Pathogenesis of Erysipelothrix piscisicarius infection in tiger barbs (Puntigrus tetrazona).

Erysipelothrix piscisicarius is an emerging bacterial pathogen and the aetiologic agent of piscine erysipelosis, a recently recognized disease of ornamental fish. However, little is known regarding the dynamics of infection in fish. The purpose of this study was to gain a better understanding of the pathogenesis of piscine erysipelosis in the tiger barb (Puntigrus tetrazona) by investigating tissue tropisms and responses to bacterial dissemination following immersion challenge with a virulent strain recovered from diseased fish. The challenge resulted in 83% mortality by day 16. Erysipelothrix piscisicarius DNA was first detected in the skin using quantitative PCR, and bacteria were visualized in association with microscopic lesions on day 4. By day 8, E. piscisicarius DNA was further detected in intestines, hearts, spleens, gills and skin; parenchymal organs were largely spared. The data suggest a primary cutaneous portal of entry and tropism for collagenous tissues, particularly those within vascular walls. Initial spread occurs directly from the dermis into interstitial areas of skeletal muscle, then centrally to the peritoneum and coelomic cavity following collagenous tissue pathways. Although histopathology revealed widespread bacterial dissemination over time, the severity of skin and muscle lesions with high levels of bacterial DNA identifies these tissues as primary targets of infection.

Description of Erysipelothrix piscisicarius sp. nov., an emergent fish pathogen, and assessment of virulence using a tiger barb (Puntigrus tetrazona) infection model.

A recently described emergent disease of ornamental fish has been associated with an Erysipelothrix species positive for the surface protective antigen (spa) C gene. Whole genome sequencing was performed on five spaC Erysipelothrix isolates from diseased ornamental fish. In addition, these spaC Erysipelothrix isolates were compared to spaA-, spaB- and other spaC-positive Erysipelothrix species isolated from terrestrial and marine mammals, birds and fish using multi-locus sequence analysis (MLSA). The genomes of fish pathogenic spaC isolates were genetically distinct from Erysipelothrix rhusiopathiae, sharing 86.61-86.94 % average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of 31.6-32.2 %, but 99.01-99.11 % ANI and 90.8-91.9 % dDDH values with the uncharacterized spaC-positive Erysipelothrix sp. strain 2 isolated from swine. The findings indicate the spaC-positive fish and swine isolates are conspecific and represent a previously unrecognized taxon. While phylogenies inferred from MLSA sequences confirm this conclusion, slight genetic differences between the spaC fish isolates and swine strain 2 were indicated. Bath immersion challenge trials were conducted using tiger barbs (Puntigrus tetrazona) exposed by immersion to 107 c.f.u. ml-1 of three fish pathogenic spaC Erysipelothrix species, and three spaA and two spaB E. rhusiopathiae isolates as a model of infection. Thirty days post-challenge, cumulative mean percentage survival was 37 % for the spaA, 100 % for the spaB and 13 % for the spaC isolates, revealing differences in virulence among the various spa genotypes in fish. Genetic findings and observed differences in virulence demonstrate the fish pathogenic spaC isolates represent a novel species, for which the name Erysipelothrix piscisicarius sp. nov. is proposed. The type strain is E. piscisicarius 15TAL0474T (=NRRL B-65533T=ATCC-TSD-175T=DSM 110099T).

Proteomic and Transcriptomic Analyses of Swine Pathogen Erysipelothrix rhusiopathiae Reveal Virulence Repertoire.

E. rhusiopathiae is the causative agent of erysipelas in animals and erysipeloid in humans, but its pathogenicity is poorly understood. To identify virulence factors associated with E. rhusiopathiae and screen engineered vaccine candidates, we used proteomics and transcriptomics to compare the highly virulent strain HX130709 with an isogenic avirulent derivative, HX130709a. 1,299 proteins and 1,673 transcribed genes were identified and 1,292 of the proteins could be associated with genes. In a comparison between HX130907 and HX130709a, 168 proteins and 475 genes exhibited differences in regulation level. Among these, levels for 61 proteins and transcripts were positively or negatively correlated. Gene Ontology (GO) analysis suggests that many of the down-regulated proteins in the attenuated strain have catalytic or binding functions. Potential protein-protein interactions suggest that some of the down-regulated proteins may regulate PTS, GMP synthase and ribosomal proteins. Morphological results showed that HX130709 and HX130709a have similar colony and capsule morphology. Growth curves and pyruvate measurements suggest that TCA cycle and saccharide phosphorylation levels were decreased and gluconeogenesis was increased in HX130709a. Our study confirms that SpaA and neuraminidase, but not hyaluronidase and capsule, are associated with virulence in E. rhusiopathiae. We conclude that the virulence of E. rhusiopathiae may be associated with slow reactions of the TCA cycle and down-regulation of selected proteins.

Erysipelas Outbreaks in Flocks of Geese in Poland--Biochemical and Genetic Analyses of the Isolates.

Erysipelothrix rhusiopathiae causes erysipelas in many vertebrate species. Severe disease outbreaks have been noted in many poultry species--chickens, ducks, emus, pheasants, pigeons, and geese. This article describes the biochemical and genetic analyses of six E. rhusiopathiae strains isolated from geese for meat production. The isolates came from birds kept in different poultry houses on one farm, and were collected during two erysipelas outbreaks. We analyzed and compared the isolates by random amplified polymorphic DNA with the use of NK6 primer and pulsed-field gel electrophoresis, with the restriction enzyme SmaI. Biochemical examination was performed with API Coryne test. Analyses showed that the three strains isolated during the first outbreak differed, whereas the three isolates from the second outbreak were identical to one another, but distinct from the isolates from the first outbreak. The results of biochemical and genetic analyses of E. rhusiopathiae strains isolated from geese suggest different sources of infection for the erysipelas outbreaks.

Fatal Erysipelothrix rhusiopathiae septicemia in two Atlantic dolphins (Stenella frontalis and Tursiops truncatus).

We describe gross, histopathologic, ultrastructural, immunohistochemical, and microbiologic features of acute septicemia by Erysipelothrix rhusiopathiae in an Atlantic spotted dolphin Stenella frontalis and an Atlantic bottlenose dolphin Tursiops truncatus. Generalized lymphadenomegaly and widespread hemorrhages were the most consistent macroscopic findings. Tricavitary effusion and icterus were noted in one individual. Histologically, all organs examined showed numerous variably sized bacillary bacterial emboli (Gram-positive; Ziehl-Neelsen-negative), typically associated with systemic congestion, edema, hemorrhages, and fibrinocellular thrombi. These bacteria were frequently intravascular, either extracellular or intramonocytic/macrophagic, and to a lesser extent, free within the interstitium of parenchymal organs. In both cases, microbiological analysis yielded E. rhusiopathiae. A primary anti-E. rhusiopathiae antibody created in mice from one of the strains isolated allowed positive immunohistochemical detection. Electron microscopy and dual immunohistochemistry with lysozyme and MAC387 antibodies confirmed the intramacrophagic location of the bacilli. E. rhusiopathiae, a known multispecies and zoonotic agent, should be considered as a potential etiologic agent in septicemia cases in free-ranging individuals of these dolphin species.

Acute and Chronic Erysipelothrix rhusiopathiae Infection in Lambs.

Polyarthritis caused by Erysipelothrix rhusiopathiae is a relatively common infection in lambs characterized by low mortality and high morbidity. E. rhusiopathiae is a ubiquitous Gram-positive bacterium that is both a commensal and a pathogen of vertebrates. The disease was studied during an outbreak in a Norwegian Spæl sheep flock. In the acute phase, 48 of 230 (20%) lambs developed clinical signs and 4 died (1.7%). One acute case was necropsied and E. rhusiopathiae was cultured from all major organs investigated and from joints. There was a fibrinous polyarthritis, increased presence of monocytes in vessels, and necrosis of Purkinje cells. Sixteen of the diseased animals (33%) developed a chronic polyarthritis. Eight of these lambs were necropsied; all had lesions in major limb joints, and 3 of 8 also had lesions in the atlanto-occipital joint. At this stage, E. rhusiopathiae was cultured only from the joints in 7 of 8 (87.5%) lambs, but by real-time polymerase chain reaction, we showed persistence of the bacterium in several organs. Pulsed-field gel electrophoresis typing of the bacterial isolates indicated that the same strain caused the acute and chronic disease. Five of 6 (83%) chronically affected animals had amyloidosis of the spleen, and 6 of 8 (75%) had amyloidosis of the liver. All chronically affected animals had a glomerulonephritis, and 6 of 8 (75%) had sparse degeneration in the brain. Ceruloplasmin and haptoglobin were significantly increased in the chronically diseased lambs. These results show that chronic ovine erysipelas is not restricted to joints but is a multisystemic disease.

Erysipelas in a numbat (Myrmecobius fasciatus).

An adult, female numbat (Myrmecobius fasciatus) was submitted to the Perth Zoo Veterinary Department for postmortem examination in November 2011. This radio-collared wild numbat had been found dead in the Dryandra Woodland, 191 km southeast of Perth, Western Australia. On external examination, the body condition was good. Three ticks (Ixodes spp.) were found on the thoracic region. The external pouch was contaminated with dirt and palpably flocculent, and the nipples oozed a purulent material. Histopathology showed widespread fibrin thrombi containing bacterial microcolonies within interstitial vessels of the mammary gland with surrounding necrotic tissue. Bacterial microcolonies were present throughout the kidney, intestine, lung, and mammary tissue, and culture produced a moderate growth of Erysipelothrix rhusiopathiae. Although erysipelas has been reported as a cause of morbidity and mortality in marsupials, this is the first report of erysipelas in the order Dasyuromorphia (marsupial carnivores) and highlights the need for ongoing surveillance for causes of disease in wild numbats and species recovery programs.

Erysipelothrix rhusiopathiae isolates recovered from fish, a harbour seal (Phoca vitulina) and the marine environment are capable of inducing characteristic cutaneous lesions in pigs.

In order to determine the diversity and pathogenicity of Erysipelothrix spp. isolates recovered from marine fish, a harbour seal (Phoca vitulina) and the marine environment, 14 isolates were characterized by genotyping, serotyping, determination of the surface protective antigen (spa) gene type and assessment of virulence in a pig bioassay. All 14 isolates were Erysipelothrix rhusiopathiae. Isolates were determined to be of serotypes 2 (n = 3), 3 (n = 1), 4 (n = 1), 12 (n = 1), 15 (n = 1) or 21 (n = 6), and one isolate cross-reacted with serotypes 5 and 21. The spa gene analysis determined that 64.3% (n = 9) were spaA and 35.7% (n = 5) were spaB1. In pigs, 10/14 isolates induced small plaques to diamond-shaped cutaneous lesions consistent with Erysipelothrix spp. infection. The results of this study indicate that the marine E. rhusiopathiae isolates have greater genetic and antigenic diversity than pig isolates and are capable of inducing classical skin lesions in pigs.

Erysipelothrix rhusiopathiae pneumonia in an immunocompetent patient.

Erysipelothrix rhusiopathiae is a Gram-positive bacillus that causes infections primarily in animals. In humans, this bacterium usually causes localized cutaneous infections called erysipeloid. Here we report a case of pneumonia with isolation of E. rhusiopathiae from bronchoalveolar lavage and sputum. To our knowledge, this is the first report of a pneumonia case caused by E. rhusiopathiae confirmed by culture.

A child with Erysipelothrix arthritis-beware of the little known.

Erysipelothrix rhusiopathiae is an established animal pathogen while the zoonotic infections in humans are rarely reported. Infections occur after exposure to animals or animal products that are mostly occupational in adults. Here we report in a child for the first time septic arthritis and osteomyelitis without an identifiable risk factor. A 5-year-old male child was admitted with pain in the left hip joint and inability to bear weight on the limb. Clinical examination followed by radiological and magnetic resonance imaging was suggestive of septic arthritis. Erysipelothrix rhusiopathiae grew from peroperative joint specimen. The infection was resolved following arthrotomy, joint lavage and antibiotic therapy.

Histologic and bacteriologic findings in valvular endocarditis of slaughter-age pigs.

Endocarditis lesions from 117 slaughter pigs were examined pathologically and etiologically in addition to 90 control hearts with cardiac valves. Lesions were located on the valves; however, the lesions had extended to the walls in 21 cases (18%). Lesions predominated on the mitral valve (59%). A total of 28 cases, from which no growth was obtained or a contamination flora was grown, were screened by fluorescence in situ hybridization (FISH) for bacteria (general bacterial probe) and probes specific for Streptococcus suis and Erysipelothrix rhusiopathiae, respectively. Using FISH, an additional 10 cases of endocarditis due to S. suis and E. rhusiopathiae were disclosed. Within lesions, streptococci predominated (53%) followed by E. rhusiopathiae (30%). Distinct features of both the lesions and the shape and localization of bacterial colonies were related to streptococci and E. rhusiopathiae. The propensity for streptococci to be localized on more than 1 valve in single hearts may be because S. suis-infected pigs tend to have been infected for a longer period compared with E. rhusiopathiae. Mineralization of endocarditis lesions was significantly associated with infection by streptococci, and was seen in 71% of the cases, whereas it was present in only 28% of lesions caused by E. rhusiopathiae. In addition, areas with mineralization were significantly correlated to the presence of a granulomatous reaction. Granulomatous endocarditis is likely a result of a foreign body reaction due to dystrophic mineralization. Local proliferation of valvular endothelial cells, found in 9 hearts in the current study, may increase the risk of developing thrombosing endocarditis in pigs.

Immunization with truncated recombinant protein SpaC of Erysipelothrix rhusiopathiae strain 715 serovar 18 confers protective immunity against challenge with various serovars.

Previously, we showed that surface protective antigen (Spa) proteins of Erysipelothrix rhusiopathiae can be classified into three molecular species-SpaA, SpaB, and SpaC-and that SpaC is the most broadly cross-protective antigen among the three Spa proteins. In this study, we examined the ability of the α-helical domain, which comprises the N-terminal half of SpaC, to elicit cross-protective immunity in mice and pigs. Mice actively immunized with the full-length protein (rSpaC664) or the α-helical domain (rSpaC427), but not the C-terminal domain (rSpaC253), were protected against challenge with E. rhusiopathiae serovars 1a, 2, 6, 19, and 18 expressing heterologous (SpaA or SpaB) and homologous (SpaC) Spas. The α-helical domain seemed to provide better protection than rSpaC664, although the differences did not reach statistical significance. Similarly, mice passively immunized with rabbit anti-rSpaC664 or anti-rSpaC427 sera, but not anti-rSpaC253 serum, were protected from challenge with various serovars. Pigs immunized with SpaC427 also developed specific antibodies against Spa proteins and were protected from challenge with the highly virulent heterologous E. rhusiopathiae strain Fujisawa (serovar 1a). Taken together, these results demonstrate for the first time the striking protective efficacy of the α-helical domain-mediated immunization in both mice and pigs, thereby highlighting its utility as the most promising candidate for the development of a safe and effective vaccine against erysipelas.

Erysipelothrix rhusiopathiae.

Erysipelothrix rhusiopathiae is a facultative, non-spore-forming, non-acid-fast, small, Gram-positive bacillus. The organism was first established as a human pathogen late in the nineteenth century. Three forms of human disease have been recognised since then. These include a localised cutaneous lesion form, erysipeloid, a generalised cutaneous form and a septicaemic form often associated with endocarditis. The organism is ubiquitous and able to persist for a long period of time in the environment, including marine locations. It is a pathogen or a commensal in a wide variety of wild and domestic animals, birds and fish. Swine erysipelas caused by E. rhusiopathiae is the disease of greatest prevalence and economic importance. Diseases in other animals include erysipelas of farmed turkeys, chickens, ducks and emus, and polyarthritis in sheep and lambs. Infection due to E. rhusiopathiae in humans is occupationally related, principally occurring as a result of contact with contaminated animals, their products or wastes, or soil. Erysipeloid is the most common form of infections in humans. While it has been suggested that the incidence of human infection could be declining due to technological advances in animal industries, infection still occurs in specific environments. Additionally, infection by the organism is possibly under-diagnosed due to the resemblance it bears to other infections, and problems encountered in isolation and identification. Various virulence factors have been suggested as being involved in the pathogenicity of E. rhusiopathiae. The presence of a hyaluronidase and neuraminidase has been recognised, and it was shown that neuraminidase plays a significant role in bacterial attachment and subsequent invasion into host cells. The role of hyaluronidase in the disease process is controversial. The presence of a heat labile capsule has been reported as important in virulence. Control of animal disease by sound husbandry, herd management, good sanitation and immunization procedures is recommended.

An autopsy case of Erysipelothrix rhusiopathiae endocarditis.

A 58-year-old man was admitted to our hospital with fever. The vegetation was confirmed by echocardiography on the tricuspid valve and Erysipelothrix rhusiopathiae was isolated by blood culture. The patient died due to heart failure, and tricuspid valve vegetation was confirmed on autopsy and the sample of Gram's staining showed gram-positive microcolonies. Although about 60 cases of E. rhusiopathiae endocarditis have been reported, Japanese cases are extremely rare.

Erysipelothrix rhusiopathiae septicemia in a Laughing kookaburra (Dacelo novaeguineae).

Erysipelothrix rhusiopathiae (E. rhusiopathiae) septicemia was demonstrated in a captive Laughing kookaburra (Dacelo novaeguineae). The bird died after a 2-week period of weakness and weight loss. At necropsy, the bird was emaciated and had reddened and wet lungs. Microscopic lesions were limited to hepatic and pulmonary congestion with focal thrombosis. Erysipelothrix rhusiopathiae was isolated by routine bacterial culture from several organs. Further characterization of the isolate by pulsed-field gel electrophoresis indicated that the isolate has a new genotype pattern 3A(III), which is 91.7% homologous to an E. rhusiopathiae that was isolated from a pig in 2001 and 88% homologous to an isolate recovered in 2000 from a turkey with septicemia. This is the first report of E. rhusiopathiae-induced septicemia in a kookaburra.

Investigations on the pathogenicity of Erysipelothrix rhusiopathiae in laying hens.

Erysipelas was diagnosed in 1998 from 34-wk-old laying hens in a free range flock in Germany. Erysipelothrix rhusiopathiae of serotype 1 was cultured from internal organs of the affected birds. This article describes the pathogenicity of the field isolate of E. rhusiopathiae in experimentally infected specific pathogen-free (SPF) laying hens. Three experiments were performed with SPF chickens inoculated at 17, 27, and 37 wk of age by either intramuscular (IM) or oral route. Inoculated birds were observed for 14 days. The highest mortality rates occurred in older birds, with 100% mortality observed in the 37-wk-old birds inoculated IM, 60% mortality reported in the younger 27-wk-old birds, and no mortality in the 17-wk-old age group. In the orally infected 27-wk-old birds, 40% mortality was detected, whereas no mortality was observed in the oldest birds by the same route. The results of the experiments support the contention that older birds are more sensitive to infection than younger birds and that mortality in laying hens is age related and dependent on the route of infection.