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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↗

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.

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↗

Erysipelothrix rhusiopathiae bacteremia in a horse.

Erysipelothrix rhusiopathiae serotype 5 was isolated from blood obtained antemortem from a horse with presenting problems of laminitis, uveitis, acute blindness, localized ventral edema and depression. The patient failed to respond to therapy and died 96 hours after the onset of clinical signs. Cultures of the lung postmortem yielded Erysipelothrix rhusiopathiae serotype 5, Beta-hemolytic Streptococcus sp., Escherichia coli, Proteus sp., and Klebsiella sp.

Animals↗

Erysipelothrix rhusiopathiae bacteremia presenting as septic shock.

Erysipelothrix rhusiopathiae rarely causes bacteremia, and it has never been reported to cause septic shock. A 58-year-old man with fulminant hyperdynamic shock associated with E. rhusiopathiae bacteremia is described. Erysipelothrix must be considered when a patient with an appropriate history presents with septic shock.

Animals↗

Characterisation of Erysipelothrix rhusiopathiae isolates from pigs associated with vaccine breakdowns.

Swine erysipelas vaccines are routinely used to protect pigs against peracute and acute/urticarial forms of Erysipelothrix. Between 1995 and 1998, 34 swine herds across four Australian states experienced vaccine failure. Forty-four isolates of Erysipelothrix rhusiopathiae of serovars 2, 1a, 1b and 1bx21 were recovered from 15 of these 34 vaccine breakdown herds. These isolates were characterised by restriction fragment length polymorphism (RFLP) analyses using RsaI and AluI on whole cell DNA and for the presence of plasmid DNA. Results were compared with those of 20 isolates from 16 herds unaffected by vaccine breakdown and 13 isolates representing 10 reference strains. The majority of breakdown herds possessed isolates of serovar 2 (9/15 herds), followed by serovar 1a (5 herds). No geographic predominance of a single serovar was evident. The identification of 10 RsaI profiles from whole cell DNA among the 44 isolates from 15 breakdown herds indicated that a single, new clonal lineage of E. rhusiopathiae was not responsible for vaccine failure. RsaI RFLP analyses detected a further 14 distinct profiles among 20 field strains unassociated with vaccine breakdowns, and none matched profiles of the 10 serovar reference strains for serovars 1a, 1b, 2 or 21. This technique is recommended for epidemiological studies of E. rhusiopathiae strains.

Animals↗

A novel protein of Erysipelothrix rhusiopathiae that confers haemolytic activity on Escherichia coli.

Erysipelothrix rhusiopathiae, the cause of swine erysipelas and human erysipeloid, produces a haemolysin. A recombinant plasmid, pHLY, conferring haemolytic activity on Escherichia coli was isolated from a genomic library of Ery. rhusiopathiae strain Tama-96. This plasmid was stable in RecA- E. coli, but unstable in a RecA+ strain. A spontaneous deletion plasmid, pMini-HLY, also conferring haemolytic activity was derived from pHLY. Two ORFs were detected in pHLY. Analysis of pMini-HLY and other deletion clones established that ORF2 was associated with haemolytic activity. The sequence of ORF1 was highly homologous to those of transposases in the IS30 family. The deletion which generated pMini-HLY was between two short direct repeat (DR) sequences flanking the ORF1 sequence, and there were inverted repeat sequences inside the two DR sequences, suggesting an insertion element. No sequence homology to the deduced amino acid sequence of ORF2 was detected in the databases, but its sequence was characteristic of a surface lipoprotein. Western blot analysis, using antiserum against the 16 kDa protein produced from ORF2, found the protein to be commonly distributed in all Erysipelothrix species.

Amino Acid Sequence↗

Surface antigen, SpaA, of erysipelothrix rhusiopathiae binds to Gram-positive bacterial cell surfaces.

In a previous study, we isolated the spaA gene encoding the surface protective antigen A, SpaA, of Erysipelothrix rhusiopathiae, and found that the N-terminal region of SpaA was responsible for protective immunity against erysipelas and that the C-terminal region contained eight repeat units consisting of 20 amino acids comprising the binding domain on the Erysipelothrix cell surface. In this study, using recombinant SpaA proteins, we showed that the repeat region bound to the cell surfaces of various Gram-positive bacterial cells, SpaA was a membrane-associated protein, this association depended on the interaction with choline residues in teichoic acid, and SpaA bound to lipoteichoic acid (LTA) of Bacillus subtilis and Staphylococcus aureus. These results showed that LTA was required for the surface association of SpaA in E. rhusiopathiae and that such an association might be common among Gram-positive bacterial cells. We suggested that an LTA-SpaA complex might have an important role in the E. rhusiopathiae infection process.

Antigens, Bacterial↗

[One case with Erysipelothrix rhusiopathiae endocarditis].

We report case of a 67-year-old alcoholic fisherman who developed infective endocarditis caused by Erysipelothrix rhusiopathiae. The initial manifestations were fever and back pain of approximately three months' duration. Auscultation of the heart revealed a loud systolic murmur at the apex and a diastolic murmur over the aortic valve area. Echocardiographic studies showed vegetations on both the aortic valve and mitral valve. Blood cultures grew Erysipelothrix rhusiopathiae, and it was sensitive to aminobenzyl penicillin. No other bacteria grew aut. A diagnosis of infective endocarditis caused by the Erysipelothnx rhusiopatniae was made, and the patient was treated with aminobenzyl penicillin 12g/day for 6 weeks. His clinical course was complicated by heart failure, multiple cerebral embolism, and renal infarctions. However, he recovered without valve replacement. Although the exact route of infection remains unknown, erosions of the skin, of his palms at the time of tho initial examination appeared to be one possible source of the systemic infection in this case. The serotype of the bacteria was Ib. To our knowledge this is the first case of serotyping of bacterium that caused endocarditis in humans.

Aged↗

Fermentation reactions of Erysipelothrix rhusiopathiae.

White, Thomas G. (U. S. Department of Agriculture, Ames, Iowa), and Richard D. Shuman. Fermentation reactions of Erysipelothrix rhusiopathiae. J. Bacteriol. 82:595-599. 1961.-A study was made to determine the effect of four different basal media, to which fermentable carbon compounds had been added, upon 22 selected strains of Erysipelothrix rhusiopathiae (insidiosa). Acid production was measured by (i) chemical indicator, (ii) change in pH, and (iii) production of titrable acidity. At least two determinations, usually four, were made for each test on each strain. The fermentation pattern varied according to the medium, the indicator, and the method of measuring acid production. Andrade's base plus serum was the most dependable medium because it permitted the least variation in the total number of different patterns. Of the three methods used to measure acid production, the chemical indicator gave the most valid and reproducible results. The within-strain variation was not extreme and most strains persisted in a given fermentation pattern under like conditions of growth and acid production. Results of the study indicated that, regardless of the medium and indicator routinely used, one should be familiar with the fermentation pattern of known strains of the erysipelas organism.

Bioreactors↗

Randomly amplified polymorphic DNA analysis of Erysipelothrix spp.

The usefulness of randomly amplified polymorphic DNA method (RAPD) to identify each species of genus Erysipelothrix and for epidemiological analysis of this genus was studied. Eighty-one strains and 18 random primers were tested. Among the tested primers, the primers NK51 (GGTGGTGGTATC) and NK6 (CCCGCGCCCC) produced noticeable results. The primer NK51 revealed four species-specific RAPD patterns. Of the 66 strains of E. rhusiopathiae, 64 had the same unique band of 884 bp. Of the 12 strains of E. tonsillarum, 11 produced a 1,265-bp band. In addition, two strains, previously thought to be E. rhusiopathiae, produced the 1,265-bp band, suggesting that they had been misclassified. One strain of E. tonsillarum produced the 884-bp band, suggesting that it too was E. rhusiopathiae. The E. rhusiopathiae strain of serovar 13 produced a 650-bp band, and the strain of serovar 18 produced a clear 420-bp band as well as three weak bands of 1,265, 918, and 444 bp. The primer NK6 revealed 14 RAPD patterns that were not serovar specific. However, different patterns were produced among strains of the same serovar showing that the RAPD method is able to identify the genetic variations of strains of this genus and can rapidly and easily differentiate strains of the same serovar. Based on these results, we concluded that the RAPD method with primers NK51 and NK6 is a rapid and reliable method to identify the species of this genus; we also concluded that this method might be a useful tool for the epidemiological analysis of the Erysipelothrix species.

Erysipelothrix↗

Subacute endocarditis due to Erysipelothrix rhusiopathiae.

Erysipelothrix rhusiopathiae is a rare cause of endocarditis. Most cases were observed in people working with animals. We report a case in a 45-year old man without any exposure to animals. He was admitted to our hospital because of dyspnoea. Blood cultures were drawn following fever on day 8 of hospitalisation. Erysipelothrix rhusiopathiae was cultured and echocardiography showed a vegetation on the mitral valve. Appropriate antibiotic therapy and surgical treatment led to a good outcome of the infection.

Alcoholism↗

[Immunity against Erysipelothrix rhusiopathiae infection by means of active immunization using homologous neuraminidase (author's transl)].

Neuraminidase may play a role as a pathogenic factor in Erysipelothrix rhusiopathiae infection. The protective effect of active immunization with purified neuraminidase was therefore tested in an infection experiment in white mice. Mice were immunized 2, 4, 6, 8 or 10 times i.p. with Erysipelothrix neuraminidase. A control group received 10 injections with physiological saline. The infective dose varied between 7 and 7 x 10(7) cells. While all control animals infected with 7 x 10(1) germs died, the lethal effect could be reduced to 50% and 25% in animals immunized twice and 4 times, respectively. Only animals immunized 8 and 10 times were still partly protected against germ numbers of 10(3)-10(4). Germ numbers of 10(5) and more were almost always fatal even in highly immunized animals. Even a high immunization with neuraminidase could only lower the lethal by a maximum factor of 10(4) germs used.

Animals↗

Occurrence of different serotypes of Erysipelothrix rhusiopathiae in retail pork and fish.

Retail pork (38 samples), cod (10 samples) and herring (10 samples) were obtained from 12 stores in the area of Lund in southern Sweden during September and October 1990. Erysipelothrix rhusiopathiae was isolated from 50% of the pork samples, 60% of the cod samples and from 30% of the samples from herring. Serotype 2 dominated on retail pork as well as on fish samples constituting 53% of the pork isolates (10 strains) and 33% of the cod isolates (2 strains). All E. rhusiopathiae isolates originating from herring were serotype 2 (3 strains). Serotypes 1b, 6, and 8 were isolated from retail pork only (6, 2 and 1 strains, respectively). Serotype 5 was isolated from cod only (3 strains) and so was serotype 9 (1 strain). The public health hazards with the occurrence of virulent strains of Erysipelothrix rhusiopathiae in retail pork and fish are discussed.

Animals↗