Fastidious Gram-negative Bacteria Research Articles

Synthesis and antibacterial activity of novel oxazolidinones with methylene oxygen- and methylene sulfur-linked substituents at C5-position

Novel oxazolidinone derivatives of the lead compound RBx 8700, containing methylene oxygen- and methylene sulfur-linked substituents at the C5-position, were synthesized. Antibacterial screening of these compounds against a panel of resistant and susceptible Gram-positive and fastidious Gram-negative bacteria gave compounds 2 and 4 as new antibacterial agents.

In vitro and in vivo antibacterial evaluation of DRF 8417, a new oxazolidinone

DRF 8417, a novel oxazolidinone, has been evaluated against Gram-positive and fastidious Gram-negative bacteria. In vitro activity of DRF 8417 was determined by broth microdilution method and in vivo efficacy studies were carried out in different murine systemic infection models. DRF 8417 exhibited potent activity against Gram-positive pathogens with MIC(50) and MIC(90) values ranging from 0.06 to 1 mg/L. MICs against Haemophilus influenzae and Moraxella catarrhalis were one to two dilutions lower than those of linezolid. The in vivo efficacy, by oral route, in different susceptible and resistant Gram-positive systemic bacterial infection models ranged from 2.0 to 2.9 mg/kg. These studies displayed the excellent in vitro and in vivo activity of DRF 8417 against Gram-positive pathogens and lower MICs when compared with linezolid against H. influenzae and M. catarrhalis.

Bartonella Infection in Domestic Cats and Wild Felids

Bartonella are vector-borne, fastidious Gram-negative bacteria causing persistent bacteremia in their reservoir hosts. Felids represent a major reservoir for several Bartonella species. Domestic cats are the main reservoir of B. henselae, the agent of cat-scratch disease. Prevalence of infection is highest in warm and humid climates that are optimal for the survival of cat fleas, as fleas are essential for the transmission of the infection. Flea feces are the likely infectious substrate. Prevalence of B. henselae genotypes among cat populations varies worldwide. Genotype Houston I is more prevalent in the Far East and genotype Marseille is dominant in western Europe, Australia, and the western United States. Cats are usually asymptomatic, but uveitis, endocarditis, neurological signs, fever, necrotic lesions at the inoculation site, lymphadenopathy, and reproductive disorders have been reported in naturally or experimentally infected cats. Domestic cats are also the reservoir of B. clarridgeiae and co-infection has been demonstrated. B. koehlerae has been isolated from domestic cats, and was identified in cat fleas and associated with a human endocarditis case. B. bovis was isolated from a few cats in the United States and B. quintana DNA was recently identified in a cat tooth. Bartonella spp. have also been isolated from free-ranging and captive wild felids from North America and Africa. Whereas, B. henselae was identified in African lions and a cheetah, some strains specific to these wild cats have also been identified, leading to the concept of a B. henselae group including various subspecies, as previously described for B. vinsonii.

In vitro activity of ertapenem against anaerobes isolated from the respiratory tract

Ertapenem is a novel parenteral carbapenem with a long serum half-life. Its spectrum of activity is similar to that of imipenem and meropenem against Gram-positive bacteria, Enterobacteriaceae and fastidious Gram-negative bacteria but it is less active against Pseudomonas aeruginosa and Acinetobacter spp. Several studies were performed in the United States but only one European study has shown that ertapenem has an excellent activity against anaerobes. The objectives of the present study were to test the activity of ertapenem against anaerobes isolated prospectively from the lower and upper respiratory tracts, and to compare their susceptibility with that of anaerobic isolates from other body sites. Fifty-three isolates from the respiratory tract, as well as 50 isolates from various other body sites were tested with E-tests against six antibiotics. For respiratory isolates and for isolates from other sites, MIC 90 values (mg/l) were, respectively, >32 and >32 for penicillin, 0.38 and 0.75 for amoxicillin/clavulanate, 48 and >256 for ceftriaxone, 0.12 and 0.75 for ertapenem, 12 and >256 for clindamycin and 2 and 12 for moxifloxacin. The higher susceptibility of respiratory tract isolates was mainly due to the different distribution of isolated species: only three respiratory isolates but 22 other isolates belonged to the Bacteroides fragilis group. This study confirms the excellent anti-anaerobic activity of ertapenem against anaerobic isolates from the respiratory tract.

A Man with a Prosthetic Aortic Valve and Subacute Calf Pain

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].

Carbon–carbon-linked (pyrazolylphenyl)oxazolidinones with antibacterial activity against multiple drug resistant gram-positive and fastidious gram-negative bacteria

In an effort to expand the spectrum of activity of the oxazolidinone class of antibacterial agents to include Gram-negative bacteria, a series of new carbon–carbon linked pyrazolylphenyl analogues has been prepared. The α- N-substituted methyl pyrazole ( 10α) in the C3-linked series exhibited very good Gram-positive activity with MICs ≤0.5–1 μg/mL and moderate Gram-negative activity with MICs=2–8 μg/mL against Haemophilus influenzae and Moraxella catarrhalis. This analogue was also found to have potent in vivo activity with an ED 50=1.9 mg/kg. β-Substitution at the C3-linked pyrazole generally results in a loss of activity. The C4-linked pyrazoles are slightly more potent than their counterparts in the C3-linked series. Most of the analogues in the C4-linked series exhibited similar levels of activity in vitro, but lower levels of activity in vivo than 10α. In addition, incorporation of a thioamide moiety in selected C4-linked pyrazole analogues results in an enhancement of in vitro activity leading to compounds several times more potent than eperezolid, linezolid and vancomycin. The thioamide of the N-cyanomethyl pyrazole analogue ( 34) exhibited an exceptional in vitro activity with MICs of ≤ 0.06–0.25 μg/mL against Gram-positive pathogens and with MICs of 1 μg/mL against fastidious Gram-negative pathogens.

In vitro activities of ertapenem (MK-0826) against recent clinical bacteria collected in Europe and Australia.

Ertapenem (MK-0826, L-749,345) is a 1-beta-methyl carbapenem with a long serum half-life. Its in vitro activity was determined by broth microdilution against 3,478 bacteria from 12 centers in Europe and Australia, with imipenem, cefepime, ceftriaxone, and piperacillin-tazobactam used as comparators. Ertapenem was the most active agent tested against members of the family Enterobacteriaceae, with MICs at which 90% of isolates are inhibited (MIC(90)s) of < or =1 microg/ml for all species. Ertapenem also was more active than imipenem against fastidious gram-negative bacteria and Moraxella spp.; on the other hand, ertapenem was slightly less active than imipenem against streptococci, methicillin-susceptible staphylococci, and anaerobes, but its MIC(90)s for these groups remained < or =0.5 microg/ml. Acinetobacter spp. and Pseudomonas aeruginosa were also much less susceptible to ertapenem than imipenem, and most Enterococcus faecalis strains were resistant. Ertapenem resistance, based on a provisional NCCLS MIC breakpoint of > or =16 microg/ml, was seen in only 3 of 1,611 strains of the family Enterobacteriaceae tested, all of them Enterobacter aerogenes. Resistance was also seen in 2 of 135 anaerobes, comprising 1 Bacteroides fragilis strain and 1 Clostridium difficile strain. Ertapenem breakpoints for streptococci have not been established, but an unofficial susceptibility breakpoint of < or =2 microg/ml was adopted for clinical trials to generate corresponding clinical response data for isolates for which MICs were as high as 2 microg/ml. Of 234 Streptococcus pneumoniae strains tested, 2 required ertapenem MICs of 2 microg/ml and one required an MIC of 4 microg/ml, among 67 non-Streptococcus pyogenes, non-Streptococcus pneumoniae streptococci, single isolates required ertapenem MICs of 2 and 16 microg/ml. These streptococci also had diminished susceptibilities to other beta-lactams, including imipenem as well as ertapenem. The Etest and disk diffusion gave susceptibility test results in good agreement with those of the broth microdilution method for ertapenem.

Isolation of new fastidious α Proteobacteria and Afipia felis from hospital water supplies by direct plating and amoebal co-culture procedures

As water is a source of nosocomial infections in hospitals, the presence of fastidious Gram-negative bacteria in water samples taken in a university hospital was investigated. Water samples were inoculated onto agar plates and into amoebal microplates for co-culture. Sixty-eight a proteobacteria isolates were obtained and characterized using phenotypic methods and 16S rRNA gene sequence comparison. The latter approach divided the strains into seven clusters. Of these, one corresponded to previously recognized Afipia felis and it is likely that six were closely related new species. As these bacteria are fastidious and can not be cultivated on standard microbiological media, their possible role in hospital-acquired human infections should be investigated.

Isolation of new fastidious α Proteobacteria and Afipia felis from hospital water supplies by direct plating and amoebal co-culture procedures

As water is a source of nosocomial infections in hospitals, the presence of fastidious Gram-negative bacteria in water samples taken in a university hospital was investigated. Water samples were inoculated onto agar plates and into amoebal microplates for co-culture. Sixty-eight α proteobacteria isolates were obtained and characterized using phenotypic methods and 16S rRNA gene sequence comparison. The latter approach divided the strains into seven clusters. Of these, one corresponded to previously recognized Afipia felis and it is likely that six were closely related new species. As these bacteria are fastidious and can not be cultivated on standard microbiological media, their possible role in hospital-acquired human infections should be investigated.

Culture of Bartonella quintana and Bartonella henselae from human samples: a 5-year experience (1993 to 1998).

Bartonella quintana and Bartonella henselae are fastidious gram-negative bacteria responsible for bacillary angiomatosis, trench fever, cat scratch disease, and endocarditis. During a 5-year period, we received 2,043 samples for culture of Bartonella sp. We found Bartonella sp. to be the etiologic agent in 38 cases of endocarditis, 78 cases of cat scratch disease, 16 cases of bacteremia in homeless people, and 7 cases of bacillary angiomatosis. We correlated the results of positive cultures with the clinical form of the disease, type of sample, culture procedure, PCR-based genomic detection, and antibody determination. Seventy-two isolates of B. quintana and nine isolates of B. henselae from 43 patients were obtained. Sixty-three of the B. quintana isolates and two of the B. henselae isolates, obtained from patients with no prior antibiotic therapy, were stably subcultured. The sensitivity of culture was low when compared with that of PCR-based detection methods in valves of patients with endocarditis (44 and 81%, respectively), skin biopsy samples of patients with bacillary angiomatosis (43 and 100%, respectively), and lymph nodes of cat scratch disease (13 and 30%, respectively). Serological diagnosis was also more sensitive in cases of endocarditis (97%) and cat scratch disease (90%). Among endocarditis patients, the sensitivity of the shell vial culture assay was 28% when inoculated with blood samples and 44% when inoculated with valvular biopsy samples, and the sensitivity of both was significantly higher than that of culture on agar (5% for blood [P = 0.045] and 4% for valve biopsy samples [P < 0.0005]). The most efficient culture procedure was the subculture of blood culture broth into shell vials (sensitivity, 71%). For patients with endocarditis, previous antibiotic therapy significantly affected results of blood culture; no patient who had been administered antibiotics yielded a positive blood culture, whereas 80% of patients with no previous antibiotic therapy yielded positive blood cultures (P = 0.0006). Previous antibiotic therapy did not, however, prevent isolation of Bartonella sp. from cardiac valves but did prevent the establishment of strains, as none of the 15 isolates from treated patients could be successfully subcultured. For the diagnosis of B. quintana bacteremia in homeless people, the efficiency of systematic subculture of blood culture broth onto agar was higher than that of direct blood plating (respective sensitivities, 98 and 10% [P < 10(-7)]). Nevertheless, both procedures are complementary, since when used together their sensitivity reached 100%. All homeless people with positive blood cultures had negative serology. The isolation rate of B. henselae from PCR-positive lymph nodes, in patients with cat scratch disease, was significantly lower than that from valves of endocarditis patients and skin biopsy samples from bacillary angiomatosis patients (13 and 33%, respectively [P = 0.084]). In cases of bacillary angiomatosis for which an agent was identified to species level, the isolation rate of B. henselae was lower than the isolation rate of B. quintana (28 and 64%, respectively [P = 0.003]). If culture is to be considered an efficient tool for the diagnosis of several Bartonella-related diseases, methodologies need to be improved, notably for the recovery of B. henselae from lymph nodes of patients with cat scratch disease.

Multicenter survey of the in vitro activity of four expanded-spectrum β-lactams against consecutive contemporary clinical isolates

A survey of the in vitro susceptibility of consecutive clinical bacterial isolates to cefotaxime, ceftazidime, cefpodoxime, and aztreonam was conducted at 10 North American medical centers during the first quarter of 1997. All four drugs had good activity against Enterobacteriaceae and fastidious Gram-negative bacteria and poor activity against enterococci and methicillin-resistant staphylococci. No significant trends in susceptibility to cefotaxime or ceftazidime were observed when compared with a previous similar survey. The use of cefpodoxime MIC >2.0 μg/mL as a marker for extended-spectrum β-lactamase production by Escherichia coli and Klebsiella spp. had good sensitivity, but the specificity was very poor for E. coli. Thus, at least for E. coli, this screening test seems to be of questionable value unless used to indicate the need for additional tests such as clavulanic acid inhibition. Disk diffusion zone diameters correlated well with the MICs for all four drugs and support the current interpretive criteria.

Capnocytophaga canimorsus infections in human: review of the literature and cases report.

Capnocytophaga canimorsus, formerly designated Dysgonic fermenter 2 (DF-2) was first described in 1976; it is a commensal bacterium of dogs and cats saliva, which can be transmitted to man by bite (54% of cases), scratch (8.5%), or mere exposure to animals (27%). We present a review of the clinical and microbiological characteristics of the Capnocytophaga canimorsus infections and 12 cases of infection in France. Over 100 cases of human infections have been reported, mainly septicemia in patients with diminished defences, due to splenectomy (33%), alcohol abuse (24%), immunosuppression (5%). However 40% of septicemia occur in patients with no predisposing conditions. Other infections are less frequent: meningitis, endocarditis, arthritis, pleural and localized eye infections. These infections range from mild to fulminating disease, with shock, respiratory distress, disseminated intravascular coagulation. Dermatological lesions (macular or maculopapular rash, purpura) or gangrene are common. This fastidious Gram-negative bacterium grows slowly on chocolate agar or on heart infusion agar with 5% rabbit blood incubated in 5% CO2. In spite of a great susceptibility of bacteria to antibiotics, the mortality is of 30%. Because of the severity of these infections, taking into account this organism in the management of bites is necessary, especially in patients with predisposing factors.

Fastidious gram-negative bacteria: meeting the diagnostic challenge with nucleic acid analysis.

The extended panorama of fastidious Gram-negative bacteria (FGNB) as opportunistic etiological agents of infectious diseases in immunocompromised patients is largely due to improved medical expertise and technology. The heightened awareness of infectious diseases due to FGNB species mandates comprehensive classification and identification systems as a basis for rapid and reliable diagnostics. The most useful approaches are combinations of nucleic acid techniques such as hybridization, genetic transformation, amplification and base sequence analysis with selected conventional criteria. Among these approaches, the widely distributed feature of natural competence in these organisms facilitates the use of the biological method of genetic transformation as a valuable addition to the more common nucleic acid techniques. We describe the development of the taxonomy of FGNB through the last four decades, with particular emphasis on the families Neisseriaceae, Moraxellaceae, and Pasteurellaceae.

Bartonella (Rochalimaea) quintana bacteremia in inner-city patients with chronic alcoholism.

Bartonella (Rochalimaea) quintana is a fastidious gram-negative bacterium known to cause trench fever, cutaneous bacillary angiomatosis, and endocarditis. Between January and June 1993 in Seattle, we isolated B. quintana from 34 blood cultures obtained from 10 patients not known to be infected with the human immunodeficiency virus (HIV). After identifying the isolates as B. quintana by direct immunofluorescence and DNA-hybridization studies, we determined strain hybridization with studies of restriction-fragment-length polymorphisms (RFLPs) of the intergenic spacer (noncoding) region of ribosomal DNA amplified by the polymerase chain reaction (PCR). To characterize the epidemiologic and clinical features of bartonella infections in these patients, we performed a retrospective case-control study using as controls 20 patients with blood cultures obtained at approximately the same time as those obtained from the index patients. B. quintana isolates from the 10 patients were indistinguishable by PCR-RFLP typing. All 10 patients had chronic alcoholism, and 8 were homeless (P = 0.001 for both comparisons with controls). The six patients who underwent HIV testing were seronegative. At the time of their initial presentation, seven patients had temperatures of at least 38.5 degrees C. Six patients had three or more blood cultures that were positive for B. quintana, and in four of these patients B. quintana was isolated from blood cultures obtained 10 or more days apart. Subacute endocarditis developed in two patients and required surgical removal of the infected aortic valve in one of them. Nine patients recovered; one died of sepsis from Streptococcus pneumoniae infection. B. quintana is a cause of fever, bacteremia, and endocarditis in HIV-seronegative, homeless, inner-city patients with chronic alcoholism.

Induction of protective immunity to Haemophilus ducreyi in the temperature-dependent rabbit model of experimental chancroid.

The temperature-dependent rabbit model for chancroid, a sexually transmitted disease caused by the fastidious Gram-negative bacterium Haemophilus ducreyi, was used to investigate the abilities of previous infection and immunization with an acellular preparation of H. ducreyi to induce protective immunity. In the first set of experiments, animals were infected intradermally with either the 35000 or Cha-1 strains of H. ducreyi and then rechallenged 30 days later with both the homologous and heterologous strains. In animals infected with the 35000 strain, statistically significant protective immunity occurred only against the homologous strain, whereas protection against both homologous and heterologous challenge was obtained in rabbits previously infected with strain Cha-1. In a separate series of experiments, rabbits were immunized with cell envelopes from either strain 35000 or strain Cha-1 and then challenged with both the homologous and heterologous strains. In rabbits immunized with strain 35000 cell envelopes, significant protective immunity was observed only against challenge with the homologous strain. In animals immunized with strain Cha-1 cell envelopes, protection was obtained against both homologous and heterologous challenge. Histopathologic analysis of sites inoculated with strain 35000 (10(5) CFU) demonstrated that the inflammatory response in control animals was predominantly suppurative (i.e., heterophilic), whereas that of immunized animals was predominantly mononuclear and, at later time points, largely histiocytic. ELISA and Western blot analyses revealed that immunization produced a better humoral immune response than did infection and provided evidence for antigenic cross-reactivity between these two strains. These results provide the experimental basis for continued efforts to identify potential H. ducreyi vaccinogens.

Beta-Lactam resistance in gram-negative bacteria: global trends and clinical impact.

Microbial drug resistance is an inescapable consequence of the utilization of antimicrobial agents in a given environment. Nowhere is the importance of resistance more evident than among agents of the beta-lactam family. Trends toward increased resistance can be seen among fastidious gram-negative bacteria like Haemophilus influenzae, where ampicillin resistance varies from 1% to 64% globally. For Escherichia coli, ampicillin resistance has risen to > or = 50% in high-risk populations, and resistance to third-generation cephalosporins is now being seen in certain areas. Inducible beta-lactamases have been responsible for increasing multiple beta-lactam resistance among certain Enterobacteriaceae and Pseudomonas aeruginosa, and this has been associated with increased use of newer cephalosporins. Xanthomonas maltophilia with its two inducible beta-lactamases is becoming an increasingly important nosocomial pathogen, especially in areas of heavy imipenem utilization. Only through the recognition of factors associated with increasing resistance and the mechanisms responsible can strategies be designed for minimizing beta-lactam resistance.

Review of preclinical studies with ofloxacin.

Most Enterobacteriaceae, enteropathogens, and fastidious gram-negative bacteria are highly susceptible to ofloxacin, a new tricyclic fluoroquinolone. Aerobic gram-negative bacilli and gram-positive bacteria are generally not as susceptible to ofloxacin. Obligate anaerobes are generally resistant to ofloxacin, while many mycobacteria, chlamydiae, legionellae, and mycoplasmas are susceptible. Ofloxacin is generally less active than ciprofloxacin against gram-negative bacteria, is similarly active against gram-positive bacteria, mycobacteria, legionellae, and mycoplasmas, and is more active against chlamydiae. However, numerous animal studies have shown these two fluoroquinolones to be similar. Ofloxacin inhibits DNA synthesis, is rapidly bactericidal, and is 1,000-2,400 times more potent against prokaryotic gyrase than against eukaryotic gyrase. The bactericidal effect of ofloxacin is not completely neutralized by inhibitors of protein or RNA synthesis. Resistance to ofloxacin arises from mutations within chromosomal genes involved with DNA gyrase and drug permeation. Selection of resistant mutants by ofloxacin is not as frequent as that seen with nalidixic acid. However, due to the cross-resistance between ofloxacin and other fluoroquinolones, all of these drugs should be used judiciously to preserve their clinical utility.

Capillary Gas Chromatography of Cellular Carbohydrates as a Means for the Differentiation of Fastidious, Slow-growing or Anaerobic Gram-negative Bacteria — A Review

The phenotypic differentiation of microaerophilic or anaerobic Gram-negative bacteria of human origin is traditionally based on the analysis of metabolic characteristics. Due to outstanding growth requirements of many kinds of parasitic bacteria, e.g. of the facultatively and strictly anaerobic members of the human periodontal flora, conventional biochemical tests are time-consuming and sometimes lead to doubtful results. Commercial diagnostic test kits often produce misidentifications because of insufficient databases or inappropriate test combinations. In this situation, cytochemical methods can provide an alternative. Especially in the case of Gram-negative bacteria which contain considerable amounts of carbohydrates in their cell wall lipopolysaccharide layers, capillary gas chromatography of peracetylated carbohydrate derivatives prepared from bacterial whole cell hydrolysates proved to be useful to characterize individual species, and sometimes even subspecies. As soon as a pure culture is available, the derivatization of the cellular carbohydrates to their peracetylated aldononitriles and peracetylated O-methyloximes, respectively, needs only about four hours and the gas chromatographic spectra are easy to evaluate.

Antimicrobial susceptibility testing of veterinary clinical isolates with the Sceptor System

The Sceptor System (Becton Dickinson) was compared with an agar dilution method for antimicrobial susceptibility testing of veterinary clinical isolates. The results indicate that the Sceptor System may be used to test gram-positive and fastidious gram-negative bacteria.

Identification of Neisseria spp., Haemophilus spp., and other fastidious gram-negative bacteria with the MicroScan Haemophilus-Neisseria identification panel

The Haemophilus-Neisseria identification (HNID) panel (American MicroScan, Sacramento, Calif.) is a 4-h microdilution format system for identification of Haemophilus and Neisseria spp., Branhamella (Moraxella) catarrhalis, and Gardnerella vaginalis. The HNID panel was evaluated by using 423 clinical isolates and stock strains of these organisms, and HNID identifications were compared with those obtained by conventional methods. In addition, 32 isolates representing six genera not included in the HNID data base were tested to determine whether these organisms would produce unique biotype numbers for possible inclusion in the data base. The HNID panel correctly identified 95.3% of 86 Neisseria gonorrhoeae strains, 96% of 25 G. vaginalis strains, and 100% of 28 Neisseria lactamica strains and 48 B. catarrhalis strains. Only 64.7% of 68 Neisseria meningitidis isolates were identified correctly owing to false-negative or equivocal carbohydrate and/or aminopeptidase reactions. Among the Haemophilus spp., 98.8% of 83 H. influenzae strains, 97.1% of 34 H. parainfluenzae strains, and 80% of 15 H. aphrophilus and H. paraphrophilus strains were correctly identified. Eight strains of Neisseria cinerea, a species not included in the data base, produced profiles identical with those for B. catarrhalis and N. gonorrhoeae. Isolates of other species not included in the data base, including Eikenella corrodens, Kingella spp., and Cardiobacterium hominis, produced unique biochemical reaction patterns on the panel. Modification of interpretative criteria for certain tests, expansion of the data base to include other species, and suggestions for additional confirmatory tests will increase the accuracy and utility of the HNID panel.