Actinobacillus actinomycetemcomitans, a rare human pathogen, was repeatedly isolated from the blood of a 20-year-old male patient with patent ductus arteriosus who developed subacute bacterial endocarditis. Difficulties in isolating and identifying the organism are discussed. The bacterial isolate was found to be susceptible to various antimicrobial agents.
Actinobacillus/isolation & purification* ; Adult ; Blood/microbiology* ; Case Report ; Endocarditis, Subacute Bacterial/microbiology* ; Human ; Male ; Septicemia/microbiology

The potential of succinic acid as an important chemical intermediates had been realized and fermentation is one of the best ways to make it possible in economical aspect. Fermentation organism is the key part of the fermentation method. The updated research developments of fermentation organisms and the fermentation characteristics and problems of them were reviewed and analyzed in this paper. Finally,the development future of fermenation organism was forecasted.
Actinobacillus ; metabolism ; Anaerobiospirillum ; metabolism ; Bioreactors ; microbiology ; Escherichia coli ; metabolism ; Fermentation ; Genetic Engineering ; methods ; Industrial Microbiology ; methods ; trends ; Succinic Acid ; isolation & purification ; metabolism

Swine respiratory diseases induce severe economic losses in the swine industry worldwide. Several methods have been developed and applied to control these diseases. However, there are still problems of disease control in the swine industry. Recently, egg yolk antibodies have been found to offer several advantages for disease control in animals and humans. In a previous study (24), antibodies to several causative pathogens of swine respiratory diseases were developed. However, several problems remained, especially in terms of reduced laying rates. Therefore, experimental vaccines were reformulated with various bacterial antigens of the swine respiratory diseases. After immunizing hens with the antigens, antibody profiles and other effects including laying rates were investigated and compared to those of the previous study. Profiles of antibody titers were very similar with those of the previous study. However, side effects, such as depression, weakness, reduction of laying rates and mortality, were dramatically lowered and laying rates were increased in hens injected with certain experimental vaccines. In particular, laying rates of hens injected with vaccines against atrophic rhinitis were increased up to 84% by injecting a vaccine composed of only the DNTs of B. bronchiseptica and P. multocida D:4. Efficacies of the vaccines against swine pneumonic pasteurellosis and pleuropneumonia were very similar with those of the previous study. These results suggest that new vaccines could be effective in the production of egg yolk antibodies against the causative agents of swine respiratory diseases.
Actinobacillus pleuropneumoniae/classification/genetics/*immunology ; Animals ; Antibodies, Bacterial ; Antibody Formation ; Bacterial Outer Membrane Proteins/genetics/isolation & purification ; Bordetella bronchiseptica/classification/genetics/*immunology ; Egg Yolk/microbiology ; Female ; Immunoglobulins/*genetics ; Oviposition ; Pasteurella multocida/classification/genetics/*immunology ; Serotyping ; Swine

OBJECTIVETo establish a 16S rDNA multiplex polymerase chain reaction (PCR) for simultaneously detecting P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens of chronic periodontitis and to study the correlation between different modes of infection and severity of the disease.

METHODSPeriodontal pocket specimens from 152 patients with mild, moderate or advanced chronic periodontitis and gingival sulcus specimens from 30 periodontally healthy individuals were collected and placed in 200 microl lysis buffer. The specimens were incubated in 100 degrees C for 10 min and 10 microl of the supernatant was directly used as PCR template. DNAs from P. gingivalis strain ATCC33277, A. actinomycetemcomitans strain Y4, T. denticola strain FM and E. coli strain DH5alpha were used as positive and negative controls in PCR with all of which were prepared by routing phenol-chloroform method. A multiplex PCR assay, using three sets of primers specific to 16S rDNA genes of the three anaerobes, was developed to detect the specimens. The target amplification fragments from 3 cases of PCR products positive for all the three anaerobes were sequenced after T-A cloning. Chi-square test was applied to analyze the correlation between different coinfection of the three anaerobes and severity of the disease.

RESULTSThe established 16S rDNA multiplex PCR assay was able to detect P. gingivalis, A. actinomycetemcomitans and T. denticola at a minimum of 10, 20 and 20 cells, respectively. In comparison with the reported corresponding sequences, similarities of the nucleotide sequences from the three anaerobes 16S rDNA amplification fragments were as high as 99.45%, 97.08% and 96.59%, respectively. Of the 30 periodontally healthy gingival sulcus specimens, only one (3.3%) positive for P. gingivalis and two (6.7%) for A. actinomycetemcomitans could be identified but all of the rest were negative. In the 152 CP periodontal pocket specimens, 147 cases (96.7%) were positive for P. gingivalis, A. actinomycetemcomitans and/or T. denticola, respectively, and 5 cases (3.3%) were negative for all the three anaerobes. The positive rate of P. gingivalis detection (91.5%, 139/152) was significantly higher than those of A. actinomycetemcomitans (72.4%, 110/152) and T. denticola (80.9%, 123/152) (chi(2) = 7.07, 18.67; P < 0.01). 89.8% of the specimens from patients showed coinfections with two (26.5%) or three anaerobes (63.3%), and the coinfection rates in the specimens from moderate and advanced CP were remarkably higher than that from mild CP (chi(2) = 10.43, P < 0.01).

CONCLUSIONThe 16S rDNA multiplex PCR established in this study showed high sensitivity and specificity, which could be used to detect P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens. CP was an disease caused by multiple pathogenic microbes while the synergistic pathopoiesis of the three microbes was closely related to the severity of the disease.

Actinobacillus Infections ; epidemiology ; microbiology ; Adult ; Aged ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; epidemiology ; microbiology ; China ; epidemiology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; microbiology ; Polymerase Chain Reaction ; methods ; Porphyromonas gingivalis ; isolation & purification ; RNA, Ribosomal, 16S ; Treponema denticola ; isolation & purification ; Treponemal Infections ; microbiology

Oral lichen planus (OLP) is a chronic inflammatory disease that is frequently detected in oral tissues. The aim of our study was to identify the prevalence of the detection of periodontopathogenic microorganisms (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Treponema denticola in OLP patients and to compare with this prevalence of periodontopathogenic microorganisms in healthy non-OLP patients. Our study included 27 (18 chronic periodontitis (OLPP) and 9 gingivitis (OLPG)) patients diagnosed with OLP along with 26 (13 chronic periodontitis (HP) and 13 gingivitis (HG)) healthy non-OLP patients. The multiplex polymerase chain reaction (PCR) with subsequent reverse hybridization method (micro-IDent) was used for identifying periodontopathogenic microorganisms present in subgingival plaque samples. The percentages of detection for A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola in subgingival plaque samples taken from OLP patients (OLPG and OLPP) were 18.5%, 85.1%, 81.4%, 88.8% and 74%, respectively. Meanwhile, in the non-OLP patients (HG and HP), these values were 7.6%, 50%, 46.1%, 73% and 57.7%, respectively. Thus, comparing the non-OLP groups with the OLP groups, the periodontopathogens' percentages of detection in the OLP groups were higher than those in the non-OLP groups. According to our study results, OLP patients have higher levels of infection with A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola than non-OLP patients. We argue that the high percentages in patients with OLP may help identify the importance of periodontopathogenic microorganisms in the progress of periodontal diseases of OLP.
Actinobacillus Infections ; diagnosis ; Adult ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; diagnosis ; Bacteroides ; isolation & purification ; Bacteroides Infections ; diagnosis ; Chronic Periodontitis ; microbiology ; Dental Plaque ; microbiology ; Dental Plaque Index ; Female ; Gingivitis ; microbiology ; Gram-Negative Bacteria ; isolation & purification ; Humans ; Lichen Planus, Oral ; microbiology ; Male ; Middle Aged ; Periodontal Attachment Loss ; microbiology ; Periodontal Index ; Periodontal Pocket ; microbiology ; Porphyromonas gingivalis ; isolation & purification ; Prevotella intermedia ; isolation & purification ; Treponema denticola ; isolation & purification ; Treponemal Infections ; diagnosis

OBJECTIVEThe aim of this study was to investigate subgingival infection frequencies of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans strains with genetic variation in Chinese chronic periodontitis (CP) patients and to evaluate its correlation with clinical parameters.

METHODSTwo multiplex polymerase chain reaction (PCR) assays were developed to detect the 16SrDNA, collagenase (prtC) and fimbria (fimA) genes of P. gingivalis and the 16SrDNA, leukotoxin (lktA) and fimbria-associated protein (fap) genes of A. actinomycetemcomitans in 60 sulcus samples from 30 periodontal healthy subjects and in 122 subgingival plaque samples from 61 patients with CP. The PCR products were further T-A cloned and sent for nucleotide sequence analysis.

RESULTSThe 16SrDNA, prtC and fimA genes of P. gingivalis were detected in 92.6%, 85.2% and 80.3% of the subgingival plaque samples respectively, while the 16SrDNA, lktA and fap genes of A. actinomycetemcomitans were in 84.4%, 75.4% and 50.0% respectively. Nucleotide sequence analysis showed 98.62%~100% homology of the PCR products in these genes with the reported sequences. P. gingivalis strains with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ were predominant in deep pockets (>6 mm) or in sites with attachment loss > or =5 mm than in shallow pockets (3~4 mm) or in sites with attachment loss < or =2 mm (P<0.05). P. gingivalis strains with prtC+/fimA+ also showed higher frequency in gingival index (GI)=3 than in GI=1 group (P<0.05).

CONCLUSIONInfection of P. gingivalis with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ correlates with periodontal destruction of CP in Chinese. Nonetheless P. gingivalis fimA, prtC genes and A. actinomycetemcomitans lktA gene are closely associated with periodontal destruction, while A. actinomycetemcomitans fap gene is not.

Actinobacillus Infections ; epidemiology ; microbiology ; Adult ; Aged ; Aggregatibacter actinomycetemcomitans ; classification ; genetics ; isolation & purification ; Bacteroidaceae Infections ; epidemiology ; microbiology ; China ; epidemiology ; Chronic Disease ; Dental Plaque ; epidemiology ; microbiology ; Female ; Gingivitis ; epidemiology ; microbiology ; Humans ; Male ; Middle Aged ; Periodontitis ; epidemiology ; microbiology ; Porphyromonas gingivalis ; classification ; genetics ; isolation & purification ; Prevalence ; Risk Assessment ; methods ; Risk Factors ; Species Specificity ; Statistics as Topic

Actinobacillus pleuropneumoniae is an important primary pathogen in pigs, in which it causes a highly contagious pleuropneumoniae. In our previous study, apxIA gene amplified from A. pleuropneumoniae Korean isolate by PCR with primer designed based on the N- and C-terminal of the toxin was cloned in TA cloning vector and sequenced. The nucleotide sequences of apxIA gene was reported to GeneBank with the accession numbers of AF363361. Identity of the Apx IA from the cloned gene in E. coli was proved by SDS-PAGE and Western blot. Yeast has been demonstrated to be an excellent host for the expression of recombinant proteins with uses in diagnostics, therapeutics and vaccine productions. Therefore, to use the yeast as a delivery system in new oral subunit vaccine, apxIA gene was subcloned into Saccharomyces cerevisiae, and ientified the expression of Apx IA protein. First, apxIA gene was amplified by PCR with the primers containing BamHI and SalI site at each end. Second, the DNA digested with BamHI and SalI was ligated into YEpGPD-TER vector, and transformed into S. cerevisiae 2805. Third, after identification of the correctly oriented clone, the 120-kDa of Apx IA protein expressed in S. cerevisiae 2805 was identified by SDS-PAGE and Western blot.
Actinobacillus pleuropneumoniae/*genetics/isolation & purification/metabolism ; Animals ; Bacterial Proteins/*biosynthesis/genetics ; Blotting, Western/veterinary ; Cloning, Molecular ; DNA, Bacterial/chemistry/genetics ; Electrophoresis, Polyacrylamide Gel/veterinary ; Hemolysin Proteins ; Pleuropneumonia, Contagious/microbiology ; Polymerase Chain Reaction/veterinary ; Saccharomyces cerevisiae/genetics/*metabolism ; Swine ; Swine Diseases/microbiology