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PubMed2005

Clostridium bacteraemia characterised by 16S ribosomal RNA gene sequencing.

BACKGROUND: Owing to problems in accurate species identification of the diverse genus clostridium, the epidemiology and pathogenicity of many species are not fully understood. Moreover, previous studies on clostridium bacteraemia have been limited and relied only on phenotypic species identification. AIMS: To characterise the epidemiology, disease spectrum, and outcome of clostridium bacteraemia using 16S ribosomal RNA (rRNA) gene sequencing. METHOD: During a four year period (1998-2001), all cases of clostridium bacteraemia were prospectively studied and all "non-perfringens" clostridium isolates identified to the species level by 16S rRNA gene sequencing. RESULTS: Fifty one blood culture isolates were identified as Clostridium perfringens and 17 belonged to 11 other clostridium species. The first case of C disporicum infection and two cases of clostridium bacteraemia in children with intussusception were also described. Of the 68 clostridium isolates from 68 different patients, 38 were associated with clinically relevant bacteraemia. The gastrointestinal and hepatobiliary tracts were common sites of both underlying disease and portal of entry in these patients. Clostridium perfringens accounted for 79% of all clinically relevant bacteraemia, with the remainder caused by a diversity of species. The attributable mortality rate of clinically relevant clostridium bacteraemia was 29%. Younger age and underlying gastrointestinal/hepatobiliary tract disease were associated with mortality (p < 0.05). CONCLUSIONS: Patients with clinically relevant clostridium bacteraemia should be investigated for the presence of underlying disease processes in the gastrointestinal or hepatobiliary tracts. 16S rRNA gene analysis will continue to be useful in further understanding the pathogenicity of various clostridium species.

Adult↗
PubMed1995

Fibronectin and laminin binding of eighteen Clostridium species.

The ability of Clostridium difficile, Clostridium perfringens, Clostridium sporogenes and fifteen other Clostridium species to bind to human serum fibronectin or laminin was tested by using protein-coated latex particles. Three groups of Clostridium species were formed, namely the pseudomembranous colitis-causing species Clostridium difficile, the gas gangrene-causing Clostridium species and other Clostridium species, which are infrequently found in human infections. Significantly more strains of gas gangrene-causing Clostridium species, and strains of Clostridium species other than Clostridium difficile recognized fibronectin or laminin than did Clostridium difficile. Experiments with monoclonal antibodies revealed the specificity of the bacterial binding to fibronectin or laminin.

Agar↗
PubMed2001

Clostridium bacteremia: emphasis on the poor prognosis in cirrhotic patients.

Bacteremic episodes caused by anaerobes are unusual and the clinical importance of Clostridium bacteremia remains unclear. This retrospective case study examined the risk factors among a group of patients who developed Clostridium bacteremia. Medical records from 73 episodes of clostridial bacteremia in 73 patients treated in a medical center during an 11-year period were reviewed. Of all episodes, 96% were community-acquired. Twelve percent of patients had polymicrobial bacteremia, with Escherichia coli being the most common accompanying bacterium. Diabetes mellitus (26%) and liver cirrhosis (25%) were the most common underlying diseases. The most common etiological organisms were Clostridium perfringens (77%), Clostridium bifermentans (9%), and Clostridium septicum (4%). Only one patient with C. septicum bacteremia had a histocytotoxic infection, which was a fatal gas gangrene. Univariate analysis of data from patients with monomicrobial Clostridium bacteremia revealed that younger age (age < 65 years), underlying liver cirrhosis, and presence of septic shock at initial presentation were associated with fatality; but only the latter two variables were independently associated with fatality in multivariate logistic regression analysis. Appropriate antimicrobial therapy for monomicrobial Clostridium bacteremia did not significantly affect clinical outcomes, which might suggest that Clostridium species in the bloodstream can be regarded as merely contaminants or transient bacteremia. This suggestion was not supported by the finding that seven of 13 cirrhotic patients with monomicrobial Clostridium bacteremia died of sepsis, of whom six had not receive appropriate antimicrobial therapy. Therefore, the clinical importance of Clostridium bacteremia should be interpreted with caution because of its high risk of mortality in susceptible hosts, particularly cirrhotic patients, who do not receive appropriate therapy timely.

Adult↗
PubMed1997

Clostridium pascui sp. nov., a new glutamate-fermenting sporeformer from a pasture in Pakistan.

Four strains of an obligately anaerobic spore-forming bacterium were isolated from soil samples from a donkey pasture in Pakistan. Comparative 16S rRNA sequence analysis demonstrated that the strains are members of phylogenetic cluster I of the genus Clostridium (Collins et al. 1994). The strains are mesophilic, nonsaccharolytic, and nonproteolytic, utilize glutamate and histidine, and produce indole. Acetate, butyrate, ethanol, hydrogen, and carbon dioxide are the products of fermentation. Although the strains phenotypically resemble the classical glutamate-fermenting clostridia, such as Clostridium cochlearium, Clostridium tetanomorphum, Clostridium tetani, and especially Clostridium malenominatum, they differ from these organisms in sugar utilization, cellular fatty acid composition, and cellular protein pattern and by a 16S rRNA sequence divergence value of approximately 4 to 8%. Phylogenetically, the strains are more closely related to Clostridium estertheticum (sequence divergence, approximately 5%) and Clostridium subterminale (sequence divergence, approximately 5%) but are phenotypically readily distinguished from these species. On the basis of phenotypic and genotypic criteria, we conclude that the four strains are members of a new species of the genus Clostridium, for which the name Clostridium pascui is proposed. The type strain is strain Cm19 (= DSM 10365).

Acetates↗
PubMed1999

Should all stool specimens be routinely tested for Clostridium difficile?

OBJECTIVE: To determine the frequency with which Clostridium difficile was detected in stool specimens from outpatients and patients hospitalized for less than 4 days to assess the usefulness of routine laboratory screening for detecting this enteric pathogen. METHODS: Seven hundred and forty-one specimens from 398 patients were cultured over a 6-month period for Salmonella, Shigella, Yersinia, Escherichia coli O157:H7, Campylobacter and Clostridium difficile. Clostridium difficile culture-positive samples were further tested for cytotoxin production. RESULTS: Campylobacter, Salmonella, Shigella and E. coli O157:H7 were isolated in 50 (6.7%) specimens from 35 (8.8%) patients. Clostridium difficile was cultured from 88 (11.9%) specimens from 35 (8.8%) patients and its cytotoxin detected in 35 (4.7%) specimens of 12 (3%) patients. Clostridium difficile was the second most frequent enteric pathogen after Campylobacter. Of 178 (24%) specimens submitted with a specific request for Clostridium difficile testing, 13 (7.3%) were cytotoxin positive (three patients); of 563 specimens for which Clostridium difficile was not requested, 22 (3.9%) were cytotoxin positive (nine patients). CONCLUSIONS: Nine of 12 patients with cytotoxin-positive specimens would have gone undiagnosed in the laboratory had all stool samples submitted not been tested. These results suggest that Clostridium difficile disease is under-recognized and that testing all stool samples for Clostridium difficile may be warranted in our community of patients.

Journal Article↗
PubMed1979

[Studies of necrotizing enteritis of suckling piglets (Clostridium perfringens type C enterotoxemia) in industrialized sow breeding units. 4. Epizootiology].

Necrotising enteritis had been the cause of death of 4.9 per cent in 5,177 nursed piglets, which was established by pathological examination. The number of piglets, in that context, which had come from industrialised sow breeding units was equivalent to 92 per cent. The nursed piglet held the third position, next to smaller ruminants (19.4 per cent) and fowl (6.0 per cent), with regard to the occurrence of Clostridium perfringens enterotoxemia or necrotising enteritis in 112,218 animals which were pathologically examined after death. Necrotising enteritis so far has been rare in the GDR. No regional accumulation has been observed. Several outbreaks on industrialised sow breeding units actually remained stationary. The occurrence of the disease may be favoured by a number of factors which are conducive to accumulation of Clostridium perfringens Type C in a given stock. Group keeping of pregnant sows, simultaneous farrowing of larger groups of sows, group treatment of nursed piglets, using neomycin, chloramphenicol, oxytetracycline, and other antibiotics to which Clostridium perfringens is primarily resistant or has acquired resistance in the course of time are some of those contributive factors. Transmission of Clostridium perfringens Type C through feedstuff is possible, though it would lead to a real outbreak only by high intensity of the contamination, and it played a minor role in proliferation of the disease. 3479 Clostridium perfringens strains were isolated from 9,481 animals, both clinically intact and after death, with 30 species being included. Type classification revealed 2454 strains of Type A (70 per cent), 204 of Type D (5.88 per cent), 164 of Type C (four per cent), and 48 of Type B (1.34 per cent). There were 688 atoxic strains (17 per cent). Swine is the major carrier of Clostridium perfringens Type C, with 87 per cent of all Clostridium perfringens Type C strains having been isolated from swine. Swine was followed by fowl (four per cent), sheep (four per cent), cattle, rabbit, and dog (1.27 per cent each). Clostridium perfringens Type C was obtained from the faeces of clinically intact sows in seven instances, including two cases with sows (0.46 per cent) from farms with no previous record of necrotising enteritis.

Animals↗
PubMed2007

Clostridium saccharogumia sp. nov. and Lactonifactor longoviformis gen. nov., sp. nov., two novel human faecal bacteria involved in the conversion of the dietary phytoestrogen secoisolariciresinol diglucoside.

Two anaerobic bacteria involved in the conversion of the plant lignan secoisolariciresinol diglucoside were isolated from faeces of a healthy male adult. The first isolate, strain SDG-Mt85-3Db, was a mesophilic strictly anaerobic Gram-positive helically coiled rod. Based on 16S r RNA gene sequence analysis, its nearest relatives were Clostridium cocleatum (96.7% similarity) and Clostridium ramosum (96.6%). In contrast to these species, the isolate was devoid of alpha-galactosidase and -glucosidase and did not grow on maltose, melibiose, raffinose, rhamnose and trehalose. The hypothesis that strain SDG-Mt85-3Db represents a new bacterial species of the Clostridium cluster XVIII was confirmed by DNA-DNA hybridisation experiments. The G+C content of DNA of strain SDG-Mt85-3Db (30.7+/-0.8 mol%) was comparable with that of Clostridium butyricum, the type species of the genus Clostridium. The name Clostridium saccharogumia is proposed for strain SDG-Mt85-3Db (=DSM 17460T=CCUG 51486T). The second isolate, strain ED-Mt61/PYG-s6, was a mesophilic strictly anaerobic Gram-positive regular rod. Based on 16S rRNA gene sequence analysis, its nearest relatives were Clostridium amygdalinum (93.3%), Clostridium saccharolyticum (93.1%) and Ruminococcus productus (93.0%). The isolate differed from these species in its ability to dehydrogenate enterodiol. It also possessed alpha-arabinosidase and -galactosidase and had a higher G+C content of DNA (48.0 mol%). According to these findings, it is proposed to create a novel genus, Lactonifactor, and a novel species, Lactonifactor longoviformis, to accommodate strain ED-Mt61/PYG-s6. The type strain is DSM 17459T (=CCUG 51487T).

4-Butyrolactone↗
PubMed2000

Identification and characterization of a bile acid 7alpha-dehydroxylation operon in Clostridium sp. strain TO-931, a highly active 7alpha-dehydroxylating strain isolated from human feces.

Clostridium sp. strain TO-931 can rapidly convert the primary bile acid cholic acid to a potentially toxic compound, deoxycholic acid. Mixed oligonucleotide probes were used to isolate a gene fragment encoding a putative bile acid transporter from Clostridium sp. strain TO-931. This DNA fragment had 60% nucleotide sequence identity to a known bile acid transporter gene from Eubacterium sp. strain VPI 12708, another bile acid-7alpha-dehydroxylating intestinal bacterium. The DNA (9.15 kb) surrounding the transporter gene was cloned from Clostridium sp. strain TO-931 and sequenced. Within this larger DNA fragment was a 7.9-kb region, containing six successive open reading frames (ORFs), that was encoded by a single 8.1-kb transcript, as determined by Northern blot analysis. The gene arrangement and DNA sequence of the Clostridium sp. strain TO-931 operon are similar to those of a Eubacterium sp. strain VPI 12708 bile acid-inducible operon containing nine ORFs. Several genes in the Eubacterium sp. strain VPI 12708 operon have been shown to encode products required for bile acid 7alpha-dehydroxylation. In Clostridium sp. strain TO-931, genes potentially encoding bile acid-coenzyme A (CoA) ligase, 3alpha-hydroxysteroid dehydrogenase, bile acid 7alpha-dehydratase, bile acid-CoA hydrolase, and a bile acid transporter were similar in size and exhibited amino acid homology to similar gene products from Eubacterium sp. strain VPI 12708 (encoded by baiB, baiA, baiE, baiF, and baiG, respectively). However, no genes similar to Eubacterium sp. strain VPI 12708 biaH or baiI were found in the Clostridium sp. strain TO-931 bai operon, and the two putative Eubacterium sp. strain VPI 12708 genes, baiC and baiD, were arranged in one continuous ORF in Clostridium sp. strain TO-931. Intergene regions showed no significant DNA sequence similarity, but primer extension analysis identified a region 115 bp upstream from the first ORF that exhibited 58% identity to a bai operator/promoter region identified in Eubacterium sp. strain VPI 12708. These results indicate that the gene organization, gene product amino acid sequences, and promoters of the bile acid-inducible operons of Clostridium sp. strain TO-931 and Eubacterium sp. strain VPI 12708 are highly conserved.

3-Hydroxysteroid Dehydrogenases↗
PubMed2000

Postpartum Clostridium sordellii infection associated with fatal toxic shock syndrome.

Clostridium bacteria are anaerobic Gram positive spore-form-ing bacilli, known to cause distinct clinical syndromes such as botulism, tetanus, pseudomembranous colitis and myonecrosis. The natural habitats of Clostridium species are soil, water and the gastrointestinal tract of animals and humans. In 5-10% of all women, Clostridium species are also found to be normal inhabitants in the microbial flora of the female genital tract. In case of a non-sexually transmitted genital tract infection, Clostridium species are isolated in 4-20%, and clostridium welchii seems to be the most common isolate. Clostridium sordellii is rarely encountered in clinical specimens (1% of Clostridium species), but it has been described as a human pathogen with fatal potential. Two toxins, a lethal and a hemorrhagic (that antigenically and pathophysiologically appear similar to Clostridium difficile toxins B and A, respectively) are responsible for this potential. Reviewing the obstetric literature, only six cases of postpartum endometritis caused by C. sordellii, are described - all being fatal. In addition, one lethal case of spontaneous endometritis resulting from C. sordellii is reported. The clinical aspects of these cases include: - sudden onset with influenza-like symptoms in previously healthy women - progressive refractory hypotension - local and spreading tissue edema - absence of fever Laboratory findings include: - marked leukocytosis - elevated hematocrit. This paper reports the seventh fatal postpartum C. sorlellii associated toxic shock syndrome - the first recognized in Scandinavia.

Adult↗
PubMed1989

[A rapid test for detecting Clostridium difficile].

Staphylococcus aureus cells, strain Cowan 1, were coated with an anti-Clostridium difficile serum, produced in rabbits. This reagents were used in the coagglutination (coa-test) for detection of Clostridium difficile. A total of 275 bacterial cultures of different species were tested for detection of Clostridium difficile by coa-test and bacterial culture. Of 183 strains positive by isolation of the organism, 181 were positive by coa-test. The sensitivity and specificity of the coa-test relative to bacterial culture was 98.9% and 90.2%. A total of 206 stool specimens of children were tested also for detection of Clostridium difficile with both methods. From 16 specimens which were positive in bacterial culture 12 were positive in coa-test. The total number of samples which were positive with the coa-test was 46. The sensitivity and specificity of the coa-test in stool specimens relative to the isolation of the organism were 75.0% and 75.8%, respectively. The high predictive value for a negative result of the coa-test relative to the culture from 97.6% (bacterial cultures) and 97.3% (stool specimens) suggest that the coa-test will be a useful, rapid and simple screening test for Clostridium difficile. Specimens positive by the coa-test must be confirmed by the bacterial culture. Of some other Clostridium spp. tested, strains of Clostridium bifermentans and Clostridium sordelli also gave a positive result.

Agglutination Tests↗