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Detection of Lawsonia intracellularis by real-time PCR in the feces of free-living animals from equine farms with documented occurrence of equine proliferative enteropathy.
Abstract: The objective of this study was to determine whether Lawsonia intracellularis was present in the feces of free-living animals collected on two equine premises with documented occurrence of equine proliferative enteropathy (EPE). Fresh feces from black-tailed jackrabbits (Lepus californicus, n=100), striped skunks (Mephitis mephitis, n=22), feral cats (Felis catus, n=14), Brewer's Blackbirds (Euphagus cyanocephalus, n=10), Virginian opossums (Didelphis virginiana, n=9), raccoons (Procyon lotor, n=4), California ground squirrels (Spermophilus beecheyi, n=3), and coyotes (Canis latrans, n=2) were collected from August 2006 to January 2007 either from the ground while walking the premises or after trapping the animals using live traps. Nucleic acid purified from feces was directly processed for polymerase chain reaction (PCR) analysis using a real-time PCR assay targeting the aspartate ammonia lyase gene of L. intracellularis. Purified DNA samples were also precipitated, preamplified for L. intracellularis, and analyzed using the same real-time PCR assay, to increase the detection limit to one L. intracellularis organism per extracted sample. Feces from jackrabbits, striped skunks, Virginian opossums, and coyotes tested PCR positive for L. intracellularis, whereas all feces from feral cats, Brewer's Blackbirds, raccoons, and ground squirrels tested PCR negative for L. intracellularis. PCR testing on DNA extracted directly from feces was positive for L. intracellularis in six of 164 fecal samples. When DNA purification from feces was followed by a precipitation and preamplification step, five additional fecal samples tested PCR positive for L. intracellularis (11/164). The largest number of PCR positive L. intracellularis fecal samples was observed in striped skunks, followed by Virginian opossums, jackrabbits, and coyotes. This is the first description of L. intracellularis in these four species. Because the fecal samples were collected at equine farms with confirmed cases of EPE, striped skunks, Virginian opossums, jackrabbits, and coyotes may act as potential sources of infection to susceptible weanlings.
Publication Date: 2008-10-30 PubMed ID: 18957657DOI: 10.7589/0090-3558-44.4.992Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article focuses on identifying the presence of the Lawsonia intracellularis bacteria in the feces of various free-ranging animals on two horse farms with recorded cases of Equine Proliferative Enteropathy (EPE). The trials saw feces from a selection of animals undergo a polymerase chain reaction (PCR) process, which revealed that some samples tested positive for the bacteria.
Study Outline
- The study was conducted to confirm whether Lawsonia intracellularis (L. intracellularis), a bacterium that causes Equine Proliferative Enteropathy (EPE) in horses, could be found in the feces of various free-ranging animals on horse farms.
- The research was conducted on two horse farms that had documented cases of EPE.
- The researchers collected fresh feces from animals including black-tailed jackrabbits, striped skunks, feral cats, Brewer’s Blackbirds, Virginian opossums, raccoons, California ground squirrels, and coyotes.
Methods Used and Results
- The collected fecal samples underwent nucleic acid purification, a process that readies the material for a Polymerase Chain Reaction (PCR) analysis. This analysis was designed to specifically target L. intracellularis.
- The PCR test revealed the presence of L. intracellularis in some samples. Positive results were found in the feces of jackrabbits, striped skunks, Virginian opossums, and coyotes.
- Meanwhile, samples from feral cats, Brewer’s Blackbirds, raccoons, and ground squirrels all tested negative for the bacteria.
- Additional testing involving DNA purification from the feces, followed by a precipitation and preamplification process, resulted in the identification of five more positive samples.
Conclusions and Implications
- The research marks the first recorded detection of L. intracellularis in the species confirmed to be carrying the bacteria.
- This implies that these infected free-ranging animals could potentially become sources of infection to susceptible weanling (young, recently weaned) animals on the farms.
- The detection of L. intracellularis in the diverse range of free-ranging animals corroborates that they may be a contributing factor in the spread of EPE in horse farms.
Cite This Article
APA
Pusterla N, Mapes S, Rejmanek D, Gebhart C.
(2008).
Detection of Lawsonia intracellularis by real-time PCR in the feces of free-living animals from equine farms with documented occurrence of equine proliferative enteropathy.
J Wildl Dis, 44(4), 992-998.
https://doi.org/10.7589/0090-3558-44.4.992 Publication
Researcher Affiliations
- Department of Medicine and Epidemiology, University of California, Davis, California 95616, USA. npusterla@ucdavis.edu
MeSH Terms
- Animals
- Animals, Wild / microbiology
- Coyotes / microbiology
- Desulfovibrionaceae Infections / epidemiology
- Desulfovibrionaceae Infections / microbiology
- Desulfovibrionaceae Infections / transmission
- Desulfovibrionaceae Infections / veterinary
- Disease Reservoirs / microbiology
- Disease Reservoirs / veterinary
- Feces / microbiology
- Hares / microbiology
- Horse Diseases / epidemiology
- Horse Diseases / microbiology
- Horse Diseases / transmission
- Horses
- Lawsonia Bacteria / isolation & purification
- Mephitidae / microbiology
- Opossums / microbiology
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Species Specificity
Citations
This article has been cited 16 times.- Wu W, Wang L, Xie R, Peng C, Yang H, Zhu H, Luo Y, Hua L, Chen H, Wu B, Peng Z. Development and application of a triplex real-time PCR method for the detection of Lawsonia intracellularis, Brachyspira hyodysenteriae, and Clostridium perfringens. Microbiol Spectr 2025 Jun 3;13(6):e0323724.
- Nishigaki A, Previdelli R, Alexander JL, Balarajah S, Roberts L, Marchesi JR. Identification of a Sub-Clinical Salmonella spp. Infection in a Dairy Cow Using a Commercially Available Stool Storage Kit. Animals (Basel) 2023 Sep 4;13(17).
- Yeh JY. Prevalence and associated risk factors for Lawsonia intracellularis infection in farmed rabbits: A serological and molecular cross-sectional study in South Korea. Front Vet Sci 2023;10:1058113.
- Leite FL, Winfield B, Miller EA, Weber BP, Johnson TJ, Sylvia F, Vasquez E, Vannucci F, Beckler D, Isaacson RE. Oral Vaccination Reduces the Effects of Lawsonia intracellularis Challenge on the Swine Small and Large Intestine Microbiome. Front Vet Sci 2021;8:692521.
- Arnold M, Crienen A, Swam H, Berg SV, Jolie R, Nathues H. Correlation of Lawsonia intracellularis positivity in quantitative PCR and herd factors in European pig herds. Porcine Health Manag 2021 Jan 22;7(1):13.
- Pusterla N, Gebhart C. Equine proliferative enteropathy caused by Lawsonia intracellularis. Equine Vet Educ 2009 Aug;21(8):415-419.
- Bengtsson RJ, Wee BA, Yebra G, Bacigalupe R, Watson E, Guedes RMC, Jacobson M, Stadejek T, Archibald AL, Fitzgerald JR, Ait-Ali T. Metagenomic sequencing of clinical samples reveals a single widespread clone of Lawsonia intracellularis responsible for porcine proliferative enteropathy. Microb Genom 2020 Apr;6(4).
- Hwang JM, Seo MJ, Yeh JY. Lawsonia intracellularis in the feces of wild rodents and stray cats captured around equine farms. BMC Vet Res 2017 Aug 11;13(1):233.
- Sampieri F, Vannucci FA, Allen AL, Pusterla N, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ. Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals. Can J Vet Res 2013 Oct;77(4):261-72.
- Sampieri F, Allen AL, Pusterla N, Vannucci FA, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ. The rabbit as an infection model for equine proliferative enteropathy. Can J Vet Res 2013 Apr;77(2):110-9.
- Pusterla N, Gebhart C. Lawsonia intracellularis infection and proliferative enteropathy in foals. Vet Microbiol 2013 Nov 29;167(1-2):34-41.
- Vannucci FA, Kelley MR, Gebhart CJ. Comparative genome sequencing identifies a prophage-associated genomic island linked to host adaptation of Lawsonia intracellularis infections. Vet Res 2013 Jul 4;44(1):49.
- Vannucci FA, Foster DN, Gebhart CJ. Laser microdissection coupled with RNA-seq analysis of porcine enterocytes infected with an obligate intracellular pathogen (Lawsonia intracellularis). BMC Genomics 2013 Jun 24;14:421.
- Pusterla N, Gebhart CJ. Equine proliferative enteropathy--a review of recent developments. Equine Vet J 2013 Jul;45(4):403-9.
- Vannucci FA, Pusterla N, Mapes SM, Gebhart C. Evidence of host adaptation in Lawsonia intracellularis infections. Vet Res 2012 Jun 20;43(1):53.
- Gross CL, Nelson PA, Haddadchi A, Fatemi M. Somatic mutations contribute to genotypic diversity in sterile and fertile populations of the threatened shrub, Grevillea rhizomatosa (Proteaceae). Ann Bot 2012 Feb;109(2):331-42.
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