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Human diseases due to A. paraphrophilus aren’t usual. The following case report describes the first isolated case of A. paraphrophilus brain abscess in our laboratory. A 9-year-old boy presented to pediatric emergencies for frontal headache, vomiting, blurred vision and left hemiparesis. Radiological diagnosis consists with a frontal abscess. Gram staining of purulent samples showed abundant neutrophils with gram negative bacilli. Culture was made on blood agar, chocolate agar, Chapman’s agar and incubated in 5% CO2 at 37°C for 24 hours. Positive culture was detected only on chocolate agar. It was monomicrobial with small yellowish non-hemolytic colonies. Gram stain of colonies showed pleomorphic gram-negative coccobacilli. The strain required V factor for growth. The isolated strain was sensitive to all antibiotics tested. The interest of this case is that it shows the emergence of A. paraphrophilus as a causative agent of brain abscesses on pediatric population without associated congenital heart disease. It may also help identifying risk factors of these infections and how to prevent them.

We present a case of endocarditis with embolic stroke and digital infarction due to the recently renamed Aggregatibacter aphrophilus. The isolation and identification of this organism can be problematic but was achieved in this case using both older phenotypic and newer genotypic methods. A benign tongue lesion is suggested as the likely portal of entry for this oropharyngeal organism. The patient made a good recovery with six weeks of intravenous ceftriaxone but will need cardiac valvular surgery at some point in the future.

We report on a 3-year-old boy presenting with left-sided eyelid myocloni due to an intracranial abscess harboring Haemophilus paraphrophilus. This is the first description of an intracranial infection with this pathogen in a child.

The aim of this study was to reinvestigate the relationships and the generic affiliations of the species Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis. The nicotinamide phosphoribosyltransferase gene (nadV) conferring V factor-independent growth was identified in Haemophilus aphrophilus. The gene encodes a polypeptide of 462 amino acids that shows 74.5 % amino acid sequence identity to the corresponding enzyme from Actinobacillus actinomycetemcomitans. Ten isolates of Haemophilus paraphrophilus all carried a nadV pseudogene. DNA from Haemophilus aphrophilus was able to transform Haemophilus paraphrophilus into the NAD-independent phenotype. The transformants carried a full-length nadV inserted in the former locus of the pseudogene. The DNA–DNA relatedness between the type strains of Haemophilus aphrophilus and Haemophilus paraphrophilus was 77 %. We conclude that the division into two species Haemophilus aphrophilus and Haemophilus paraphrophilus is not justified and that Haemophilus paraphrophilus should be considered a later heterotypic synonym of Haemophilus aphrophilus. Forty strains of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus and Haemophilus segnis were investigated by multilocus sequence analysis. The 40 strains form a monophyletic group clearly separate from other evolutionary lineages of the family Pasteurellaceae. We propose the transfer of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus and Haemophilus segnis to a new genus Aggregatibacter gen. nov. as Aggregatibacter actinomycetemcomitans comb. nov. (the type species; type strain ATCC 33384T=CCUG 13227T=CIP 52.106T=DSM 8324T=NCTC 9710T), Aggregatibacter aphrophilus comb. nov. (type strain ATCC 33389T=CCUG 3715T=CIP 70.73T=NCTC 5906T) and Aggregatibacter segnis comb. nov. (type strain HK316T=ATCC 33393T=CCUG 10787T=CCUG 12838T=CIP 103292T=NCTC 10977T). The species of the genus Aggregatibacter are independent of X factor and variably dependent on V factor for growth in vitro.

Chronic granulomatous disease (CGD) is a rare inherited disorder in which phagocytes are incapable of generating bactericidal-reactive oxygen derivatives. Typically these patients are susceptible to life-threatening infections with catalase-producing organisms. Haemophilus species, particularly H. paraphrophilus, are not associated with CGD infections, because these organisms rarely if ever produce catalase. Haemophilus species are part of the indigenous oral microbial flora and, other than H. influenzae, are rarely recognized as pathogens. They are fastidious and require additional growth factors and capnophilic culture conditions for optimal growth and identification. Here we describe three cases of infection with non-H. influenzae (NHI) Haemophilus species in CGD patients. These organisms were catalase-negative and therefore not expected to be virulent in CGD patients, but they were also H(2)O(2) production-negative, thereby negating the putative loss of virulence of being catalase-negative. These are the first reports of NHI Haemophilus species in CGD and reinforce the critical need for careful microbiologic evaluation of infections in CGD patients.

The closely related species Actinobacillus actinomycetemcomitans , Haemophilus aphrophilus , and Haemophilus paraphrophilus are common findings in oral microbiota. The aims of this study were to evaluate the applicability of the Rapid NH and API ZYM kits and arbitrarily primed PCR (AP-PCR) in the identification and differentiation of the three species from each other. The material included 62 clinical isolates and three reference strains of A. actinomycetemcomitans representing the 5 serotypes and 18 AP-PCR genotypes. Haemophilus species included 12 clinical isolates and 11 reference strains of H. aphrophilus , H. paraphrophilus , and 5 other species. For the PCR amplification, the oligonucleotide 5′-CAGCACCCAC-3′ was used as a primer. Contrary to the consistent performance of API ZYM, the Rapid NH system was able to identify only 10 of 65 (15%) A. actinomycetemcomitans isolates, whereas all Haemophilus species were correctly identified. The API ZYM test differentiated A. actinomycetemcomitans from H. aphrophilus and H. paraphrophilus by negative β-galactosidase and α-glucosidase reactions and a positive esterase lipase reaction. However, the API ZYM test was unable to differentiate H. aphrophilus from H. paraphrophilus , it also could not differentiate A. actinomycetemcomitans serotypes from each other. Among the H. aphrophilus isolates three AP-PCR genotypes and among H. paraphrophilus isolates only one AP-PCR genotype, distinct from those of A. actinomycetemcomitans , were found. The Rapid NH test showed poor ability to identify clinical isolates of all A. actinomycetemcomitans serotypes. Moreover, AP-PCR genotyping proved to be a rapid method for the species differentiation of A. actinomycetemcomitans , H. aphrophilus , and H. paraphrophilus .

To define the clinical, microbiological, and therapeutic characteristics of haemophilus endocarditis, we reviewed the charts of 42 adults with haemophilus endocarditis (native valve disease, 37; prosthetic valve disease, five) who were followed up between 1983 and 1995 in France. The mean duration of symptoms before diagnosis was 34 days. The causative Haemophilus species were as follows: H. parainfluenzae (26 adults), H. aphrophilus (9), H. paraphrophilus (4), and H. influenzae (3). According to the Duke criteria, 38 cases of endocarditis were definitive and four were possible. Thirty-nine patients received combination antibacterial therapy and three received therapy with a beta-lactam agent alone (mean duration, 46 days). Arterial embolism occurred in 15 patients. Cardiac surgery was indicated for 18 patients; 16 of these surgeries were performed within 3 months. Two patients died of heart failure. In conclusion, haemophilus endocarditis is rare and is mainly due to H. parainfluenzae. Although surgery is often necessary, haemophilus endocarditis has a favorable prognosis.

Human diseases caused by Haemophilus paraphrophilus (H.paraphrophilus) are unusual. The following case report describes a 67-year-old man who presented with pain and swelling of the right side of the face. Fine needle aspiration suggested a parotid gland abscess. Microbiological studies identified H.paraphrophilus. This is the first time a parotid abscess has been found to be caused by this organism.

Acute appendicitis is a common surgical emergency. The etiology and pathophysiology of appendicitis have been well investigated. Aggregatibacter aphrophilus is a fastidious gram-negative coccobacilli. Detection of this organism in clinical samples and its differentiation from Haemophilus aphrophilus or from Aggregatibacter actinomycetemcomitans in routine microbiology settings could be difficult.In this rare case, we report the isolation of Aggregatibacter aphrophilus from the appendix of a 14-year-old boy presented with acute appendicitis. The genotypic method using 16S rRNA sequencing was used for identification of the organism at species level.This case highlights the importance of detecting fastidious and rare microorganisms such as Aggregatibacter aphrophilus that could be associated with acute appendicitis.

Endophthalmitis due to gram-negative bacilli has been associated with a high degree of vision loss. We report three cases due to the nonenteric gram-negative bacilli Moraxella nonliquefaciens, Haemophilus paraphrophilus, and multidrug-resistant Haemophilus influenzae. The features of these cases are compared with those of other reported cases of endophthalmitis due to unusual nonenteric gram-negative bacilli. Fifty-eight percent of patients had no vision in the affected eye after treatment. Early surgical intervention with vitrectomy and intravitreous antibiotics in addition to parenteral antibiotics should be included in the treatment of endophthalmitis due to gram-negative bacilli.

Chemical analysis by us has indicated that Actinobacillus actinomycetemcomitans is not a homogeneous species. The present study used chemometric methods and a multitude of chemical characters to examine this further. Strains were characterized by cell sugar and fatty acid contents, lysis kinetics during EDTA and EDTA plus lysozyme exposure, methylene blue reduction, and API ZYM enzymatic assessment of whole cells and outer membrane vesicles/fragments. In total, 41 quantitative variables were analyzed from each of 9 strains and treated with principal component analysis and soft independent modeling of class analogy. These methods divided A. actinomycetemcomitans into 2 strain groups. One group contained ATCC 33384, ATCC 29522, FDC 2112 and FDC 2043; the other comprised ATCC 29524, ATCC 29523, FDC 2097, FDC 511 and FDC Y4. With an F-test, the groups (classes) of A. actinomycetemcomitans strains could be distinguished at 95% confidence limits. Both groups were distinct from members of the genera Haemophilus and Pasteurella (Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Pasteurella multocida and Pasteurella haemolytica).

Patients with alcoholic cirrhosis who have ascites have a high risk of developing spontaneous bacterial peritonitis (SBP). The authors report a case of SBP caused by Haemophilus paraphrophilus, the first-reported SBP in literature with this pathogen. Later on, the patient also developed tuberculous (TB) peritonitis associated with thoracic Pott's disease, a combination never reported before. The diagnoses were confirmed by positive mycobacterium cultures of both omental tissues and vertebral tissues. This report also illustrates prominent computed tomography findings of TB peritonitis and magnetic resonance imaging of spinal cord compression of Pott's disease. Tuberculosis is a treatable and curable disease and should be considered as a potential offending pathogen on differential diagnosis in SBP of alcoholic cirrhotic patients. Timely biopsy and surgical intervention with these kinds of TB are needed to lead early diagnosis and result in an excellent outcome.

Aggregatibacter aphrophilus was previously known as Haemophilus aphrophilus and is a rare cause of disease in humans. A recent reclassification of these organisms has placed them in the new genus of Aggregatibacter species. The organism seems to be a normal component of oral flora and has been reported to cause endocarditis, sinusitis, pneumonia, empyema, soft tissue abscess, meningitis, vertebral discitis, and septic arthritis. Brain abscess due to Aggregatibacter is rare. We report a case of cerebral abscess due to Aggregatibacter aphrophilus and discuss the characteristics of this organism.

Introduction Aggregatibacter paraphrophilus (former name Haemophilus paraphrophilus) is a normal commensal of the oral flora. It is a rare cause of hepatobiliary or intracerebral abscesses. Case presentation We report a case of a 53-year-old Caucasian man with a liver abscess and subsequent brain abscesses caused by Aggregatibacter paraphrophilus. The probable source of the infection was the oral flora of our patient following ingestion of a dental filling. The presence of a large patent foramen ovale was a predisposing factor for multifocal abscesses. Conclusion In this case report, we describe an unusual case of a patient with both liver and brain abscesses caused by an oral commensal Aggregatibacter paraphrophilus that can occasionally show significant pathogenic potential.

Haemophilus species are rarely associated with hepatobiliary infections. We report a case of hepatic abscess caused by Haemophilus paraphrophilus and review the English-language literature for reports of infections of the liver and biliary system caused by Haemophilus species. Most patients identified had predisposing conditions. The pathogenesis of hepatobiliary infections due to Haemophilus species may involve ascending spread from the gastrointestinal tract or hematogenous seeding following oropharyngeal colonization.

Outer membrane vesicles and fragments from Actinobacillus actinomycetemcomitans, Actinobacillus lignieresii, Actinobacillus ureae, Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Haemophilus parainfluenzae, Pasteurella haemolytica, and Pasteurella multocida were isolated and examined semiquantitatively for 19 enzyme activities by using the API ZYM micromethod. The enzyme contents of vesicles and fragments were compared with the enzyme contents of whole cells of the same organisms. Enzymic data were analyzed by using principal-component analysis and soft independent modeling of class analogy. This technique allowed us to distinguish among the closely related organisms A. actinomycetemcomitans, H. aphrophilus, and H. paraphrophilus. A. actinomycetemcomitans was divided into two groups of strains. A. lignieresii fell outside or on the border of the A. actinobacillus class. A. ureae, H. influenzae, H. parainfluenzae, P. haemolytica, and P. multocida fell outside the A. actinomycetemcomitans, H. aphrophilus, and H. paraphrophilus classes.

The authors report a case of orbital cellulitis with abscess formation that developed after surgical management of canaliculitis. The patient developed pain, erythema, proptosis, and limited eye movements 3 days after canalicular debridement. She required urgent drainage of an orbital abscess and was treated with intravenous antibiotics for 2 weeks. Complications from surgical management of canaliculitis are unusual. Clinicians should be aware of the rare but serious risk of spread of the infection to the orbit.

Spondylodiscitis caused by Aggregatibacter aphrophilus, formerly known as Haemophilus paraphrophilus, is an unusual condition and can be very difficult to diagnose. We report a case of cervical spondylodiscitis complicated by spinal epidural abscess in a 63-year-old woman, without underlying predisposing conditions. The source of infection was identified as a periodontal infection. The patient was successfully treated with systemic antibiotics.

The taxonomic distinction between Actinobacillus (Haemophilus) actinomycetemcomitans and Haemophilus aphrophilus and the taxonomic distinction between H. aphrophilus and Haemophilus paraphrophilus have been questioned. This study was done to determine whether multivariate statistical analyses of carbohydrate data from lipopolysaccharides could be used to distinguish between these closely related species. Lipopolysaccharides were extracted with phenol-water and purified. Carbohydrates were assessed by using gas chromatography and gas chromatography-mass spectrometry after methanolysis and derivatization with trifluoroacetic acid anhydride. The lipopolysaccharides from all of the species contained rhamnose, fucose, galactose, glucose, L-glycero-D-mannoheptose, and glucosamine plus galactosamine, but in varying amounts. A. actinomycetemcomitans and H. paraphrophilus also contained D-glycero-D-mannoheptose, while H. aphrophilus did not. Sample- and variable-oriented principal-component analyses of the carbohydrate data clearly distinguished among A. actinomycetemcomitans, H. aphrophilus, and H. paraphrophilus. Soft independent modelling of class analogy showed that no sample in the A. actinomycetemcomitans class fell within the 95% confidence limits of the H. aphrophilus class. H. paraphrophilus fell outside both classes.

Many clinical laboratories have difficulty in identifying a group of organisms which are catalase negative, oxidase positive Gram negative rods. We describe a case of purulent sacroiliitis due to Haemophilus paraphrophilus where the organism was initially misidentified as Eikenella corrodens leading to inappropriate antimicrobial chemotherapy. We review the strains of H. paraphrophilus and E. corrodens that were identified by the National Collection of Type Cultures over the last ten years. Only 21 of 100 strains identified as E. corrodens were submitted as E. corrodens . Seven strains submitted as possible E. corredens were identified as H. paraphrophilus . Several different species of Gram negative rods may produce pitting on agar and this seems to be poorly recognised. However, further tests are available to facilitate correct identification of these strains.

We report a case of prosthetic, aortic valve, infective endocarditis caused by Haemophilus paraphrophilus. There are no other cases described in the available literature where this microorganism has caused prosthetic valve endocarditis and no other case reported involving only the aortic valve.

OBJECTIVE To report the first case of Haemophilus paraphrophilus vertebral osteomyelitis--the second reported case of osteomyelitis of any site caused by this organism. CLINICAL FEATURES A 41-year-old male bus driver with no significant previous medical history presented with severe abdominal and back pain, which was eventually localised to the eleventh thoracic vertebra (T11). H. paraphrophilus was isolated from pus aspirated from the vertebral body. INTERVENTIONS AND OUTCOME The patient was treated with penicillin given intravenously for four weeks, then with antibiotics given orally for a further three weeks, with good clinical response. CONCLUSION H. paraphrophilus is an infrequent pathogen which may be difficult to identify and test for antibiotic susceptibility, but can cause serious infection, including primary haematogenous osteomyelitis.

Actinobacillus actinomycetemcomitans, Actinobacillus ureae, Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Haemophilus parainfluenzae, Pasteurella haemolytica and Pasteurella multocida strains were examined by transmission electron microscopy for the presence of bacteriophages. Phages were detected in serotype a (SUNY 75) and e (UOH 1705) and in the fresh clinical isolates UOH Q1243 and UOH Q1247 of A. actinomycetemcomitans. Phages were not found in serotype b, c and d strains of A. actinomycetemcomitans, in the fresh clinical isolate UOH Q1244 of this species or in old strains (including reference strains) of related species from the Actinobacillus-Haemophilus-Pasteurella group.

STUDY DESIGN A case of paraspinal abscess formation from Haemophilus paraphrophilus is presented. OBJECTIVES To describe a case of paraspinal abscess formation from H. paraphrophilus, a fastidious commensal organism of the mouth and pharynx. A precise bacteriologic identification can be difficult; techniques for such identification are discussed. SUMMARY OF BACKGROUND DATA Spinal abscess caused by H. paraphrophilus is unusual and can be very difficult to diagnose. METHODS The etiology, clinical presentation, technical examinations, and treatment are reviewed. RESULTS Prolonged antibiotic treatment was curative, although surgery was considered. CONCLUSIONS Bacteriologic diagnoses in these rare infections are difficult. Antibiotic therapy was curative in the patient described.

Restriction enzyme analysis of PCR-amplified 16S rRNA genes was used to distinguish among clinical isolates of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus which were originally identified by conventional phenotypic methods. This PCR-based method is a reliable and rapid alternative to conventional methods for identification of these bacterial species.

Summary A case of brain abscess involving Haemophilus paraphrophilus and Actinomyces odontolyticus is presented. This combination of organisms has not previously been described. All brain abscess specimens should routinely be processed rapidly and cultured for a prolonged period to ensure recovery of fastidious organisms which may have implications for antibiotic therapy.

The 64-kDa protein to which about half the sera from patients with localized juvenile periodontitis and rapidly progressive periodontitis reacted strongly was purified from Actinobacillus actinomycetemcomitans Y4. Determination of the TV-terminal sequence of the protein revealed that it was a GroEL-like protein. The DNA fragment containing the groEL gene of A. actinomycetemcomitans was amplified by polymerase chain reaction, and the groESL operon was cloned by using colony hybridization with the amplified fragment from A. actinomycetemcomitans chromosomal DNA. Sequence analysis revealed that structures of the operon and its products were typical in gram-negative bacteria. Rabbit polyclonal antibodies to the 64-kDa protein cross-reacted with approximately 65-kDa proteins of Haemophilus aphrophilus, Haemophilus influenzae, Haemophilus paraphrophilus, Escherichia coli and Eikenella corrodens but not with any cellular proteins of Porphyromonas gingivalis, Prevotella intermedia and Fusobacteriuni nucleatum. It is possible that antibodies reactive to the 64-kDa protein in periodontitis patients are induced by the cross-reactivity with the hsp60 proteins of other bacteria.

Endocarditis due to Haemophilus paraphrophilus is an uncommon disease. We report a case of H. paraphrophilus endocarditis with embolic complications in which the causative organism was resistant to beta-lactam antibiotics. Before April 1994, 16 cases of H. paraphrophilus endocarditis had been reported. Infection by this organism usually affects a previously damaged mitral valve. We emphasize the fastidiousness of the organism and the high incidence of embolic complications, which determine the outcome. To our knowledge we describe the first patient with endocarditis due to beta-lactam-resistant H. paraphrophilus.

Diagnosis: Cardiobacterium hominis prosthetic valve endocarditis. C. hominis is one of the fastidious gram-negative bacteria that belong to the HACEK group of organisms (Haemophilus parainfluenzae, Haemophilus aphrophilus, Haemophilus paraphrophilus, Actinobacillus actinomycetemcomitans, C. hominis, Eikenella corrodens, and Kingella kingae). Tucker et al. [1] first reported it as a cause of endocarditis in 1962. The HACEK group are part of the normal oropharyngeal flora. Poor dentition and/ or having undergone dental work within the past 6 months have been described as sources of infection [2]. HACEK organisms cause -3% of cases of infective endocarditis [3]. One study found that it takes, on average, 3.4 days (range, 1-10 days) to grow these organisms with automated blood culture systems [3]. On Gram stain, C. hominis appears as teardropand dumbbell-shaped gram-negative rods. Figure 1 demonstrates the filamentous form of this organism. Classically, one can see a rosette shape on the Gram stain. The other gram-negative HACEK organisms have their own characteristics on Gram stain. Both A. actinomycetemcomitans and Haemophilus species are small coccobacillary rods, but, with the latter, one can occasionally see filamentous forms. E. corrodens typically appears as slender straight rods with round ends. K. kingae appears as short straight rods with square ends that form pairs and chains [4]. Biochemical testing of C. hominis usually reveals it to be oxidase positive, indole positive, and catalase negative; E corrodens and K. kingae are oxidase positive, whereas Haemophilus species and A. actinomycetemcomitans are oxidase negative. A. actinomycetemcomitans is the only catalase-positive organism in the HACEK group [4].

Summary Strains of H. aphrophilus , H. paraphrophilus , and A. actinomycetemcomitans are phenotypically very similar. Ribotyping of 24 strains of H. aphrophilus , 22 strains of H. paraphrophilus , 8 strains of A. actinomycetemcomitans , and one strain each of the species Pasteurella aerogenes, H. parahaemolyticus , and the genus Capnocytophaga was studied using a non-radioactive digoxigenin labelled probe based on E. coli 16S- and 23S-ribosomal RNA. Restriction fragments were generated using restriction enzyme EcoRl. The ribotypes were analysed by a numerical approach using UPGMA clustering. Two major clusters were seen: One contained all A. actinomycetemcomitans strains, the other all H. aprophilus and all except one H. paraphrophilus strain intermingled between each other. The H. paraphrophilus strain not found in the H. aphrophilus / H. paraphrophilus cluster, the H. parahaemolyticus, P. aerogenes , and the Capnocytophaga strains clustered separetely from each other and the two major clusters. The H. paraphrophilus strain with the deviating ribotype was atypical in other respects: it neither did ferment lactose nor mannose and it was isolated from a deer in contradiction to the remaining H. paraphrophilus strains, which were human isolates. This study supports the view that H. aphrophilus and H. paraphrophilus should be regarded as one species separated from A. actinomycetemcomitans .

Summary: Genetic relationships among isolates assigned to Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus and H. paraphrophilus were determined by analysis of electrophoretically demonstrable allelic variation in 14 structural genes encoding metabolic enzymes. Among the 51 isolates analysed there were 25 electrophoretic types (ETs), among which mean genetic diversity per locus was 0.753. Cluster analysis of ETs demonstrated one well-defined group of 11 ETs representing solely the genotypes of all 17 isolates assigned to A. actinomycetemcomitans. The remaining 14 ETs represented the genotypes of the 34 isolates of H. aphrophilus and H. paraphrophilus. With the exception of ATCC 13252, all strains of H. aphrophilus were closely related, whereas strains assigned to H. paraphrophilus included distantly related lineages, some of which were similar to those of H. aphrophilus and should be assigned to this species. Thus, the study showed that there is no significant overall genetic similarity between A. actinomycetemcomitans and the two Haemophilus spp.

We report a case of a 42-year-old man man who presented with neurological symptoms and was found to have an intracranial abscess. A stereotactic aspiration of the abscess yielded a pure growth of Haemophilus paraphrophilus. The patient responded to treatment with cefotaxime. We postulate the mechanism of infection in this patient.

Haemophilus paraphrophilus was recovered in pure culture from purulent material collected at surgery from a patient presenting with a spinal epidural abscess and a severe neurological deficit. This is the first report of such an occurrence.

Infective endocarditis by Hemophilus species is very rare;there are only 22 reported-cases of Hemophilus paraphrophilus endocarditis. We report a case of Hemophilus paraphrophilus endocarditis in a middle-aged woman with cerebral embolism and hemorrhage. (Korean Circulation J 1998;28(4):642-646)

Summary: Analysis of the subgingival microflora has recently implicated Actinobacillus (Haemophilus) actinomycetemcomitans and several black Bacteroides species in the aetiology of juvenile, adult and rapidly progressing periodontitis. Rapid bacteriological diagnosis has been hampered by the slow growth and fastidious nature of these bacteria. To construct diagnostic probes, dideoxy sequencing of the 16S rRNA molecules from A. (H.) actinomycetemcomitans, Haemophilus aphrophilus, Bacteroides gingivalis, Bacteroides intermedius subgroup II, Bacteroides asaccharolyticus and several closely related species was performed. Next, oligodeoxynucleotides, complementary to defined regions of the 16S rRNA exhibiting considerable evolutionary divergence, were synthesized for use as molecular probes. In a dot-blot hybridization assay, all strains from each of the species for which probes were constructed were correctly identified, with a detection limit of less than 5 × 103 organisms. No cross-hybridization to closely related species (except for H. aphrophilus and Haemophilus paraphrophilus) or contaminating bacteria was observed. Using a modified DNA/RNA hybridization technique, the detection could be performed in less than 12 h, as compared to 2-3 weeks using conventional bacteriological procedures.

Selected human oral and nonoral strains of the genera Actinobacillus and Haemophilus were examined by transmission and scanning electron microscopy. The strains examined were morphologically identical to recognized Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus. By transmission electron microscopy, the cells were typically gram negative in morphology, with several strains possessing some extracellular ruthenium red-staining polymeric material. Numerous vesicular structures, morphologically identical to lipopolysaccharide vesicles, were seen to originate from and be continuous with the surface of the outer membrane. Large numbers of these vesicles were also found in the external environment. Scanning electron microscopic observations revealed that both actinobacilli and haemophili possessed surface projections and an amorphous surface material which connected and covered adjacent cells.

A case of Haemophilus paraphrophilus endocarditis successfully treated with ampicillin is described. The patient, a 24-year-old woman, had a prolapsed mitral valve. The organism was initially misidentified as H. parainfluenzae , which it closely resembles. H. paraphrophilus is distinguished by its requirement of 10% CO2 for growth on NaCl-free medium and its inability to ferment xylose.

The clinical course of a patient with brain abscess due to Haemophilus paraphrophilus is described. The organism was recovered in pure culture from purulent material collected at surgery. The role of this organism as a human pathogen is reviewed.

Genetic transformation studies were used to determine relatedness within the family Pasteurellaceae. Among strains with < 60% relatedness to Haemophilus influenzae based on deoxyribonucleic acid hybridization, two groups were identified; one, showing competition for homospecific transformation with H. influenzae, contained Haemophilus parainfluenzae, Haemophilus parasuis, Haemophilus aphrophilus, Haemophilus paraphrophilus, Pasteurella pneumotropica, Pasteurella multocida, and Actinobacillus actinomycetemcomitans, and the other, showing little or no competition for homospecific transformation with H. influenzae, contained Haemophilus ducreyi, Haemophilus parahaemolyticus, Actinobacillus lignieresii, Actinobacillus equuli, Actinobacillus pleuropneumoniae, and Pasteurella ureae. Such groupings support existing studies which have used only deoxyribonucleic acid hybridization or numerical analysis.

Fifteen gram-negative, saccharolytic rods isolated from advanced periodonlal pockeis were compared to type and reference strains of Arlinohacillus actinomycetemcomitans, Haemaphilus aphrophilus, and reference strains of Haemophilus paraphrophilus. Phenotypic features of organisms were used in cluster analysis and multidimensional scaling. Deoxyribonucleic acid (DNA) was extracted from all strains and moles percent guanine plus cytosine content determined. DNA/DNA homology experiments were performed between reference strains of each species and oral isolates. Five strains from periodontosis lesions, two strains from Papillon-Lefevre periodontal lesions, and four strains from periodontitis pockeis of adults were similar to A. actinomycetemcomitans by cluster analyses and by DNA/DNA homologies. Except for a single H. paraphrophilus isolate, the H. aphrophilus and H, paraphrophilus isolates could not be distinguished as two distinct species as there was overlap in biochemical characteristics and DNA/DNA homology. Two strains isolated from periodontosis lesions, two strains from adult periodontilis sites, and three oral reference strains were very similar to each other and to the H. aphrtiphilus type strain using phenotypic features. Hydrogen peroxide decomposition was not found to be a consistent feature of A. actinomycetemcumitans strains isolated from periodontal pockets.