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Endocarditis caused by Actinobacillus actinomycetemcomitans.

Actinobacillus actinomycetemcomitans is a gram-negative coccobacillus which is a very rare cause of bacterial endocarditis. Preexisting cardiac lesions are a main contributing factor, and antibiotic prophylaxis has long been felt necessary before dental or other manipulation to prevent endocarditis. Penicillin in combination with an aminoglycoside has been the most often used treatment regimen. We present a case of endocarditis caused by this organism which developed after antibiotic prophylaxis for dental cleaning. Streptomycin and rifampin therapy resulted in the cure of the infection. The treatment and epidemiology of Actinobacillus endocarditis are reviewed.

Actinobacillus Infections↗

Qualitative and quantitative determination of enterobacterial common antigen (ECA) with monoclonal antibodies: expression of ECA by two Actinobacillus species.

The presence and quantity of the enterobacterial common antigen (ECA) in several species belonging to the family Enterobacteriaceae as well as to other gram-negative families were determined by a solid-phase enzyme-linked immunosorbent assay system and Western blotting by using mouse monoclonal antibodies specific for ECA. Except for Erwinia chrysanthemi, previously known to be an exception, all species known or presumed to belong to Enterobacteriaceae produced ECA (89 of 90 species). Most species not belonging to Enterobacteriaceae did not produce ECA (25 of 28 species), with one already known (Plesiomonas shigelloides) and two hitherto unknown (Actinobacillus equuli and Actinobacillus suis) exceptions. Interestingly, all strains of P. shigelloides produced ECA, regardless of the presence of the Shigella sonnei cross-reacting O antigen. Quantitation of the amount of ECA in members of the family Enterobacteriaceae revealed a remarkable heterogeneity among genera and species as well as within one species. We conclude that the rapid, sensitive, and reliable determination of ECA is a useful aid in taxonomic classification and may help to characterize the relatedness of the family Enterobacteriaceae to other families. However, a quantitative analysis of ECA appears to be without value for these purposes.

Actinobacillus↗

Prevalence of seroreactors to the 104-kilodalton hemolysin of Actinobacillus pleuropneumoniae in swine herds.

A rabbit homologous polyclonal antiserum to the 104-kilodalton hemolysin of Actinobacillus pleuropneumoniae serotype 1 strain CM-5 was specifically produced and used in an antigen capture enzyme-linked immunosorbent assay (ELISA) to detect swine serum antibodies to this potentially important virulence factor. Sera from pigs experimentally infected with the most common disease-producing serotypes (serotypes 1, 2, 5, and 7) of A. pleuropneumoniae produced positive results in this ELISA. Of 144 serum samples collected from 10 herds free of pleuropneumonia and 155 serum samples from 11 herds with a history of the disease, 68 (47%) and 148 (95%), respectively, were found positive by the ELISA. In addition, pigs naturally infected with Actinobacillus suis produced antibodies which seroreacted in this ELISA. The results indicated that a high proportion of swine have antibodies seroreactive with the 104-kilodalton hemolysin produced by A. pleuropneumoniae.

Actinobacillus↗

Molecular cloning and sequencing of the aroA gene from Actinobacillus pleuropneumoniae and its use in a PCR assay for rapid identification.

The gene (aroA) of Actinobacillus pleuropneumoniae, serotype 2, encoding 5-enolpyruvylshikimate-3-phosphate synthase was cloned by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829, and the nucleotide sequence was determined. A pair of primers from the 5' and 3' termini were selected to be the basis for development of a specific PCR assay. A DNA fragment of 1,025 bp was amplified from lysed A. pleuropneumoniae serotypes 1 to 12 of biovar 1 or from isolated DNA. No PCR products were detected when chromosomal DNAs from other genera were used as target DNAs; however, a 1,025-bp DNA fragment was amplified when Actinobacillus equuli chromosomal DNA was used as a target, which could be easily differentiated by its NAD independence. The PCR assay developed was very sensitive, with lower detection limits of 12 CFU with A. pleuropneumoniae cells and 0.8 pg with extracted DNA. Specificity and sensitivity make this PCR assay a useful method for the rapid identification and diagnosis of A. pleuropneumoniae infections.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗

Evaluation of a multiplex PCR test for simultaneous identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6.

Serotype-specific DNA regions involved in the biosynthesis of capsular polysaccharides (cps region) were used to develop a multiplex PCR test for the simultaneous species identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6. Primers specific for serotypes 2, 5, and 6 were combined with the already existing species-specific primers used in a PCR test based on the omlA gene. The PCR test was evaluated with serotype reference strains of A. pleuropneumoniae as well as 182 Danish field isolates previously serotyped by latex agglutination or immunodiffusion. For all serologically typeable strains, a complete correspondence was found between the results obtained by the multiplex PCR test and the results obtained by the traditional serotyping methods. Six of eight serologically nontypeable strains could be allocated to a serotype on the basis of the multiplex PCR results. The species specificity of the assay was evaluated with a collection of 93 strains representing 29 different species within the family Pasteurellaceae, as well as species normally found in the respiratory tracts of swine. All of these strains were negative by the multiplex PCR test, including 50 field isolates of the phylogenetically closely related species Actinobacillus lignieresii. When the multiplex PCR test was used to test Danish field strains, it was able to identify the serotypes of approximately 94% of all strains isolated from swine with clinical disease. More than 90% of the isolates that cross-reacted by the latex agglutination test were of serotype 2, 5, or 6. Determination of the serotype by PCR represents a convenient and specific method for the serotyping of A. pleuropneumoniae in diagnostic laboratories.

Actinobacillus pleuropneumoniae↗

Actinobacillus and Pasteurella species isolated from horses with lower airway disease.

Seventy-three bacterial isolates from 65 horses with and without evidence of lower airway disease were identified to assess whether the association with disease was accounted for by a small or large number of species. Just over half (50.5 per cent were Actinobacillus equuli, 17.8 per cent were A suis-like, 11 per cent were Pasteurella pneumotropica, 8.2 per cent were A lignieresii, 6.8 per cent were P haemolytica and 5.5 per cent were P mairii. These results suggest that a range of Actinobacillus and Pasteurella species can be isolated from the lower airways of horses, with many of the isolates being A equuli. Among the horses examined, lower airway inflammation was significantly associated with A suis-like bacteria and A lignieresii. However, these two species constituted too small a proportion of the undifferentiated Actinobacillus/Pasteurella species to explain the association of this group of bacteria with lower airway disease. Since A equuli constituted just over 50 per cent of the group of bacteria it is possible that it too is playing a significant role in lower airway disease.

Actinobacillus↗

The effects of fluoride on the percentage bacterial composition of dental plaque, on caries incidence, and on the in vitro growth of Streptococcus mutans, Actinomyces viscosus, and Actinobacillus sp.

Administration of 250 mug/ml NaF in drinking water to rats reduced both caries incidence (P less than 0.001) and the percentage of S mutans (0.01 greater than P greater than 0.001) although the percentage of Actinobacillus sp. was increased (P less than 0.001). An Actinobacillus sp. and S mutans FA1 both proliferated in NaF broth only if the NaF greater than 10 mug/ml, which was associated with the inhibition of acid production.

Actinobacillus↗

Two Actinobacillus species isolated from laboratory rodents.

A study was made of the cultural and biochemical characteristics of 17 strains of Actinobacillus isolated from laboratory rats and hamsters. 2 types were recognized which differed in their urease reaction and colony colour on blood agar. The results obtained suggest that one was Actinobacillus equuli but the other could not be identified as belonging to a described species. The characteristics of both organisms are such that they could be confused with Pasteurella pneumotropica. One of the types was isolated from tapwater.

Actinobacillus↗

Pasteurella pneumotropica and the prevalence of the AHP (Actinobacillus, Haemophilus, Pasteurella)-group in laboratory animals.

134 bacterial isolates originally identified as Pasteurella pneumotropica were cultured from healthy, ill or dead mice, rats, hamsters, guinea pigs, rabbits and cats originating from 7 conventional laboratory animal facilities. They occurred seldom in pure culture and were found in a variety of organs. Thorough identification (41 criteria) revealed that only 83 isolates (62%) were P. pneumotropica and could be subdivided into 3 biotypes. 3 isolates were P. aerogenes, 1 was P. ureae, 11 (8%) were qualified as Actinobacillus spp. and 13 (10%) as Haemophilus spp. Close relationship of the 3 genera--the 'AHP-group' --made the differentiation difficult. 23 atypical cultures were discarded at the beginning of the study as not belonging to the 'AHP-group'. Two-thirds of isolates were associated with inflammation or suppuration; Haemophilus spp. seemed to be more pathogenic than Pasteurella and Actinobacillus species.

Actinobacillus↗

Recovery of members of the Pasteurella-Actinobacillus-group from guinea pigs.

In the course of post-mortem bacteriological examination of conventional guinea pigs, 88 isolates belonging to the Pasteurella-Actinobacillus-group were recovered from 69 of 279 animals (25%). Most isolates were recovered from pneumonic lung, enteritic jejunum and inflamed mammary gland. No relationship was found between biotype and source of isolation. About 50% of isolates were recovered in pure culture or as the predominant micro-organism. It is concluded that members of the Pasteurella-Actinobacillus-group must be considered potentially pathogenic for guinea pigs.

Actinobacillus↗

In vitro migratory responses of swine neutrophils to actinobacillus pleuropneumoniae.

Swine neutrophils were quantitatively examined for the direct and indirect migratory responses to Actinobacillus pleuropneumoniae (APP) in vitro and the effects of pseudorabies virus (PrV), frequently co-infecting with APP, were also observed. About 30% of swine neutrophils responded to viable APP, while 3.2% of the neutrophils responded to 0.1% casein which served as the control. The migration of APP was not affected by preincubation of neutrophils with PrV, which inhibited the random migration. When the random migration was normalized to 1, the chemotactic indices for APP, opsonized-APP and casein were 64, 70 and 8.5, respectively. Heat-killed APP or E. coli lipopolysaccharide stimulated the production of interleukin-8 activity by adherent peripheral blood mononuclear cells (PBMC). Preincubation of PBMC with PrV inhibited the production of neutrophil attractant activity when stimulated with heat-killed APP. The results suggested that the direct chemotaxis of neutrophils to viable APP might contribute to early infiltration in Actinobacillus pleuropneumonia, and that PrV might inhibit indirect recruitment of neutrophils to infected lungs by compromising the functions of PBMC.

Actinobacillus pleuropneumoniae↗

Serum and gingival fluid antibodies as adjuncts in the diagnosis of Actinobacillus actinomycetemcomitans-associated periodontal disease.

Serum antibody titers to Actinobacillus actinomycetemcomitans were measured in 200 subjects by an enzyme-lined immunosorbent assay (ELISA) using whole microorganisms as antigen. Comparisons were made between titers found in periodontally normal subjects and titers in subjects with localized juvenile periodontitis (LJP), postlocalized juvenile periodontitis, generalized juvenile periodontitis or adult periodontitis. It was found that titers to all three serotypes of A. actinomycetemcomitans were elevated in LJP patients' sera, while serum antibody levels in other diseased groups were not significantly elevated to any of the serotypes. Patient sera were also examined for serum antibody to oral Haemophili previously shown to cross-react with A. actinomycetemcomitans. Similar antibody titers were found in both normal subjects and in patients with various forms of periodontal disease to Haemophilus aphrophilus, H. influenzae and H. parainfluenzae. The A. actinomycetemcomitans antibodies which were elevated in LJP patients could not be correlated with antibody titers to cross-reacting Haemophili, suggesting that these antibodies are A. actinomycetemcomitans-specific. Serum antibody responses in six of the LJP patients were assessed to autologous strains of A. actinomycetemcomitans. Each patient was found to be infected with only a single serotype of A. actinomycetemcomitans, and specific antibodies to the infecting serotype were found in the patients' sera. In families, the LJP patients had significantly elevated IgG, IgA and IgM serum antibody titers to A. actinomycetemcomitans, while the IgG and IgA antibody titers in periodontally normal siblings were at levels comparable to those found in normal subjects. However, IgM serum antibodies were elevated in the periodontally normal siblings of LJP patients suggesting that the formation of IgM antibodies to A. actinomycetemcomitans may precede the clinical appearance of localized juvenile periodontitis. Gingival crevicular fluid and serum antibody levels to A. actinomycetemcomitans were compared in LJP patients. Comparable titers of IgG, IgA and IgM antibodies were found in serum and gingival fluid in most subjects; however, gingival fluid samples sometimes showed higher titers than serum, likely resulting from local antibody synthesis. The value of serum antibody determinations to A. actinomycetemcomitans in the diagnosis of Actinobacillus-associated periodontitis was also assessed. The predictive value of a positive test (significantly elevated anti-A. actinomycetemcomitans IgG) was 86%, while the specificity was 89%.(ABSTRACT TRUNCATED AT 400 WORDS)

Actinobacillus↗

Evidence for the role of highly leukotoxic Actinobacillus actinomycetemcomitans in the pathogenesis of localized juvenile and other forms of early-onset periodontitis.

BACKGROUND: Actinobacillus actinomycetemcomitans leukotoxin is thought to be an important virulence factor in the pathogenesis of localized juvenile and other forms of early-onset periodontitis. Some highly leukotoxic A. actinomycetemcomitans strains produce 10 to 20 times more leukotoxin than other minimally leukotoxic strains. The distribution, clonality, and intrafamilial transmission of highly leukotoxic A. actinomycetemcomitans were examined in order to determine the importance of leukotoxin in the pathogenesis of periodontitis. METHODS: The polymerase chain reaction (PCR) was used to differentiate highly leukotoxic from minimally leukotoxic strains in examining 1,023 fresh A. actinomycetemcomitans isolates and strains from our culture collection. These were obtained from 146 subjects including 71 with localized juvenile periodontitis (LJP), 4 with early-onset periodontitis, 11 with post-localized juvenile periodontitis, 41 with adult periodontitis, and 19 periodontally normal subjects. The arbitrarily primed polymerase chain reaction (AP-PCR) analysis of 30 oral isolates from each of 25 subjects was used to determine the intraoral distribution of A. actinomycetemcomitans clones. AP-PCR was also used to examine the transmission of A. actinomycetemcomitans in 30 members of 6 families. The clonality of 41 highly leukotoxic A. actinomycetemcomitans strains was evaluated by both AP-PCR and ribotyping. RESULTS: Highly leukotoxic A. actinomycetemcomitans was found only in subjects with localized juvenile and early-onset periodontitis. Fifty-five percent of the LJP subjects harbored highly leukotoxic A. actinomycetemcomitans isolates. Seventy-three percent of the A. actinomycetemcomitans isolates in these subjects were highly leukotoxic. Highly leukotoxic A. actinomycetemcomitans infected younger subjects (mean age 13.95 years, range 5 to 28 years) than minimally leukotoxic (mean age 35.47 years, range 6 to 65 years). Most subjects were infected with only one A. actinomycetemcomitans genotype. However, PCR of whole dental plaques and subsequent analysis of up to 130 individual oral isolates suggested a possible shift in A. actinomycetemcomitans over time in that a few subjects harbored both highly leukotoxic and minimally leukotoxic strains. AP-PCR analysis was consistent with intrafamilial A. actinomycetemcomitans transmission. Ribotyping and AP-PCR analysis confirmed a previous report that highly leukotoxic A. actinomycetemcomitans consists of a single clonal type. CONCLUSIONS: This study suggests that localized juvenile and other forms of Actinobacillus-associated periodontitis are primarily associated with the highly leukotoxic clone of A. actinomycetemcomitans.

Actinobacillus Infections↗

Isolation of a hemolytic Actinobacillus from waterfowl.

A previously undescribed species of hemolytic Actinobacillus was isolated from six waterfowl, three with periocular serous exudation and two with airsacculitis and bronchopneumonia. Cultural and biochemical characteristics were compared with those of Actinobacillus and Pasteurella spp, using a numerical technique.

Actinobacillus↗

Isolation of Actinobacillus suis from a Canada goose (Branta canadensis).

Blindness from conjunctivitis caused by Actinobacillus suis was found in a Canada goose (Branta canadensis) from a wildlife refuge in Kentucky. Lesions were not observed elsewhere and other body organs were grossly normal. Actinobacillus spp. in birds is rare and this is apparently the first report of conjunctivitis resulting from this infection in waterfowl.

Actinobacillus Infections↗

Efficacy of florphenicol premix in weanling pigs experimentally infected with Actinobacillus pleuropneumoniae.

The efficacy of a florfenicol premix was studied in weaning pigs experimentally inoculated with Actinobacillus pleuropneumoniae. Twenty five clinically healthy pigs were distributed into 3 groups; group A non-medicated, groups B and C orally medicated with 20 and 40 ppm of florfenicol respectively. The pigs were fed during 12 consecutive days and on day 5 all the groups were challenged with A. pleuropneumoniae serotype 1. All the animals in Group A developed clinical signs. Most of the pigs in the medicated groups maintained a good health status. Postmortem examination revealed severe pleuropneumonia in pigs from the control group and pneumonic lesions in 40% of the animals treated with 20 ppm of florfenicol. Development of pleuropneumonia was prevented in all the pigs medicated with 40 ppm of florfenicol. Actinobacillus pleuropneumoniae was recovered from the lungs of all control animals and from one pig of each of the medicated groups, however, the avidin biotin peroxidase (ABC-P) method detected the presence of the microorganism in all the animals. We demonstrated that medication with feed containing 40 ppm of florfenicol blocked efficiently the signs and lesions caused by A. pleuropneumoniae and increased the daily body weight gain.

Actinobacillus pleuropneumoniae↗

Identification of genes involved in biosynthesis of Actinobacillus pleuropneumoniae serotype 1 O-antigen and biological properties of rough mutants.

Actinobacillus pleuropneumoniae is an important pathogen of swine. Lipopolysaccharide (LPS) has been identified as the major adhesin of A. pleuropneumoniae and it is involved in adherence to porcine respiratory tract cells. We previously generated seven rough LPS mutants of A. pleuropneumoniae serotype 1 by using a mini-Tn10 transposon mutagenesis system [Rioux S, Galarneau C, Harel J et al. Isolation and characterization of mini-Tn10 lipopolysaccharide mutants of Actinobacillus pleuropneumoniae serotype 1. Can J Microbiol 1999; 45: 1017-1026]. The purpose of the present study was to characterize these mutants in order to learn more about LPS O-antigen biosynthesis genes and their organization in A. pleuropneumoniae, and to determine the surface properties and virulence in pigs of these isogenic mutants. By mini-Tn10 insertions in rough mutants, four putative genes (ORF12, ORF16, ORF17, and ORF18) involved in O-antigen biosynthesis in A. pleuropneumoniae serotype 1 were found within a region of 18 ORFs. This region is homologous to the gene cluster of serotype-specific O-polysaccharide biosynthesis from A. actinomycetemcomitans strain Y4 (serotype b). Two mutants showed homology to a protein with identity to glycosyltransferases (ORF12); two others had the mini-Tn10 insertion localized in genes encoding for two distinct proteins with identity to rhamnosyltransferases (ORF16 and ORF17) and three showed homology to a protein which is known to initiate polysaccharide synthesis (ORF18). These four ORFs were also present in A. pleuropneumoniae serotypes 9 and 11 that express an O-antigen that serologically cross-reacts with serotype 1. Evaluation of some biological properties of rough mutants seems to indicate that the absence of O-chains does not appear to have an influence on the virulence of the bacteria in pigs and on the overall surface hydrophobicity, charge and hemoglobin-binding activity, or on LAL activation. An acapsular mutant was included in the present study in order to compare the influence of O-chains and capsule polysaccharides on different cell surface properties. Our data suggest that capsular polysaccharides and not O-chains polysaccharides have a major influence on surface properties of A. pleuropneumoniae serotype 1 and its virulence in pigs.

Actinobacillus pleuropneumoniae↗

Stimulation and suppression of the oxygenation activity of porcine pulmonary alveolar macrophages by Actinobacillus pleuropneumoniae and its metabolites.

The effects of Actinobacillus (Haemophilus) pleuropneumoniae and its metabolites on the oxygenation activity of porcine pulmonary alveolar macrophages (PAM) were studied, using a chemiluminescence technique. Actinobacillus pleuropneumoniae strains of serotypes 2, 3, and 9 in a dose of 1, 10, and 100 colony-forming units/macrophage first stimulated the oxygen radical production of PAM. After having reached a peak value, oxygenation activity decreased, finally resulting in total suppression of PAM. All these effects were neutralized by homologous convalescent pig sera that had been adsorbed onto inactivated A pleuropneumoniae strains. Moreover, cross-neutralization was shown between serotypes 2 and 3. Inactivated A pleuropneumoniae strains did not influence the oxidative activity of PAM. Undiluted and lower dilutions of sterile A pleuropneumoniae culture supernatants were toxic for PAM, whereas higher dilutions of the supernatants stimulated oxygen radical production of the macrophages. These effects were heat-sensitive and were neutralized by homologous convalescent pig sera. Cross-neutralization was shown between serotypes 2 and 3. These findings indicated that stimulation and inhibition of the oxygenation activity of PAM are attributable to heat-sensitive metabolites produced by A pleuropneumoniae.

Actinobacillus pleuropneumoniae↗