Application of Real Time PCR for Diagnosis of Swine Dysentery

Akase, Satoru; Uchitani, Yumi; Sohmura, Yoshiko; Tatsuta, Keikichi; Sadamasu, Kenji; Adachi, Yoshikazu

doi:10.1292/jvms.71.359

Cited by 11 publications

(12 citation statements)

References 17 publications

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“…field Minimum inhibitory concentration patterns of Brachyspira species isolates 1141 isolates to commonly prescribed antimicrobials. 1,7,14,16,18 Similar evaluations of recent Brachyspira spp. isolates in the United States are lacking.…”

Section: Introductionmentioning

confidence: 88%

“…Brachyspira-associated disease, which includes swine dysentery (SD), characterized by severe mucohemorrhagic diarrhea with high morbidity caused by Brachyspira hyodysenteriae, and porcine intestinal spirochetosis (PIS), associated with mucoid, cement-gray diarrhea with depressed growth caused by Brachyspira pilosicoli, represents a severe threat to the health of growing pigs. 1,4,6,7,16,18 Diagnosis can be complicated by the specialized media and growth conditions necessary to identify these species in vitro as well as the presence of other Brachyspira species described as minimally or nonpathogenic. 4,12,17,21 While the incidence of Brachyspira-associated disease decreased in the 1990s, possibly due to management changes associated with confinement rearing, detection of this agent from clinically ill swine has increased in recent years.…”

Section: Introductionmentioning

confidence: 99%

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Species characterization and minimum inhibitory concentration patterns of Brachyspira species isolates from swine with clinical disease

et al. 2011

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Abstract. Typhlocolitis and dysentery due to Brachyspira hyodysenteriae infection represent an economically important disease syndrome in growing pigs. Largely disappearing from U.S. swine herds in the late 1990s and early 2000s, Brachyspiraassociated disease and bacterial isolation from swine with clinical disease has increased in the last several years, and non-B. hyodysenteriae isolates are commonly identified. Antimicrobial resistance has been demonstrated in Brachyspira spp. isolates from Europe and Asia, and may be the reason for the resurgence in U.S. herds. Seventy-nine clinical isolates identified at the Iowa State University Veterinary Diagnostic Lab were tested with multiple polymerase chain reaction assays to establish species identity, and evaluated for minimum inhibitory concentrations (MICs) using an agar dilution method against lincomycin, gentamicin, valnemulin, tiamulin, salinomycin, and carbadox. Only 38.0% of isolates could be confirmed as the known pathogens B. hyodysenteriae (30.4%) or Brachyspira pilosicoli (7.6%). Twenty of the 79 isolates (25.3%) were identified as Brachyspira murdochii, and 13.9% could not be identified to species. The MIC values were consistently high against lincomycin and moderately high against gentamicin. The remaining antimicrobials had MICs that were at the low end of the test ranges. Brachyspira murdochii and Brachyspira spp. had significantly greater MIC values against several of these drugs than other Brachyspira spp. examined. The increased incidence of these less definitively characterized Brachyspira species with increased MIC values to commonly prescribed antimicrobials may, at least in part, explain the increased prevalence and severity of this disease complex in recent years. Further research is necessary to understand these changes.

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Section: Introductionmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Species characterization and minimum inhibitory concentration patterns of Brachyspira species isolates from swine with clinical disease

et al. 2011

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“…The primers and the Taqman probe for B. hyodysenteriae were based on the sequence of NADH oxidase gene (GenBank accession no. U19610), previously described by Akase et al (2009). The primers and the probe for L. intracellularis used in the study were previously designed by Lindecrona et al (2002) and based on the sequence of 16S rDNA gene of L. intracellularis available in GenBank (accession No.…”

Section: Methodsmentioning

confidence: 99%

Application of real-time PCR for detection of Lawsonia intracellularis and Brachyspira hyodysenteriae in fecal samples from pigs

Żmudzki¹,

Szczotka²,

Podgórska³

et al. 2012

Polish Journal of Veterinary Sciences

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The aim of the study was to develop and validate real-time PCR method for the quantification of Lawsonia intracellularis and Brachyspira hyodysenteriae in porcine feces. Before the optimization process was performed two different extraction methods were compared to select the more efficient one. Based on the results achieved at this stage the boiling procedure was rejected and a commercially available silica-membrane based method was chosen for further analysis. The primers and the Taqman probe for B. hyodysenteriae and L. intracellularis were based on the sequence of NADH oxidase gene and 16S rDNA gene, respectively. The detection limit of the real-time PCR for suspension of feces inoculated with B. hyodysenteriae and L. intracellularis was determined to be 1.5x10 3 CFU/ml and 6.5x10 1 CFU/ml, respectively. The results of this study demonstrate that our real-time PCR is able to detect low number of B. hyodysenteriae and L. intracellularis cells which is satisfying in routine diagnosis of swine dysentery and proliferative enteropathy. Therefore, it is possible to identify both subclinically infected pigs and those representing an acute form of mentioned diseases. In summary, the quantitative real-time PCR is useful for routine diagnosis of L. intracellularis and B. hyodysenteriae. Compared to conventional PCR, the new validated quantification method based on real-time PCR is fast and with reduced risk of laboratory contamination. The novel technique is specific and even more sensitive than the previously used one. Furthermore, the new real-time PCR enables quick detection and quantification of both pathogens in fecal samples, which helps to estimate the health status of a pig herd.

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“…Primers were obtained from a commercial source (Genomed S.A., Poland). The sequences of primers and probes are as follows: for B. hyodysenteriae (forward primer: 5′-TATGAAGAAGGCAGC AGACGTTTAT-3′, reverse primer: 5′-GTAGGAAGAAGAAATC TGACAATGCA-3′, probe: 5′-FAM-ACACAATCATGCTGAA GC-TAMRA-3′) (Akase et al, 2009); for B. pilosicoli (forward primer: 5′-GTAGTCGATGGGAAACAGGT-3′, reverse primer: 5′-TTACTCACCACAAGTCTCGG-3′, probe: 5′-FAM-TATT CGACGAGGATAACCATCACCT-BHQ-1-3′) (Ståhl et al, 2011); for L. intracellularis (forward primer: 5′-GCGCGCGTAGGTG GTTATAT-3′, reverse primer: 5′-GCCACCCTCTCCGATACTC A-3′, probe: 5′-FAM-CACCGCTTAACGGTGGAACAGCCTT-TA MRA-3′) (Lindecrona et al, 2002). All assays were carried out using the Rotor-Gene Q real-time PCR system (Qiagen, Hilden, Germany).…”

Section: Dna Extraction and Pcrmentioning

confidence: 99%

A survey on the occurrence of Brachyspira pilosicoli and Brachyspira hyodysenteriae in growing-finishing pigs

2019

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Background: The major pathogenic intestinal spirochetes affecting pigs during the growing- finishing stage of production include Brachyspira hyodysenteriae and Brachyspira pilosicoli. Infections by these pathogens, which affect the economics of pig production, can result in mortality, growth rate losses and substantial antibiotic costs. The aim of this study was to assess the current occurrence of B. hyodysenteriae and B. pilosicoli in Polish pig herds. Moreover, associations between the presence of diarrhea or other intestinal pathogens and occurrence of B. hyodysenteriae and B. pilosicoli in pigs were investigated. Methods: Between January 2017 and August 2019, a total of 401 samples of pig feces from 95 different herds were submitted to the National Veterinary Research Institute of Poland. These samples were obtained from pigs older than 7 weeks. All the received fecal samples were examined for the presence of B. hyodysenteriae, B. pilosicoli and Lawsonia intracellularis by real-time PCR. Results: For B. pilosicoli, 4.5% (95% CI, 2.5–7.0%) of samples and 13.7% (95% CI, 7.5–22.3%) of herds were positive. Out of 12 samples, B. pilosicoli was detected simultaneously with L. intracellularis, B. hyodysenteriae and B. pilosicoli were detected alone in two samples each. In terms of B. hyodysenteriae, 7.0% of samples (95% CI, 4.7–9.9%) from 18.9% of herds (95% CI, 11.6–28.3%) were positive in real time PCR. The presence of B. hyodysenteriae in fecal samples was associated with the presence of diarrhea in pigs. Conclusions: This study confirmed that B. pilosicoli infections occur in Polish pig herds, but the prevalence is at a low level and the presence of B. pilosicoli is not associated with the development of diarrhea in pigs. B. hyodysenteriae is still a common cause of diarrhea among pigs from Polish herds.

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Application of Real Time PCR for Diagnosis of Swine Dysentery

Cited by 11 publications

References 17 publications

Species characterization and minimum inhibitory concentration patterns of Brachyspira species isolates from swine with clinical disease

Species characterization and minimum inhibitory concentration patterns of Brachyspira species isolates from swine with clinical disease

Application of real-time PCR for detection of Lawsonia intracellularis and Brachyspira hyodysenteriae in fecal samples from pigs

A survey on the occurrence of Brachyspira pilosicoli and Brachyspira hyodysenteriae in growing-finishing pigs

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