Kim, Sung Jae
(Department of Veterinary Microbiology, College of Veterinary Medicine, Seoul National University)
,
Jung, Woo Kyung
(Department of Veterinary Microbiology, College of Veterinary Medicine, Seoul National University)
,
Hong, Joonbae
(Food & Microbiology Team, Test & Research Department, Consumer Safety Center, Korea consumer Agency)
,
Yang, Soo-Jin
(Department of Animal Science and Technology, Chung-Ang University)
,
Park, Yong Ho
(Department of Veterinary Microbiology, College of Veterinary Medicine, Seoul National University)
,
Park, Kun Taek
(Department of Biotechnology, Inje University)
Enterotoxigenic Escherichia coli는 신생 및 이유기 돼지설사의 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 3 가지의 다른 multiplex PCR을 수행하여 총 13 종의 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) non-fimbrial 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.
Enterotoxigenic Escherichia coli는 신생 및 이유기 돼지설사의 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 3 가지의 다른 multiplex PCR을 수행하여 총 13 종의 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) non-fimbrial 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.
Enterotoxigenic Escherichia coli is one of the major causative infectious agents of diarrhea in newborn and post-weaning pigs and leads to a large economic loss worldwide. However, there is limited information on the distribution and characterization of virulence genes in E. coli isolated from diarr...
Enterotoxigenic Escherichia coli is one of the major causative infectious agents of diarrhea in newborn and post-weaning pigs and leads to a large economic loss worldwide. However, there is limited information on the distribution and characterization of 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 and characterization of virulence genes in E. coli related to diarrhea in piglets, the rectal swab samples of diarrheic piglets (aged 2 d to 6 w) were collected from 163 farms between 2013 and 2016. Five to 10 individual swab samples from the same farm were pooled and cultured on MacConkey agar plates, and E. coli were identified using the API 32E system. Three sets of multiplex PCRs were used to detect 13 E. coli virulence genes. As a result, a total of 172 E. coli isolates encoding one or more of the virulence genes were identified. Among them, the prevalence of individual virulence gene was as follows, (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) non-fimbrial adhesin (19.6%): EAE (14.0%), AIDA-1 (11.6%) and PAA (8.7%), respectively. Taken together, various pathotypes and virotypes of E. coli were identified in diarrheic piglets. These results suggest a broad array of virulence genes is associated with coliform diarrhea in piglets in Korea.
Enterotoxigenic Escherichia coli is one of the major causative infectious agents of diarrhea in newborn and post-weaning pigs and leads to a large economic loss worldwide. However, there is limited information on the distribution and characterization of 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 and characterization of virulence genes in E. coli related to diarrhea in piglets, the rectal swab samples of diarrheic piglets (aged 2 d to 6 w) were collected from 163 farms between 2013 and 2016. Five to 10 individual swab samples from the same farm were pooled and cultured on MacConkey agar plates, and E. coli were identified using the API 32E system. Three sets of multiplex PCRs were used to detect 13 E. coli virulence genes. As a result, a total of 172 E. coli isolates encoding one or more of the virulence genes were identified. Among them, the prevalence of individual virulence gene was as follows, (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) non-fimbrial adhesin (19.6%): EAE (14.0%), AIDA-1 (11.6%) and PAA (8.7%), respectively. Taken together, various pathotypes and virotypes of E. coli were identified in diarrheic piglets. These results suggest a broad array of virulence genes is associated with coliform diarrhea in piglets in Korea.
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제안 방법
coli strains recently isolated from diarrheic piglets in Korea through the polymerase chain reaction and (2) investigated the prevalence and distribution of these virulence genes. The results of the present study will provide information, regarding the distribution of different virulence factors in E. coli causing diarrhea in pigs, which is required to control diarrheic disease in piglets due to pathogenic E. coli.
대상 데이터
Five to 10 samples were collected during diarrhea outbreak in each farm. The samples were submitted to the Research and Development Unit of the Green Cross Veterinary Products (Young-in, Korea), where they were pooled and cultured directly on eosin methylene blue (EMB) agar (BD-Difco, Franklin Lakes, NJ, USA). Subsequently, metallic green sheen colonies on EMB agar were selected, and then cultured on MacConkey agar (BDDifco).
이론/모형
The primer pairs for the 13 virulence factors were used for three sets of multiplex PCR (Table 1). The PCR was performed according to the protocol described by Zhang et al.11) using BioMetra TOne PCR machine (Analytik Jena AG, Jena, Germany). The reference E.
성능/효과
Most of the isolates investigated carried the toxin-encoding genes. Among the 172 isolates, 155 isolates (90.1%) carried at least one toxin-encoding gene, and the number of isolates with genes encoding different toxins was as follows: 33 (19.2%), 36 (20.9%), 44 (25.6%), 27 (15.7%), and 83 (48.3%) for LT, STa, STb, Stx2e, and EAST1, respectively. The most common toxinencoding genes were EAST1 and STb.
Among the 172 isolates, 99 (57.6%), 28 (16.3%), 34 (19.8%), 20 (11.6%), and 83 (58.4%) isolates carried the genes for ETEC, STEC, EPEC, DAEC, and EAST1, respectively. Among the 99 ETEC, 28 STEC, 34 EPEC, 20 DAEC, and 83 EAST1 isolates, 38 (38.
Among the 172 isolates, except EAST1, most of investigated toxins strongly associated with the fimbrial adhesins. Among the 34, 36, 44, and 27 isolates with the genes encoding LT, STa, STb, and Stx2e, 30 (90.9%), 26 (72.2%), 35 (79.5%), and 19 (73.1%) isolates carried the genes for fimbrial adhesin, respectively (Table 2). Among the fimbrial adhesins, LT and STb more strongly associated with F4 than F18, respectively.
Among the 99 ETEC, 28 STEC, 34 EPEC, 20 DAEC, and 83 EAST1 isolates, 38 (38.4%), 4 (14.3%), 15 (44.1%), 1 (5%), and 41 (49.4%) isolates carried the genes for only ETEC, STEC, EPEC, DAEC, and EAST1, respectively (Table 2).
. In the present study, 90.9%, 72.2%, and 79.5% of the fimbrial isolates carried the LT, STa, and STb genes, respectively. Several studies have indicated that STb is the most prevalent in the E.
The prevalence of the AIDA-1, pAA, and EAE genes, which encode non-fimbrial adhesins, was approximately 6%–7%.
4%) carried the genes for more than two pathotype-representative factors, and can be classified into the following 14 pathotypes. Twenty (11.6%), 16 (9.3%), 7 (4.1%), 6 (3.5%), and 4 (2.3%) isolates carried the genes for ETEC/EAST1, ETEC/STEC, EPEC/EAST1, ETEC/DAEC/EAST1, and ETEC/EPEC/EAST1, respectively. The number of isolates with the following four pathotypes ETEC/DAEC, ETEC/ EPEC, DAEC/EAST1, and EPEC/DAEC/EAST1 was 2 (1.
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