Occurrence of Erysipelothrix rhusiopathiae on pork and in pig slurry, and the distribution of specific antibodies in abattoir workers

Strains of Erysipelothrix rhusiopathiae isolated at 19 pig farms serving a certain abattoir, and on pork and in workers of this abattoir were studied. Mouse-pathogenic E. rhusiopathiae was found in pig slurry from two farms (11%). The strains belonged to serotypes 7 and 16 (both from the same farm) or were untypable. In pig slurry from the abattoir lairage only serotype 2 strains were found and all were pathogenic to mice. Mouse-pathogenic E. rhusiopathiae strains of serotype 2 were also recovered from 25 pork loins (25%). A mouse-pathogenic E. rhusiopathiae (serotype 2) strain was isolated from one of the 16 hand infections of slaughterhouse workers. The E. rhusiopathiae strains were phenotypically grouped by the API 50 CH system. Variations were demonstrated for the different serotypes. In 20 of 138 workers antibodies against E. rhusiopathiae were found; 14 had increased levels of IgG antibodies, seven had increased levels of IgM antibodies and one had an increased level of both.     

Occurrence of Erysipelothrix rhusiopathiae on pork and in pig slurry, and the distribution of specific antibodies in abattoir workers

Strains of Erysipelothrix rhusiopathiae isolated at 19 pig farms serving a certain abattoir, and on pork and in workers of this abattoir were studied. Mouse-pathogenic E. rhusiopathiae was found in pig slurry from two farms (11%). The strains belonged to serotypes 7 and 16 (both from the same farm) or were untypable. In pig slurry from the abattoir lairage only serotype 2 strains were found and all were pathogenic to mice. Mouse-pathogenic E. rhusiopathiae strains of serotype 2 were also recovered from 25 pork lions (25%). A mouse-pathogenic E. rhusiopathiae (serotype 2) strain was isolated from one of the 16 hand infections of slaughterhouse workers. The E. rhusiopathiae strains were phenotypically grouped by the API 50 CH system. Variations were demonstrated for the different serotypes. In 20 of 138 workers antibodies against E. rhusiopathiae were found; 14 had increased levels of IgG antibodies, seven had increased levels of IgM antibodies and one had an increased level of both.     

Comparison of methods for detection of Erysipelothrix spp. and their distribution in some Australasian seafoods

For many years, Erysipelothrix rhusiopathiae has been known to be the causative agent of the occupationally related infection erysipeloid. A survey of the distribution of Erysipelothrix spp. in 19 Australasian seafoods was conducted, and methodologies for the detection of Erysipelothrix spp. were evaluated. Twenty-one Erysipelothrix spp. were isolated from 52 seafood parts. Primary isolation of Erysipelothrix spp. was most efficiently achieved with brain heart infusion broth enrichment followed by subculture onto a selective brain heart infusion agar containing kanamycin, neomycin, and vancomycin after 48 h of incubation. Selective tryptic soy broth, with 48 h of incubation, was the best culture method for the detection of Erysipelothrix spp. with PCR. PCR detection was 50% more sensitive than culture. E. rhusiopathiae was isolated from a variety of different fish, cephalopods, and crustaceans, including a Western rock lobster (Panulirus cygnus). There was no significant correlation between the origin of the seafoods tested and the distribution of E. rhusiopathiae. An organism indistinguishable from Erysipelothrix tonsillarum was isolated for the first time from an Australian oyster and a silver bream. Overall, Erysipelothrix spp. were widely distributed in Australasian seafoods, illustrating the potential for erysipeloid-like infections in fishermen.     

Improved method for the isolation of Campylobacter jejuni and Campylobacter coli bacteriophages

A centrifugation and filtration method of isolating Campylobacter phages has been developed. Forty-nine Campylobacter phages were isolated from 272 effluent samples of which 42 produced lysis with Campylobacter jejuni strains and seven with C. coli strains. Phages were recovered from pig manure, abattoir effluents, human faeces, sewage and poultry manure. Phages were not isolated from water samples, cattle and sheep faeces or farm pasture soil.     

DNA relatedness among Erysipelothrix rhusiopathiae strains representing all twenty-three serovars and Erysipelothrix tonsillarum.

The levels of relatedness among strains of Erysipelothrix rhusiopathiae (serovars 1 through 23 and type N) were estimated by performing DNA-DNA hybridization experiments with the type strains of E. rhusiopathiae and Erysipelothrix tonsillarum, which are the two Erysipelothrix species that have been described. Two distinct DNA relatedness groups were identified. The group 1 strains, representing serovars 1, 2, 4 through 6, 8, 9, 11, 12, 15 through 17, 19, and 21 and type N, exhibited more than 73% hybridization with the type strain of E. rhusiopathiae but less than 24% hybridization with the type strain of E. tonsillarum. Group 2 included serovar 3, 7, 10, 14, 20, 22, and 23 strains, and these strains exhibited more than 66% hybridization with the type strain of E. tonsillarum but less than 27% hybridization with the type strain of E. rhusiopathiae. Strains representing serovars 13 and 18 exhibited low levels of hybridization (16 to 47%) with both of the type strains, indicating that these serovars may be members of a new genomic species. The members of the E. rhusiopathiae and E. tonsillarum groups resembled each other in many phenotypic characteristics, but differed in their ability to produce acid from saccharose and in their pathogenicity for swine. Our results confirm that the genus Erysipelothrix contains two main genomic species, E. rhusiopathiae and E. tonsillarum, which can be differentiated into serovars.     

Antibodies to selected viral and bacterial pathogens in European wild boars from southcentral Spain

Serum samples from 78 European wild boars (Sus scrofa) harvested during the 1999-2000 hunting season were tested for antibodies to Brucella spp., classical swine fever virus, Erysipelothrix rhusiopathiae, Haemophilus parasuis, Leptospira interrogans serovar pomona, Mycoplasma hyopneumoniae, pseudorabies virus (PRV), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus, Salmonella serogroups B, C, and D, Streptococcus suis, and swine influenza virus (SIV) serotypes H1N1 and H3N2. Samples were collected from Sierra Morena and Montes de Toledo in southcentral Spain. Antibodies were detected to PRV (36%), L. interrogans serovar pomona (12%), PPV (10%), E. rhusiopathiae (5%), SIV serotype H1N1 (4%), Salmonella serogroup B (4%), and Salmonella serogroup C (3%). Our results suggest that more research is needed to describe the epidemiology of infectious diseases of Spanish wild boars.     

Evaluation of the use of pooled serum, pooled muscle tissue fluid (meat juice) and pooled faeces for monitoring pig herds for Salmonella

AIMS: Monitoring for Salmonella in slaughter pigs is important to enable targeted control measures to be applied on problem farms and at the abattoir. The aim of this study was to determine whether pooled serum and meat juice could be used to identify finishing pig herds with a high prevalence of infection. METHODS AND RESULTS: Samples of meat juice, serum, caecal contents, carcase swabs and pooled faeces from pig pens were taken from 20 commercial pig finishing farms and comparisons were made between the results of Salmonella culture, individual ELISA tests on serum and meat juice and pooled samples of serum and meat juice. Salmonella was isolated from samples from 19 of 20 farms. None of the ELISA tests showed a statistically significant correlation with caecal carriage of Salmonella or contamination of carcases. Serum mean optical density (O.D.) from pools of five, 10 or 20 sera showed a significant correlation with the Salmonella status of farm pen faeces. All pooled serum O.D. and sample/positive control ratio results correlated significantly with the results of the conventional individual sample ELISA. There was a statistically significant correlation between the incidence of Salmonella in farm pen pooled faeces and the prevalence of Salmonella in caeca of slaughter pigs. CONCLUSIONS: The results show a generally poor correlation between serological and bacteriological results but pooled serum or meat juice samples could be used as a cheaper substitute for serological screening of farms for Salmonella than individual samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The availability of a cheaper test should allow the costs of Salmonella monitoring of pig farms to be reduced or allow more regular testing to enhance the designation of farm Salmonella risk status.     

Naturally occurring erysipelas in rats

Erysipelothrix rhusiopathiae was isolated from the main limb joints of two Sprague Dawley rats affected by spontaneous lesions of chronic fibrinopurulent polyarthritis, endocarditis and mycocarditis.     

Erysipelothrix rhusiopathiae neuraminidase and its role in pathogenicity

The role of the enzyme neuraminidase in pathogenicity of the bacillus Erysipelothrix rhusiopathiae was studied. Different substances with low and high molecular weight were tested as inducers of E. rhusiopathiae neuraminidase biosynthesis. It was found that macromolecular complexes induce the secretion of the enzyme. K(M) values for different substrates showed that the affinity of the E. rhusiopathiae neuraminidase increases in parallel with the enlargement of the molecular weight of glycoproteins. Results from the rabbits skin test confirmed the role of E. rhusiopathiae neuraminidase as a factor of pathogenicity with spreading functions.     

Development of a novel fluorescent microbead-based immunoassay and comparison with three enzyme-linked immunoassays for detection of anti-Erysipelothrix spp. IgG antibodies in pigs with known and unknown exposure

A novel fluorescent microbead immunoassay (FMIA) using the recombinant polypeptide SpaA415 was developed for detection of anti-Erysipelothrix spp. IgG in pig sera. The diagnostic performance of the FMIA was evaluated on samples from pigs with known and unknown Erysipelothrix spp. exposure and compared to an in-house enzyme-linked immunosorbent assay (ELISA-1) based on the same capture antigen, and two commercially available ELISAs (ELISA-2 and ELISA-3). Sera from pigs experimentally infected with Erysipelothrix rhusiopathiae serotype 1a (n=60) or 19 (n=12), sera from pigs vaccinated with a commercial attenuated-live vaccine based on serotype 1a (n=12) or a commercial bacterin based on serotype 2 (n=12), and 90 field samples were utilized. The sensitivity on 22 true positive samples collected in the later stages of infection/post-vaccination was 100% for the FMIA and ELISA-1, 63.6% for ELISA-2 and 81.8% for ELISA-3. The earliest antibody response was detected 7days post inoculation with the FMIA (77.8%) and ELISA-1 (11.1%), and at 14days post-vaccination (dpv) with FMIA (50%) and ELISA-1 (50%). On field samples, a higher seroprevalence was found in pigs older than 21days with all four assays. Kappa analysis indicated that the FMIA and ELISA-1 had almost complete agreement whereas the agreement was slight with ELISA-2 and fair with ELISA-3. The sensitivity of both immunoassays based on the rSpaA415 antigen was higher compared to that of the two commercial ELISAs. The rSpaA415 FMIA has great potential as an inexpensive ELISA alternative for detection of antibodies against E. rhusiopathiae in the future.     

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.     

Intercellular adhesion molecule-1 (ICAM-1) and MHC Class II on chondrocytes in arthritic joints from pigs experimentally infected with Erysipelothrix rhusiopathiae

Abstract This study set out to investigate the in vivo expression and distribution of the porcine homologues of the intercellular adhesion molecule-1 (ICAM-1) and MHC Class II as markers of chondrocyte activation during the development of chronic polyarthritis, which was experimentally induced in Landrace pigs by intra-articular injection of Erysipelothrix rhusiopathiae . ICAM-1 was found to be strongly expressed in vivo on chondrocytes and synovial cells in arthritic joints but was not detected in cartilage from unaffected joints. Although the majority of ICAM-1 positive chondrocytes did not co-express MHC Class II, chondrocyte-type cells expressing both molecules were detected in the transition zone as the disease progressed, particularly at 5 months post-infection. At this stage infiltration of CD4⁺ T lymphocytes into the damaged cartilage was also apparent. ICAM-1 and MHC Class II are not constitutively expressed on porcine chondrocytes but appeared to be induced as arthritis progressed. The detection of these markers in the pig helps to establish the validity of this animal model for immunopathological studies.     

Microbiological status of piggery effluent from 13 piggeries in the south east Queensland region of Australia

AIMS: To assist in the development of safe piggery effluent re-use guidelines by determining the level of selected pathogens and indicator organisms in the effluent ponds of 13 south-east Queensland piggeries. METHODS AND RESULTS: The numbers of thermotolerant coliforms, Campylobacter jejuni/coli, Erysipelothrix rhusiopathiae, Escherichia coli, Salmonella and rotavirus were determined in 29 samples derived from the 13 piggeries. The study demonstrated that the 13 final effluent ponds contained an average of 1.2 x 10(5) colony-forming units (CFU) 100 ml(-1) of thermotolerant coliforms and 1.03 x 10(5) CFU 100 ml(-1) of E. coli. The Campylobacter level varied from none detectable (two of 13 piggeries) to a maximum of 930 most probable number (MPN) 100 ml(-1) (two of 13 piggeries). Salmonella was detected in the final ponds of only four of the 13 piggeries and then only at a low level (highest level being 51 MPN 100 ml(-1)). No rotavirus and no Erysip. rhusiopathiae were detected. The average log10 reductions across the ponding systems to the final irrigation pond were 1.77 for thermotolerant coliforms, 1.71 for E. coli and 1.04 for Campylobacter. CONCLUSIONS: This study has provided a baseline knowledge on the levels of indicator organisms and selected pathogens in piggery effluent. SIGNIFICANCE AND IMPACT OF THE STUDY: The knowledge gained in this study will assist in the development of guidelines to ensure the safe and sustainable re-use of piggery effluent.     

Persistence and Distribution of Erysipelothrix rhusiopathiae and Bacterial Indicator Organisms on Land Used for Disposal of Piggery Effluent

Erysipelothrix rhusiopathiae was isolated from 15 of 40 effluents collected from commercial piggeries. The organism was isolated from soil and pasture of experimental disposal sites for up to two weeks after application of effluent naturally infected with Ery. rhusiopathiae. The organism was more commonly isolated from soil than pasture. Times for 90% reduction (T₉₀ values) of indicator organisms over a six week period following the effluent applications where Ery. rhusiopathiae was detected for 7 d or more, were 8–19 d for faecal coliforms in top soil and 5–12 d on pasture. T₉₀ for faecal streptococci was 10–14 d in soil and 8–11 d on pasture. Laboratory investigations indicated that the death rate of Ery. rhusiopathiae was up to six times greater than Escherichia coli in soil at field capacity. In dry soil the differences in die-out rate of the two organisms was less marked.     

Detection of Candida species by polymerase chain reaction (PCR) in blood samples of experimentally infected mice and patients with suspected candidemia

In this study, we have established and evaluated a genus-specific polymerase chain reaction (PCR) and species-specific nested PCRs for the detection of Candida species in blood samples of neutropenic mice and patients suspected of candidemia. DNA segments of the gene encoding cytochrome P450 L1A1 were targeted for amplification by using genus and species-specific primers. As compared to the genus-specific PCR, the species-specific nested PCRs improved the sensitivity by 10 times with the detection limit < 10 yeast cells. Of the 18 blood samples tested daily over a period of 8 days following Candida albicans infection in neutropenic mice, four samples were positive by genus-specific PCR and 11 were positive by species-specific nested PCR. The PCR results were correlated with culture findings obtained on blood samples. Two of the three blood culture-positive samples were positive by genus-specific PCR and all the three with species-specific nested PCR. Among 15 mice, which were negative by blood culture but had C. albicans isolated from visceral organs, 2 and 8 mice yielded positive results by genus-specific PCR and species-specific nested PCR, respectively. Consistent with the results of the animal study, species-specific nested PCR yielded much higher positivity as compared to culture (52.2% versus 21.2%) in patients suspected for candidemia. Moreover, 8 specimens which were negative for Candida by genus-specific PCR became positive by species-specific nested PCR. No correlation was apparent between PCR positivity and Candida antigen titers. The results suggest that nested PCR is a sensitive technique for the detection of Candida species from blood samples, and thus it may have application in the diagnosis of suspected cases of candidemia and candidiasis.     

Isolation of Erysipelothrix rhusiopathiae from apparently healthy pigs reared under intensive and free range systems of management

The carriage of Erysipelothrix rhusiopathiae by pigs reared under free range and intensive systems of management was compared. Large white (exotic) breeds kept under an intensive system and local (native) breeds kept under a free range system were studied. Of the 29 pigs kept under the intensive system, 44.8% carried E. rhusiopathiae in their tonsils while 51.7% of pigs kept under free range system also carried the organism in their tonsils. In each of the systems 6.9% of the pigs carried E. rhusiopathiae in their bile and muscles. Blood samples from all the animals surveyed were negative for the organism. There was no significant difference in the carriage of E. rhusiopathiae in pigs reared under the two systems of management. The ubiquity of E. rhusiopathiae in the environment, its resistance to environmental influences, poor standards of hygiene and the problem of detecting apparently healthy animals, are possible epidemiological factors which may have increased the carrier rate in the intensive system where E. rhusiopathiae was supposed to be less. The total number of carrier-pigs in the intensive system was 44.8% while in the free range system it was 51.7%.     

2012–2015年江淮地区猪丹毒杆菌分离株血清型、spaA基因遗传进化及PFGE基因型分析

【目的】了解2012–2015年江淮地区猪丹毒杆菌分离株血清型分布、spaA基因遗传进化关系和基因分型特征。【方法】收集临床分离鉴定的42株猪丹毒杆菌,应用琼脂扩散沉淀实验、PCR扩增和序列分析技术、脉冲场凝胶电泳分型技术(PFGE)分别测定分离株的血清型、spaA基因遗传变异性及PFGE基因型。【结果】42株猪丹毒杆菌分离株血清型均为1a型;spaA基因与猪丹毒杆菌国内外参考株核苷酸序列相似性为98.5%–100%,分离株在第609 bp处出现T突变为G、769 bp处C突变为A,对应的氨基酸第203位Ile突变为Met、第257位Leu突变为Ile,为Met-203、Ile-257型;分离株形成8个PFGE基因型,相似度达88.8%–100%,优势基因型为ER2 (54.8%),弱毒疫苗G4T10和GC42株独立为同一个基因型。【结论】江淮地区致病猪丹毒杆菌流行血清型为1a型,spaA基因相似性高,分离株变异小、源于同一克隆系,Met-203、Ile-257型菌株致病力强,是江淮地区猪丹毒发生与流行的主要致病菌型。     

Serological and pathogenic characterization of Erysipelothrix rhusiopathiae isolates from two human cases of endocarditis in Japan

We characterized the serological and pathogenic properties of two Erysipelothrix rhusiopathiae isolates from human cases of infective endocarditis in Japan. One isolate was recovered from a fisherman, and was identified as serovar 3, which is known to be prevalent among fish isolates. This strain exhibited high virulence in mice but was avirulent in swine. Another was untypable, and avirulent in both mice and swine. Our results suggest that various serological and athogenical types of E. rhusiopathiae can induce human endocarditis. This is the first report to characterize the pathogenicity of E. rhusiopathiae isolates from human endocarditis.     

Localization of viable bacteria and bacterial antigens in arthritic joints of Erysipelothrix rhusiopathiae-infected pigs

Abstract Chronic polyarthritis was induced in pigs by infection with Erysipelothrix rhusiopathiae (serovar 2, strain T28). Viable bacteria could be reisolated as long as 5 months post-infection from synovial fluid, synovial tissue and from isolated chondrocytes. The number of viable bacteria could be increased by hypotonic shock of the chondrocytes indicating a substantial intracellular amount of bacteria. Bacterial antigens were shown by immunohistochemistry to be present on the surface of both chondrocytes and synovial cells in arthritic joints. Neither viable bacteria nor bacterial antigen were detected in unaffected joints.     

The aerobic psychrotrophic populations on meat and meat contact surfaces in a meat production system and on meat stored at chill temperatures

At a city abattoir, a wholesaler and 10 different supermarkets, surface microbiological samples were taken of carcasses, hands and apron fronts of members of staff and equipment (mincers and saws). In addition, minced meat, packaged and displayed in chilled cabinets, was also sampled. Carcasses, personnel surfaces and equipment were monitored by a modified agar sausage technique. From each of the highest dilution psychrotrophic plate counts, five colonies were selected randomly, isolated and identified (1265 in total). Microbes developing on chilled meat were also isolated from other surfaces in the production chain. On chilled meat (51%) and at the abattoir (36%) pseudomonads were the predominant organisms followed by the Gram-positive cocci on chilled meat and by Acinetobacter, Moraxella and Alcaligenes spp. at the abattoir. At the wholesaler Gram-positive cocci (32%) predominated, followed by Alcaligenes, Moraxella and Alcaligenes spp. Pseudomonas, Alcaligenes, Neisseriaceae and related genera, Gram-positive cocci, species from the coryneform groups of bacteria and yeasts were identified from all the surfaces monitored. Identification with the API NE20 was unsatisfactory. Enterbacteriaceae, lactobacilli and endospore-forming bacteria were identified occasionally, but their significance as contaminating organisms seems low. No Salmonella spp. were identified. and accepted 25 July 1989