• 제목/요약/키워드: Salmonella spp.

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오리 도체에서 등온유전자증폭기법을 이용한 Salmonella spp. 신속 고감도 검출 기법 연구 (Rapid and Sensitive Detection of Salmonella spp. by Using a Loop-Mediated Isothermal Amplification Assay in Duck Carcass Sample)

  • 조애리;동희진;조성범
    • 한국축산식품학회지
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    • 제33권5호
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    • pp.655-663
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    • 2013
  • 본 연구에서는 Salmonella invA 유전자를 마커로 Salmonella 특이 LAMP primer를 제작하였고, inclusivity 및 exclusivity 실험 결과는 각각 100%로 관찰되었다. 순수 배양된 Salmonella 희석액에서의 검출한계는 18.17분에서 $3.2{\times}10^3$ CFU/mL ($R^2$ = 0.9446)으로 관찰되어 신속 간단하고 민감도가 높아 Salmonella 검출을 위한 방법으로 효과적일 뿐만 아니라 간접적인 정량기법으로서도 활용 가능함을 알 수 있었다. 1차 증균 배지인 BPW에 S. Enteritidis 접종 후 0시간 및 12시간 배양 후 LAMP의 검출한계는 각각 $3.2{\times}10^3$ CFU/mL 및 $3.2{\times}10^0$ CFU/mL으로 관찰되었고, 오리도체 시료가 포함된 BPW에 S. Enteritidis 접종 후 0시간 및 12시간 증균액에서의 LAMP 검출한계는 각각 $3.2{\times}10^3$ CFU/mL 및 $3.2{\times}10^0$ CFU/mL으로 관찰되어 PCR과 비교하였을 때 오리도체시료 성분에 의한 영향이 비교적 낮았음을 알 수 있었다. 또한, 실제 도압장에서 채취한 오리도체시료를 BPW에 6시간 배양한 1차 증균액에 개발한 LAMP기법을 적용한 결과, 96%의 민감도 및 84%의 특이도를 보여 오리도체에서의 Salmonella 신속 스크리닝 기법으로서 효과적으로 활용될 수 있음을 알 수 있었다.

Lactobacillus spp의 Salmonella enteritidis KU 101에 대한 보호 효과와 L. casei YIT 9018의 16S-23S rRNA Intergenic Spacer Region 염기배열 특성 (Protective Activities of Lactobacillus casei YIT 9018 against Salmonella enteritidis KU101 and Characteristics of 16S-23S rRNA Intergenic Spacer Region Sequence)

  • 배진성;윤영호
    • Journal of Animal Science and Technology
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    • 제45권3호
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    • pp.473-482
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    • 2003
  • Lactobacillus spp. 급여에 의하여 Salmonella enteritidis 감염에 의한 치사율의 저하 성향, 시험관내 억제활성, 장액 내의 총 IgA 농도변화와 Lactobacillus casei.의 16S-23S rRNA intergenic spacer region의 sequence를 측정 비교한 결과는 다음과 같다. Lactobacillus spp. 5균주는 시험관내 Salmo- nella enteritidis 억제활성 측정 결과 모든 시험균주가 억제활성을 나타내었고 그 억제활성의 차이가 통계적인 유의성을 보였으며, 억제활성의 차이는 L. helveticus CU 631 > L. rhamnosus GG ATCC 53103 > L. johnsonii C-4 > L. acidophilus ATCC 4356 > L. casei YIT 9018 인 것으로 나타났다. Lactobacillus spp. 급여에 의하여 Salmonella enteritidis challenge에 의한 치사율 저하효과는 Lb. helveticus CU 631은 대조구의 생존률 62%에 대하여 100%로서 가장 높은 치사율 저하효과를 나타내었고, L casei YIT 9018, L johnsonii C-4의 생존률이 70%와 50%를 나타내었다. Lactobacillus spp.의 16S-23S rRNA intergenic spacer region의 sequence를 측정하고 gene bank에 등록된 균주의 sequence와 비교한 결과 homology의 차이를 나타내었다.

TaqMan 실시간 중합 효소 연쇄반응에 의한 살모넬라속의 검출 및 ompC 항원단백 유전자의 비교 (Detection of Salmonella spp. by TaqMan real-time PCR and comparison of nucleotide sequences of ompC gene among Salmonella)

  • 이영성;최경성;김명철;한재철;채준석
    • 대한수의학회지
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    • 제42권4호
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    • pp.513-522
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    • 2002
  • Antigenic ompC genes of S. gallinarum, S. pullorum and S. dublin were characterized among Salmonella spp. isolated from chickens and other animals to identify genetic variation. Salmonella ompC gene fragment (1,027 bp) was amplified by PCR and the amplicons were cloned for comparison of nucleotide sequences. The identity of the sequences between S. gallinarum and S. pullorum, S. gallinarum and S. dublin, S. pullorum and S. dublin was 99.8%, 97.6% and 97.8%, respectively. Also, we found that ompC has some diversity between S. gallinarum and S. pullorum, and other Salmonella spp. which may be useful to type the organisms. Similar to diagnosis in other organisms, the TaqMan PCR method can be applied to rapid and accurate diagnosis of salmonellosis in chickens and other animals. We designed PCR primers and TaqMan probe for flagellin gene (fliC) for detection of Salmonella spp. by TaqMan PCR. The TaqMan PCR method was 10,000 times more sensitive than conventional PCR.

양돈장 살모넬라 혈청 양성율과 도축돈 살모넬라균 분리율의 상관관계 및 분리 균주의 항생제 내성 (Correlation between the Salmonella seroprevalence on farms and the isolation rate from slaughtered pigs and antimicrobial resistance from the isolates)

  • 김경언;안종민;양병훈;박영희;박미영;정준용
    • 한국동물위생학회지
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    • 제37권4호
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    • pp.233-240
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    • 2014
  • Salmonella spp. are one of the most common bacteria that causes heavy losses in swine industry and have implications for public health. In this study, the correlation between Salmonella seroprev-alence on farms and the isolation rate from slaughtered pigs was analyzed and the antimicrobial resistance of the isolated Salmonella spp. was investigated. A total of 3,001 serum samples for ELISA were collected from 17 farms during two consecutive years (2012-2013). The mean values of ELISA OD% for each 8 age groups were as follows; gilt 27.83 (n=472), sow 23.75 (n=367), 150 days (d) of pig 16.53 (n=278), 130 d 11.87 (n=366), 100 d 9.46 (n=378), 20 d 9.17 (n=394), 70 d 6.56 (n=382), 40 d 3.72 (n=364). From July 2013 to January 2014, a total of 53 (8.0%) Salmonella strains were isolated from 665 slaughtered pigs shipped from those 17 farms. The mean values of ELISA OD% for each age groups serum samples that were collected in the second half of 2013 showed a positive correlation at 100 d (0.61, P<0.05), 130 d (0.45, P<0.1) with the isolation rate of Salmonella spp. in the slaughtered pigs. All the isolates were identified by a real-time PCR and tested for antimicrobial susceptibility. As a result, the predominant serovar was S. Typhimurium (52.8%) and there were 15 strains showing their own antimicrobial resistance pattern. All strains were susceptible to amoxicillin, cefepime, ciprofloxacin and some of them were resistant to streptomycin and tetracycline (60%), ampicillin (53.3%), chloramphenicol (33.3%), respectively.

경남지역 가금류 도축장 신선육에서 분리한 Salmonella spp.와 Enterococcus faecalis의 독성인자 보유 패턴 분석 (Analysis of virulence gene profiles of Salmonella spp. and Enterococcus faecalis isolated from the freshly slaughtered poultry meats produced in Gyeong-Nam province)

  • 하도윤;차휘근;한권식;장은희;박하영;배민진;조아름송이;이후근;고병효;김도경;황보원;김상현
    • 한국동물위생학회지
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    • 제41권3호
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    • pp.157-163
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    • 2018
  • In order for monitoring of pathogenic bacterial contamination in the freshly slaughtered poultry meats produced in Gyeong-Nam province, we first isolated 4 strains of Salmonella spp. and 32 strains of Enterococcus faecalis from the total 164 samples, then we analyzed potential virulence gene profiles of the bacterial isolates by PCR using species-specific primer. The potential virulence genes we selected in this study were stn, invA, fimA, spvR, and spvC for the isolates of Salmonella spp. and those of esp, cylM, cylA, cylB, gelE, fsrA, fsrB, and fsrC were for the isolates of E. faecalis. The PCR results showed that all 5 virulence genes were detected simultaneously in the all isolates of Salmonella spp. However, there was a diverse occurrence pattern of the virulence genes in the case of E. faecalis. The gene for enterococcal surface protein (esp) was not detected among the isolates (0/32), and the haemolysin gene prevalence rate of cylA, cylB, and cylM were 3.1% (1/32), 9.3% (3/32), and 9.3% (3/32), respectively. Moreover, the genes of gelE, fsrA, fsrB, and fsrC that associated with gelatinase activity were detected in the rate of 53.1% (17/32), 53.1% (17/32), 53.1% (17/32), and 53.1% (17/32), respectively. In conclusion, in the isolates of Salmonella spp., all possessed 5 virulence genes tested, suggesting that they are all related with each other clonally. However, in the case of E. faecalis isolates, the occurrence of the haemolysin genes (cylM, cylA, cylB) and the gelatinase genes (gelE, fsrABC) was highly variable among the isolates.

Detection of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in Kimchi by Multiplex Polymerase Chain Reaction (mPCR)

  • Park, Yeon-Sun;Lee, Sang-Rok;Kim, Young-Gon
    • Journal of Microbiology
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    • 제44권1호
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    • pp.92-97
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    • 2006
  • We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

In vivo Antagonistic Effect of Lactobacillus helveticus CU 631 against Salmonella enteritidis KU101 infection

  • Bae, Jin-Seong;Byun, Jung-Ryul;Yoon, Yung-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권3호
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    • pp.430-434
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    • 2003
  • In vivo antagonistic effect of Lactobacillus helveticus CU 631 and Lactobacillus spp. against typical enteritis causing pathogen Salmonella enteritidis KU 101 have been determined, which showed an increase in survival rate and the decline in viable cell numbers of pathogen in liver and spleen at sacrifice. A signifcant difference in the antagonistic effect against KU 101 were observed, which was species and/or strain dependent of Lactobacillus (p<0.01), the survival rate of the mice in the Salmonella infection by feeding L. helveticus CU 631 has been shown to be 157%, whereas those of L. rhamnosus GG ATCC 53103, L. acidophilus ATCC 4356, L. johnsonii C-4 were 137%, 132%, 119% respectively on the basis of lactobacilli non-associated control KU101 fed mice to be 100%. Viable cells of S. enteritidis KU101 in the liver and in the spleen at sacrifice were decreased in Lactobacillus spp. fed group with no significant difference. The higher level of total secretory IgA concentration in the intestinal fluid of lactobacilli fed mice than control mice have been observed. In vitro antagonistic activity of Lactobacillus spp. against KU101 have been determined, a prominent antagonistic activity of CU 631 against KU 101 were demonstrated.

인천지역 도축장에서 생산된 돼지고기의 미생물 오염도 조사 (Survey on the contamination of microorganisms in pork from slaughterhouse in Incheon area)

  • 황원무;이성모;황현순;한정희
    • 한국동물위생학회지
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    • 제27권1호
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    • pp.7-15
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    • 2004
  • For measuring general bacterial count and Escherichia coli count, the standard plate count was used to conduct a test on 113 cases of pork carcass surfaces from slaughterhouses in Incheon area from January to February 2003. And Salmonella spp, Staphylococcus aureus and Listeria monocytogenes was used to conduct a test on 68 cases of pork carcass surfaces and 76 cases of feces, that is to say, a total of 144 cases. The results were obtained as follows: In the case of general bacterial count, 29 cases(25.7%) were in the range of 100∼999 and 62 cases(54.9%) were in the range of 1,000∼9,999 and 22 cases(19.4%) were in the range of 10,000∼99,999. Meanwhile as regards E coli count, 22 cases(19.4%) were in the range of 1∼9 and 69 cases(61.2%) were in the range of 10∼99 and 22 cases(19.4%) were in the range of 100∼999. On 68 cases of pork carcass surfaces, 7 strains(10.2%) of Salmonella spp, and 12 strains(17.6%) of S aureus were detected and 27 strains(39.7%) of L monocytogenes, respectively. As for the detected Salmonella spp, 6 strains of the B group, 3 strains of S enterica subsp salame and 2 strains of S typhimurium were detected, respectively. On 76 cases of feces, 14 strains(18.4%) of Salmonella spp, and 15 strains(19.7%) of L monocytogenes and 14 strains(18.4%) of S aureus were detected respectively. As for the detected Salmonella spp, 6 strains of the B group, 4 strains of S derby and 8 strains of the C group, 5 strains of S rissen were detected, respectively. All of 42 strains of L monocytogenes were type 1. As a result of conducting a toxin test on the detected S aureus, all of 26 strains were found to be non-toxin.

Prevalence and antimicrobial resistance patterns of Salmonella spp. isolated from different aged pigs in Korea

  • Kim, Eun Mi;Kim, Hye Kwon;Park, Seong Jun;Lee, Chul Seung;Luo, Yuzi;Moon, Hyoung Joon;Yang, Jeong Sun;Park, Bong Kyun
    • 대한수의학회지
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    • 제47권4호
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    • pp.395-398
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    • 2007
  • A total of 114 Salmonella spp. (17.90%) were isolated from 637 fecal swab samples collected in October 2005 and they were from 25 (78.13%) of 32 farms investigated nation-widely in Korea. Salmonella spp. from age group of 30-, 60-, 90-, 120-day olds were 17.61%, 16.98%, 15.72%, and 21.25%, respectively. Nine serovars of Salmonella spp. were identified. The predominant serovars of isolates were S. Typhimurium (including var Copenhagen), S. Agona, S. Derby, and S. Heidelberg by turns. Almost isolates belonged to serogroup B (69.30%). All isolates were resistant to penicillin G and oxytetracycline, and this was considered due to be used as feed additive through the most of pig farms for decades. And also, frequent resistance observed for ampicillin, sulfamethoxazole-trimethoprim, and lincomycin-spectinomycin, which are commonly used in veterinary medicine for decades, indicates an urgent need to utilize these antimicrobials more prudently if their benefits are to be preserved.

국내 유통 돈분 액비의 미생물 함량 모니터링 (Monitoring of Microorganisms in Commercial Liquid Pig Manures in Korea)

  • 임성묵;이지호;고우리;;김원일
    • 한국토양비료학회지
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    • 제44권6호
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    • pp.1181-1184
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    • 2011
  • Total aerobic bacteria, Esherichia coli O157:H7, and Salmonella spp. were examined in commercial liquid pig manures. Commercial liquid pig manures (n=33) were collected from muck joint resource recovery plant at April, June, August, October 2009, Korea. Total aerobic bacteria were incubated at $37^{\circ}C$ for 24-48 hrs, and quantified as a colony-forming unit (CFU) $mL^{-1}$. Analysis of Esherichia coli O157:H7 and Salmonella spp. were followed by Korean Food Standards Codex method. Colony of Salmonella spp. was confirmed by API kit and real time polymerase chain reaction (PCR). Total aerobic bacteria isolated from fermented commercial liquid pig manures ranged from 2.8 to $24.3{\times}10^4\;CFU\;mL^{-1}$. Esherichia coli O157:H7 was not detected, and Salmonella spp. showed the low detection frequency at only 1 sample. This study suggests that continuous monitoring in commercial liquid pig manures is required to improve the agricultural food through management of agricultural land contaminated with liquid pig manures.