Study of the in vitro activities of rifaximin and comparator agents against 536 anaerobic intestinal bacteria from the perspective of potential utility in pathology involving bowel flora.

Rifaximin, ampicillin-sulbactam, neomycin, nitazoxanide, teicoplanin, and vancomycin were tested against 536 strains of anaerobic bacteria. The overall MIC of rifaximin at which 50% of strains were inhibited was 0.25 microg/ml. Ninety percent of the strains tested were inhibited by 256 microg/ml of rifaximin or less, an activity equivalent to those of teicoplanin and vancomycin but less than those of nitazoxanide and ampicillin-sulbactam.

In vitro activities of DX-619 and four comparator agents against 376 anaerobic bacterial isolates.

The activity of DX-619 was evaluated against 376 anaerobic isolates using the reference CLSI agar dilution method. Overall, 90% of the strains were susceptible to DX-619 at < or =1 microg/ml. It was more active than the other four compounds tested except for meropenem, which showed virtually identical overall activity.

Activities of ABT-773 against microaerophilic and fastidious organisms.

ABT-773 was tested against 317 fastidious isolates; it inhibited 99% of organisms at a concentration of 4.0 microg/ml. With ampicillin-sulbactam and levofloxacin, only 2 and 6% of these strains, respectively, were resistant at the breakpoint. With clindamycin, penicillin G, and metronidazole, 22, 26, and 58% of the strains, respectively, were resistant.

Characterisation of aerobic gram-negative bacteria from subgingival sites of dogs--potential bite wound pathogens.

Ninety-eight aerobic, gram-negative bacterial isolates from subgingival samples from family-owned dogs with naturally occurring periodontitis were characterised phenotypically by conventional biochemical testing, by cellular fatty acid profiling and by the use of commercial identification systems. The majority (48, 81%) of the fermentative isolates but only 18% of the non-fermenters were identified by conventional biochemical testing alone. With additional cellular fatty acid profiling, another 7 (12%) fermentative and 23 (59%) non-fermentative isolates were identified to genus or group level. Cellular fatty acid analysis was essential for the identification of most non-fermenters, many of which are difficult to identify due to a paucity of positive reactions in routine biochemical tests. Commercial identification systems were less useful and did not contribute to further identification of these problematic isolates. This study underlines the difficulties encountered in the identification of canine oral bacteria--a group of potential bite wound pathogens--and presents schemes for microbiology laboratories to characterise such isolates.

Short-term benefit from oral vancomycin treatment of regressive-onset autism.

In most cases symptoms of autism begin in early infancy. However, a subset of children appears to develop normally until a clear deterioration is observed. Many parents of children with "regressive"-onset autism have noted antecedent antibiotic exposure followed by chronic diarrhea. We speculated that, in a subgroup of children, disruption of indigenous gut flora might promote colonization by one or more neurotoxin-producing bacteria, contributing, at least in part, to their autistic symptomatology. To help test this hypothesis, 11 children with regressive-onset autism were recruited for an intervention trial using a minimally absorbed oral antibiotic. Entry criteria included antecedent broad-spectrum antimicrobial exposure followed by chronic persistent diarrhea, deterioration of previously acquired skills, and then autistic features. Short-term improvement was noted using multiple pre- and post-therapy evaluations. These included coded, paired videotapes scored by a clinical psychologist blinded to treatment status; these noted improvement in 8 of 10 children studied. Unfortunately, these gains had largely waned at follow-up. Although the protocol used is not suggested as useful therapy, these results indicate that a possible gut flora-brain connection warrants further investigation, as it might lead to greater pathophysiologic insight and meaningful prevention or treatment in a subset of children with autism.

Beta-lactamase production in Prevotella intermedia, Prevotella nigrescens, and Prevotella pallens genotypes and in vitro susceptibilities to selected antimicrobial agents.

The present study investigated the beta-lactamase production of 73 Prevotella intermedia, 84 Prevotella nigrescens, and 14 Prevotella pallens isolates and their in vitro susceptibilities to six antimicrobial agents. The P. intermedia and P. nigrescens isolates were recovered from oral and extraoral samples obtained from subjects in two geographic locations from 1985 to 1995. The clonality of the beta-lactamase-positive and beta-lactamase-negative isolates and the clustering of the genotypes were studied by arbitrarily primed-PCR fingerprinting. beta-Lactamase production was detected in 29% of P. intermedia isolates, 29% of P. nigrescens isolates, and 57% of P. pallens isolates. No difference in the frequencies of beta-lactamase production by P. intermedia and P. nigrescens between isolates from oral and extraoral sites, between isolates obtained at different time periods, or between P. intermedia isolates from different geographic locations was observed. However, the P. nigrescens isolates from the United States were significantly more frequently (P = 0.015) beta-lactamase positive than those from Finland. No association between the genotypes and beta-lactamase production or between the genotypes and the sources of the isolates was found. The penicillin G MICs at which 90% of the isolates were inhibited were 8 microg/ml for P. intermedia, 8 microg/ml for P. nigrescens, and 16 microg/ml for P. pallens. For the beta-lactamase-negative isolates, the corresponding values were 0.031, 0.031, and 0.125 microg/ml, and for the beta-lactamase-positive isolates, the corresponding values were 16, 8, and 32 microg/ml. All isolates were susceptible to amoxicillin-clavulanate, cefoxitin, metronidazole, azithromycin, and trovafloxacin. The MICs of amoxicillin-clavulanate and cefoxitin were relatively higher for the beta-lactamase-positive population than for the beta-lactamase-negative population.

Genetic relatedness between oral and intestinal isolates of Porphyromonas endodontalis by analysis of random amplified polymorphic DNA.

Genomic fingerprints from the DNA of 27 strains of Porphyromonas endodontalis from diverse clinical and geographic origins were generated as random amplified polymorphic DNA (RAPD) using the technique of PCR amplification with a single primer of arbitrary sequence. Cluster analysis of the combined RAPD data obtained with three selected 9- or 10-mer-long primers identified 25 distinct RAPD types which clustered as three main groups identifying three genogroups. Genogroups I and II included exclusively P. endodontalis isolates of oral origin, while 7/9 human intestinal strains of genogroup III which linked at a similarity level of 52% constituted the most homogeneous group in our study. Genotypic diversity within P. endodontalis, as shown by RAPD analysis, suggests that the taxon is composed of two oral genogroups and one intestinal genogroup. This hypothesis remains to be confirmed.

Comparison of recovery of anaerobic bacteria using the Anoxomat, anaerobic chamber, and GasPak jar systems.

The Anoxomat system provides an automated evacuation-replacement technique to create an anaerobic or microaerophilic environment in a jar. We evaluated the Anoxomat system for the growth of obligate anaerobes and for the recovery of anaerobic organisms from clinical specimens, and compared its performance to that of an anaerobic chamber and the GasPak System. Of the 54 stock strains tested, the Anoxomat, the chamber, and the GasPak recovered 95%, 95% and 93% at 24 h, respectively. On 29 occasions (51%), the colonies on the Anoxomat plates were slightly larger than those in the chamber and on 17 (30%) occasions larger than the colonies on the GasPak jar plates. At 48 h, the Anoxomat, the chamber, and the GasPak recovered 93.5%, 94.4% and 88.9%, respectively; of 108 anaerobes isolated from 31 clinical specimens. Methylene blue indicators became decolorized (average of 10 tests) within 2 h inside the Anoxomat jars, 2 h 10 min inside the anaerobic chamber, and 2 h 30 min inside the GasPak jars.

Characteristics of an unusual anaerobic pigmented gram-negative rod isolated from normal and inflamed appendices.

During our studies of the bacterial etiology of appendicitis, we often isolated a previously undescribed anaerobic gram-negative rod. This organism resembled the Bacteroides fragilis group because it was resistant to bile and because of its special-potency-disk pattern (resistant to vancomycin, kanamycin, and colistin), but unlike the B. fragilis group, this bacterium produced brown pigment on media containing hemolysed blood. The cellular fatty acid pattern, with iso-C15:0 being the predominant acid, was most closely related to the fatty acid profile of Porphyromonas species; however, this organism differed from Porphyromonas species by being bile-resistant and by not producing butyrate as a metabolic endproduct. Enzymatic activities of 31 isolates were determined with use of the API ZYM system and Rosco diagnostic tablets. These profiles were different from those of Prevotella, Porphyromonas, and related species. This organism was isolated from 40% of appendiceal tissue samples; no obvious qualitative or quantitative difference in rates of isolation from patients with inflamed or normal appendices was observed.

Role of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens in extraoral and some odontogenic infections.

Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens were isolated from 138 subjects with various infections (intraabdominal, skin and soft-tissue, head and neck, pleuropulmonary, and odontogenic infections and bacteremia). The phenotypic identification of 173 isolates was completed by molecular methods. Arbitrarily primed polymerase chain reaction (AP PCR) analysis was used to determine the genetic similarity of intraindividual P. intermedia/P. nigrescens group isolates recovered from 12 subjects. All 19 P. gingivalis isolates (16 intraabdominal isolates and three odontogenic isolates) hybridized with the P. gingivalis-specific DNA probe. Of the 154 P. intermedia/ P. nigrescens group isolates, 74 were identified as P. intermedia; 78, as P. nigrescens; and 2, as P intermedia/P. nigrescens-like isolates. P. intermedia and P. nigrescens were isolated with equal frequency from patients with all other infections except those with bacteremia, from whom only P. nigrescens isolates were recovered. There were 12 cases in which multiple P. intermedia/ P. nigrescens group isolates were recovered; in nine, only one of the species was isolated, whereas in three, two different species were detected. The intraindividual isolates representing the same species always exhibited identical AP PCR genotypes.

Decrease in the CGGn trinucleotide repeat mutation of the fragile X syndrome to normal size range during paternal transmission.

The fragile X syndrome, the most common inherited form of mental retardation, is caused by the expansion of a CGGn trinucleotide repeat in the FMR-1 gene. Although the repeat number usually increases during transmission, few cases with reduction of an expanded CGGn repeat back to the normal size range have been reported. We describe for the first time a family in which such reduction has occurred in the paternal transmission. The paternal premutation (delta = 300 bp) was not detected in one of the five daughters or in the son of this daughter, although he had the grandpaternal RFLP haplotype. Instead, fragments indicating the normal CGGn repeat size were seen on a Southern blot probed with StB12.3. PCR analysis of the CGGn repeat confirmed this; in addition to a maternal allele of 30 repeats, an allele of 34 repeats was detected in the daughter and, further, in her son. Sequencing of this new allele revealed a pure CGGn repeat configuration without AGG interruptions. No evidence for a somatic mosaicism of a premutation allele in the daughter or a normal allele in her father was detected when investigating DNA derived from blood lymphocytes and skin fibroblasts. Another unusual finding in this family was lack of the PCR product of the microsatellite marker RS46 (DXS548) in one of the grandmaternal X chromosomes, detected as incompatible inheritance of RS46 alleles. The results suggest an intergenerational reduction in the CGGn repeat from premutation size to the normal size range and stable transmission of the contracted repeat to the next generation. However, paternal germ-line mosaicism could not be excluded as an alternative explanation for the reverse mutation.

Survival of anaerobes in original specimens transported by overnight mail services.

Overnight mail delivery was evaluated for its effect on the recovery of facultative and anaerobic microbes in cultures of clinical specimens from patients. Ten clinical specimens, which were collected at different geographic locations and during different weather conditions, were cultured at the site and after overnight delivery to a distant laboratory. Forty-five facultative anaerobic isolates and 48 anaerobes were recovered. There was no significant difference in numbers of strains or relative quantities recovered in cultures of transported and nontransported specimens. With proper collection, transport, and inoculation of specimens, overnight delivery did not compromise recovery of clinically relevant microbes.

Bacteriology of skin and soft-tissue infections: comparison of infections in intravenous drug users and individuals with no history of intravenous drug use.

The bacteriology of cutaneous or subcutaneous abscesses (86 specimens) among intravenous drug users (IVDUs) was compared with the bacteriology of abscesses (74 specimens) in patients with no history of intravenous drug use (non-IVDUs). The IVDU abscesses yielded 173 aerobes and 131 anaerobes. Staphylococcus aureus was the most common aerobe isolated (50% of specimens yielded this isolate), followed by "Streptococcus milleri" (46%). The commonly encountered anaerobes were Fusobacterium nucleatum (17%), pigmented Prevotella species (22%), Peptostreptococcus micros (17%), Actinomyces odontolyticus (15%), and Veillonella species (13%). The non-IVDU isolates included 116 aerobes and 106 anaerobes. S. aureus was isolated from 53% of these specimens, followed by coagulase-negative staphylococci (19%), "S. milleri" (19%), and Streptococcus pyogenes (16%). The main groups of anaerobes were Peptostreptococcus species (35%), Bacteroides species (19%), and gram-positive bacilli (31%). Overall, 67% of the IVDU isolates were of oral origin, compared with 25% of the non-IVDU isolates. Of the specimens from IVDUs and non-IVDUs, 48% and 67%, respectively, yielded only aerobes, and 2% and 4%, respectively, yielded only anaerobes. Sixty-four percent of the patients had one or more beta-lactamase-producing organisms.

The fragile X syndrome in Finland: demonstration of a founder effect by analysis of microsatellite haplotypes.

Microsatellite markers RS46 (DXS548) and FRAXAC2 flanking the fragile X mutation, an expansion of a (CGG)n repeat within the FMR-1 gene, were typed in 60 unrelated northern and eastern Finnish fragile X families and in a control population from the same geographical region. A significant difference was found in allelic and haplotypic distributions between the normal X and fragile X chromosomes. Evidence for a strong founder effect was detected, with the haplotype 196-153 being present on 80% of the fragile X chromosomes, but on only 8% of the normal X chromosomes. In addition to this major haplotype, four minor haplotypes were found on the fragile X chromosomes. These results suggest that the majority of present-day fragile X mutations in Finland may have a common initial ancestor, probably from the 16th century.

Diagnosis of fragile X syndrome by direct mutation analysis.

A total of 27 fragile X pedigrees consisting of over 100 nuclear families were analyzed by Southern blotting methods and probes StB12.3 and StB12.3xx to detect the expansion of the (CGG)n repeat within the FMR-1 gene and the abnormal methylation pattern of the adjacent DNA region responsible for the fragile X syndrome. Clinical expression was found to be associated with the presence of a full mutation (delta > 500 bp, associated with abnormal methylation) in all the males and 50% of the females studied, whereas individuals carrying a premutation (delta = 100-700 bp) were normal. A preferential size increase in the enlarged (CGG)n repeat was detected in successive generations, the instability being stronger when transmitted from a female than from a male. No expansion of the premutation to the full mutation occurred in the paternal transmissions, and the size increase was significantly smaller than in the maternal transmissions. This could partly explain the stability of the premutation through several generations in families with transmitting males. In the maternal transmissions, the risk of expansion of a premutation to a full mutation appeared to depend on its size. The critical maternal premutation size leading invariably to the full mutation was between delta = 175-200 bp. This is important for genetic counseling and also explains the commonly observed clustering of affected individuals in fragile X families.

Peritonsillar abscess, retropharyngeal abscess, mediastinitis, and nonclostridial anaerobic myonecrosis: a case report.

Peritonsillar abscess is a potentially life-threatening complication of acute tonsillitis. On occasion, peritonsillar abscess can extend to neck spaces and/or to the mediastinum. We describe a case of a patient with a peritonsillar abscess that extended to the neck, producing bilateral retropharyngeal abscesses and myonecrosis of the strap muscles. Culture of a specimen of the necrotic muscle yielded Prevotella intermedia, Prevotella buccae, Lactobacillus catenaforme, another Lactobacillus species, Peptostreptococcus anaerobius, and some nonanaerobes. Culture of the peritonsillar abscess yielded P. intermedia and P. buccae plus P. anaerobius, Peptostreptococcus asaccharolyticus, Bifidobacterium dentium, viridans and group F streptococci, and Citrobacter diversus. Culture of the retropharyngeal abscess yielded Fusobacterium nucleatum and Actinomyces odontolyticus in addition to most of the aforementioned organisms. The patient underwent repeated drainage and debridement procedures and was treated with various antimicrobial agents and ultimately recovered. This case highlights the polymicrobial nature of peritonsillar abscess and the serious complications that this infection may lead to.

Comparison of the Accu-CulShure system and a swab placed in a B-D Port-a-Cul tube for specimen collection and transport.

We compared the Accu-CulShure guarded specimen collection device and a swab inserted into a B-D Port-a-Cul transport tube in terms of their efficacy under ideal conditions for recovery of bacteria from 10 decubitus ulcer specimens. Cultures yielded 57 aerobes and 21 anaerobes; 76 isolates were recovered with use of Accu-CulShure, and 72 isolates were recovered with use of Port-a-Cul. Both systems were comparable for recovery of organisms in terms of quantitative and qualitative results.

Porphyromonas-like gram-negative rods in naturally occurring periodontitis in dogs.

A total of 259 Gram-negative Porphyromonas-like rods isolated from subgingival plaque samples of 16 family-owned dogs with naturally occurring periodontitis were characterized phenotypically by biochemical reactions, metabolic end products and enzymatic activities (API-ZYMTM, RoscoTM). Four distinct groups were found. Group A isolates (63) were asaccharolytic, lipase negative, trypsin positive and produced phenylacetic acid (PAA) from peptone-yeast extract glucose broth. Unlike P. gingivalis strains they were catalase positive. Group B isolates (42) differed from those of group A by a positive lipase reaction and from those of group D by failing to ferment sugars. Group C isolates (88) were asaccharolytic and did not produce PAA. They were alpha-fucosidase, N-acetyl-beta-glucosaminidase (beta-NAG) and trypsin negative, resembling P. endodontalis, but unlike human isolates, they were catalase positive. Subgroup C.1 isolates (6) differed from those of parent group C by producing minor amounts of PAA, and subgroup C.2 isolates (12) were beta-NAG positive. Group D isolates (46) were weakly fermentative, lipase, catalase and trypsin positive, and produced PAA. They resembled the B (P.) salivosus type strain which, in our hands, fermented weakly glucose, lactose and mannose. Two isolates could not be assigned to any of the previous groups.