Reproduction of mucohaemorrhagic diarrhea and colitis indistinguishable from swine dysentery following experimental inoculation with "Brachyspira hampsonii" strain 30446

PLoS One. 2013;8(2):e57146. doi: 10.1371/journal.pone.0057146. Epub 2013 Feb 27.

Abstract

Background: Mucohaemorrhagic diarrhea caused by Brachyspira hyodysenteriae, swine dysentery, is a severe production limiting disease of swine. Recently, pigs in western Canada with clinical signs indistinguishable from swine dysentery were observed. Despite the presence of spirochetes on fecal smears, recognized Brachyspira spp. including B. hyodysenteriae could not be identified. A phylogenetically distinct Brachyspira, called "B. hampsonii" strain 30446, however was isolated. The purpose of this study was to experimentally reproduce mucohaemorrhagic colitis and characterize strain 30446 shedding following inoculation.

Methods and findings: Eighteen 13-week-old pigs were randomly assigned to inoculation (n = 12) or control (n = 6) groups in each of two trials. In trial 1, pigs were inoculated with a tissue homogenate collected from clinically affected field cases. In trial 2, pigs were inoculated with a pure broth culture of strain 30446. In both trials, mucohaemorrhagic diarrhea was significantly more common in inoculated pigs than controls, all of which remained healthy. In animals with mucohaemorrhagic diarrhea, significantly more spirochetes were observed on Gram stained fecal smears, and higher numbers of strain 30446 genome equivalents were detected by quantitative PCR (qPCR). Strain 30446 was cultured from colon and/or feces of all affected but no control animals at necropsy.

Conclusions: "Brachyspira hampsonii" strain 30446 causes mucohaemorrhagic diarrhea in pigs following a 4-9 day incubation period. Fecal shedding was detectable by day 4 post inoculation, and rarely preceded the onset of mucoid or haemorrhagic diarrhea by more than 2 days. Culture and 30446-specific qPCR are reliable methods of detection of this organism in feces and tissues of diarrheic pigs. The emergence of a novel Brachyspira spp., such as "B. hampsonii", creates diagnostic challenges including higher risk of false negative diagnostic tests. We therefore recommend diagnostic laboratories routinely use Brachyspira culture, nox-based and species-specific PCR, and DNA sequencing to diagnose Brachyspira-associated colitis in pigs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Husbandry
  • Animals
  • Brachyspira / physiology*
  • Colitis / diagnosis
  • Colitis / microbiology*
  • Colitis / pathology
  • Colon / microbiology
  • Colon / pathology
  • Diarrhea / diagnosis
  • Diarrhea / microbiology*
  • Diarrhea / pathology
  • Feces / microbiology
  • Gastrointestinal Hemorrhage / diagnosis
  • Gastrointestinal Hemorrhage / microbiology*
  • Gastrointestinal Hemorrhage / pathology
  • Gram-Negative Bacterial Infections / diagnosis
  • Gram-Negative Bacterial Infections / microbiology*
  • Gram-Negative Bacterial Infections / pathology
  • Mucus / metabolism*
  • Phylogeny
  • Swine / microbiology
  • Swine Diseases / diagnosis
  • Swine Diseases / microbiology*
  • Swine Diseases / pathology

Grants and funding

This research was supported by grants from the Canadian Swine Health Board (www.swinehealth.ca) and by internal funding from the University of Saskatchewan Industry Liaison Office, Western College of Veterinary Medicine Research Trust (www.usask.ca). Salary support for HK was provided by the Merck-Merial Veterinary Scholars Summer Research Program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.