Clinical Significance and Characterization of Streptococcus tigurinus Isolates in an Adult Population.

Streptococcus tigurinus is a newly described member of the Streptococcus mitis group. Due to the difficulty in distinguishing viridans group streptococci (VGS) by phenotype, analysis of 16S rRNA sequences is necessary for the accurate identification of most species. Through a laboratory policy of analyzing all clinically significant isolates from the VGS group by16S rRNA gene sequencing, we identified 14 S. tigurinus isolates from 11 patients. The Vitek 2 system most commonly gave an excellent rating to an incorrect identification (e.g., Streptococcus mitis), as did matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (e.g., Streptococcus oralis). S. tigurinus strains were recovered from numerous body sites, including the blood, peritoneal fluid, bone, synovial fluid, a perianal abscess, and an arm wound. Retrospective chart review indicated that most isolates were clinically significant, with bacteremia (n = 5), soft tissue infections (n = 3) osteomyelitis (n = 2), infected joint prosthesis (n = 2), and peritonitis (n = 2) being the most common, thus expanding the spectrum of disease associated with S. tigurinus.

Comparison of agar based media for primary isolation of Helicobacter pylori.

AIMS: To determine the best medium for the primary isolation of Helicobacter pylori. METHODS: Sixty six gastric mucosal biopsy specimens frozen in 1 ml Cysteine Albimi media with 20% glycerol from 22 histologically proven H pylori infected patients were cultured on brain heart infusion agar (BHIA) with 7% fresh whole defibrinated horse blood, egg yolk agar (EYA), Columbia blood agar-cyclodextrin agar (CBA-Cd), and commercial trypticase soy agar (TSA) supplemented with 5% sheep blood. RESULTS: Successful primary isolation of H pylori was 96% with BHIA, 78% with TSA, 64% for EYA, and 32% with CBA-Cd. Colonies appeared earlier on BHIA (4.7 +/- 0.1 days, 5.3 +/- 0.4 days, 5.3 +/- 0.4 days, and 7.1 +/- 0.9 days for BHIA, TSA, EYA, and CBA-Cd) and there were more colonies on BHIA than on CBA-Cd, EYA or TSA (599 +/- 88, 104 +/- 66, 260 +/- 107, and 358 +/- 89, respectively). CONCLUSIONS: Success of a medium for passage of isolates apparently does not reliably predict usefulness for primary isolation. Freshly made BHIA with 7% horse blood medium is recommended for primary isolation. However, the easily obtainable TSA media would be the best alternative for routine clinical laboratories with no access to BHIA.

Recovery of Coccidioides immitis from blood and abscess fluid using the BacT/alert system.

We report the recovery of Coccidioides immitis from the blood and abscess fluid of two separate patients by using two automated blood culture systems. In the first case, an aspirate from a neck abscess containing C. immitis spherules was serially diluted and inoculated into liquid media used by the BacT/Alert and the Bactec NR660 blood culture systems. BacT/Alert bottles inoculated with 10(5), 10(4), 10(3), 10(2), 10 and two spherules produced a positive signal at 19, 24, 35, 42, 57, and 62 h postinoculation, respectively. Bactec NR660 bottles containing > 10(2) shperules and 10 spherules produced a positive signal after approximately 72 and 96 h of incubation, respectively. In the second case, a blood specimen incubated in BacT/Alert blood culture both was signaled positive after 82 h of incubation. No organisms were detected by Gram stain of the broth, but C. immitis grew after blind subculture. Our observations demonstrate that these rapid blood culture systems are capable of supporting growth of C. immitis. To our knowledge, this report is the first to detect C. immitis by these blood culture systems.

Antimicrobial susceptibility of clinical isolates of Brucella.

A number of antimicrobial agents have been used in the treatment of human brucellosis with varying effectiveness. The purpose of this study was to test the in vitro susceptibility of isolates of four Brucella species to a variety of antimicrobial agents, and to study in vitro synergy of combinations of agents. Minimal inhibitory concentrations (MICs) were determined using conventional broth microdilution methods and commercially available systems. Conventional checkerboard synergy microdilutions were prepared for gentamicin or streptomycin plus tetracycline, and rifampicin plus tetracycline. Synergy or antagonism was determined by the fractional inhibitory concentration index. Penicillin G and ampicillin showed in vitro activity against Brucella (MIC90 4 micrograms/ml), whereas the antipseudomonal penicillins were less active (carbenicillin MIC90 12 micrograms/ml, piperacillin MIC90 32 micrograms/ml). Among the third generation cephalosporins tested, cefotaxime (MIC90 2 micrograms/ml) demonstrated greatest activity. As a class, aminoglycosides were equivalent (MIC90 1-4 micrograms/ml). All strains were sensitive to tetracycline (MIC90 0.25 microgram/ml), trimethoprim-sulfamethoxazole (MIC90 1/19 micrograms/ml), and rifampin (MIC90 1 microgram/ml). Erythromycin (MIC90 greater than 8 micrograms/ml) and vancomycin (MIC90 greater than 16 micrograms/ml) demonstrated no activity. In vitro synergy (fractional inhibitory concentration index less than 0.5) was demonstrated with tetracycline plus rafampin in six of eight isolates tested.

Antimicrobial susceptibility testing of Helicobacter pylori. Comparison of E-test, broth microdilution, and disk diffusion for ampicillin, clarithromycin, and metronidazole.

The optimal method for the determination of the minimum inhibitory concentration (MIC) of antimicrobials against Helicobacter pylori has not been established. The epsilometer agar diffusion gradient test (E-Test; AB Biodisk, Solna, Sweden) was compared with broth microdilution, the reference method, and disk diffusion for the antimicrobial susceptibility testing of 122 clinical isolates of H. pylori to ampicillin, clarithromycin, and metronidazole. Isolates were considered to be resistant when the MIC values was > 8 micrograms/ml for either ampicillin or metronidazole and > 2 micrograms/ml for clarithromycin. For an individual isolate, the MICs for ampicillin and clarithromycin determined by broth microdilution and the E-test were highly reproducible, with replicate results being within +/- 1 log2 dilution. The correlation between the MICs determined by E-test and broth microdilution was excellent for both ampicillin and clarithromycin (90.1% and 88.5% were within +/- log2 dilution, and 98.3% and 96.7% of the values were within +/- 2 log2 dilution, respectively). In no instance did the interpretation of "sensitive" or "resistant" differ. Conversely, only 70.5% of the E-test results of metronidazole were within +/- 1 log2 dilution of the broth microdilution results. In addition, 15 (12.3%) of the H. pylori isolates interpreted as resistant by the E-test were sensitive by the broth microdilution method. All discrepancies occurred when the E-test MIC values fell between 8 and 32 micrograms/ml. The results of the ampicillin and clarithromycin disk diffusion assay correlated 100% with the results of the broth microdilution. However, these data suggest that when the E-test MIC results of metronidazole yield values between 8 and 32 micrograms/ml, the MIC should be reevaluated by another method.

A new Brevibacterium sp. isolated from infected genital hair of patients with white piedra.

A new aerobic gram-positive non-sporeforming bacillus has been isolated from infected genital hair of patients with white piedra in association with Trichosporon beigelii. This species has been characterised morphologically, nutritionally, by DNA base composition, cell-wall analysis and cellular fatty-acid profile on the basis of 14 isolates. The G+C content of DNA is 63.05 mol%. Cell walls possess meso-diaminopimelic acid (Type IV) and the sugars glucose, galactose, xylose and ribose; mycolic acids are not present. The species has a distinct colonial and microscopic morphology, is strongly proteolytic and produces methanethiol. These findings and the cellular fatty-acid profile are compatible with the genus Brevibacterium. A new species is proposed based on the following characters: colonial and microscopic growth and morphology; conditions for rod-to-coccus cycle; ribose utilisation; and tellurite reduction. The type strain has been named Brevibacterium mcbrellneri E2cr (ATCC 49030). The strong proteolytic properties may be the mechanism of pathogenesis.

Antibiotic resistance patterns during aminoglycoside restriction.

When amikacin first became available its use was restricted to prevent the emergence of resistant strains of gram-negative bacilli to this new agent. Gentamicin was the aminoglycoside most widely used at this time, and the incidence of gentamicin-resistant bacteria was 14%, while only 2.4% were resistant to amikacin. For a period of 15 months gentamicin use was restricted, and amikacin was used almost exclusively. Amikacin use was associated with a fall in the incidence of gentamicin-resistant bacteria to 9.2% (p less than .00005), while amikacin resistance remained unchanged at 2.2% (NS). During a period of 21 months after all aminoglycoside restrictions were lifted, gentamicin use again increased, and was accompanied by a return of gentamicin resistance to the baseline level of 15.3%. During this period, amikacin resistance also increased to 4.0% (p less than .0000001) but was due primarily to an increase in resistant Pseudomonas aeruginosa. Escherichia coli was the most frequently isolated gram-negative bacillus during all three periods, and it remained sensitive to both antibiotics regardless of the drug in use. In contrast, P. aeruginosa showed a high level of resistance to gentamicin, which fell when this antibiotic was restricted, only to return to a high level with reinstitution of gentamicin. While there was also an increase in amikacin resistant strains of P. aeruginosa with unrestricted aminoglycoside use, there was no apparent shift in the pattern of aminoglycoside modifying enzymes among a small random selection of amikacin-resistant bacteria. Impaired uptake of antibiotic was the predominant mechanism responsible for P. aeruginosa resistance among strains that did not produce aminoglycoside acetyltransferase (AAC)(6').

Genotypic and phenotypic characterization and clinical significance of 'Haemophilus quentini' isolated from the urinary tract of adult men.

'Haemophilus quentini' has been proposed as the name for a distinct and homogeneous Haemophilus genospecies associated with urogenital tract and neonatal-related infections. Reports of 'H. quentini' isolation from adult men are rare and the disease potential in this population is unknown. We report six cases where 'H. quentini' was isolated from the genito-urinary tract in males. The isolation of 'H. quentini' during routine urine and urethral culture in adult men may aid in the determination of unresolved urethritis and possible urinary tract infections.

Isolation and characterization of two hemolytic phenotypes of Vibrio damsela associated with a fatal wound infection.

Two hemolytic phenotypes of Vibrio damsela, isolated from the tissue of a patient with a fatal wound infection, were characterized. The patient had underlying disease, and the wound was associated with an injury inflicted during the handling of a catfish. The phenotypes were morphologically and biochemically similar except for their lecithinase, lipase, and hemolytic activities. When grown on rabbit blood agar, one phenotype (LZ) produced a large zone of hemolysis (10 mm) around the colony, whereas the other type (SZ) produced only a small zone (1 to 2 mm). On sheep blood agar, the differences in hemolytic activity were more subtle. By a modified CAMP test in which V. damsela was streaked perpendicularly to Staphylococcus aureus, it was determined that a factor elaborated by the LZ phenotype (but not the SZ phenotype) protected sheep erythrocytes from the hemolysis normally caused by S. aureus toxins. Cell-free filtrates of broth cultures of each phenotype had the same effects on erythrocytes as did the organisms themselves.

Streptococcus bovis septicemia and large bowel neoplasia.

Streptococcus bovis septicemia is a relatively uncommon entity that is associated with an increased incidence of colonic neoplasms. Three of four patients with S. bovis endocarditis subsequent to septicemia underwent colonoscopy. The fourth patient underwent a barium enema and a proctoscopic examination. Polyps were found in three patients, and adenocarcinoma of the colon in one. Patients with S. bovis endocarditis should be considered at high risk for colonic neoplasms. Screening colonoscopy is recommended for these patients, and follow-up colonoscopy may be warranted.

Potential errors in recognition of Erysipelothrix rhusiopathiae.

Here we describe four isolations of Erysipelothrix rhusiopathiae associated with polyarthralgia and renal failure, septic arthritis, classic erysipeloid, and peritonitis. Although the biochemical identification was straightforward in each case, recognition presented a challenge to the clinical microbiologist, since in three cases E. rhusiopathiae was not initially considered due to unusual clinical presentations, in two cases the significance might not have been appreciated because growth was in broth only, and in one case the infection was thought to be polymicrobic. Because the Gram stain can be confusing, abbreviated identification schemes that do not include testing for H(2)S production could allow E. rhusiopathiae isolates to be misidentified as Lactobacillus spp. or Enterococcus spp. in atypical infections.

Nonpathogenic nematodes in gastrointestinal aspirates obtained during endoscopy.

The gastroenterologist must be alert to diagnosing parasitic diseases when collecting intestinal fluid samples or cytologies during endoscopy. Between 1979 and 1982, we identified nematodes in 10 endoscopic specimens. None were Strongyloides stercoralis. We emphasize the importance and technical aspects of distinguishing nonpathogenic nematodes from Strongyloides.

Use of the RapID-ANA system and sodium polyanetholesulfonate disk susceptibility testing in identifying Haemophilus ducreyi.

Haemophilus ducreyi has traditionally been difficult to identify. We have utilized simple test methods to identify 19 fresh isolates obtained during a recent outbreak of chancroid in Houston and six strains of H. ducreyi from other outbreaks. Tests were performed from growth on chocolate agar after 48 h of incubation at 35 degrees C with increased humidity and CO2. All isolates exhibited typical colonial morphology and Gram stain. Isolates were catalase negative and oxidase and nitrate positive (in enriched broth). The RapID NH system failed to identify these strains because of negative reactions with alkaline phosphatase and nitrate reductase. However, by using the RapID-ANA system, all strains were positive for alkaline phosphatase and arginine, glycine, and serine aminopeptidases. Their biochemical profiles were distinct from those obtained with 66 strains representing 13 species similar to H. ducreyi. We also investigated the use of sodium polyanetholesulfonate (SPS) disk susceptibility to identify and differentiate H. ducreyi from other species. All H. ducreyi isolates were susceptible, as evidenced by the presence of a zone of inhibition with an average size of 15 mm around the SPS disk. With the exceptions of Neisseria gonorrhoeae, Gardnerella vaginalis, and Capnocytophaga spp., no other strain showed any evidence of inhibition. The latter three organisms can be easily differentiated from H. ducreyi by various features including reactions in the RapID-ANA. We conclude that, by considering simple growth and biochemical characteristics, SPS susceptibilities, and reactions in RapID-ANA, it is possible for more clinical laboratories to definitively identify this organism.

Limited vs full microbiological investigation for the management of symptomatic polymicrobial urinary tract infection in adult spinal cord-injured patients.

Spinal cord-injured (SCI) patients often suffer from symptomatic polymicrobial urinary tract infection (UTI). The objective of this study was to evaluate the clinical outcome and cost-savings associated with antibiotic therapy based on limited vs full microbiological investigation of urine cultures in adult SCI patients with symptomatic polymicrobial UTI (> or = 2 organisms growing in urine cultures). In the first part of the study, a total of 40 evaluable patients were prospectively randomized in a single-blinded fashion to receive antibiotic therapy based on either limited (21 patients) or full microbiologic investigation (19 patients) of urine cultures. The practicality of a limited microbiological investigation was further examined in the second part of the study where 12 consecutive patients with symptomatic polymicrobial UTI initially had only limited microbiological investigation of urine cultures and received antibiotic therapy accordingly. When analyzing all patients in the study, the likelihood of adequate clinical response was not significantly different between those who received antibiotic therapy based on limited (28/33 = 85%) vs full (18/19 = 95%) microbiological investigation of urine cultures (P = 0.40). An average of 183 US dollars could be saved per patient by managing symptomatic polymicrobial UTI based on a limited vs a full microbiological investigation. These results suggest that in adult SCI patients with symptomatic polymicrobial UTI antibiotic therapy guided by a limited microbiological investigation may be practical, adequate and cost-saving.

Assessment of morphology for rapid presumptive identification of Mycobacterium tuberculosis and Mycobacterium kansasii.

Mycobacterium tuberculosis often exhibits serpentine cording when grown in liquid medium, whereas Mycobacterium kansasii can be larger and cross-barred. We assessed the use of these morphologic characteristics as a cost-effective method for rapid presumptive identification of isolates from BACTEC bottles. Without specific training, using the Kinyoun acid-fast stain, definitive cording was found in 237 of 373 specimens positive for M. tuberculosis (64%) and cross-barring was recognized within 63 of 76 (83%) of the specimens positive for M. kansasii, giving sensitivities specificities, positive predictive values, and negative predictive values of 63.5, 96, 92, and 79%, respectively, for M. tuberculosis and 83, 95, 59, and 98%, respectively, for M. kansasii. With training and experience, these results improved to 74.5, 98, 96, and 84% and 93, 98, 79, and 98%, respectively. The major improvements were in distinguishing the pseudocording, or loose aggregation of Mycobacterium avium complex from M. tuberculosis and the long beaded forms of Mycobacterium gordonae from M. kansasii. Mycobacterium asiaticum and Mycobacterium szulgai, which rarely occur, are genetically related to M. kansasii and morphologically difficult to distinguish. In defined circumstances, serpentine cording and cross-barring can be used for rapid presumptive identification of M. tuberculosis and M. kansasii, respectively, and as guides for initial probe selection to reduce costs.

Aerococcus urinae in urinary tract infections.

Aerococcus urinae is a rarely reported pathogen, possibly due to difficulties in the identification of the organism. A. urinae is a gram-positive coccus that grows in pairs and clusters, produces alpha-hemolysis on blood agar, and is negative for catalase and pyrrolidonyl aminopeptidase. Some of these characteristics and its being absent from the databases of most commercial identification systems could allow A. urinae to be misidentified as a streptococcus, enterococcus, or staphylococcus. We report two cases of urinary tract infection (UTI) caused by A. urinae and characterize these isolates by morphology, biochemical testing, whole-cell fatty acid analysis, 16S rRNA gene sequencing, and antibiotic susceptibilities. Most patients infected with A. urinae are elderly males with predisposing conditions who present initially with UTI. Because A. urinae is resistant to sulfonamides, treatment could be inappropriate, with infections resulting in serious complications, including death. It is important for the clinician and the microbiologist to consider A. urinae a potential pathogen and proceed with thorough microbiological identification.

Streptococcus bovis infection of the central nervous system: report of two cases and review.

Streptococcus bovis is an uncommon cause of meningitis and subdural empyema. We report one case each of meningitis and subdural empyema in which S. bovis biotype II was isolated from both the spinal fluid and blood. In one case, the organisms were seen on a gram-stained preparation of cerebrospinal fluid. The first patient presented with gastrointestinal symptoms of unknown etiology, was immunosuppressed, and recovered. The second patient presented with syncope, developed a subdural empyema, and died; at autopsy, a colonic adenoma was found. A review of the English-language literature revealed only 14 previously reported cases of meningitis due to S. bovis and no cases of subdural empyema due to S. bovis. These cases indicate the importance of complete laboratory identification of specific organisms and confirm the need for a thorough neurological examination and search for underlying gastrointestinal disease in cases of S. bovis infection.

Extraintestinal human infection caused by Edwardsiella tarda.

Edwardsiella tarda is an uncommon enteric bacterium which has been found generally in animal hosts and occasionally in human feces. Three cases of extraintestinal infection caused by E. tarda which are described herein include a typhoid-like illness, peritonitis with sepsis, and cellulitis from a wound acquired while fishing. The microbiology of E. tarda and the previous reports of infection due to this organism are reviewed.