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Oxygen tolerance of fresh clinical anaerobic bacteria.

The oxygen tolerance and sensitivity of 57 freshly isolated anaerobic bacteria from clinical specimens was studied. All the organisms tolerated 8 h or more of exposure to oxygen in room air. Growth of the isolates in increasing oxygen concentrations demonstrated that the 57 isolates varied in oxygen sensitivity from strict to aerotolerant anaerobes. Comparison of the oxygen tolerance and sensitivity showed that the most tolerant organisms (best survival after prolonged exposure) included anaerobes capable of growth at only 0.4% or less O2 (strict) as well as those able to grow in as much as 10% O2. The least tolerant were predominately strict anaerobes. Decrease in the inoculum size from a concentration of 10(8) to 10(6) colony-forming units per ml had only a minor effect. The data indicate that the brief oxygen exposure with bench techniques in clinical laboratories would not be deleterious to the anaerobic bacteria present in clinical specimens.

Anaerobiosis↗

Gelatin agar medium for detecting gelatinase production by anaerobic bacteria.

A new medium, Lombard-Dowell gelatin agar, was developed for detecting gelatinase activity by anaerobic bacteria. The medium contained: Trypticase (BBL Microbiology Systems), 5.0 g; yeast extract (Difco Laboratories), 5 g; sodium chloride, 2.5 g; sodium sulfite, 0.1 g; L-tryptophan, 0.2 g; L-cystine, 0.4 g; hemin, 10.0 mg; vitamin K1, 10.0 mg; agar, 20.0 g; D-glucose, 1.0 g; gelatin, 4.0 g; and distilled water to 1 liter. The pH was adjusted to 7.5. The medium was dispensed in 100- by 15-mm quadrant plastic dishes (5 ml per quadrant). To test for gelatinase activity, we inoculated the medium with a young enriched thioglycolate or chopped meat glucose broth culture or a turbid cell suspension in Lombard-Dowell broth, using a sterile cotton swab, and incubated it under anaerobic conditions for 48 h at 35 degrees C. The quadrants were then flooded with Frazier solution, and clear zones around the bacterial growth were recorded as positive for gelatinase activity. The new medium was tested with a variety of anaerobic bacteria, and the results were compared with data obtained with the conventional technique for detecting gelatinase activity. Overall, there was satisfactory agreement between the two tests in the detection of gelatinase activity, but the Lombard-Dowell gelatin agar tests was more rapid and somewhat more sensitive than the conventional test.

Agar↗

Anaerobic specimen transport device.

A device is described and evaluated for the anaerobic transport of clinical specimens. The device limits the amount of oxygen entering with the sample to a maximum of 2%, which is rapidly removed by reacting with hydrogen in the presence of a palladium catalyst. The viability on swabs of 12 species of anaerobes, four strains of facultative anaerobes and a strain of Pseudomonas aeruginosa, was maintained during the length of the tests (24 or 48 h). The results demonstrated that this device protected even the more oxygen-sensitive clinical anaerobes from death due to oxygen exposure. This device can be used for swabs as well as for anaerobic collection and liquid and solid specimens.

Anaerobiosis↗

Evaluation of anaerobic microdilution MIC results with the Prompt Inoculation System.

The Prompt Inoculation System (3M Co.) was compared with the overnight suspension inoculation procedure used with a broth microdilution anaerobic commercial system (Micro-Media Systems) for differences in MIC. MIC results from both suspension methods using six National Committee for Clinical Laboratory Standards-recommended quality control organisms were identical in 18 instances (75%) and within +/- 1 log2 dilution in 96% of the comparisons. Results with 45 anaerobic clinical isolates also compared satisfactorily; 83% of the results were identical and 97% were within +/- 1 log2 dilution. In addition, the direct (Prompt)-inoculated microdilution trays produced better growth and more valid MIC results; 92% of the clinical isolates produced MIC results versus 79% by the overnight suspension procedure.

Anti-Bacterial Agents↗

Anaerobic endocarditis caused by Staphylococcus saccharolyticus.

The first case of infective endocarditis caused by the anaerobe Staphylococcus saccharolyticus is reported. The infection occurred in a previously healthy 61-year-old male with no known predisposing valvular heart disease. The patient was successfully treated with a combination of 2 g of nafcillin every 4 h and 90 mg of gentamicin every 8 h for 6 weeks.

Drug Therapy, Combination↗

Numbers and types of anaerobic bacteria isolated from clinical specimens since 1960.

Between 1960 and 1974, 826 specimens, excluding stool, urine, sputum, and blood, yielded 689 (83%) positive cultures, of which 403 (58.5%) contained anaerobic bacteria. This represents 48.8% of the total specimens cultured. Isolates from 153 specimens obtained and stocked from 1965 to 1974 were reidentified by current criteria. Gram-negative bacilli, primarily bacteroides, were the most frequently isolated anaerobes, being found in 70% of 153 anaerobe-positive specimens and accounting for 42% of the total anaerobes isolated. Gram-positive cocci were second in occurrence, being found in 66% of 153 specimens and accounting for 40% of the total isolates. Bacteroides fragilis was by far the most frequently isolated species. Compairson of 14 years of cumulative data with data from current studies covering 1- to 2-year periods indicated that the anaerobes isolated from clinical material have not changed significantly in type or relative numbers.

Anaerobiosis↗

Presumptive diagnosis of anaerobic bacteremia by gas-liquid chromatography of blood cultures.

By quantitative gas-liquid chromatography of glucose-containing blood cultures at the moment of first signs of growth, a presumptive diagnosis of anaerobic bacteremia could be made in 24 out of 26 cultures yielding obligate anaerobes upon subsequent culture. With Bacteroides sp. (20 strains isolated), elevated levels of isovaleric acid (greater than or equal to 0.1 mumol/ml) and/or succinic acid (greater than or equal to 5 mumol/ml) were detected in the medium used. An exception to this were two strains of Bacteroides thetaiotaomicron that did not produce sufficient quantities of these acids. In the case of butyrate-producing gram-positive cocci (four strains), butyric acid in amounts of greater than or equal to 0.8 mumol/ml was detected. Propionibacteria (five strains) produced propionic acid in amounts of greater than or equal to 3.9 mumol/ml. No false positive results were found in 103 blood cultures with growth of aerobic or facultatively anaerobic bacteria only.

Bacterial Infections↗

Anaerobic axillary abscess.

Fifty-two patients with axillary abscesses were seen during two years. Staphylococcus aureus was isolated from 34, anaerobic bacteria from 12, and skin flora from five; in one case the pus was sterile. Seven patients with hidradenitis suppurativa had recurrent infection with abscess formation, which was bilateral in three. Anaerobes were isolated in five of these cases and skin flora alone in two. Anaerobes are secondary invaders in this condition, and histological examination shows that the primary abnormality is obstruction of pilosebaceous follicles and apocrine glands, associated with keratin plugging of the follicles. Chemotherapy offers little hope of cure, although metronidazole removes the offensive smell of the discharge. Radical surgery is usually indicated.

Abscess↗

Anaerobic bacteria in routine urine culture.

One hundred and fifty-eight anaerobic organisms from 147 patients were isolated from 15,250 consecutive clean mid-stream or catheter urine specimens. The pathogenicity of the anaerobic genera commonly isolated from urine is reviewed and discussed. Failure to establish anaerobic isolates as pathogens and a paucity of reported cases proving anaerobic bacteria as significant causes of urinary tract infection permitted discontinuation of anaerobic culture as part of the routine screening procedure for investigation of urinary tract infections.

Adult↗

The isolation of anaerobic bacteria from wound swabs.

The isolation of anaerobic bacteria from routine wound swabs by three procedures was evaluated. Recovery of anaerobic organisms was doubled by immediate incubation of seeded plates, and the recovery could be further dramatically improved by the use of prereduced media, in conjunction with an anaerobic chamber. Recommendations for the treatment of swabs and cultures for anaerobic investigation are made.

Anaerobiosis↗

Bacteriological findings in cultures of clinical material from Bartholin's abscess.

Purulent exudate from 28 cases of Bartholin's abscess were examined for aerobic, anaerobic, and microaerophilic bacteria. Three cases gave no bacterial growth, five cases grew facultative bacteria only, and a further three grew a mixture of facultative and anaerobic bacteria. One case gave a pure growth of a microaerophilic streptococcus. Anaerobic bacteria were the only isolates in 16 cases and Bacteroides species were the most common organisms isolated; they were present as 62.5% of the total anaerobes cultured and accounted for 45.4% of the total bacteria cultured. Escherichia coli was the most common of the facultative organisms, accounting for 18% of the total bacteria. Most infections seemed to be caused by a single organism, anaerobic types predominating.

Abscess↗

Novobiocin and the differentiation of peptococci and peptostreptococci.

Paper discs containing 5 microng of novobiocin were used as a presumptive test to differentiate peptococci and peptostreptococci. Zone diameters were measured and minimum inhibitory concentrations (MIC) of the antibiotic for each group were performed to ascertain the activity of the antibiotic against these genera. All strains of peptococci showed no zone of inhibition in the disc test together with an MIC of 25 microng/ml or greater. All strains of peptostreptococci showed zones of inhibition of at least 15 mm diameter together with an MIC of 1-6 microng/ml or less.

Microbial Sensitivity Tests↗

Survival of gram positive anaerobic cocci on swabs and their isolation from the mouth and vagina.

The survival of Gram positive anaerobic cocci on plain cotton wool and albumin coated swabs held in various transport media was investigated. Results suggested that in most cases Amies', Stuart's and VMGII media do not offer any more protection to the bacteria than storing swabs dry in their containers. A technique was developed for the isolation and identification of Gram positive anaerobic cocci from the mouth and vagina, incorporating bicozamycin in the medium as a selective agent. Few strains were recovered from the oral cavity, but larger numbers were isolated from the vagina. Using a minimum number of antibiotic sensitivity and biochemical tests, including analysis of end products by gas-liquid chromatography, most isolates were identified to species level.

Albumins↗

Heparinase production by anaerobic bacteria.

The production of heparinase by a wide range of anaerobic bacteria isolated from clinical specimens was investigated. None of the 29 strains of Bacteroides fragilis produced heparinase. Of 62 other Bacteroides tested, only two of four strains of B ovatus, two of three strains of B thetaiotaomicron, and two of four strains of B uniformis were heparinase producers. None of the 48 strains of fusobacteria or seven strains of Veillonella produced heparinase. The anaerobic cocci (19 peptococci and seven peptostreptococci) were also negative for heparinase production as were 46 Clostridium spp tested. It was concluded that heparinase production by anaerobic bacteria was unlikely to play a part in the regional thrombophlebitis that sometimes occurs in anaerobic infections.

Bacteria, Anaerobic↗

Gardnerella vaginalis and anaerobic bacteria in genital disease.

In a study of Gardnerella vaginalis and anaerobic bacteria in non-specific vaginitis (NSV) and other genital disease 89 patients attending a genital medicine clinic had vaginal samples examined for conventional pathogens and for quantitative analysis of G vaginalis and aerobic and anaerobic bacterial flora. The overall incidence of G vaginalis was 20%; G vaginalis (mean concentration 7.0 log10/g of secretion) occurred predominantly in patients with NSV (57%) but also in sexual contacts of non-specific urethritis (NSU) (37.5%) and in patients with other conditions (11.8%). G vaginalis is therefore a relatively common isolate in patients with vaginal discharge. The concentration of aerobic and anaerobic bacteria ranged from 4.9-11.0 log10/g of secretion with an anaerobe-to-aerobe ratio of 10:1. Anaerobic bacteria, particularly anaerobic Gram-positive cocci (mean concentrations 7.7 log10/g), were present in patients with NSV and in association with G vaginalis, but they also occurred in other clinical groups and with other pathogens, particularly Trichomonas vaginalis. Anaerobic bacteria may therefore play an important role in the pathogenesis of vaginal infections.

Anaerobiosis↗