiotic to some endoparasitoid wasps. Genome analyses of different PDVs provide a number of genes putatively associated with alteration of host insect physiological processes. Especially, PDV gene products assist host wasp development by suppressing immune responses and delaying larval development of parasitized lepidopteran hosts. Thus, PDV genes can be applied to control insect pests by incorporating them into crop genomes. This talk illustrates two examples of PDV genes: CpBV-CST1 and CpBV-ELP1. CpBV-CST1 has been known to inhibit insect cysteine proteases to suppress immune and development, while CpBV-ELP1 exhibits a high cytotoxicity to insect cells. Transgenic tobaccos expressing CpBV-CST1 or CpBV-ELP1 were constructed by using Agrobacterium-mediated transgenic system and exhibited antixenosis against chewing and sucking insect pests on tobacco.

Transcriptome analysis was conducted for the identification of genes associated with insecticide resistance in Frankliniella occidentalis. Resistant strain (FO_RDAHC) exhibited 39.2- ~ 533-fold resistance to acrinathrin, spinosad, emmamectin benzoate and thiamethoxam compared with a susceptible FO_RDA strain. Average 7.6 million reads (± 5,068,895 reads) were obtained from the pyrosequencing and were assembled into the draft CDS database. Gene annotation was conducted by BLAST (UniProt), Pfam, FUNCAT and COG analysis. In the deferentially expressed gene (DEG) analysis, 838 genes were up-regulated and 815 genes were down-regulated over 2-fold ratio in FO_RDAHC strain. Highly up-regulated genes included genes encoding several cuticle-related proteins, cytochrome P450s, esterases and transporter genes. An autotransporter protein gene exhibited the highest up-regulation (596 fold) whereas a GMC oxido-reductase revealed the highest down-regulation (12 fold). Further study would be necessary to validate the actual transcript levels of DEGs and to investigate their functional roles in insecticide resistance.

Streptococci are among the normal human microflora that populate the oral cavity. However, oral streptococci are known as a major causative agent for dental caries and bacterial endocarditis. Tetracycline is a broad-spectrum antibiotic that is used for oral infections but two mechanisms of tetracycline resistance in streptococci have been reported. The tet(K) and tet(L) genes in these bacteria are related to the active efflux of tetracycline, whereas tet(M) and tet(O) confer ribosomal protection from this antibiotic. It has been reported that the tetracycline resistance of streptococci is related mainly to the activity of tet(M) and tet(O). In our present study, we examined the prevalence of tet(M) and tet(O) in oral streptococci isolated from Korean dental plaques using PCR. One hundred and forty eight of 635 isolates (23.3%) were tetracycline resistant; 68 of these strains (46%) harbored tet(M) and 3 strains (2%) were positive for tet(O). However, tet(M) and tet(O) did not co-exist in any of the resistant strains. Seventy seven of the 148 tetracycline resistant strains (52%) were negative for both the tet(M) and tet(O) genes.

Erythromycin is a macrolide antibiotic and inhibits bac- terial protein synthesis by stimulating the dissociation of the peptidyl-tRNA molecule from the ribosomes during elon- gation. The use of macrolides has increased dramatically over the last few years and has led to an increase in bac- terial resistance to these antibiotics. Bacterial resistance to erythromycin is generally conferred by the ribosome methy- lation and/or transport (efflux) protein genes. Among the identified erythromycin-resistant genes, erm(B) (erythromy- cin methylation) and mef(A) (macrolide efflux) are gene- rally detectable in erythromycin-resistant streptococcal spe- cies. The distribution of these genes in oral streptococcal iso- lates has been reported in studies from other countries but has not been previously examined in a Korean study. We here examined by PCR the presence of erm(B) and mef(A) in oral streptococci isolated from Korean dental plaques. Among the 57 erythromycin-resistant strains tested, 64.9% harbored erm(B) whereas 40.4% were positive for mef(A). Eleven isolates had both the erm(B) and mef(A) genes. Twenty six isolates had only erm(B) and 12 isolates had only mef(A). Eight of the 57 strains examined were negative for both genes.