[Symposium: Corynebacterium kutscheri infection in mouse-rat colonies. 4. Detection of corynebacterium contamination by cortisone treatment and fluorescent antibody technic].
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The fatty and mycolic acid profiles of 52 strains of clinical origin belonging to Corynebacterium urealyticum were subjected to numerical analysis along with those of representative members of Corynebacterium ammoniagenes, Corynebacterium bovis, Corynebacterium glutamicum, Corynebacterium jèikeium, Corynebacterium minutissimum, Corynebacterium pseudodiphtheriticum, Corynebacterium pseudotuberculosis, Corynebacterium xerosis, Corynebacterium renale, Corynebacterium cystitidis, "Corynebacterium ulcerans" and one strain of the Corynebacterium F1 group. Strains were divided into eight clusters at an amalgamation distance of 7.4. Corynebacterium urealyticum appeared as an homogeneous cluster clearly distant from others, that included several members of the genus Corynebacterium, and it was characterized by its content on unsaturated mycolic acids of mainly 28 (28:1) and 30 (30:3) carbon atoms. On the basis of these results the taxonomic "status" of Corynebacterium urealyticum, a new species within the genus Corynebacterium "sensu stricto", is further justified.
A high-performance liquid chromatography (HPLC) study of 307 strains of Corynebacterium species and related taxa revealed that strains classified as "Corynebacterium aquaticum"; "Corynebacterium asperum"; and Centers for Disease Control (CDC) groups 1, 2, A-3, A-4, A-5, B-1, B-3, E, F-2, and I-2 as well as some unidentified coryneforms do not contain any corynomycolic acids; therefore, they should not be included in the genus Corynebacterium. Such an HPLC method of identification permitted the correct assignment to the genus Rhodococcus of two unpigmented strains of coryneform bacteria whose mycolic acid profiles were comparable to those of Rhodococcus equi. Bacteria belonging to CDC groups ANF-1, ANF-3, F-1, G-1, G-2, and I-1, as well as some other Corynebacterium sp. strains, yielded corynomycolic acid HPLC patterns related to those of Corynebacterium species. Either similarities or differences were observed in the corynomycolic acid profiles of Corynebacterium species tested after culture on sheep blood agar and/or sheep blood agar supplemented with Tween 80, which demonstrated that identification at the species or group level is possible. However, Corynebacterium striatum and CDC group I-1 bacteria as well as CDC group G-1 and group G-2 bacteria had indistinguishable HPLC patterns. Conversely, some variations were observed within some species as Corynebacterium xerosis, C. striatum, and Corynebacterium minutissimum. The evaluation procedure of this HPLC method by mass spectrometry analysis of isolated eluted peaks revealed that analytical reverse-phase HPLC alone does not provide any structural information, since isomers with identical polarities coeluted as a single peak. Nevertheless, HPLC is a rapid and reliable method for identification of corynomycolic acid-containing bacteria in the clinical microbiological laboratory.
An enzyme-linked immunosorbent assay (ELISA) which detects rat antibody to Corynebacterium kutscheri was developed and compared with a tube agglutination assay in two infectivity studies. In four groups of rats inoculated intratracheally with 100-fold dilutions of Corynebacterium kutscheri , mortality, lesions, and recovery of Corynebacterium kutscheri were confined to the two highest dose groups. Five of 17 (29%) test rats seroconverted within 14 to 21 days post-inoculation as assayed by ELISA; four of these five (80%) seroconverted within 42 to 98 days post-inoculation as assayed by tube agglutination. ELISA titers were one to 16 times greater than tube agglutination titers. In 15 rats challenged intranasally with 6.9 X 10(6) Corynebacterium kutscheri , clinical signs were observed in one of 15 (6.7%), and bacteriologic and histopathologic evaluations detected exposure to Corynebacterium kutscheri in three of 14 (21.4%) and one of 14 (7.1%) of the rats, respectively. Fourteen days post-inoculation, seroconversion was observed in 14 of 15 (93.3%) by ELISA, as compared to six of 15 (40%) by tube agglutination (p less than 0.001). The ELISA was more sensitive than tube agglutination through 21 days post-inoculation, while sera assayed 33 to 53 days post-inoculation had comparable titers in both assays. Specificity studies using four other Corynebacterium sp indicated that the Corynebacterium kutscheri ELISA cross reacted with two of four (50%) sets of corynebacterial antisera (Corynebacterium ulcerans and Corynebacterium equi) prepared in rats, whereas no cross reactivity was observed by tube agglutination.
BACKGROUND: Corynebacterium species are part of the normal skin flora. The incidence of nosocomial infections caused by Corynebacterium species have increased substantially over the past two decades. However, the clinical course of Corynebacterium peritonitis complicating peritoneal dialysis remains unclear. METHOD: We reviewed all the Corynebacterium peritonitis in our dialysis unit from 1995 to 2002. During this period, there were 1485 episodes of peritonitis recorded; 27 (1.8%) of which were caused by Corynebacterium species. RESULTS: The underlying renal diagnosis and prevalence of comorbid conditions of the 27 patients were similar to our whole dialysis population. The bacteria isolated were resistant to penicillin in 8 cases (29.6%). Three cases (11.1%) had concomitant exit-site infection. The overall primary response rate was 74.1%; the complete cure rate was 37.0%. Episodes that received vancomycin as initial antibiotic had a marginally higher primary response rate (9 in 10 vs 11 in 17 episodes, P = 0.2) and complete cure rates (7 in 10 vs 3 in 17 episodes, P = 0.12) than the episodes that received cephalosporins, although neither of the differences was statistically significant. Thirteen cases (48.1%) had recurrent peritonitis after antibiotic therapy, 8 of which had the recurrent episode at least 30 days after stopping antibiotics (median 54 days, range 43-60 days). Eight recurrent cases (61.5%) were successfully cured by another 3 week course of intra-peritoneal vancomycin. CONCLUSIONS: Recurrent Corynebacterium peritonitis is common after a 2 week course of antibiotics. Recurrent Corynebacterium peritonitis may be delayed up to 2 months after the antibiotic is stopped. Recurrent peritonitis can usually be cured with a 3 week course of intra-peritoneal vancomycin, which is probably the preferred antibiotic regimen for Corynebacterium peritonitis.
An in vitro microassay was used to evaluate growth of five strains of Corynebacterium bovis in mammary secretions collected from quarters of five Holstein cows at 0, 14, and 28 d of involution, at parturition, and 14 d after parturition. Variation in growth among different strains of Corynebacterium bovis was observed. Corynebacterium bovis grew well in mammary secretions obtained at the last milking of lactation, at parturition, and 14 d after parturition. However, growth of four strains of Corynebacterium bovis in mammary secretions obtained at 14 and 28 d of involution was reduced significantly. In contrast, a streptomycin-resistant strain of Corynebacterium bovis grew well in mammary secretions obtained during involution. These data suggest that mammary secretions support growth of Corynebacterium bovis during lactation but inhibit growth during the nonlactating period. Inhibition of growth in secretions collected during the nonlactating period may be associated with the high rate of spontaneous elimination of Corynebacterium bovis intramammary infection from cessation of milking to parturition. Conversely, enhanced growth in milk may be related to persistent Corynebacterium bovis intramammary infections during lactation.
Corynebacterium urealyticum has been well documented as a urinary pathogen in Europe. The purpose of this study was to investigate the incidence and clinical relevance of Corynebacterium urealyticum as a urinary pathogen in a predominantly Third World population (South Africa) and to attempt to increase the isolation rate by culturing urine specimens on a selective medium. Two methods were used to isolate Corynebacterium urealyticum from urine specimens. Blood agar plates from routine urine cultures of 7,912 urine specimens were incubated for 48 hours and 1,281 specimens were cultured on a selective medium as well as on routine media. The antimicrobial susceptibility of all isolates of Corynebacterium urealyticum was tested. The yield of Corynebacterium urealyticum on blood agar was three isolates in three patients (0.038% of 7,912), all of whom had pyuria, alkaline urine and risk factors for Corynebacterium urealyticum infection. Use of selective media increased the yield of Corynebacterium urealyticum to 15 of 1,281 specimens (1.17%), however 73% of these urine samples yielded other pathogens and none had an alkaline pH which could not be attributed to the presence of another urealytic pathogen. All isolates were susceptible to vancomycin and 92.6% susceptible to norfloxacin. The pathogenic potential of Corynebacterium urealyticum was confirmed in South African patients, but the incidence of infection was low. The use of a selective medium is therefore not cost-effective in all cases but could be used selectively on the basis of typical urine findings and patient criteria.
Nineteen strains of Gram-positive, non-motile, non-spore-forming, catalase-positive, rod-shaped bacteria isolated from pigs were characterized by using biochemical, molecular chemical and molecular genetic methods. Two distinct groups of organisms were discerned, based on their colonial morphology, CAMP (Christie-Atkins-Munch-Petersen) reaction and numerical profile by using the API Coryne system. The first group (13 strains) gave a doubtful discrimination between Corynebacterium striatum and Corynebacterium amycolatum, whilst the second group (six strains) were identified tentatively as Corynebacterium urealyticum. Comparative 16S rRNA gene sequencing studies demonstrated that all of the isolates belonged phylogenetically to the genus CORYNEBACTERIUM: The first group of organisms was highly similar to Corynebacterium testudinoris with respect to 16S rRNA gene sequences and physiological characteristics, whereas the remaining six isolates formed a hitherto unknown subline within the genus, associated with a small subcluster of species that included Corynebacterium auriscanis and its close relatives. The unknown Corynebacterium sp. was distinguished readily from these and other species of the genus by biochemical tests. Based on both phenotypic and phylogenetic evidence, it is proposed that the new isolates from pigs should be classified as a novel species, Corynebacterium suicordis sp. nov. The type strain is P81/02(T) (=CECT 5724(T)=CCUG 46963(T)).
The relationship between infection stone and Corynebacterium species was investigated in vitro and in vivo. Urease activity of urease-splitting Corynebacterium species was evaluated by 2 methods; an increase in pH of human urine after inoculation of Corynebacterium species and direct measurement of urease activity of 10(7) CFU organisms from amounts of ammonia by indophenol method. Formation of infection bladder stone was induced in male Wistar rats by implanting a zinc disc and inoculating 10(6) CFU organisms surgically into the bladder. Urine was alkalinized by the inoculation of Corynebacterium renale, C. pilosum and group D2 Corynebacterium. C. renale and C. pilosum had strong urease activity, and group D2 Corynebacterium had moderate activity. C. pseudodiphtheriticum did not produce the elevation of urinary pH and had little urease activity. Infection stones were formed in 100% of rats by inoculation of C. renale and C. pilosum and 88% of rats by group D2. Urinary pH was elevated in all inoculated rats. In conclusion, C. renale, C. pilosum and group D2 Corynebacterium may play a role in formation of infection stones.
DNA relatedness experiments were performed with 38 clinical isolates and 13 reference strains of coryneform taxa exhibiting a lipid requirement for optimal growth. Forty-five of these strains split into five genomic groups at the species level, whereas six other strains remained unclustered. Genomospecies II fits Corynebacterium accolens, but the other genomospecies were not genetically related to any of the defined Corynebacterium species. Phylogenetic analyses of genes coding for small-subunit rRNA sequences revealed that two genomospecies (I and III) and C. accolens form a tight cluster within the robust branch that groups all Corynebacterium species presently sequenced. Reference strains of biotypes C-1, C-2, and C-3 of "Corynebacterium pseudogenitalium" were found to fall into genomospecies I, as well as "Corynebacterium tuberculostearicum," Centers for Disease Control and Prevention (CDC) coryneform group G-1, and CDC coryneform group G-2 reference strains. Biochemical tests allowed differentiation between genomospecies except between genomospecies IV and V and between six unclustered strains and genomospecies I. We propose a new classification for these lipid-requiring diphtheroids within the genus Corynebacterium with the delineation of some CDC coryneform group G-1 strains (genomospecies III) as a new species for which the name Corynebacterium macginleyi is proposed. The type strain is strain JCL-2 (CIP 104099), isolated from a human corneal ulcer.
The 16S rRNA gene sequences of 30 strains representing 23 validated Corynebacterium species and 7 currently non-valid Corynebacterium species were determined. These sequences were aligned with the sequences of other Corynebacterium species and related actinomycete taxa. A comparative sequence analysis revealed that there is considerable phylogenetic depth and internal structure in the genus Corynebacterium. Turicella otitidis and the amycolate species Corynebacterium amycolatum were located at the periphery of the genus Corynebacterium. It was evident that the species of the genus Corynebacterium form a monophyletic association and, together with other chemotype IV and mycolic acid-containing taxa (including the genera Dietzia, Gordona, Mycobacterium, Nocardia, Rhodococcus, and Tsukamurella), form a natural suprageneric group.
Six unidentified gram-positive, rod-shaped organisms recovered from the cloacae of apparently healthy wild penguins were characterized by phenotypic and molecular taxonomic methods. Chemotaxonomic investigations revealed the presence of a cell wall based on meso-diaminopimelic acid and long-chain cellular fatty acids of the straight-chain saturated and monounsaturated types, consistent with the genus Corynebacterium. Corynomycolic acids, which are characteristic of the genus, were also detected, albeit in small amounts. Comparative 16S rRNA gene sequencing studies showed that the unidentified organisms were phylogenetically related to corynebacteria and represent a novel subline associated with a small subcluster of species that includes Corynebacterium xerosis, Corynebacterium amycolatum and Corynebacterium freneyi. The unknown isolates were readily distinguished from their closest phylogenetic relatives and all other Corynebacterium species with validly published names by using a combination of biochemical and chemotaxonomic criteria. Based on both phenotypic and 16S rRNA gene sequence considerations, it is proposed that the unknown isolates recovered from penguins be classified as a novel species in the genus Corynebacterium, Corynebacterium sphenisci sp. nov. The type strain is CECT 5990T (= CCUG 46398T).
Experimental challenge procedures were used to study infectivity and virulence of Corynebacterium bovis. Challenge procedures using Staphylococcus aureus (Newbould 305) and Streptococcus agalactiae (McDonald 44) were used to study effects of Corynebacterium bovis infections on superinfection with major pathogens. Rate of infection under experimental challenge conditions was significantly greater with Corynebacterium bovis than previously observed for Staphylococcus aureus or Streptococcus agalactiae. Principal location of Corynebacterium bovis colonization appeared to the teat canal region, although the organism was isolated from 75% of the teat cisterns by puncture technique. Quarters with Corynebacterium bovis infections were more resistant to infection by Staphylococcus aureus than bacteriologically negative quarters. Quarters infected with Corynebacterium bacterium bovis were approximately 8.5-fold more susceptible to Streptococcus agalactiae infection than negative quarters. Somatic cell counts were doubled in negative quarters that developed Corynebacterium bovis infections; the geometric mean was 2.4 X 10(5).
A coryneform bacteria known as Corynebacterium group D2 pending definitive taxonomic classification, has emerged as a human pathogen. Although first described 16 years ago, the organism has only recently been implicated in clinical infections. It differs from all previously recognized species of Corynebacterium, and in common with Corynebacterium jeikeium exhibits a broad spectrum of antimicrobial resistance, being sensitive to only vancomycin, teicoplanin and some quinolones. The microorganisms is mainly involved in urinary tract infections, and its strong urease activity seems to play an important role in its pathogenicity. Corynebacterium jeikeium and group D2 are very different from the clinical and microbiological point of view. Corynebacterium group D2, like other coryneform bacteria, may be isolated from healthy skin in hospitalized patients, mainly females, while Corynebacterium jeikeium is more prevalent in males. Microbiologists should be aware that such a slow-growing microorganism may be responsible for clinical infections and should not be overlooked in routine cultures of clinical specimens.
An unknown Gram-positive rod-shaped bacterium was isolated from skin scrapings from the infected head of a sheep and subjected to a polyphasic taxonomic analysis. Chemical analysis revealed the presence of straight-chain and monounsaturated fatty acids and short-chain (C32-C36) mycolic acids consistent with the genus Corynebacterium. Comparative 16S rRNA gene sequencing confirmed that the unknown rod was a member of the genus Corynebacterium, with the organism forming a distinct sub-line and displaying greater than 3% sequence divergence with established species. The unknown Corynebacterium isolate was readily distinguished from recognized species of the genus by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium from a sheep be classified in the genus Corynebacterium, as Corynebacterium capitovis sp. nov. The type strain of Corynebacterium capitovis is CCUG 39779T (= CIP 106739T).