• Title/Summary/Keyword: Lawsonia intracellularis

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Detection of Lawsonia intracellularis in swine feces by polymerase chain reaction (돼지분변에서 PCR에 의한 Lawsonia intracellularis 검색)

  • 장성준;김정화;김영태;김기향;김중규;김영욱;최일영
    • Korean Journal of Veterinary Service
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    • v.24 no.1
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    • pp.43-50
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    • 2001
  • Swine proliferative enteritis(SPE) caused by inかsoma intracellularis is a common enteric disaese of grower and finisher pig. Swine affected with SPE show variable clinical signs including diarrhea, weight loss, aberrant growth and death. The characteristic lesion of ileitis at necropsy is marked thickening of the last section of the small intestine. The inner lining of the thickened intestine proliferates almost like a cancer and curved rod bacteria(L intracellularis) are always seen inside the intestinal wall. Infected swine shed the organism in the feces. Isolation and growth of pure L intracellularis in vitro requires a suitable cell culture. This procedure is difficult and not a practical means of diagnosis, thus the polymerase chain reaction(PCR) test of feces can be used to determine whether a pig is shedding the infective organism. A sensitive assay based on amplification of a 319bp DffA fragment of the L intracellularis of Swine proliferative enteritis was attempted for the detection of the organism in the 62 feces of swine. L intracellularis was identified on three herds and detected in 6 fecal samples, representing a infection rate of 9.7%. The PCR was very sensitive and specific on the individual level. The PCR technique could be very useful for the diagnosis of this disease.

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Distribution of Lawsonia intracellularis in livestock transport car of slaughterhouse, Korea (도축장 출하차량에서 Lawsonia intracellularis 분포율 조사)

  • Lee, Su-Ji;Lee, Hee-Seon;Seo, Ji-Soo;Kim, Tae-Gyeom;Jeong, Jae-Kyo
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.245-250
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    • 2018
  • Lawsonia intracellularis is the pathogenic agent of porcine proliferative enteritis (PPE). The bacterial pathogen infects the intestinal crypt cells which causes hyperplasia of the infected cells and leads to the process of intestinal pathogenesis. PPE includes some clinical maninfestations, including acute hemorrhagic diarrhea with sudden death in growing pigs and porcine intestinal adenomatosis, to a chronic diarrhea with reduced productivity of the infected pigs. The purpose of the present studies were carried out to determine L. intracellularis in livestock transport car of slaughterhouse. Distribution of L. intracellularis in livestock transport car were conducted using real-time polymerase chain reaction (real-time PCR) testing method, total 300 samples. Of 300 samples, 119 (39.7%) were detected as positive to L. intracellularis in livestock transport car. In seasonal analysis, 42 (28.0%) out of 150 samples in spring and summer season. 77 (51.3%) out of 150 sample in autumn and winter season. In regional analysis, 53 (88.3%) out of 60 cars and the detection ratio showed that regional variation in livestock transport car.

Development of rapid diagnosis technology for porcine proliferative enteropathy (1) - Preparation of the samples and antibody for rapid detecting the lawsonia in pig feces - (돼지증식성회장염 신속검진 기술개발(1) - 돼지 분변에서의 로소니아균 검출을 위한 항원, 항체 준비 -)

  • Kim, Hyuck Joo;Hong, Jong Tae;Yu, Byeong Kee;Kim, Gi Young;Lee, Jin Ju;Kim, Suk
    • Journal of Biosystems Engineering
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    • v.37 no.6
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    • pp.420-428
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    • 2012
  • Purpose: Porcine proliferative enteropathy(PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. The bacterial pathogen invades the intestinal epithelial cells which causes hyperplasia of the infected cells and leads to the process of disease pathogenesis. For diagnosing PPE in a pig farm in earlier stage, a rapid diagnosing test equipment is needed for farmers. To test the equipment appropriately, we prepare the samples and antibodies for rapid detecting the Lawsonia intracellularis in pig feces. Methods : To prepare the PPE infected samples, we sampled PPE suspected pig feces in a pig farm. To manufacture a anti-Lawsonia intracellularis antibody for capturing the Lawsonia intracellularis, the rabbit-anti LsaA synthetic peptide polyclonal antibody was inoculated to rabbits. To select the couple of antibodies which is most well sandwiched with the bacteria, ELISA test was done with PPE infected ileum samples. Finally, to verify the PPE infected feces which would be used to test the rapid kit, PCR test was done on the sampled PPE suspected feces Results : The rabbit-anti LsaA synthetic peptide polyclonal antibody is developed, and is verified to capture the bacterial well through the fluorescence antibody test. Also, we found that the monoclonal antibody and the polyclonal antibody could be used as couples for sandwiching the bacteria. Finally, through the PCR test for samples of pig feces, we could prepare the 150 PPE positive samples and 50 PPE negative samples. Conclusions : The manufactured polyclonal antibody and the imported monoclonal antibody could be used to capture the bacteria using the sandwich techniques. Also, the prepared PPE infected negative and positive samples could be used to test the performance of the rapid kit to capture the bacterium Lawsonia intracellularis.

Seroprevalence of porcine proliferative enteropathy before initiating vaccine marketing in Korea

  • Yeh, Jung-Yong
    • Korean Journal of Veterinary Research
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    • v.55 no.1
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    • pp.61-63
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    • 2015
  • Proliferative enteropathy caused by Lawsonia intracellularis is one of the most common enteric diseases in pigs. The objective of this study was to determine the prevalence of serum antibodies against L. intracellularis in the general swine population of Korea from 2005 to 2008. In total, 8,008 swine serum samples obtained from 1,001 herds were tested. The samples were analyzed with an immunoperoxidase monolayer assay to detect anti-L. intracellularis antibodies. The overall 4-year average true prevalence was 40.0% (CI: 39.4 - 40.6%) at the individual animal level and 71.9% (CI: 70.3-73.4%) at the herd level.

Evaluation of host and bacterial gene modulation during Lawsonia intracellularis infection in immunocompetent C57BL/6 mouse model

  • Kirthika, Perumalraja;Park, Sungwoo;Jawalagatti, Vijayakumar;Lee, John Hwa
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.41.1-41.15
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    • 2022
  • Background: Proliferative enteritis caused by Lawsonia intracellularis undermines the economic stability of the swine industry worldwide. The development of cost-effective animal models to study the pathophysiology of the disease will help develop strategies to counter this bacterium. Objectives: This study focused on establishing a model of gastrointestinal (GI) infection of L. intracellularis in C57BL/6 mice to evaluate the disease progression and lesions of proliferative enteropathy (PE) in murine GI tissue. Methods: We assessed the murine mucosal and cell-mediated immune responses generated in response to inoculation with L. intracellularis. Results: The mice developed characteristic lesions of the disease and shed L. intracellularis in the feces following oral inoculation with 5 × 107 bacteria. An increase in L. intracellularis 16s rRNA and groEL copies in the intestine of infected mice indicated intestinal dissemination of the bacteria. The C57BL/6 mice appeared capable of modulating humoral and cell-mediated immune responses to L. intracellularis infection. Notably, the expression of genes for the vitamin B12 receptor and for secreted and membrane-bound mucins were downregulated in L. intracellularis -infected mice. Furthermore, L. intracellularis colonization of the mouse intestine was confirmed by the immunohistochemistry and western blot analyses. Conclusions: This is the first study demonstrating the contributions of bacterial chaperonin and host nutrient genes to PE using an immunocompetent mouse model. This mouse infection model may serve as a platform from which to study L. intracellularis infection and develop potential vaccination and therapeutic strategies to treat PE.

Antimicrobial susceptibility pattern of Lawsonia intracellularis recently isolated from pig with proliferative hemorrhagic enteropathy in Korea

  • Seo, Byoung-Joo;Koh, Sang-Eog;Oh, Yeonsu;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.42 no.4
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    • pp.301-304
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    • 2019
  • The objective of this study was to determine the in vitro intracellular and extracellular minimum inhibitory concentrations (MICs) of 13 antimicrobials against one recently isolate Lawsonia intracellularis, the etiological agent of proliferative enteropathy (PE). The final MICs were assessed by counting the number of heavily infected cells (HICs;>30 bacteria per cell) using an immunoperoxidase monolayer assay. Enrofloxacin (InMIC; 1~2 ㎍/mL and ExMIC; 16 ㎍/mL) still presented the most notable antimicrobial susceptibility, and marbofloxacin (2 ㎍/mL and 8 ㎍/mL) was followed. Colistin (0.25 ㎍/mL and 2 ㎍/mL) presented a susceptibility followed by tylvalosin (1 ㎍/mL and 2 ㎍/mL). Florfenicol and lincomycin had the weakest susceptibility and amoxicillin, penicillin G, chlortetracycline, oxytetracycline, tiamulin, tilmicosin, and tylosin displayed weak susceptibility. Although some antibiotics showed decreased susceptibility patterns, they showed similar patterns to recent antibiotic susceptibility patterns in Korea. In addition, these results could be one of contributions in clinical fields.

Development of Rapid Diagnostic Technology for Pig Disease (2) - Rapid detection of PPE in the pig feces -

  • Kim, Hyuck-Joo;Hong, Jong-Tae;Yu, Byeong-Kee;Kim, Giyoung;Kim, Suk
    • Journal of Biosystems Engineering
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    • v.38 no.2
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    • pp.121-128
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    • 2013
  • Purpose: Porcine proliferative enteropathy (PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. In order to diagnose PPE rapidly, the rapid kit was developed and tested. Methods: In this study, a rapid kit was developed to screen the PPE rapidly at the pig farm. Also, occult blood test with fecal occult blood (FOB) kit was done for detecting the blood in pig feces which might be the evident of hemorrhagic PPE. For developing the kit, we tested fecal samples of PPE infected pigs diagnosed by polymerase chain reaction (PCR) method. Results: With the developed rapid kit, Lawsonia intracellularis was detected in high density emulsion of ileum. On the other hand, the test result of detecting Lawsonia in feces showed too high non-specific response. In addition, nevertheless the FOB test result showed that blood evident could be founded in pig feces, the diagnosing result was not fit to PCR test result, which shows blood in pig feces could be from not only hemorrhagic PPE but also many reasons. Conclusions: To deal with the PPE effectively, it will be better for farmers to screen the PPE in earlier stage with easy and rapid diagnosing tool on farm. This study found out that the rapid kit could detect the Lawsonia intracellularis and hemoglobin in pig feces. However, the non-specific response to negative samples of PPE was too high to use at a pig farm. Further research is needed for lowering the non-specific response with the rapid kit.

Detection and genetic characterization of Lawsonia intracellularis from swine in Korea

  • Chu, Jia-Qi;Hu, Xu-Min;Kim, Myung-Cheol;Park, Chang-Sik;Jun, Moo-Hyung
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.223-231
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    • 2010
  • A total of 191 samples collected from the commercial swine farms located in Chungnam province were investigated by PCR to estimate the prevalence of Lawsonia (L.) intracellularis infection. In the group of the pigs with proliferative enteritis, 14 (93.3%) of 15 intestinal samples and 12 (80.0%) of 15 feces were positive in PCR. In contrast, a relatively low positive rate (18.0%, 29 of 161 samples) was determined in the group of normal healthy pigs. The group of pigs over 120 days showed the highest positive rates (26.8%, 15 of 56 samples). In the comparison of the sequences of 210bp for species specific fragments and 301bp for outer membrane protein, the isolates (L1. L2) showed almost 100% identity with the reference L. intracellularis (L08049, USA). For the sequences of partial 16s rDNA, the homologies among the 5 isolates (L1-L5) were 97.4% to 99.3%, and those of 5 sequences (L1-L5) versus 5 overseas reference strains of L. intracellularis ranged from 98.6% to 99.8%. In the comparison of the nucleotide sequences among 5 isolates and other species in Desulfovibrionales showed 82.4 to 99.5% identities. The 5 isolates shared relatively low identities (76.9% to 84.4%) with the species of alpha-proteobacteria. In phylogenetic analysis based on the 16s rDNA sequences, all of the 5 isolates (L1-L5) were located in the same branch with the strains of L. intracellularis that were previously isolated from the pigs in USA and China. Seven strains of Desulfovibrio sp. were clustered in the neighboring branches, whereas alpha and gamma Proteobacteria showed distant relationship with L. intracellularis strains. The present findings suggest that L. intracellularis infection is endemic in the swine farms in the regions, and that the domestic isolates maintained very limited genetic variation.

Survey of porcine proliferative enteritis for the pig farms in Gyeongnam district (경남지역 양돈장의 돼지증식성회장염 감염률에 대한 연구)

  • Park, Dong-Yeop;Park, Ae-Ra;Jung, Eun-Hee;Bae, Jae-Hyeong;Lee, Guk-Cheon;Hwang, Bo-Won;Lee, Min-Kwon
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.361-368
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    • 2009
  • Porcine proliferative enteritis (PPE) is a transmissible gastroenteric disease caused by Lawsonia intracellularis. Clinically, PPE causes hemorrhagic diarrhea and sometimes death in growing pigs, but when the disease progresses to a chronic phase, the infected pig no longer displays significant symptoms. The purpose of the present studies were carried out to determine L. intracellularis in the pig farms and slaughter house, in Gyeongnam area. A survey of proliferative enteritis in pig was conducted using polymerase chain reaction (PCR) testing method, total 1,495 samples. PCR products showed a specific band at the 210bp, 329bp in the specimens of feces and mucosal scraping. Of 420 fecal specimens, 113 (26.9%) were identified as positive to PPE. Of 1,075 mucosal scraping specimens, 109 (10.1%) were identified as positive to PPE. Of total 1,495 specimens, 222 (14.8%) were identified as positive to PPE.