Evaluation of selective media for primary isolation of Treponema hyodysenteriae and Treponema innocens

Szynkiewicz, Z.; Binek, M.

doi:10.1016/0147-9571(86)90077-9

Cited by 7 publications

(6 citation statements)

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“…The criteria for identification of this isolate and the isolates recovered during the study included colony morphology, growth characteristics, hemolytic pattern, and cell morphology. Cultures of T. hyodysenteriae and T. innocens were grown in an autoclaved liquid medium, F/S or F/C (14), supplemented with 5% fetal bovine serum, and incubated for 24 to 48 h at 37°C prior to inoculation onto agar medium. All cultures were determined to be pure by dark-field microscopy and inoculation onto TSA incubated aerobically and anaerobically.…”

Section: Materials and Metho»smentioning

confidence: 99%

“…Isolation methods for T. hyodysenteriae have included the use of membrane filter plates (20), serial filtration through filters of decreasing pore size (5), serial dilution and plating (11), and selective media (8,22,24). The most widely used selective medium, Trypticase soy agar (TSA; BBL Microbiology Systems, Cockeysville, Md.)…”

mentioning

confidence: 99%

“…Other selective media have been reported, including a medium made by Jenkinson and Wingar and containing colistin, vancomycin, and spectinomycin (CVSBA) (8); Szynkiewicz and Binek have reported a medium containing spectinomycin and vancomycin (24).…”

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confidence: 99%

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Improved selective medium for the isolation of Treponema hyodysenteriae

Kunkle

Kinyon

1988

J Clin Microbiol

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An agar medium with improved selection for Treponema hyodysenteriae was developed. Cultures of T. hyodysenteriae and T. innocens, feces from 11 clinically normal pigs, and colonic contents from 6 pigs with gross lesions consistent with swine dysentery were diluted in phosphate-buffered saline and plated on Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.) with 5% citrated bovine blood (TSA), TSA with 400 ,ug of spectinomycin per ml (TSA-S400), TSA-S400 with 25 ,ug each of colistin and vancomycin per ml, and TSA with 5% pig feces extract and five antimicrobial agents (spiramycin, rifampin, vancomycin, colistin, and spectinomycin) (BJ). Viable numbers of T. hyodysenteriae grown on BJ were virtually identical to those for TSA, TSA-S400, and TSA-S400 with colistin and vancomycin. Pure cultures of four isolates of T. hyodysenteriae and three isolates of T. innocens were sustained through six subcultures on BJ. Fecal floras were completely inhibited on BJ for 14 of 17 fecal samples from both groups of pigs. A total of 461 colonic specimens from naturally occurring cases of porcine intestinal disease were plated on TSA-S400 and BJ. T. hyodysenteriae was isolated on both TSA-S400 and BJ for 69 specimens and on BJ alone for an additional 19 specimens.

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Section: Materials and Metho»smentioning

confidence: 99%

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confidence: 99%

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Improved selective medium for the isolation of Treponema hyodysenteriae

Kunkle

Kinyon

1988

J Clin Microbiol

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“…The type strain: B78 T (ATCC 27164T) and the reference strain B204 (ATCC 31212) were included in the analysis as positive controls. All the faeces and mucosal scrapings from the large intestines were streaked on TSA plates, supplemented with 5-10% fresh defibrinated ovine blood, L-cysteine (50 mg/l), spectinomycin (40 mgl), vancomycin (7 mg/l), yeast extract (500 mg/l) (Szynkiewicz and Binek 1986) and incubated in anaerobic jars (Merck, USA), in an atmosphere of 8-10% CO 2 and 90% H 2 (BD GasPak TM EZ Anaerobe Container System) for 3 days at 39.5 o C. In order to obtain a pure culture of B. hyodysenteriae, after a first incubation a re-cultivation (overall 3 to 4 passages) on TSA medium was performed. For the further studies, field isolates were frozen at -70 o C. After thawing the isolates were passaged at least twice before susceptibility testing.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Antimicrobial susceptibility of Brachyspira hyodysenteriae isolated from 21 Polish farms

Żmudzki

Szczotka²,

Nowak³

et al. 2012

Polish Journal of Veterinary Sciences

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Swine dysentery (SD) is a common disease among pigs worldwide, which contributes to major production losses. Antimicrobial susceptibility testing of B. hyodysenteriae, the etiological agent of SD, is mainly performed by the agar dilution method. This method has certain limitations due to difficulties in interpretation of results. The aim of this study was the analysis of antimicrobial susceptibility of Brachyspira hyodysenteriae (B. hyodysenteriae) Polish field isolates by broth microdilution procedure. The study was performed on 21 isolates of B. hyodysenteriae, collected between January 2006 to December 2010 from cases of swine dysentery. VetMIC TM Brachyspira panels with antimicrobial agents (tiamulin, valnemulin, doxycycline, lincomycin, tylosin and ampicillin) were used for susceptibility testing of B. hyodysenteriae. The minimal inhibitory concentration (MIC) was determined by the broth dilution procedure. The lowest antimicrobial activity was demonstrated for tylosin and lincomycin, with inhibition of bacterial growth using concentrations >128 μg/ml and 32 μg/ml, respectively. In the case of doxycycline, the MIC values were ≤ 2.0 μg/ml. No decreased susceptibility to tiamulin was found among the Polish isolates and MIC values for this antibiotic did not exceed 1.0 μg/ml. The results of the present study confirmed that Polish B. hyodysenteriae isolates were susceptible to the main antibiotics (tiamulin and valnemulin) used in treatment of swine dysentery. Further studies are necessary to evaluate a possible slow decrease in susceptibility to tiamulin and valnemulin of B. hyodysenteriae strains in Poland.

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“…This method is the only alternative to PCR for detection of B. hyodysenteriae in fecal specimens (Szynkiewicz and Binek 1986). Laboratory identification of both bacteria is laborious and often takes several days or even weeks to receive the definitive result.…”

Section: Introductionmentioning

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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Evaluation of selective media for primary isolation of Treponema hyodysenteriae and Treponema innocens

Cited by 7 publications

References 7 publications

Improved selective medium for the isolation of Treponema hyodysenteriae

Improved selective medium for the isolation of Treponema hyodysenteriae

Antimicrobial susceptibility of Brachyspira hyodysenteriae isolated from 21 Polish farms

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

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