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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.

Abattoirs↗

Endocarditis of the aortic valve in the dog.

The clinical features of endocarditis of the aortic valve in 24 dogs were reviewed. This condition was found most commonly in large-breed, middle-aged male dogs. Evidence of antecedent infection or immunosuppression was usually not historically verified or found at necropsy. However, an association with congenital heart disease, especially discrete subaortic stenosis, was demonstrated. The most frequent clinical findings were systolic and diastolic murmurs and bounding arterial pulses, with or without signs of congestive heart failure. The most commonly isolated organisms were Corynebacterium sp, Erysipelothrix rhusiopathiae, and Streptococcus sp. In addition to antibiotic therapy, treatment for congestive heart failure often was required. Despite aggressive therapy, most affected dogs died as a result of congestive heart failure, arrhythmias, infarction, sepsis, or renal failure.

Animals↗

[The virulence of erysipelothrix rhusiopathiae strains and their neuraminidase production (author's transl)].

We studied the virulence of 20 Erysipelothrix rhusiopathiae strains in albino mice and investigated the interdependence between virulence, growth rate, and neuraminidase production. The minimal number of germs, lethal to mice (LD100) varied among the strains between 10(1) and mor ethan 10(7) per animal. The course of the disease was also very different. Some strains killed the mice rapidly within 3 days. In other strains, however, even a very high number of bacteria caused death only 7 days after infection or later, in which cases the disease became manifest long in advance by ruffled fur and glutinous eyes. A significant correlation was found between the average neuraminidase production in different media and the virulence (p equal to 99%; Fig. 2). We found a less marked dependence between growth rate and virulence (Fig. 3). Neuraminidase activity and growth rate seem to influence virulence rather independently. This is concluded from the fact, that some virulent strains are characterized by quick growth and by producing either no neuraminidase or in small quantities only. On the other hand, neuraminidase production cannot be separated completely from growth rate. An induction of neuraminidas activity in vivo was not observed, even when the bacteria persisted in the mice over a long period of time. We therefore conclude that a short term enzyme induction is not probable.

Enzyme Induction↗

[100 years of erysipelas prophylaxis: significance and reduction of animal experiments].

The history of erysipelas prophylaxis began in 1882 when Pasteur first discovered the attenuating effect of rabbit passages on the erysipelas bacterium. Ten years later, the German veterinarian Lorenz demonstrated the protecting effect of erysipelas antiserum. He developed a method of serovaccination which was successfully used in Germany for more than 50 years. Both scientists employed laboratory animals for the development of their live vaccines. Lorenz additionally recommended an animal model with grey mice to control the potency of erysipelas sera. In 1944, Fortner and Dinter published the results of their investigation on a skin scarification test in swine. This modus of infection was the basis of the first reliable model for efficacy testing of erysipelas vaccines in domestic animals. Shortly after World War II, the first inactivated erysipelas vaccines were being developed. At that time, also a strict quality control was introduced for this product group which required extensive animal experiments in laboratory mice and pigs for the determination of efficacy. WHO established International Standards for erysipelas vaccines and antisera concerning potency testing in mice. These animal models were finally incorporated in pharmacopoeia monographs. Animal experiments have played an important role in the development and quality control of erysipelas vaccines. And the success of this quality control based on animal experiments has had a significant impact on the quality control systems for veterinary vaccines in general. Today, we have a far more detailed knowledge about pathogenesis and immunology of swine erysipelas. This knowledge now allows the introduction of alternative methods according to the 3R concept. With these new methods, animal numbers can be decreased and suffering caused by challenge infection can be reduced. The ultimate goal, i.e. quality control of erysipelas vaccines carried out without routine performance of animal experiments, should be achieved in the near future.

Animal Testing Alternatives↗

HOG CHOLERA. II. RELIABILITY OF THE AGAR DOUBLE DIFFUSION PRECIPITATION TEST FOR THE DIFFERENTIATION OF H.C. VIRUS FROM OTHER INFECTIOUS AGENTS IN SWINE TISSUE.

The specificity in the agar diffusion precipitation test of the reaction between the antigen of hog cholera virus diffusing from infected tissues and its homologous antibody was verified. Alternate freezing and thawing of infected tissues was found to give optimum release of the antigen from fresh tissue frozen for 18 hours. A study of the effect of the size and age of pigs upon the diffusion of the antigen from tissues showed that tissues from pigs of less than 250 lbs. gave good results provided the tissues were from animals showing gross clinical manifestations. Specimens from infected breeding sows and dead animals usually did not give a reaction.

Agar↗

Properties of repeat domain found in a novel protective antigen, SpaA, of Erysipelothrix rhusiopathiae.

Erysipelothrix rhusiopathiae is a small gram-positive rod bacterium that causes erysipelas in swine and a variety of diseases in other animals and humans. Although live-attenuated or bacterin vaccines are effective in protecting against erysipelas, the genetic construction of their active antigen has not been identified. To clarify the surface antigen(s) involved in protective and arthritic response, using monoclonal antibody I2A against the surface proteins of E. rhusiopathiae, we identified a protective antigen, which consists of 606 amino acids. Analysis of deletion derivatives of the gene, spaA(surface protective antigen), showed that the SpaA protein binds tightly to the bacterial cell surface via eight repeat units with a GW-module consisting of 20 amino acids at the C-terminus. Although DeltaSpaA lacking their repeat units lost its ability to induce protection against E. rhusiopathiae infection, intact SpaA protein showed the protection. We conclude that the presence of repeat units is essential both for the binding of SpaA to the bacterial cell surface and for protection. We believe that the repeat region at the C-terminus should be a candidate for a subunit vaccine against erysipelas.

Amino Acid Sequence↗

Home intravenous antibiotic therapy for patients with infective endocarditis.

Although home intravenous antibiotic therapy (HIAT) is increasingly being used for various infectious diseases, outpatient treatment of infective endocarditis (IE) is still uncommon. Recently, the American Heart Association recommended outpatient treatment of endocarditis only for infections with streptococci that are highly susceptible to penicillin. Herein, the experience with HIAT in patients with IE due to a diversity of pathogens is presented. During a 3-year period, 37 patients with IE who were in a stable condition and were cooperative were enrolled in a service for HIAT after completion of diagnostic procedures. Of the 37 patients, 21 were male; mean age was 64.3 years (range 20-87 years); in most cases (26/37), IE involved a native valve. Causative organisms were Streptococcus spp. (20), Staphylococcus spp. (10), Enterococcus spp. (2), Enterobacter spp. (1), and Erysipelothrix rhusiopathiae (1), while three were unknown. The most common antibiotics used were ceftriaxone and vancomycin. Almost three-quarters of the intravenous lines were peripheral. The mean duration of HIAT was 26.2 +/- 8.5 days, with 92% of the patients cured by it. Most complications were minor. Six patients were rehospitalised and two of them required valve replacement. In half of the rehospitalised patients, the complication was unrelated to HIAT. Surprisingly, almost all of the complications necessitating rehospitalisation occurred in patients with streptococcal IE and most involved native valves. HIAT may be suitable for IE due to a diversity of pathogens and involving prosthetic as well as native heart valves, provided there are proper patient and antibiotic selections, good follow-up, and vigilant monitoring of complications.

Adult↗

Alpha-mannosidase: a rapid test for identification of Arcanobacterium haemolyticum.

A 4-h alpha-mannosidase test for identification of Arcanobacterium haemolyticum strains (n = 139) and differentiation of A. haemolyticum from Actinomyces pyogenes strains (n = 30) and other gram-positive rods was evaluated. Practically all A. haemolyticum strains (138 of 139) and the Listeria monocytogenes type strain were alpha-mannosidase positive, while all A. pyogenes strains and Corynebacterium (n = .25) strains as well as the Rhodococcus equi and Erysipelothrix rhusiopathiae type strains were negative. The rapid alpha-mannosidase test, in conjunction with a Gram stain and catalase and reverse CAMP tests, was useful in identification of A. haemolyticum and in differentiation of A. haemolyticum from A. pyogenes and Corynebacterium spp.

Actinomyces↗

Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates: initial development of in vitro susceptibility test methods.

Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine. This report summarizes studies to establish standardized susceptibility testing methods, interpretive criteria, and reagent details for use in veterinary methods recently developed by the National Committee for Clinical Laboratory Standards (NCCLS) (standards M31-A and M37-A, NCCLS, Wayne, Pa., 1999). A total of 636 fastidious and nonfastidious animal and human pathogens were processed by using media and procedures described by the NCCLS. Tiamulin disk diffusion tests used a 30-microg disk concentration, and the proposed MIC breakpoints corresponding to levels achievable in animal target tissues (lung) were < or =4 microg/ml for susceptibility and > or =32 microg/ml for resistance. Correlate zone diameters for specific nonfastidious species were as follows: for Pasteurella multocida and staphylococci tested on Mueller-Hinton agar, susceptibility at > or =19 mm and resistance at < or =11 mm, and for Actinobacillus suis, Erysipelothrix rhusiopathiae, and Streptococcus suis tested on enriched chocolate Mueller-Hinton agar, susceptibility at > or =16 mm and resistance at < or =8 mm. When Actinobacillus pleuropneumoniae was tested, a susceptibility breakpoint of < or =16 microg/ml (> or =9 mm) was suggested for veterinary fastidious medium broth and enriched chocolate Mueller-Hinton agar. Absolute categorical agreement between NCCLS dilution and disk diffusion test results with these criteria ranged from 90.5 to 96.2%. Tiamulin susceptibility testing methods appear to be accurate in their categorical classification for indicated species, and their availability will allow immediate testing of animal isolates to guide therapy via appropriate levels of dosing and to monitor the development of resistance for agents in this unique class.

Humans↗

[Erysipeloid on the islands of the Ohhotsk Sea. 2. Landscape types of the natural foci of erysipeloid].

Cases of skin (skin-artericular) form of erysipeloid were recorded in the islands of the Sea of Okhotsk. The natural foci of the causative agent of this infection were polyhostal and polyvectoral in character. The causative agent of erysipeloid exists among the animals habitating on land and sea. Mass species of animals characteristic of the island landscape served as the sources of infection. Their four landscape types (mountaineous-taiga, of sea coast and rocks, anthropurgic settlement, and of water bodies--salt and freshwater) were preliminarily distinguished by the combination of biocenological, epidemiological, and epizootological peculiarities of natural erysipeloid foci.

Animals↗

Enzyme-linked immunosorbent assay employing a recombinant antigen for detection of protective antibody against swine erysipelas.

The specificities and sensitivities of five recombinant proteins of the surface protective antigen (SpaA) of Erysipelothrix rhusiopathiae were examined by indirect enzyme-linked immunosorbent assay (ELISA) with the aim of developing a reliable serological test for the detection of protective antibody against E. rhusiopathiae. Fully mature protein and the N-terminal 416 amino acids (SpaA416) showed sufficient antigenicities, and further examination was done with SpaA416 because of its higher yield. The antibody titers of pigs experimentally immunized with commercial live vaccine and two types of inactivated vaccines clearly increased after immunization, and all pigs were completely protected against challenge with virulent strains. On the other hand, the antibody titers of nonimmunized control pigs remained very low until they were challenged, and all showed severe symptoms or subsequently died. Interference with the production of antibody against live vaccine by maternal antibody or porcine respiratory and reproductive syndrome virus infection 1 week after vaccination was also clearly detected. Because the ELISA titer correlated well with the protection results, the specificity and sensitivity of the ELISA were further evaluated with sera collected from pigs reared on 1 farm on which animals had acute septicemia, 2 farms on which the animals were infected or free from infection, and 10 farms on which the animals were vaccinated with live vaccine, among others. The ELISA titers clearly revealed the conditions of the herds. These results indicate that the SpaA416 ELISA is an effective method not only for evaluating pigs for the presence of protective antibody levels resulting from vaccination or maternal antibody but also for detecting antibody produced by natural infection. This test has important potential for the effective control of swine erysipelas.

Animals↗

An outbreak of erysipelas in 2-day-old poults.

Systemic erysipelas infection was seen in 2-to-4-day-old poults from three separate ranches owned by the same company. The affected poults were all from the same breeder source; poults from other breeder sources were seemingly unaffected. Mortality increased on days 2 and 3, ranging from 2% to 8.5%. Birds submitted were dehydrated and very weak, with one half of the poults submitted having died during transport to the lab. Gross lesions included swollen, congested livers and spleens, as well as hemorrhagic breast muscle in one case. Toes were swollen and reddish-purple in color. The poults had been toe-trimmed during hatchery processing using a commercial microwave. Histologically, periportal inflammation with heterophilic infiltration in the liver was noted. Spleens showed hyalinization of arteries, lymphoid depletion, and necrosis. Toe joints showed purulent synovitis and cellulitis. Gram stains done on impression smears of liver and spleen showed rare to moderate numbers of small gram-positive rods. Erysipelothrix rhusiopathiae was isolated from 18 of 22 livers cultured, five of six toe joints cultured, and from the yolk sac in two birds.

Animals↗

Grouping of some clinically relevant gram-positive rods by automated fatty acid analysis. Diagnostic implications.

One hundred and ninety-four strains of aerobically growing Gram-positive rods of the genera Corynebacterium, Actinomyces, Arcanobacterium, Erysipelothrix, Listeria, Oerskovia, Nocardia, Rhodococcus, and of unnamed Center for Disease Control (CDC) groups were checked for cellular fatty acid profiles with the microbial Identification System (Microbial ID, Newark, Del., USA). In order to obtain unified data usable for the clinical laboratory, 24 or 48 h sheep blood agar cultures were used. It was thought that grouping and perhaps identification could be aided by this approach. With the aid of numerical analysis, four groups (two consisting of two subgroups each) were established. The discriminatory ability of the scheme, however, was only 77.2%, indicating that grouping of an unknown isolate could be done with some accuracy, but that speciation would still require biochemical testing.

Bacterial Infections↗

Bulleidia extructa gen. nov., sp. nov., isolated from the oral cavity.

Five strains of anaerobic non-sporing Gram-positive bacilli isolated from advanced periodontitis (four strains) and a dentoalveolar abscess (one strain) that did not correspond to existing species were subjected to phenotypic and genetic characterization. Following 16S rDNA sequence analysis, they were found to constitute a novel branch of the low G+C Gram-positive division of the phylogenetic tree related to Erysipelothrix rhusiopathiae and Holdemania filiformis. A new genus Bulleidia, and the species Bulleidia extructa, are proposed. Growth of B. extructa in broth media was poor but was enhanced by the addition of fructose, glucose or maltose together with Tween 80. Glucose and maltose were fermented and arginine was hydrolysed. Acetate, lactate and trace amounts of succinate were the end products of glucose fermentation. The G+C content of the DNA of the type strain is 38 mol%. The type strain of Bulleidia extructa is DSM 13220T.

Bacteria, Anaerobic↗

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.

Animals↗

In vitro antibacterial activity of LY333328, a new semisynthetic glycopeptide.

LY333328 is a semisynthetic N-alkyl derivative of LY264826, a naturally occurring structural analog of vancomycin. LY333328 was evaluated for its in vitro inhibitory and bactericidal activities in comparison with those of the two currently available glycopeptides (vancomycin and teicoplanin). Glycopeptide-susceptible test strains included a total of 311 isolates (most of clinical origin) from the genera Staphylococcus, Enterococcus, Streptococcus, Aerococcus, Gemella, Lactococcus, Listeria, Corynebacterium, and Clostridium. Test strains resistant or intermediate to vancomycin and/or teicoplanin included 56 clinical isolates of Enterococcus (of the VanA, VanB, and VanC phenotypes) and 32 clinical isolates of Staphylococcus (S. haemolyticus, S. epidermidis, and S. aureus), 31 strains of gram-positive genera outside the spectrum of activity of vancomycin (Leuconostoc, Pediococcus, Lactobacillus, and Erysipelothrix), and laboratory-derived organisms obtained after exposure of susceptible Staphylococcus isolates to teicoplanin (6 strains) or laboratory-derived organisms with resistance determinants received from VanA enterococci (2 Enterococcus and 25 Listeria transconjugants). LY333328 was highly active against staphylococci, enterococci, and listeriae (whether they were clinical or laboratory-derived strains) resistant to the currently available glycopeptides. In particular, the MICs of LY333328 did not vary substantially between teicoplanin-susceptible and teicoplanin-resistant staphylococci and between vancomycin-susceptible and vancomycin-resistant enterococci. LY333328 demonstrated fairly good inhibitory activity even against most strains of Leuconostoc, Pediococcus, and Erysipelothrix (MIC range, 1 to 8 microg/ml), whereas it proved less active (although much more active than vancomycin or teicoplanin) against Lactobacillus strains. In minimal bactericidal concentration (MBC) and time-kill studies, LY333328 demonstrated excellent bactericidal activity; enterococci, in particular, which were largely tolerant of vancomycin and teicoplanin, were uniformly killed by LY333328, with MBC-to-MIC ratios of 4 to 8 for most vancomycin-susceptible and vancomycin-resistant strains. In attempts to select for resistant clones, no survivors stably growing in the presence of 10 microg of LY333328 per ml were obtained from the Staphylococcus and Enterococcus test strains exposed to the drug.

Anti-Bacterial Agents↗

Bacterial identity and characteristics in healthy and unhealthy respiratory tracts of sheep and calves.

The aim of this study was to compare different bacteriological aspects of the respiratory systems of healthy (H) versus unhealthy (UH) animals with respiratory signs. The prevalence of different bacterial species was determined in the upper and lower respiratory tract of H and UH Najdi sheep, Somali sheep and Holstein calves. The characteristics of Pasteurella spp. isolates, and the biotype of Pasteurella haemolytica were identified in H and UH animals, Eighteen out of 28 (64.3%) of the identified bacterial species in the upper respiratory tract were more prevalent in the nasal cavities of UH Najdi and Somali sheep and Holstein calves with respiratory signs than in apparently healthy animals; four of the most prevalent bacteria in the upper respiratory system of UH sheep were Moraxella spp., Pseudomonas pseudomallei, Erysipelothrix spp., Pasteurella multocida, while three of the most prevalent bacteria in UH calves were Pasteurella haemolytica, Actinomyces spp., and Pseudomonas aeruginosa. The prevalence of six different bacterial species was greater in the lungs of UH animals, namely Actinomyces pyogenes, Erysipelothrix spp., P. haemolytica, Pasteurella ureae, Staphylococcus aureus, and Staphylococcus epidermidis, which could be risk factors in the complexity of the prevalent respiratory diseases of the animals surveyed. Of the biochemical, cytological and colonial characteristics studied in the identified P. haemolytica and P. multocida, two characters were significantly different (p < 0.05) in organisms isolated from UH as compared to those from H animals. These were the higher loss of haemolytic power by the strains of P. haemolytica and the decreased fermentation of trehalose by all the strains of P. multocida recovered from healthy animals. The only biotype of P. haemolytica isolated from H animals was biotype A, while both biotypes A (88.0% of the isolates) and T (12.0% of the isolates) were recovered from UH animals.

Animals↗

Quality control of inactivated erysipelas vaccines: results of an international collaborative study to establish a new regulatory test.

According to the specifications of the European Pharmacopoeia (Ph. Eur.) monograph (Swine Erysipelas Vaccine (Inactivated), Monograph no. 64, European Pharmacopoeia, 3rd edn., 1997) on erysipelas vaccines for veterinary use, batch potency is estimated in a multi-dilution assay after immunisation and infection of mice. Recently, we described a serological assay system (ELISA) which has the potential to replace this challenge-based model (Beckmann R, Cussler K. Wirksamkeitsprüfung von Rotlaufimpfstoffen an der Labormaus. ELISA kontra Infektionsversuch. ALTEX 1994;Suppl. 1:39-45; Rosskopf-Streicher U, Johannes S, Hausleithner D, Gyra H, Cussler K. Suitability of an ELISA for the batch potency test in laboratory mice. Pharmeuropa BIO 1998;1:65-70). The humoral immune response is quantified in pooled sera of ten mice three weeks after immunisation. The results are expressed as relative potency (RP) in comparison to a reference serum. After a pre-validation study had been performed with success (Rosskopf-Streicher U, Johannes S, Wilhelm M, Gyra H, Cussler K. Potency testing of swine erysipelas vaccines by serology - results of a pre-validation study. ALTEX 1999;16:123-8), we initiated an international collaborative study with five European manufacturers and seven regulatory authorities to validate the assay and model. All participants were provided with blind-coded erysipelas vaccines of different potencies, the ELISA kit and test instructions. The participants had to immunise mice, to prepare serum samples and to perform the ELISA. Inter-laboratory reproducibility was reported by the pass/fail criteria of the vaccines under test. Intra-laboratory precision was assessed by comparing repeated measurements on three consecutive days. Day-to-day variation within the laboratories was statistically analysed by comparing pairs of RPs using Lin's concordance correlation coefficient. The results show that the ELISA is indeed a suitable alternative to replace the vaccination-challenge test. Furthermore, this new model reduces the number of animals required for the potency test by approximately 80%.

Analysis of Variance↗