Development and Validation of an Immunohistochemical Method for Rapid Diagnosis of Swine Erysipelas in Formalin-Fixed, Paraffin-Embedded Tissue Samples

Opriessnig, Tanja; Bender, Joseph S.; Halbur, Patrick G.

doi:10.1177/104063871002200116

Cited by 19 publications

(15 citation statements)

References 9 publications

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“…For this reason, accurate, reliable, and timely diagnostic strategies are important (4). Immunohistochemistry techniques have been shown to be highly sensitive and specific, especially when diagnostic specimens include lesions from antimicrobial-treated pigs or chronically affected pigs (16). Our previous studies confirmed that an Erysipelothrix species-selective broth technique is more sensitive than traditional direct plating of regular and contaminated specimens (1).…”

supporting

confidence: 63%

“…strain 2 (serotype 18) were analyzed, and Spa proteins can be classified into three molecular species, SpaA, SpaB, and SpaC (29). The SpaA protein was identified in E. rhusiopathiae serotypes 1a, 1b, 2,5,8,9,12,15,16,17, and N, the SpaB protein was identified in E. rhusiopathiae serotypes 4, 6, 11, 19, and 21, and the SpaC protein was identified only in serotype 18 (29). Additional work further differentiated SpaB into subtypes SpaB1 (serotypes 4,6,8,19,21) and SpaB2 (serotype 11) (21).…”

mentioning

confidence: 99%

“…have been isolated from domestic and wild species of both birds and mammals and have been identified as the causative agent of the clinical disease known as "erysipelas" in animals and "erysipeloid" in humans (2). The genus Erysipelothrix consists of four species and 25 associated serotypes: E. rhusiopathiae (serotypes 1a, 1b, 2, 4, 5,6,8,9,11,12,15,16,17,19,21, N), E. tonsillarum (serotypes 3,7,10,14,20,22,23), Erysipelothrix sp. strain 1 (serotype 13), and Erysipelothrix sp.…”

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confidence: 99%

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Characterization of Erysipelothrix Species Isolates from Clinically Affected Pigs, Environmental Samples, and Vaccine Strains from Six Recent Swine Erysipelas Outbreaks in the United States

Bender

Shen

Irwin

et al. 2010

Clin Vaccine Immunol

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The aim of this study was to characterize Erysipelothrix sp. isolates from clinically affected pigs and their environment and compare them to the Erysipelothrix sp. vaccines used at the sites. Samples were collected during swine erysipelas outbreaks in vaccinated pigs in six Midwest United States swine operations from 2007 to 2009. Pig tissue samples were collected from 1 to 3 pigs from each site. Environmental samples (manure, feed, central-line water, oral fluids, and swabs collected from walls, feed lines, air inlets, exhaust fans, and nipple drinkers) and live vaccine samples were collected following the isolation of Erysipelothrix spp. from clinically affected pigs. All Erysipelothrix sp. isolates obtained were further characterized by serotyping. Selected isolates were further characterized by PCR assays for genotype (E. rhusiopathiae, E. tonsillarum, Erysipelothrix sp. strain 1, and Erysipelothrix sp. strain 2) and surface protective antigen (spa) type (A, B1, B2, and C). All 26 isolates obtained from affected pigs were E. rhusiopathiae, specifically, serotypes 1a, 1b, 2, and 21. From environmental samples, 56 isolates were obtained and 52/56 were E. rhusiopathiae (serotypes 1a, 1b, 2, 6, 9, 12, and 21), 3/56 were Erysipelothrix sp. strain 1 (serotypes 13 and untypeable), and one was a novel species designated Erysipelothrix sp. strain 3 (serotype untypeable). Four of six vaccines used at the sites were commercially available products and contained live E. rhusiopathiae serotype 1a. Of the remaining two vaccines, one was an autogenous live vaccine and contained live E. rhusiopathiae serotype 2 and one was a commercially produced inactivated vaccine and was described by the manufacturer to contain serotype 2 antigen. All E. rhusiopathiae isolates were positive for spaA. All Erysipelothrix sp. strain 1 isolates and the novel Erysipelothrix sp. strain 3 isolate were negative for all currently known spa types (A, B1, B2, and C). These results indicate that Erysipelothrix spp. can be isolated from the environment of clinically affected pigs; however, the identified serotypes in pigs differ from those in the environment at the selected sites. At one of the six affected sites, the vaccine strain and the isolates from clinically affected pigs were of homologous serotype; however, vaccinal and clinical isolates were of heterologous serotype at the remaining five sites, suggesting that reevaluation of vaccine efficacy using recent field strains may be warranted.

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confidence: 99%

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Characterization of Erysipelothrix Species Isolates from Clinically Affected Pigs, Environmental Samples, and Vaccine Strains from Six Recent Swine Erysipelas Outbreaks in the United States

Bender

Shen

Irwin

et al. 2010

Clin Vaccine Immunol

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“…A variety of approaches can be used to demonstrate that IHC antibodies are specific, including western blot analysis combined with mass spectrometry or crystallography, staining of cells from known in vitro cell lines, and qualitative assessment of tissue samples [7]–[9]. Although these validation methods are useful first steps to ensure specificity, they do not guarantee sufficient sensitivity to ensure all cells of interest are included in the final count in histological preparations suitable for stereological estimation of cell number.…”

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confidence: 99%

Application of Immunohistochemistry in Stereology for Quantitative Assessment of Neural Cell Populations Illustrated in the Göttingen Minipig

2012

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BackgroundStereology is the study of estimating geometric quantities. When successfully applied, the combination of immunohistochemistry (IHC) and stereology eliminates intra- and interobserver variability for cell type identification.Methodology/Principal FindingsWe propose a method to validate existing antibody based cell type markers for stereological application. Comparison was made on the 100-days-old Göttingen minipig (G-mini) neocortex between estimates of total neuron number derived from Giemsa staining using morphological criteria and immunohistochemistry-based cell counting with NeuN. The mean total neuron numbers estimated by the two staining methods were not significantly different. Estimated quantities, including glial cell number, neocortical volume, cell densities and glial-to-neuron ratio were also presented. Additionally, we assessed other commonly used glial markers and discussed how to evaluate the advantages and disadvantages of these markers for stereological estimation of cell number.Conclusion/SignificanceThe concordance in quantitative estimates of total neuron number derived from NeuN- and Giemsa-stained sections provides evidence for the sensitivity and specificity of NeuN as a neuronal marker in the G-mini. Although time-consuming, quantitative validation of IHC should always be considered in stereological studies if there is doubt of the sensitivity, specificity, or reproducibility of cell type markers. Inaccurate staining may cause both over- and underestimation of the total cell number and inflict considerable limitation when analyzing the results.

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“…21 Disagreement between direct bacterial isolation and the IHC assay was found in 25 of 170 (14.7%) swine cases. 21 This result might be similar to the findings in the eclectus parrot, in which the antiinflammatory drugs used may have interfered with bacterial infection. Different pathological presentations of erysipelas infection have been described in birds.…”

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confidence: 97%

Outbreak of Mortality in Psittacine Birds in a Mixed-Species Aviary Associated With Erysipelothrix rhusiopathiae Infection

Galindo-Cardiel

Opriessnig²,

Molina

et al. 2011

Vet Pathol

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Erysipelothrix rhusiopathiae septicemia, associated with an increased mortality of captive psittacines in a mixed-species aviary, was diagnosed by histopathology, Gram staining, bacterial culture and sequencing, immunohistochemistry, and real-time polymerase chain reaction (PCR). Over a period of 23 days with no premonitory signs, 2 rainbow lorikeets and an eclectus parrot died. Of these birds, one lorikeet and the eclectus were submitted for necropsy. The main pathologic findings were thrombosis (2/2), bacterial embolism/thromboembolism (2/2), necrotizing hepatitis (2/2), necrohemorrhagic myocarditis (1/2), fibrinohemorrhagic and heterophilic visceral coelomitis (1/2), submandibular necrosuppurative dermatitis with necrotizing vasculitis and bacterial and fungal thromboembolism (1/2), and locally extensive rhabdomyonecrosis with bacterial embolism (1/2). Intralesional bacteria were positive by Gram staining and immunohistochemistry in both cases. E. rhusiopathiae was isolated by routine bacterial culture from the liver of the lorikeet, which was also positive by real-time PCR. This report is one of the rare descriptions of erysipelas in psittacines, and to the authors' knowledge, it appears to be the first in the described species using immunohistochemistry and real-time PCR on avian paraffin-embedded tissues for the diagnosis.

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Development and Validation of an Immunohistochemical Method for Rapid Diagnosis of Swine Erysipelas in Formalin-Fixed, Paraffin-Embedded Tissue Samples

Cited by 19 publications

References 9 publications

Characterization of Erysipelothrix Species Isolates from Clinically Affected Pigs, Environmental Samples, and Vaccine Strains from Six Recent Swine Erysipelas Outbreaks in the United States

Characterization of Erysipelothrix Species Isolates from Clinically Affected Pigs, Environmental Samples, and Vaccine Strains from Six Recent Swine Erysipelas Outbreaks in the United States

Application of Immunohistochemistry in Stereology for Quantitative Assessment of Neural Cell Populations Illustrated in the Göttingen Minipig

Outbreak of Mortality in Psittacine Birds in a Mixed-Species Aviary Associated With Erysipelothrix rhusiopathiae Infection

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