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PubMed1999

Direct and rapid detection by PCR of Erysipelothrix sp. DNAs prepared from bacterial strains and animal tissues.

A PCR method for rapid screening of Erysipelothrix spp. in the slaughterhouse was carried out by using four species-specific sets of oligonucleotide primers after initial amplification with the primer set MO101-MO102, which amplifies the 16S rRNA sequences of all four Erysipelothrix species. The DNA sequences coding for the rRNA gene cluster, including 16S rRNA, 23S rRNA, and the noncoding region downstream of 5S rRNA, were determined in order to design primers for the species-specific PCR detection system. The homology among the 4.5-kb DNA sequences of the rRNA genes of Erysipelothrix rhusiopathiae serovar 2 (DNA Data Bank of Japan accession no. AB019247), E. tonsillarum serovar 7 (accession no. AB019248), E. rhusiopathiae serovar 13 (accession no. AB019249), and E. rhusiopathiae serovar 18 (accession no. AB019250) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four Erysipelothrix species. The results of PCR tests performed directly with tissue specimens from diseased animals were compared with the results of cultivation tests, and the PCR tests were completed within 5 h. The test with this species-specific system based on PCR amplification with the DNA sequences coding for the rRNA gene cluster was an accurate, easy-to-read screening method for rapid diagnosis of Erysipelothrix sp. infection in the slaughterhouse.

Abattoirs↗
PubMed2001

Serovars of Erysipelothrix species isolated from the tonsils of healthy cattle in Japan.

Serovars of 79 Erysipelothrix isolates recovered from the tonsils of healthy slaughtered cattle over a 1-year period in Japan were determined by an agar double-diffusion precipitation system using typing sera representing all the known serovars, 1 through 23 and type N, of Erysipelothrix. A total of 43 out of the 79 Erysipelothrix isolates could be classified into nine serovars but the remaining 36 isolates were untypable. Of 42 isolates identified as Erysipelothrix rhusiopathiae, 4, 6, 2, 3, 1,12, 13 and 1 isolates belonged to serovars 1b, 2, 5, 9, 12, 13, 19 and 21, respectively. One isolate belonged to Erysipelothrix tonsillarum serovar 3.

Animals↗
PubMed2004

Erysipelothrix septicemia in a little blue penguin (Eudyptula minor).

On June 25, 2002, aquarium veterinarians treated a 5-year-old, male little blue penguin (Eudyptula minor) that was acutely recumbent and dull, with inappetence of 24-hour duration. The penguin died within 10 minutes of presentation despite emergency resuscitation efforts. Gross pathologic findings consisted of pulmonary congestion and intestinal hemorrhage. Histopathologic findings included necrosis of tips of intestinal villi, increased numbers of mononuclear cells in pulmonary interstitium and hepatic sinusoids, and gram-positive bacteria in systemic microvasculature. Transmission electron microscopic examination revealed short gram-positive bacilli located in lumina of glomerular capillaries and in cytoplasm of mononuclear phagocytic cells in the lung and liver. Erysipelothrix rhusiopathiae was recovered from the lung, liver, and intestine by bacteriologic culture. Amplicons from polymerase chain reaction (PCR) tests using Erysipelothrix genus-specific primers and total genomic DNA extracted from formalin-fixed, paraffin-embedded tissue sections of lung and intestine demonstrated 99% nucleotide sequence identity with 16S small-subunit ribosomal DNA of E. rhusiopathiae and E. tonsillarum. The source of infection was speculated to be fish in the diet; however, repeated attempts to detect Erysipelothrix spp. from the mucous layer of food fish using bacteriologic culture and PCR were unsuccessful. This is the first report of erysipelas in a captive aquatic bird. Details of the isolation of E. rhusiopathiae and the application of molecular testing to identify Erysipelothrix DNA in formalin-fixed, paraffin-embedded tissue sections are given.

Animals↗
PubMed1999

Serotyping and pathogenicity of Erysipelothrix strains isolated from tonsils of slaughter pigs in Thailand.

Erysipelothrix strains were isolated from the tonsils of 46 (15.0%) of 307 apparently healthy slaughter pigs in Thailand during the period of August to September, 1997. A total of 27 of the 46 Erysipelothrix isolates could be classified into 5 serovars but the remaining 19 were untypable in this study. Of the 25 isolates serologically identified as Erysipelothrix rhusiopathiae, 20, 4, and 1 isolates belonged to serovars 2, 12, and 17, respectively. Only 2 isolates from the tonsils belonged to Erysipelothrix tonsillarum and represented either serovar 7 or 10. Although the periods and the districts of the survey were limited, the information obtained in the present investigation demonstrates the presence of a variety of serovars in pigs in Thailand. Of 29 selected isolates belonging to serovars 2, 7, 10, 12, 17, and untypable, only 5 (17.2%) were virulent for both mice and pigs. Five of these virulent isolates belonging to serovars 2 and 12 killed less than 30% of mice immunized with a swine erysipelas bacterin commercially available in Thailand, suggesting that the vaccine elicited a sufficient immunity to these field isolates.

Agglutination Tests↗
PubMed2004

Detection of Erysipelothrix rhusiopathiae in clinical and environmental samples.

Erysipelothrix rhusiopathiae is pathogenic for both animals and humans, causing erysipelas in swine and erysipeloid in humans. In swine, disease may be either acute or chronic, resulting in the development of arthritis and endocarditis. In Japan, erysipelas remains an animal hygiene problem causing great economic loss as infected swine are disused. Human infection closely resembles that seen in swine, with both acute and chronic forms also. The most common presentation is erysipeloid, a localized cutaneous infection. In Western Australia, an erysipeloid-like infection referred to as "crayfish poisoning" occurs in lobster fishermen and handlers. A second type of presentation is a generalized cutaneous form involving lesions that progress from the initial site of infection or appear in remote areas. The third and most serious form of disease is a septicemia that is almost always linked to endocarditis. The mortality rate in Erysipelothrix endocarditis is still high (38%) and can be explained by the use of vancomycin (to which Erysipelothrix spp. are inherently resistant) as empirical therapy. Therefore, it is critical to have an early diagnosis of E. rhusiopathiae infection.Unfortunately, several problems exist with the diagnosis of E. rhusiopathiae infections by conventional cultural procedures, and these infections are often incorrectly diagnosed. First, because of their very small colony size and slow growth rates, it is difficult to isolate E. rhusiopathiae from heavily contaminated specimens. Various selective media have been described to improve the isolation of E. rhusiopathiae from contaminated specimens; however, not all contaminants are inhibited. The development of two polymerase chain reaction (PCR) methods has created an opportunity to greatly improve the efficiency with which these organisms are detected and identified. Makino et al. designed a PCR method that amplifies a 407-bp DNA fragment derived from the 16S rRNA coding sequence. The primers in this method are specific for the genus Erysipelothrix and do not differentiate between the species. A second set of primers designed by Shimoji et al. amplifies a 937-bp DNA fragment which is derived from a sequence associated with virulence of E. rhusiopathiae. These primers are specific for E. rhusiopathiae only. Shimoji et al. also utilized a selective enrichment medium based on tryptic soy broth containing ethidium bromide and sodium azide.

Air Microbiology↗
PubMed1998

Occurrence of Erysipelothrix spp. in chicken meat parts from a processing plant.

From March 1996 to March 1997, 153 domestic raw chicken meat samples, including 71 thigh, 50 outer breast muscle, and 32 white meat samples, from a processing plant located in a chicken abattoir in Nagano Prefecture, Japan, were examined for the presence of Erysipelothrix spp. Erysipelothrix spp. were isolated from 49 (30.0%) of the 153 chicken meat samples. Of 67 Erysipelothrix isolates, 65 and 2 isolates were identified as E rhusiopathiae and E. tonsillarum. E. rhusiopathiae and E. tonsillarum isolates were serotyped into 11 and 2 different serovars, respectively. These findings might indicate that domestic chicken meat is frequently contaminated with E. rhusiopathiae and seems to be a potential source of human Erysipelothrix infection.

Abattoirs↗
PubMed2005

Neuraminidase production by Erysipelothrix rhusiopathiae.

In order to characterise neuraminidase activity by Erysipelothrix, 85 isolates of Erysipelothrix spp. from a variety of sources including human clinical, marine and terrestrial animals, and the environment were investigated for neuraminidase production. Neuraminidase activity was detected by a peanut lectin haemagglutination method. The effects of media, incubation conditions and pH on the production and activity of neuraminidase were also investigated. Enzyme activity was detected only in the supernatants of the isolates of Erysipelothrix rhusiopathiae which had been incubated in cooked meat broth and Todd Hewitt broth supplemented with horse serum after 16 and 36 h incubation at 37 degrees C. The maximum titres were reached at 40 h in cooked meat broth and 56 h in Todd Hewitt broth supplemented with horse serum. All 58 isolates and the type strain (ATCC 19414) of E. rhusiopathiae produced detectable neuraminidase activity with titres between 10 and 320. The optimal pH for the enzyme activity varied among the isolates with a pH between 6.0 and 7.0 covering the highest enzyme activity of the most. There was no statistically significant difference in the level of neuraminidase activity between isolates from different sources (p > 0.05). Neuraminidase activity was not detected in the non-pathogenic Erysipelothrix spp. such as E. tonsillarum. Neuraminidase was detected only in E. rhusiopathiae suggesting its possible role as a virulence factor. Enzyme production and activity were medium and pH dependent. The peanut lectin haemagglutination assay is a simple, rapid and sensitive method and is particularly useful for the analysis of multiple samples.

Animals↗
PubMed2003

Pathogenicity for mice and swine of Erysipelothrix isolates from the tonsils of healthy cattle.

The pathogenicity of 79 Erysipelothrix isolates from bovine tonsils for mice and swine was determined. Five (6.3%) isolates were lethal for mice. These isolates belonged to serovars 1b (one isolate), 2 (2), 19 (1) and 21 (1). The 50% lethal dose values of the isolates ranged from 0.33 to 5x10(2) CFUs in mice. Twenty Erysipelothrix isolates (25.3%) were weakly virulent inducing only emaciation while 12 (15.2%) inducing emaciation and ruffled hair. In swine, clinical signs of varying severity were observed. Four isolates were virulent, capable of inducing localized or generalized urticarial lesions accompanied with a rise in body temperature after intradermal inoculation. One isolate each of serovars 1b, 2 and 19 was highly virulent, capable of inducing generalized urticarial lesions while another Erysipelothrix isolate of serovar 2 induced only a localized urticarial lesion at the site of inoculation. Another isolate of serovar 1b induced itching and irritation without obvious urticarial lesion at the site of inoculation. On the other hand, one isolate of serovar 21 and two other isolates of serovar 2 could not induce experimentally any clinical sign of erysipelas other than rise in body temperature. There was a rise in growth agglutination (GA) titer of serum in all the inoculated swine. These observations suggest that Erysipelothrix isolates from cattle are pathogenic for mouse and swine, and may also be pathogenic for other animals and humans.

Agglutination Tests↗
PubMed1995

Classification of Erysipelothrix strains on the basis of restriction fragment length polymorphisms.

Restriction fragment length polymorphisms of the 16S rRNA genes of Erysipelothrix strains were studied by cleavage of the chromosomal DNA with restriction endonuclease EcoRI, followed by hybridization to a 420-bp internal fragment of the 16S rRNA gene. Thirty-two Erysipelothrix type and reference strains were classified, together with seven field strains. Reference strains of all serotypes and the type strains of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum were included. Nine ribopatterns were observed. Pattern A was represented by 16 strains and included strains of serotypes 1b, 2 to 8, 11 to 13, 15 to 17, 19, and 23. Pattern B was represented by two strains (serotypes 1a and 9). Pattern C was represented by five strains (serotypes 5, 6, and 21). Pattern D was represented by one strain of serotype 4. Pattern E was represented by 11 strains of serotypes 2, 7, 10, 20, 22, 24, and 25. Patterns F, G, H, and I were each represented by a single strain of serotypes 26, 2, 18, and 3, respectively. All the different ribopatterns had some bands in common. Patterns B, C, and D were most similar to pattern A, while patterns F, G, H, and I resembled pattern E. Partial sequencing of the 16S rRNA gene of nine selected strains resulted in three different sequences, i.e., the typical E. rhusiopathiae sequence, the E. tonsillarum sequence, and a third sequence found for two strains. Strains of the same serotype were found to have different ribopatterns as well as different partial 16S rDNA sequences.

Animals↗
PubMed1996

Serovars of Erysipelothrix strains isolated from pigs affected with erysipelas in Japan.

Serovars of 1,046 Erysipelothrix strains isolated from diseased pigs during a period from 1983 to 1993 in Japan were determined by an agar gel double-diffusion precipitation system using typing sera representing all the known serovars, 1 through 23 and type N, of 2 species of Erysipelothrix. A total of 943 strains could be serotyped within the serovars. The remaining 103 strains could not be determined for their serovars and were classified as untypable. Of the 938 (99.5%) strains serologically identified as Erysipelothrix rhusiopathiae, 423 (40.4%) belonged to serovar 1a, 359 (34.3%) to serovar 2, 65 (6.2%) to serovar 1b, 35 (3.3%) to serovar 6, 19 (1.8%) to serovar 5, 11 (1.1%) to serovar 21, and the remaining 26 to serovars 4, 8, 11, 12, 15, 17, or 19. Only 5 (0.5%) strains isolated from the cases of erysipelas belonged to Erysipelothrix tonsillarum and represented serovars 7 or 23.

Animals↗
PubMed2003

Molecular identification of Erysipelothrix isolates from the tonsils of healthy cattle by PCR.

For 79 isolates from the tonsils of healthy cattle identified as Erysipelothrix by cultivation, biochemical and serological tests, genotypic identification was performed by polymerase chain reaction (PCR) using four species-specific sets of oligonucleotide primers (ER1F-ER1R, ER2F-ER2R, ER3F-ER3R and ER4F-ER4R). The results of PCR for 79 bovine isolates were compared with those of serological typing. For 19 isolates, serotyping and genotyping results were the same. PCR allowed for the identification of 36 untypable isolates as Erysipelothrix species, strain 1. Serotyping and genotyping results of the remaining 24 isolates were different. Supplemental tests are frequently needed for Erysipelothrix identification.

Animals↗
PubMed1976

[Erysipelothrix endocarditis (author's transl)].

Following erysipeloid a 46-year-old man fell ill with septicaemia and endocarditis. Treatment with high doses of antibiotics could not prevent his death. Infection with Erysipelothrix rhusiopathiae in man occurs through a skin lesion and only rarely leads to spreading of the agent in the body presenting as arthritis, meningitis, or endocarditis. Erysipelothrix endocarditis is a severe disease leading to widespread destruction of the involved cardiac valves. The aortic valve is most commonly involved. 16 out of 28 patients with erysipelothrix endocarditis reported in the literature died, eleven of them despite treatment with antibiotics.

Animals↗
PubMed2001

[Mitral valve endocarditis caused by Erysipelothrix rhusiopathiae].

HISTORY AND CLINICAL FINDINGS: A 67 year-old country woman was admitted to the hospital because of a four weeks history of continuous catarrh, arthralgia and fever. Recently, she had also developed upper abdominal pain after oral ibuprofen treatment. The clinical examination showed a patient of impaired general condition. The heart and lungs were auscultatory normal and there were no signs of dyspnea, cyanosis or inflammatory skin lesions. EXAMINATIONS: Physical examination of heart and lung, electrocardiography and transthoracic echocardiography were without pathological findings. CLINICAL COURSE: Gastroscopy revealed acute antral gastritis and duodenitis with presence of Helicobacter pylori. Eradication therapy resolved the abdominal symptoms but fever returned after the antibiotic therapy was stopped. The patient developed a severe endocarditis with progressive mitral regurgitation within a few days. Erysipelothrix rhusiopathiae was isolated from blood cultures and identified by conventional and molecular methods. The patient was treated successfully with 3 x 2 g ampicillin daily, applied parenterally for six weeks, and a mitral valve replacement. CONCLUSION: This was an unusual manifestation of systemic Erysipelothrix rhusiopathiae infection. The bacterium Erysipelothrix rhusiopathiae has still to be considered in the diagnosis and treatment of endocarditis in patients with increased risk of exposure (e.g. farmers, butchers and fishermen).

Aged↗
PubMedDate not supplied

Superoxide release in human fibroblasts upon treatment with culture supernatants of the arthritogenic bacteria Erysipelothrix rhusiopathiae and Mycoplasma arthritidis.

Culture supernatants of the arthritogenic bacteria Mycoplasma pneumonia, Mycoplasma arthritidis, Borrelia burgdorferi and Erysipelothrix rhusiopathiae stimulated primary cultures of human fibroblasts to release reactive oxygen species into the environment, whereas cell walls and membranes of these bacteria had no effects. Lipopolysaccharides of various gram-negative bacteria and lipid A, the lipid moiety of endotoxines, also failed to stimulate the release of reactive oxygen species by fibroblasts. The stimulatory fractions of the culture supernatants of Mycoplasma arthritidis and Erysipelothrix rhusiopathiae exhibited a molecular weight of about 9.5 kDa. After an induction period of 5 min the presence of the stimulant was not necessary any more. The primary radical released by the fibroblasts was the superoxide anion O2-. Radical formation took place continuously over some hours. Additionally, low-level chemiluminescence of fibroblasts was increased upon stimulation with culture supernatants of Mycoplasma arthritidis and Erysipelothrix rhusiopathiae. No irreversible injury of the fibroblast was caused upon stimulation and the cells exhibited normal proliferation pattern after replacing them to the culture medium.

Arthritis, Infectious↗
PubMed2026

Oxford Nanopore Sequencing of Clinical DNA for Identification and Comparative Genomic Analysis of Erysipelothrix piscisicarius.

The genus Erysipelothrix comprises facultative anaerobic, nonspore-forming, gram-positive bacteria that can cause skin infections and severe diseases such as septicemia and endocarditis in humans. Although E. rhusiopathiae is the primary pathogen, other species may also be involved, necessitating accurate identification. However, 16S rDNA sequencing lacks sufficient resolution to differentiate among Erysipelothrix species. In this study, we used Oxford Nanopore Technology (ONT) to directly sequence low-quality DNA extracted from heart valve tissue of a 66-year-old female patient with a fatal case of septicemia and aortic endocarditis. In contrast to 16S rDNA Illumina sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), which incorrectly identified the pathogen as E. rhusiopathiae, direct sequencing via ONT precisely identified E. piscisicarius as the cause of infection. About 1.47 Mb genome was retrieved from nanopore direct sequencing. Within the E. piscisicarius genome, we detected genes associated with virulence. Phylogenetic analysis showed that our strain clustered with a human-derived E. piscisicarius strain from China and swine-derived strains from Brazil. In conclusion, this study demonstrated that ONT can be used to sequence low-quality DNA extracted directly from patient specimens, obtain a draft bacterial genome, and reliably distinguish between pathogenic species.

Aged↗
PubMedDate not supplied

Specific detection and semiquantitation of microorganisms in tissue by nucleic acid hybridization. II. Investigation of synovia from pigs with chronic Erysipelothrix arthritis.

Synovial tissues from animals with chronic Erysipelothrix arthritis were examined by a technique based on RNA-DNA hybridization in an attempt to detect the inducing organism, Erysipelothrix insidiosa (EI). Twelve synovial specimens from 5 animals whose joints lacked culturable EI also lacked EI detectable by this technique. Because the technique is capable of detecting approximately one organism per 50 mammalian cells, it is concluded that no more than this number were present in the involved tissue. Implications of these results for proposed pathogenetic mechanisms in this disorder and in human rheumatoid arthritis are discussed.

Animals↗
PubMed2005

Visceral botryomycosis in a case of Erysipelothrix rhusiopathiae endocarditis.

Botryomycosis is a rare bacterial infection of the skin and, rarely, viscera that is characterized by the formation of characteristic hyaline grains. We encountered a patient with Erysipelothrix rhusiopathiae endocarditis who developed visceral botryomycosis. He was a 54-year-old black man who presented in sepsis with a history of progressive weakness and severe weight loss. He died 10 hours after admission. At autopsy, endocarditis was identified, along with infarcts of the spleen and kidneys. Microscopically, visceral botryomycosis was seen. With these bacteria, an animal source is usually identified, although one was not found in this man. Erysipelothrix rhusiopathiae is an organism that is becoming identified in a range of infections in humans, the manifestations and pathophysiology of which are still being discovered.

Endocarditis, Bacterial↗
PubMed2001

Induction of cross-protection in mice against dolphin Erysipelothrix rhusiopathiae isolates with a swine commercial vaccine.

Erysipelothrix rhusiopathiae is well known to cause disease in dolphins. This disease occurs either in an peracute way, leading to mortality even before clinical signs are observed or in a sub-acute way, characterized by rhomboidal skin lesions, that can be treated with penicillin or its derivatives. Commercial swine vaccines, containing inactivated serotype 2 strains, are currently used for vaccination but it is not known whether these vaccines induce protection against E. rhusiopathiae isolates from dolphins. In the present study, it was demonstrated in a mouse model that vaccination with a commercial swine vaccine (Eurovac Ery, Eurovet, Belgium) containing inactivated serotype 2 E. rhusiopathiae strains induced protection against challenge with three E. rhusiopathiae isolates from dolphins. The duration of the protection varied, depending on the challenging isolate, between 8 and >23 weeks. There was however no positive correlation between the amount of antibodies at the moment of challenge and the observed protection. In conclusion, vaccination trials in mice indicate that commercial serotype 2 swine Erysipelothrix vaccines induce protection against erysipelas caused by dolphin pathogenic isolates.

Animals↗