Enteropathogenic Escherichia coli (EPEC) is one of the etiological agents that causes diarrhea in weaning pigs. In this study, we report that mutating both relA and spoT genes in EPEC E2348/69 can promote bacterial clearance in porcine gastrointestinal tract (GIT). Our experimental analyses showed that an E2348/69 ΔrelAΔspoT mutant strain was not detected in porcine feces after 1 day post-infection (dpi), whereas its parental strain was continuously detected in porcine feces within 10 dpi. Histologic assessment revealed that the mutant strain was unable to induce moderate pathologic lesions in porcine GIT when compared to those with the wildtype strain. Taken together, our data suggest that the relA and spoT genes in EPEC play an important role in bacterial survival and pathogenesis in porcine GIT.

창원시의 3개 지점(남천, 창원천, 청운지)에서 물 시료를 채취하고, 채취한 시료로부터 genomic DNA를 분리하였다. 분리된 DNA와 class A extended-spectrum beta-lactamase(ESBL) 유전 자 5가지(blaOXA-1, bl aSHV, blaTEM, blaCTX-M-1, blaCTX-M-9)를 target으로 하는 primers를 사용하여 quantitative PCR을 수행하였다. 시료를 채취한 지점별로 ESBL 유전자들의 수와 비율에서 확연한 차이 가 있음을 알 수 있었다. 남천의 경우 DNA 30ng당 ESBL 유전자가 1.93×106 copy나 존재하는 데에 반해 창원천의 경우에는 1.47×105 copy 그리고 청운지에는 9.5×103 copy 밖에 존재하지 않았다. 흐르 는 하천인 남천과 창원천에서는 각 ESBL 유전자들의 비율에서 큰 차이가 없었다. 즉, 남천에서는 blaOXA-1 유전자가 65.3%, blaCTX-M-1 유전자가 33.6%를 차지하여서 이 두 유전자가 전체 ESBL 유전자 의 99% 가까이를 차지하고 있었다. 창원천에서도 blaOXA-1 유전자가 64.1%, blaCTX-M-1 유전자가 19.1%를 차지하므로 이 두 유전자가 전체의 83% 이상을 차지하고 있었다. 하천과 다르게 폐쇄된 환경 인 청운지 연못에서는 내성세균들의 총량 자체가 다른 두 하천에 비해서 월등하게 적었다. 또한 내성 세균의 비율도 달라서 blaCTX-M-1 유전자가 전체의 87.5%, 그리고 blaCTX-M-9 유전자가 9.8%를 차지하 고 있었다.

수은은 생물 축적과 먹이사슬을 통한 생물 농축되며, 미량에서도 유해한 영향을 나타내기 때문 에 해양 환경 내에서 중요한 문제가 되고 있다. 그러나 해양 소형 갑각류에 대한 수은의 생 물 영향은 다른 금속에 비해 연구가 미흡하다. 본 연구에서는 기수산 물벼룩 Diaphanosoma celebensis을 아치사 농도(0.2, 0.4, 0.8 μg/l)의 무기 수은(HgCl2)에 48시간 노출시킨 후, 대사 관련 유전자의 발현 양상을 조사하였다. 해독효소 유전자 5종(cytochrome P450; cyp360A1, cyp361A1, cyp4AP3, cyp4C122, cyp370C5)과 소화효소 6종(alpha amylase (AMY), alpha amylase related protein (AMY-like), trypsin (TRYP), chymotrypsin-like protein (CHY), lipase (LIP), pancreatic lipase-related protein (PLRP))의 유전자 발현을 quantitative real time reverse transcription polymerase chain reaction (qRT-PCR)을 이용하여 분석하였다. Cyp 유전자의 경우 clan2에 속 하는 cyp370C5와 clan4에 속하는 cyp4AP3 유전자의 발현이 농도 의존적으로 유의하게 증가 하였다. 한편 소화효소 유전자 중에서는 단백질 소화와 관련 있는 TRYP 유전자의 발현이 농도 의존적으로 증가하였다. 이러한 결과는 cyp370C5와 cyp4AP3가 수은 독성을 해독하는 과정 에서 중요한 역할을 담당할 것으로 보이며, 수은이 소화효소 유전자의 발현을 조절함으로써 에너지 대사에 영향을 미칠 수 있음을 제시한다. 본 연구는 해양 소형 갑각류에서 수은에 대한 분자 수준의 영향을 이해하는데 도움이 될 것이다.

Expression changes of stress-induced peroxidase (swpa2 and swpa4) and storage root-specific sporamin (spo-A and spo-B) genes were examined using qRT-PCR after treatment with wounding and bacterial pathogens on leaves of sweetpotato (Ipomoea batatas) plants. As a result of examining the expression change in the wounding treatment condition for 48 hours after treatment, which is a representative physical stress, the expression of all genes increased after 12 hours of wounding treatment, but the expression pattern of each gene group showed distinct differences thereafter. Expression levels of swpa2 and swpa4 strongly increased up to 36 or 48 hours after wounding treatment, whereas spo-A and spo-B expression levels strongly decreased after 24 or 36 hours after wounding treatment. Peroxidase and sporamin genes are involved in the early response after wounding treatment and, in particular, the peroxidase swpa2 and swpa4 genes are also involved in the late response after wounding treatment. Gene expression analysis after infection with P. chrysanthemi, which causes softness in sweetpotato, showed that the swpa2 and swpa4 genes were weakly induced after 8 hours and then strongly induced after 20 hours during pathogen infection. Expression of the spo-A gene was weakly induced in the pathogen-treated group after 20 hours, whereas spo-B showed an expression pattern similar to that of the peroxidase genes. The above results indicate that expression of the stress-induced peroxidase gene used in this study is induced not only by abiotic stress but also by biological stress caused by bacterial pathogen invasion and that peroxidase plays an important function in the initial defense response.

Fescues, which are widely cultivated as grasses and forages around the world, are often naturally infected with the endophyte, Epichloë. This fungus, transmitted through seeds, imparts resistance to drying and herbivorous insects in its host without causing any external damage, thereby contributing to the adaptation of the host to the environment and maintaining a symbiosis. However, some endophytes, such as E. coenophialum synthesize ergovaline or lolitrem B, which accumulate in the plant and impart anti-mammalian properties. For example, when livestock consume excessive amounts of grass containing toxic endophytes, problems associated with neuromuscular abnormalities, such as convulsions, paralysis, high fever, decreased milk production, reproductive disorders, and even death, can occur. Therefore, pre-inoculation with non-toxic endogenous fungi or management with endophyte-free grass is important in preventing damage to livestock and producing high-quality forage. To date, the diagnosis of endophytes has been mainly performed by observation under a microscope following staining, or by performing an immune blot assay using a monoclonal antibody. Recently, the polymerase chain reaction (PCR)-based molecular diagnostic method is gaining importance in the fields of agriculture, livestock, and healthcare given the method’s advantages. These include faster results, with greater accuracy and sensitivity than those obtained using conventional diagnostic methods. For the diagnosis of endophytes, the nested PCR method is the only available option developed; however, it is limited by the fact that the level of toxic alkaloid synthesis cannot be estimated. Therefore, in this study, we aimed to develop a triplex real-time PCR diagnostic method that can determine the presence or absence of endophyte infection using DNA extracted from seeds within 1 h, while simultaneously detecting easD and LtmC genes, which are related to toxic alkaloid synthesis. This new method was then also applied to real field samples.

In crustaceans, molting is regulated by interactions between ecdysteroid and juvenile hormone (JH) signaling pathway-related genes. Unlike the ecdysteroid signaling pathway, little information on the role of JH signaling pathway-related genes in molting is available in zooplanktonic crustaceans. In this study, three genes (juvenile hormone acid O-methyltransferase (JHAMT ), methoprene-tolerant (Met ), and juvenile hormone epoxide hydrolase (JHEH )) which are involved in the synthesis, receptor-binding, and degradation of JH were identified using sequence and phylogenetic analysis in the brackish water flea, Diaphanosoma celebensis. Transcriptional changes in these genes during the molting cycle in D. celebensis were analyzed. Sequence and phylogenetic analysis revealed that these putative proteins may be functionally conserved along with those of insects and other crustaceans. In addition, the expression of the three genes was correlated with the molting cycle of D. celebensis, indicating that these genes may be involved in the synthesis and degradation of JH, resulting in normal molting. This study will provide information for a better understanding of the role of JH signaling pathwayrelated genes during the molting process in Cladocera.

Grapevine leaf rust (GLR) caused by Phakopsora euvitis results in the reduction of fruit quality and yield loss in grape production. The purpose of this study was to investigate the expression of genes related with defense responses in the grapevines infected with rust pathogens. In two genotypes of Ampelopsis species inoculated with P. euvitis, the real-time PCR with RNAs was performed to investigate transcripts levels of nine defense-related genes, including pathogenesis-related 1 (PR1), β-1,3glutannase (Glu), chitinase (Chi), superoxide dismutases (SOD), glutathione peroxidase (GPX), phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), stilbene synthase 1 (STS1), and resveratrol O-methyltransferase (ROMT). All tested genes were upregulated in YG11030 as well as YG10075, while mostly the expression of genes was higher in YG10075 than that in YG11030. Glu and STS showed significantly high expression in YG10075 than in YG11030. Expression of ROMT was upregulated in YG11030 and downregulated in YG10075 at 24 hours after inoculation. The differential expression pattern of the tested genes seems to be related with the defense responses, considered to be involved in plant-resistant responses against the infection by P. euvitis, and further studies on resistant responses based on the expression of genes would provide valuable information in breeding grapes resistant to diseases.

This investigation aimed to assess the appetite response changes of olive flounder to starving and re-feeding conditions. Three different feeding groups (2 weeks feeding, fed; 2 weeks starving, starved; and 1 week starving and 1 week feeding, re-fed) were established to examine the changes in appetite-related genes for each group. The weight gain of the fish was highest for the fed group and lowest for the starving group. Based on the daily feed intake (DFI) and cumulative feed intake (CFI), overall food intake was found to increase in the re-fed group more than in the fed group from week 1 to week 2 of the experiment. Hypocretin neuropeptide precursor (HCRT) and galanin receptor 1 (GAL-R1) mRNA expression in the brain of olive flounder were decreased in the starved group. Corticotropin-releasing hormone (CRH) was decreased in all experimental groups, except for the fed group. However, overall leptin concentrations in the plasma did not change across groups. Considering the differences between this study and previous studies on starving and feeding, various factors (except the production and expression mechanisms of appetite-related factors in response to starving) are likely acting on the appetite responses of the fish. In this study, a 1-week re-feeding period induced substantial effects on appetite response when compared to a 2-week feeding period. These findings show that even if re-feeding is performed after starving, the unbalance caused by the re-feeding can affect various physiological changes in fish by feed intake efficiency.

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.