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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↗

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↗

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↗

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↗

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↗

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↗

[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↗

[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↗

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↗

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↗

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↗

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↗

Adjuvant properties of Propionibacterium avidum KP-40 in vaccination against endemic viral and bacterial infections. III. Swine immunized with live attenuated Erysipelothrix rhusiopathiae vaccine and experimentally infected with virulent strains R203 and R270B of E. rhusiopathiae.

Fifty 4-month old piglets were treated with immunomodulating Propionibacterium avidum KP-40 (PA) and/or immunized with live attenuated Erysipelothrix rhusiopathiae vaccine (Orvac). Four weeks after vaccination all animals were inoculated with viable Erysipelothrix rhusipathiae. The vaccine induced the appearance of high titers of specific IgG antibodies with peak values (1:115-1:200) three weeks after immunization. Administration of PA together with the vaccine did not influence antibody titers. Analysis of the course of experimental erysipelas infection in vaccinated and/or PA-treated swine revealed the prophylactic and beneficial effects of PA. PA-treated animals showed a significantly lower lethality rate than untreated controls and the course of the disease was considerably milder, with a shorter period of fever and a faster recovery. Vaccination provided good protection of swine against the development of erysipelas and therefore, the only significant difference in animals treated with PA applied together with the vaccine was a higher gain of body mass after infection.

Adjuvants, Immunologic↗

Susceptibility of Erysipelothrix rhusiopathiae to antimicrobial agents and home disinfectants.

AIM: Erysipelothrix rhusiopathiae causes the occupationally-related infection erysipeloid in humans, and may be responsible for infections in lobster fishermen in Western Australia. There are little recent data pertaining to antimicrobial susceptibility, or susceptibility to disinfectants that might be used in the environment. The aim of this study was to determine the susceptibility of E. rhusiopathiae from human, animal and environmental sources to various antimicrobial agents and disinfectants. METHODS: The susceptibility of 60 E rhusiopathiae isolates was determined using a recommended agar dilution procedure. Susceptibility to disinfectants was achieved using a broth microdilution method. RESULTS: Penicillin and ceftriaxone, with low minimum inhibitory concentrations (MICs) (MIC90 0.03 mg/l and 0.125 mg/l, respectively), remained active against E. rhusiopathiae and should continue to be recommended for treatment. Ciprofloxacin MICs were particularly low (MIC90 0.06 mg/l), offering an alternative agent for the penicillin allergic patient. Erysipelothrix rhusiopathiae is still resistant to vancomycin (MIC90 64 mg/l), highlighting the importance of early diagnosis of E. rhusiopathiae infection in cases of endocarditis. In addition, 31 E. rhusiopathiae isolates were tested against several commercially available home disinfectants. Most were effective in killing E. rhusiopathiae with minimum bactericidal concentrations of 0.001% for Pine O Cleen, and 0.03% for Domestos, Linely and the Wheelie Bin Phenyl Cleanser. CONCLUSIONS: There appeared to be no new emergence of antibiotic resistance in E. rhusiopathiae. Various disinfectants could be used following mechanical cleaning of work environments, such as fishing boats, and equipment, to reduce the risk of infection with E. rhusiopathiae.

Animals↗

Erysipelothrix rhusiopathiae: bacteriology, epidemiology and clinical manifestations of an occupational pathogen.

Erysipelothrix rhusiopathiae has been recognised as a cause of infection in animals and man since the late 1880s. It is the aetiological agent of swine erysipelas, and also causes economically important diseases in turkeys, chickens, ducks and emus, and other farmed animals such as sheep. The organism has the ability to persist for long periods in the environment and survive in marine locations. Infection in man is occupationally related, occurring principally as a result of contact with animals, their products or wastes. Human infection can take one of three forms: a mild cutaneous infection known as erysipeloid, a diffuse cutaneous form and a serious although rare systemic complication with septicaemia and endocarditis. While it has been suggested that the incidence of human infection could be declining because of technological advances in animal industries, infection still occurs in specific environments. Furthermore, infection by the organism may be under-diagnosed because of the resemblance it bears to other infections and the problems that may be encountered in isolation and identification. Diagnosis of erysipeloid can be difficult if not recognised clinically, as culture is lengthy and the organism resides deep in the skin. There have been recent advances in molecular approaches to diagnosis and in understanding of Erysipelothrix taxonomy and pathogenesis. Two PCR assays have been described for the diagnosis of swine erysipelas, one of which has been applied successfully to human samples. Treatment by oral and intramuscular penicillin is effective. However, containment and control procedures are far more effective ways to reduce infection in both man and animals.

Animals↗

Pulsed-field gel electrophoresis in differentiation of erysipelothrix species strains.

We report here the first analysis of Erysipelothrix spp. using pulsed-field gel electrophoresis (PFGE). Seventy strains of Erysipelothrix spp. were analyzed. SmaI, AscI, and NotI were tested for the ability to cleave the DNA extracted from those strains, and among them, SmaI was the most reliable enzyme. Sixty-three distinct PFGE patterns were produced, and no DNA degradation was observed, allowing the identification of all of the strains. Based on these results and on those of a previous analysis using randomly amplified polymorphic DNA and ribotyping, PFGE with SmaI might be considered to be more sensitive than those methods and to be the best method for epidemiological studies of strains of this genus.

Animals↗

Isolation of a high-molecular mass glycoprotein from culture supernatant of an arthritogenic strain of the bacteria Erysipelothrix rhusiopathiae reacting with "inductive" monoclonal antibodies derived from rats with erysipelas polyarthritis.

A glycoprotein exhibiting a relative molecular mass of about 1000 kDa was purified to homogeneity from culture supernatant of arthritogenic bacteria (Erysipelothrix rhusiopathiae, strain T28) by ultrafiltration, ammonium sulfate precipitation, molecular mass exclusion, and ion exchange chromatography. Fractions obtained were analysed for their antigenic content by an enzyme linked immunosorbent assay (ELISA) using rabbit immune serum raised against this strain of Erysipelothrix rhusiopathiae. Distinct monoclonal antibodies obtained from rats suffering from erysipelas polyarthritis display a unique property by inducing very efficiently protective and regulatory mechanisms while being unable to generate classical "passive immunity". These "inductive" monoclonal antibodies recognize most likely linear epitopes on the purified glycoprotein. This makes it a prime source for analysing the target structure of these in vivo "inductive" antibodies.

Animals↗

Characterization of Erysipelothrix rhusiopathiae isolated from an opossum (Didelphis virginiana) with septicemia.

Erysipelothrix rhusiopathiae was isolated from a wild-caught opossum (Didelphis virginiana). The opossum was quarantined in isolation and removed from contact with other animals. After a 2-mo period it was found dead in its cage, and presented for postmortem examination. Pure cultures of Erysipelothrix rhusiopathiae (SLU isolate) were recovered from heart blood, liver, spleen and lungs. To compare pathogenicity, an experimental infection was attempted in CF1 mice with a single dose of 1.5 x 10(7) organisms of both an ATCC standard strain and SLU isolate of E. rhusiopathiae. Similar signs, lesions and results of culture were found for both strains. The findings suggest that opossums can be infected with E. rhusiopathiae.

Animals↗