Recovery Of Anaerobic Bacteria Research Articles

BIOME-Preserve: A novel storage and transport medium for preserving anaerobic microbiota samples for culture recovery.

Sequencing-based protocols for studying the human microbiome have unearthed a wealth of information about the relationship between the microbiome and human health. But these microbes cannot be leveraged as therapeutic targets without culture-based studies to phenotype species of interest and to establish culture collections for use in animal models. Traditional sample collection protocols are focused on preserving nucleic acids and metabolites and are largely inappropriate for preserving sensitive anaerobic bacteria for later culture recovery. Here we introduce a novel microbiome preservation kit (BIOME-Preserve) that facilitates recovery of anaerobic bacteria from human stool. Using a combination of culture recovery and shallow whole-genome shotgun sequencing, we characterized the anaerobes cultured from fresh human stool and from human stool held at room temperature in BIOME-Preserve for up to 120 hours. We recovered several species of interest to microbiome researchers, including Bifidobacterium spp., Bacteroides spp., Blautia spp., Eubacterium halii (now Anaerobutyricum hallii), Akkermansia muciniphila, and Faecalibacterium prausnitzii. We also demonstrated that freezing at -80°C did not adversely affect our ability to culture organisms from BIOME-Preserve, suggesting that it is appropriate both as a transport medium and as a medium for longer-term ultra-cold storage. Together, our results suggest BIOME-Preserve is practical for the collection, transport, and culture of anaerobic bacteria from human samples and can help enable researchers to better understand the link between the microbiome and human health and how to leverage that link through novel microbiome-based therapeutics.

Recovery of Anaerobic Bacteria From Wounds After Lawn-Mower Injuries

Accidental injury while using lawn mowers can cause serious infectious complications in the injured extremity. Anaerobic bacteria were rarely recovered from this infection. Two children who sustained injury in their foot by a lawn mower developed severe wound infection. Culture of the wound from 1 patient had heavy growth of Clostridium bifermentans and Peptostreptococcus magnus, and the culture from the other child grew Clostridium perfringens. Antimicrobial therapy directed at the pathogens and vigorous surgical irrigation and debridement led to complete recovery from the infection. This report illustrates the recovery of anaerobic bacteria from children that had wound infection after lawn-mower injury.

Recovery of anaerobic bacteria from a case of dissecting cellulitis

International Journal of DermatologyVolume 45, Issue 2 p. 168-169 Recovery of anaerobic bacteria from a case of dissecting cellulitis Itzhak Brook MD, MSc, Itzhak Brook MD, MSc Washington, DC, USASearch for more papers by this author Itzhak Brook MD, MSc, Itzhak Brook MD, MSc Washington, DC, USASearch for more papers by this author First published: 21 January 2005 https://doi.org/10.1111/j.1365-4632.2005.02594.xCitations: 11Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume45, Issue2February 2006Pages 168-169 RelatedInformation

Microbiology and Management of Decubitus Ulcers in Children

The recent increased recovery of anaerobic bacteria from children has led to greater appreciation of the role anaerobes play in pediatric infections at all body sites, including decubitus ulcers. In studies that employed adequate method for recovery of anaerobes polymicrobial, aerobic, and anaerobic bacteria were isolated from about half of the children with decubitus ulcers. A correlation between the site of the ulcer and the microbial flora of the ulcer was found. The predominant isolates were Staphylococcus aureus, Peptostreptococcus spp., Bacteroides fragilis group, and Pseudomonas aeruginosa. Most of the S. aureus isolates were recovered from ulcers of the hand and the leg. Organisms that resided in the mucous membranes close to the ulcer predominated in the wound next to these areas. Enteric gram-negative rods, Group D enterococcus, and B. fragilis group predominated in ulcers of the buttocks. Group A streptococci, Haemophilus influenzae, pigmented Prevotella and Porphyromonas spp., and Fusobacterium spp. were most frequently recovered in ulcers of the skull. Management of decubitus ulcers should include administration of antimicrobials effective against the polymicrobial bacterial flora according to the anatomic site of the ulcer.

Erratum

There were 2 errors associated with a brief report that appeared only in the electronic edition of the 1 July 2001 issue of the journal (Brook I. Recovery of anaerobic bacteria from 3 patients with infection at a pierced body site. Clin Infect Dis 2001; 33:e12–3). First, Dr. Brook's report was inadvertently omitted from the table of contents of the print edition of the journal. Dr. Brook's report should have been listed in the table of contents as the fourth electronic-only brief report to be mentioned on page 138 of the print edition. Second, in sentence 2 of the abstract of Dr. Brook's report, the second set of parentheses should include the species “Staphylococcus aureus,” not “Streptococcus aureus.” The journal regrets these errors.

Recovery of anaerobic bacteria from four children with postthoracotomy sternal wound infection.

The cases of 4 children who developed postthoracotomy sternal wound infection caused by anaerobic bacteria are presented. The predominate anaerobes were Peptostreptococcus species and pigmented Prevotella species. Polymicrobial infection was present in all cases, and aerobic bacteria also were recovered in 2 instances. All patients responded to surgical debridement and antimicrobials effective against the isolated aerobic and anaerobic bacteria. These findings highlight the potential importance of anaerobic bacteria in postthoracotomy sternal wound infection.

Ocular infections due to anaerobic bacteria.

The increased recovery of anaerobic bacteria in clinical infection has led to greater appreciation of these organisms in ocular infections. In studies that employed adequate method for recovery of anaerobes they were isolated from about a third of patients with conjunctivitis, half of the time in pure culture. The predominant recovered anaerobes were Clostridium spp., gram-negative anaerobic bacilli, and Peptostreptococcus spp. Anaerobic bacteria were also recovered from patients who wore contact lenses and developed conjunctivitis. Anaerobic bacteria were also reported in cases of keratitis. The most frequently recovered anaerobes were Propionibacterium spp., Peptostreptococcus spp., Clostridium spp., Prevotella spp., and Fusobacterium spp. The most frequently recovered anaerobes from dacryocystitis were Peptostreptococcus spp., Propionibacterium spp., Prevotella spp., and Fusobacterium spp. This review describes the microbiology, diagnosis and management of ocular infections due to anaerobic bacteria in children.

Blood culture bottles for transportation and recovery of anaerobic bacteria from non-blood samples.

Using bacterial suspensions as simulated non-blood specimens, the capacity of three different BacT/ Alert blood culture bottles for the transportation and recovery of anaerobic bacteria with different sensitivity to air was evaluated. To better assess the performance of the BacT/Alert bottles, three other liquid media specially designed for anaerobes were included in the study. Attention was paid to recovery rates in relation to species, initial bacterial concentration, and time needed for detection. Of the BacT/Alert blood culture bottles, the anaerobic FAN bottle yielded the highest recovery rates, but its performance was limited compared with chopped meat broth in tubes. This broth allowed detection of all the tested species within 48 h. Since collection and transportation of anaerobic bacteria are of major importance for a reliable culture result, improvements are necessary.

Blood culture bottles for transportation and recovery of anaerobic bacteria from non-blood samples.

Using bacterial suspensions as simulated non-blood specimens, the capacity of three different BacT/ Alert blood culture bottles for the transportation and recovery of anaerobic bacteria with differ ...

Effect of media on transport and recovery of anaerobic bacteria.

Clinical specimens containing anaerobic bacteria require special handling to enable satisfactory recovery of these important pathogens. Problems relating to inadvertently exposing the bacteria to oxygen can arise at the time of specimen collection, during transport, and during the culture setup process. A considerable variety of transport and culture materials exists for the clinical laboratory, yet not much has been written recently on the optimal utilization of culture media or the maintenance of anaerobiasis to maximize recovery of strict anaerobic bacteria. Comparative studies are needed to define how clinical laboratories can carry out cost-efficient specimen processing for recovery of these pathogens that is timely and medically relevant.

Aerobic and Anaerobic Microbiology of Empyema: A Retrospective Review in Two Military Hospitals

The microbiology and clinical features of empyema were studied retrospectively in 197 patients whose specimens yielded bacterial growth after inoculation for aerobic and anaerobic bacteria. Three hundred forty-three organisms (216 aerobic or facultative and 127 anaerobic organisms) were isolated. Aerobic bacteria were isolated in 127 (64 percent) patients, anaerobic bacteria in 25 (13 percent), and mixed aerobic and anaerobic bacteria in 45 (23 percent). The predominant aerobic or facultative organisms were <i>Streptococcus pneumoniae</i> (70 isolates), <i>Staphylococcus aureus</i> (58), <i>Escherichia coli</i> (17), <i>Klebsiella pneumoniae</i> (16), and <i>Haemophilus influenzae</i> (12). The predominant anaerobes were pigmented Prevotella and Porphyromonas species (24), <i>Bacteroides fragilis</i> group (22), anaerobic cocci (36), and Fusobacterium species (20). β-Lactamase-producing organisms were recovered in 49 (38 percent) of 128 tested specimens. These included all 42 tested <i>S aureus</i> and 15 <i>B fragilis</i> group, 4 of 9 <i>K pneumoniae</i>, 3 of 9 <i>H influenzae</i>, 3 of 8 pigmented Prevotella and Porphyromonas species, and 2 of 6 <i>E coli</i>. Most patients from whom <i>S pneumoniae</i> and <i>H influenzae</i> were recovered had pneumonia, and most patients with <i>S aureus</i> had pneumonia, aspiration pneumonia, and lung abscesses. The recovery of anaerobic bacteria was mostly associated with the concomitant diagnosis of aspiration pneumonia, and lung, subdiaphragmatic, dental, and oropharyngeal abscesses. These data highlight the importance of anaerobic bacteria in selected cases of empyema.

Microbiology of Empyema in Children and Adolescents

The microbiology of empyema was studied in 72 children and adolescents whose specimens yielded bacterial growth after inoculation for aerobic and anaerobic bacteria. A total of 93 organisms, 60 aerobic or facultative and 33 anaerobic, were isolated. Aerobic bacteria was isolated in 48 (67%) patients, anaerobic bacteria in 17 (24%), and mixed aerobic and anaerobic bacteria in 7 (10%). The predominant aerobic or facultative bacteria were Haemophilus influenzae (15 isolates), Streptococcus pneumoniae (13), and Staphylococcus aureus (10). The predominant anaerobes were Bacteroides sp (15 isolates, including 7 Bacteroides fragilis group and 5 Bacteroides melaninogenicus group), anaerobic cocci (9), and Fusobacterium sp (6). beta-lactamase activity was detected in at least one isolate in 20 (37%) of the 54 tested patients. These included all 8 tested S aureus and 7 B fragilis group, 3 of 10 H influenzae, 2 of 4 B melaninogenicus group, and 1 of 2 Klebsiella pneumoniae. Most cases of S pneumoniae and H influenzae were associated with pneumonia. The recovery of anaerobic bacteria was mostly associated with the concomitant diagnosis of aspiration pneumonia, lung abscess, subdiaphragmatic abscess, and abscesses of dental or oropharyngeal origin. The data highlight the importance of anaerobic bacteria in selected cases of empyema in children and adolescents.

Septic Arthritis Caused by Bacteroides fragilis

As improvements in bacteriologic techniques have enhanced the recovery of anaerobic bacteria from clinical specimens, there has been an increasing awareness of the role of anaerobes in disease. Bacteroides fragilis is the most common anaerobic organism found in clinical specimens. Although it is the anaerobe most frequently associated with bacteremia and a common isolate in intraabdominal infections, infections of the female genital tract, wounds, and abscesses, B. fragilis is a rare cause of septic arthritis. The isolation of this organism from four patients with septic arthritis in three Cleveland hospitals between 1978 and 1982 suggests that septic arthritis due to B. fragilis may be a more common clinical entity than previously appreciated. In this report we describe these cases and review the pertinent literature.

Recovery of anaerobic bacteria from vented blood-culture bottles.

From January 1979, to July 1981, blood from each patient suspected of having bacteremia was collected in two bottles of commercially prepared tryptic soy broth; one bottle was vented for 24 hr to the atmosphere. Bottles were incubated at 37 degrees C for a maximum of seven days and were examined daily for signs of growth. Blind subcultures onto chocolate-agar plates were performed 18-24 hr after collection and again at 48 hr. For 63,106 bottles inoculated, a total of 4,788 strains were isolated, for a rate of 6.4% (excluding contaminants). Two hundred and thirty (4.8%) isolates were anaerobes, of which 75 (33%) were identified as Bacteroides fragilis. It is significant that 20 of these anaerobic isolates (approximately 9%) were recovered only from the vented bottles. These included eight strains of B. fragilis; three strains of Clostridium perfringens; and one strain each of Bacteroides distasonis, Fusobacterium nucleatum, Peptococcus saccharolyticus, and Eubacterium species. These results emphasize the need to check vented bottles for anaerobes, since significant numbers of these bacteria could be missed by failing to do so. These data further show the desirability of holding anaerobic blood cultures for a minimum of one week.

A cooked meat blood culture medium; shelf-life and clinical evaluation compared with other systems

The recovery of anaerobic bacteria from small inocula was examined in an in vitro simulated blood culture system. A freshly prepared cooked meat medium gave the best results while Difco Thiol medium and a commercially available cooked meat medium gave good recovery; Brewer's thioglycollate medium was unsatisfactory. Those media tested performed less well after storage. However, stored Difco Thiol medium and laboratory prepared cooked meat media could be revitalised by steaming, and restored to full efficacy. In a clinical trial of blood culture media, freshly prepared cooked meat medium was compared with an established two-bottle system of Brewer's thioglycollate and Brain Heart Infusion media. The inclusion of freshly prepared cooked meat medium increased the percentage of positive blood cultures from 10·1 to 11·8 per cent. Eighty six per cent of positive cultures could have been detected by our previous technique; 98 per cent could be isolated by our adoption of a freshly prepared cooked meat medium with Brain Heart Infusion Broth.

Recovery of anaerobic bacteria from small inocula: a model for blood culture studies.

The recovery of anaerobic, facultative anaerobic, and aerobic pathogens from very small inocula was studied under conditions that could be related to routine blood culture procedures. Results with Brain Heart Infusion broth were unsatisfactory. Freshly prepared Brewer thioglycollate medium gave apparently good results, but there are disadvantages when this medium is used for blood culture. Results with Difco Thiol broth were disappointing. A modification of Robertson's cooked-meat broth supplemented with Brain Heart Infusion gave good recovery and sustained viability with a wide range of test organisms including exacting strains. The findings raise points of practical importance.

Oxygen tolerance of anaerobic bacteria isolated from necrotic dental pulps

The oxygen tolerance of 43 anaerobic reference strains and 36 anaerobic strains from necrotic dental pulps was studied. All strains survived for two hours or more as colonies on the surface of a medium supplemented with blood, and as many as 26 of the 79 strains survived for more than seven days. The hemolysed blood in the medium significantly increased the survival time for many of the strains. Factors influencing the death rate were studied in one of the strains and it was found that the lysed red cells of the blood and not the serum had a protective effect and that catalase had the same protective effect as the hemolysed blood. The finding that hemolysed blood significantly increased the oxygen tolerance of many anaerobes may explain some of the divergent results regarding the efficiency of various methods for the recovery of anaerobic bacteria from clinical specimens. The use of media supplemented with blood during various phases of processing a specimen might be more important for a high recovery of anaerobic bacteria from clinical sources than the measures taken to minimize exposure of the specimen to air.

SURVIVAL OF ANAEROBIC BACTERIA DURING TRANSPORTATION

The effect of evacuation of atmospheric air during transportation on recovery of anaerobic bacteria was investigated. Evacuation of atmospheric air from glass tubes by flushing with pure carbon dioxide lowered the content of oxygen to about 0.4 per cent. Three B. fragilis strains and one strain of Fusobacterium mortiferum and of Peptostreptococcus anaerobius were investigated. Bacterial recovery was determined one hour and 24 hours after evacuation of atmospheric air by pure carbon dioxide and pure nitrogen, was compared to bacterial recovery from samples transported with free access to atmospheric air. Evacuation by pure carbon dioxide significantly improved the recovery of one B. fragilis strain after 24 hours of transportation and significantly impaired the recovery of Peptostreptococcus anaerobius after one hour of transportation, while evacuation by pure nitrogen significantly improved the recovery of Peptostreptococcus anaerobius after 24 hours of transportation. In all other cases, however, no statistically significant effect on bacterial recovery was found.

The recovery of anaerobic bacteria from swabs.

When a standard sample of simulated exudate containing known numbers of anaerobic bacteria was taken up on a swab and plated on solid medium, the number of colonies subsequently cultured represented a very small proportion of the original sample. Evidence is produced that the apparent loss is not primarily attributable to inactivation on the swab but rather to retention of organisms on the swab. This was demonstrable with Clostridium welchii and with Bacteroides species that have hitherto been regarded as relatively oxygen-sensitive.When stock strains of Bacteroides species were held for some hours on swabs, some progressive loss of viability was demonstrable. A measure of protection was afforded when these organisms were held aerobically on blood agar medium, but a very exacting anaerobe and some wild strains of faecal anaerobes showed gradual inactivation under these conditions.These findings may have important implications in relation to currently employed bacteriological sampling procedures with swabs in clinical practice.

Comparison of three anaerobic systems for the isolation of anaerobic bacteria from clinical specimens.

An anaerobic glove box, the GASPAK jar system, and a roll-streak tube system with prereduced, anaerobically sterilized medium were compared for recovery of anaerobic bacteria from selected clinical specimens collected in anaerobic tubes. Both prereduced plating media held in an anaerobic glove box for a minimum of 48 hr. before use and fresh plating media exposed to atmospheric oxygen for no longer than 1 hr. were used in the glove box and GASPAK systems. In general, the recoveries of anaerobic bacteria with the three systems were comparable. Bacteroides melaninogenicus strains were recovered with the glove box and GASPAK systems and were not detected with roll-streak tubes, but the PRAS medium used in the roll-streak tube system did not contain hemin or menadione, whereas some of the media used in the other systems did. Only one organism, a peptostreptococcus, was recovered in the roll-streak tube system and not in the GASPAK or glove box systems. Of 29 anaerobes isolated from the 15 specimens examined, 28 were recovered in both the GASPAK and glove box systems and 23 were isolated with the roll-streak tube system. The advantages and disadvantages of each system are discussed.