Ruminant rhombencephalitis-associated Listeria monocytogenes strains constitute a genetically homogeneous group related to human outbreak strains - PubMed
Ruminant rhombencephalitis-associated Listeria monocytogenes strains constitute a genetically homogeneous group related to human outbreak strains
Paulo Ricardo Dell'Armelina Rocha et al. Appl Environ Microbiol. 2013 May.
Erratum in
- Appl Environ Microbiol. 2013 Nov;79(22):7114
Abstract
Listeriosis is a disease that causes significant economic losses at the farm level because of high morbidity and mortality in ruminants. This study was performed to investigate the role of ruminants in the epidemiology of listeriosis in northern Italy and the possible association of animal-adapted strains of Listeria monocytogenes with strains associated with human disease. Twenty ruminant rhombencephalitis isolates previously confirmed as L. monocytogenes by bacteriology and PCR were characterized by serotyping, pulsed-field gel electrophoresis, multi-virulence-locus sequence typing (MVLST), and multiplex single nucleotide polymorphism (mSNP) typing for the detection of epidemic clones. Subtyping results were subsequently compared with those obtained from human, food, and environmental isolates of L. monocytogenes, including 311 isolates from the University of Turin, Grugliasco, Italy, and 165 isolates representing major human listeriosis outbreaks worldwide, in addition to other unrelated isolates. Both mSNP typing and MVLST showed that 60% of the isolates analyzed belonged to epidemic clone I (ECI), which has been epidemiologically linked to several human outbreaks of listeriosis. In particular, the 1981 Canada outbreak was linked to the use of sheep manure and the 1985 California outbreak was linked to the use of raw cow's milk. In our study, ECI isolates were collected from different ruminant species on geographically and temporally distinct occasions for the last 13 years. Our results support the hypothesis that ruminants represent possible natural reservoirs of L. monocytogenes strains capable of causing epidemics of listeriosis in humans.
Figures
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Histopathologic evaluations. (A) Bovine cerebellum. Multifocal-to-coalescing microabscesses (arrows) and mononuclear perivascular cuffing (asterisks) in the white matter. H&E staining. Magnification, ×25. (B) Ovine brainstem. Malacic area (arrows) composed mainly of neutrophils. H&E staining. Magnification, ×25. (C) Bovine brainstem. Microabscess composed mainly of macrophages and a few neutrophils. H&E staining. Magnification, ×400. (D) Caprine brainstem. Immunopositivity (brown areas) for Listeria species in the cytoplasm of macrophages and neutrophils within a microabscess. DAB chromogen counterstained with hematoxylin. Magnification, ×400.

Dendrogram produced by the unweighted-pair group method using average linkages showing clustering by Dice coefficient of the combined PFGE (AscI and ApaI) profiles of the 20 L. monocytogenes isolates obtained from animal cases analyzed in this study. PFGE divided the L. monocytogenes animal isolates into nine unique and four shared profiles, P1 (two isolates), P2 (three isolates), P3 (four isolates), and P4 (two isolates).

Unrooted neighbor-joining tree computed in MEGA 5.0 (52) based on the comparison of MVLST results for the L. monocytogenes ruminant isolates investigated in this study (20 clinical veterinary isolates) with 165 isolates previously characterized by MVLST (13, 21, 22). Statistical support was provided by bootstrapping with 1,000 replicates. The scale bar corresponds to the number of differences observed over the alignment of 2,606 bp.
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