Escherichia coli is one of the major causative infectious agents of diarrhea in preweaning and postweaning pigs and leads to a large economic loss worldwide. However, there is limited information on the distribution and virulence genes in E. coli isolated from diarrheic piglets, which also applies to the current status of pig farms in Korea. To investigate the prevalence of virulence-associated genes in E. coli related to diarrhea in piglets, the rectal swab samples of diarrheic piglets were collected from Seoul National University’s veterinary pathological department between 2022 and 2023. E. coli strains were identified using the VITEK II system. Two sets of multiplex PCRs and one single PCRs used to detect 10 E. coli virulence genes. As a result, a total of 145 E. coli isolates were identified encoding one or more of the virulence genes. Among them, the prevalence of individual virulence gene was as follows, piglets virulence genes were STa 58.6%(85/145), STb 22.1%(32/145), LT 15.2%(22/145), EAST1 24.8%(36/145), Stx2e 6.9%(10/145), F4 31.7%(46/145), F5 1.4%(2/145), F6 11.7%(17/145), F18 4.1%(6/145), and F41 0.7%(1/145) respectively. These results suggest that E. coli disease in piglets may not be associated with a single toxin or a major gene combination, but with more varied and complex toxins and their combinations in Korea.

Xenorhabdus와 Photorhabdus 속은 각각 곤충병원성 선충인 Steinernema와 Heterorhabditis에 공생하는 공생세 균이다. 감염성 선충의 유충은 공생세균을 표적 곤충의 혈강에 전달하고, 여기서 세균이 증식하여 숙주 선충의 발달을 돕는다. 이러한 선충과 세균 복합체의 성공적 공생관계는 세균의 이차대사산물을 통한 숙주의 면역억제 에 달려져 있다고 알려져 있다. 본 연구에서는 서로 다른 살충력을 보이는 6종의 Xenorhabdus를 확보하고 이러한 차이가 세균의 성장속도와 NRPS (Non ribosomal peptide synthease)에 의해 생성되는 세균의 이차대사산물 발현에 서 기원한다는 것을 확인하였다. 서로 다른 균주들은 콩명나방 (Tenebrio molitor)에 대한 살충력에 차이를 가지고 있었다. 이러한 세균들은 TSB 배지에서는 세균 성장 속도에 차이가 존재하지 않았지만 콩명나방 혈강 내에서는 세균의 성장 속도에 차이가 존재하는 것으로 나타났다. 또한 각 세균의 이차대사산물 추출물을 통한 곤충의 면역 억제 실험 결과 PLA2 활성 억제, 세포독성 능력들이 살충력과 상관관계가 있는 것으로 나타났다. 이러한 이차대 사산물의 경우 많은 물질이NRPS (Non ribosomal peptide synthease)에 의해 생성되므로 각 세균 별 NRPS의 유전자 발현을 보았을 때 흥미롭게도 살충력이 더 높은 스트레인의 세균이 일부 NRPS 유전자의 발현이 더 높은 것으로 나타났다. NRPS에 의해 합성되는 물질을 포함한 세균의 이차대사산물의 차이를 서로 비교하기 위하여 이차대사 산물 추출액을 GC-MS/MS를 이용하여 분석하였다. 본 연구를 통해 곤충병원세균에 살충력의 기원이 NRPS를 통해 합성되는 이차대사산물에 있다는 것을 확인하였으며 이를 이용한 다양한 NRPS 유래 물질 연구는 신규 살충 물질 개발에 들어가는 비용과 시간을 획기적으로 줄일 수 있을 것으로 기대된다.

To develop a functional probiotic that inhibits gingipain, a major virulence factor of Porphyromonas gingivlais (P. gingivalis), we screened over 30 probiotic strains for their ability to inhibit gingipian activity. We investigated the inhibition of expression of gingipain genes kgp, rgpA, and rgpB as well as gingipain activity, using freeze dried cell-free supernatants of Weisiella cibaria SPM402 (WC402) and Lactobacillus paracasei SMP412 (LP412), both of which demonstrated antibacterial activity against P. gingivalis. Thus, it was verified that kgp expression was reduced by approximately 0.71±0.02 folds and rgpB expression was reduced by approximately 0.71±0.14 folds at a concentration of WC402 10 mg/mL. Meanwhile, at the same concentration of 10 mg/mL of LP412, kgp expression was reduced by approximately 0.19±0.08 folds, rgpA expression was reduced by approximately 0.09±0.02 folds, and rgpB expression was reduced by approximately 0.24±0.03 folds. At a concentration of 10 mg/mL, Kgp activity was inhibited by approximately 78.65±3.58% (cell associated gingipain, CAG), 82.45±1.22% (cell-free gingipain, CFG) by WC402 and 80.71±2.11% (CAG), and 85.81±0.05% (CFG) by LP412 respectively. Rgp activity was also effectively inhibited by approximately 78.6±1.01% (CAG), 86.78±0.47% (CFG) and 82.93±1.26% (CAG), 88.81±0.36% (CFG) by WC402 and LP412 respectively. Based on these results, W. cibaria SPM402 and L. paracasei SPM412 can be regarded as functional probiotics with the ability to inhibit gingipain activity and exhibit antibacterial effects against P. gingivalis.

Enterotoxigenic Escherichia coli는 신생 및 이유기 돼지 설사의 주요 원인체로서 전세계적으로 양돈산업에 큰 경제적 손실을 끼치고 있다. 그러나 현재 국내에는 이러한 E. coli가 보유하는 다양한 병원성유전자의 분포 및 특성에 대한 정보가 부족한 실정이다. 이에 본 연구에서는 2013년부터 2016년까지 국내 163개 양돈농장에서 이유기 설사증 개체로부터 면봉스왑 샘플을 채취하여 동일 농장의 개체일 경우 5개에서 10개 정도를 혼합한 후, MacConkey agar에 배양하여 최종 API 32E system을 통하여 동정하였다. 분리된 모든 균주에 대해서 3가지의 다른 multiplex PCR을 수행하여 총 13종의 병원성유전자의 분포를 확인하였다. 이를 통하여 총 172개의 최소 한가지 이상의 병원성 유전자를 가지는 E. coli 균주를 확인하였고, 그 결과 병원성 유전자의 분포는 (1) fimbrial adhesins (43.0%): F4 (16.9%), F5 (4.1%), F6 (1.7%), F18 (21.5%), and F41 (3.5%); (2) toxins (90.1%): LT (19.2%), STa (20.9%), STb (25.6%), Stx2e (15.1%), EAST1 (48.3%); and (3) nonfimbrial adhesin (19.6%): EAE (14.0%), AIDA-1 (11.6%) and PAA (8.7%)로 나타났다. 결론적으로 본 연구결과는 국내 양돈농장의 이유기 설사증에 관연하는 E. coli는 다양한 종류의 병원성 유전자를 가지고 있으며 그러한 병원성 유전자의 조합도 매우 다양하게 분포하고 있음을 나타낸다.

Streptococcus mutans is one of the important bacteria that forms dental biofilm and cause dental caries. Virulence genes in S. mutans can be classified into the genes involved in bacterial adhesion, extracellular polysaccharide formation, biofilm formation, sugar uptake and metabolism, acid tolerance, and regulation. The genes involved in bacterial adhesion are gbps (gbpA, gbpB, and gbpC) and spaP. The gbp genes encode glucan-binding protein (GBP) A, GBP B, and GBP C. The spaP gene encodes cell surface antigen, SpaP. The genes involved in extracellular polysaccharide formation are gtfs (gtfB , gtfC , and gtfD ) and ftf , which encode glycosyltransferase (GTF) B, GTF C, and GTF D and fructosyltransferase, respectively. The genes involved in biofilm formation are smu630, relA , and comDE. The smu630 gene is important for biofilm formation. The relA and comDE genes contribute to quorumsensing and biofilm formation. The genes involved in sugar uptake and metabolism are eno, ldh , and relA . The eno gene encodes bacterial enolase, which catalyzes the formation of phosphoenolpyruvate. The ldh gene encodes lactic acid dehydrogenase. The relA gene contributes to the regulation of the glucose phosphotransferase system. The genes related to acid tolerance are atpD, aguD, brpA, and relA . The atpD gene encodes F1F0-ATPase, a proton pump that discharges H+ from within the bacterium to the outside. The aguD gene encodes agmatine deiminase system and produces alkali to overcome acid stress. The genes involved in regulation are vicR, brpA, and relA .

Spodoptera litura Fabricius is the serious, polyphagous pest of various crops. Due to the high resistant to most of chemical pesticides, it is hard to control S. litura in the fields. We investigated the virulence of four entomopathogenic nematodes (EPNs) in each larval stage. All EPNs were more effective to younger larvae than older larvae. Mortality, larval penetration rate and reproduction rate were significantly higher in H. indica, S. carpocapsae and S. longicaudum than the H. bacteriophora. Three high effective EPNs induced complete mortality of first instar larvae within 48 h, but of fifth instar larvae within 72 h. These EPNs also highly effective to control S. litura in pot assay. Thus, H. indica, S. carpocapsae and S. longicaudum, can be used as efficient biological control agents of S. litura.

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.

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.

The diamondback moth, Plutella xylostella is one of the world’s major pests. Economic cost to control this pest wasestimated between US$1.3 billion and US$2.3 billion based on management costs. Conservative estimate included yieldloss caused by 5% diamondback moth was estimated US$4 billion-US$5 billion. P. xylostella was managed by chemicalinsecticide such as organophosphates, carbamates and pyrethroids. But insecticide resistance which is caused by repeatedapplication makes it difficult to control this pest. For environmental friendly control of diamondback moth, entomopathogenicfungi could be used as alternative. We conducted bioassay to select high virulent isolate to larva of diamondback mothwith forty six entomopathogenic fungi which were isolated from soil samples by insect-bait method. As a result of bioassaytwelve isolates was selected as candidate. We investigated control efficacy of these twelve isolates with potted Chinesecabbage at laboratory and greenhouse.