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1.
Vet Microbiol ; 93(3): 261-73, 2003 May 29.
Article in English | MEDLINE | ID: mdl-12695049

ABSTRACT

The aims of the study were to characterize isolates of Bartonella henselae and to determine the prevalence of bacteremic domestic cats in urban and suburban parts of Prague, Czech Republic. Five (18%) gram-negative fastidious bacterial single-cat isolates were recovered from 27 hemocultures incubated without previous freezing. Four of these isolates originated from flea infested stray cats (n=6) and one from a shelter cat without any ectoparasites (n=21). None of the 34 previously frozen specimens from flea free pet cats yielded any bacteria. All five isolates were catalase and oxidase negative. Their enzymatic activity, RFLP profile of citrate synthetase gene (gltA) and DNA-DNA hybridization results were typical of B. henselae. According to their PvuII and BglI ribotypes the isolates could be allocated to two homogeneous groups. Ribotype HindIII and RFLP of 16S-23S rRNA spacer region analysis gave unique profiles different from those of Bartonella quintana, Bartonella elizabethae and Bartonella clarridgeiae. The 16S rRNA type-specific amplification revealed an identical profile typical of B. henselae genotype II for all the cat isolates studied. Pulsed-field gel electrophoresis (PFGE) assigned a different profile to each of the isolates studied. Determination of the enzymatic activity, RFLP of gltA gene, RFLP of 16S-23S rRNA spacer region, and HindIII ribotype could be efficient tools for identification of B. henselae isolates. Ribotyping (PvuII, BglI), 16S rRNA typing and PFGE may be useful methods to prospect ecology and epidemiology of the agent.


Subject(s)
Bartonella Infections/veterinary , Bartonella henselae/isolation & purification , Cat Diseases/microbiology , Animals , Bartonella Infections/enzymology , Bartonella Infections/epidemiology , Bartonella Infections/microbiology , Bartonella henselae/enzymology , Bartonella henselae/genetics , Cat Diseases/epidemiology , Catalase/metabolism , Cats , Citrate (si)-Synthase/chemistry , Citrate (si)-Synthase/genetics , Czech Republic/epidemiology , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Electrophoresis, Gel, Pulsed-Field/veterinary , Female , Male , Nucleic Acid Hybridization , Oxidoreductases/metabolism , Polymerase Chain Reaction/veterinary , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 16S/chemistry , RNA, Ribosomal, 16S/genetics , Ribotyping/veterinary
2.
ILAR J ; 44(1): 20-7, 2003.
Article in English | MEDLINE | ID: mdl-12473828

ABSTRACT

In recent years, the emergence or re-emergence of critical issues in infectious disease and public health has presented new challenges and opportunities for laboratory animal care professionals. The re-emergence of bioterrorism as a threat activity of individuals or small groups has caused a heightened awareness of biosecurity and improved biosafety. The need for animal work involving high-risk or high-consequence pathogens and for arthropod-borne diseases has stimulated renewed interest in animal biosafety matters, particularly for work in containment. Application of these principles to animals retained in outdoor environments has been a consequence of disease eradication programs. The anticipated global eradication of wild poliovirus has prompted the promulgation of new biosafety guidelines for future laboratory and animal work. Increased concern regarding the use of biologically derived toxins and hazardous chemicals has stimulated a new categorization of facility containment based on risk assessment. Recognition that prion disease agents and other high-consequence pathogens require safe handling and thorough destruction during terminal decontamination treatment has led to the development of new biosafety guidelines and technologies. The implementation of these guidelines and technologies will promote state-of-the-art research while minimizing risk to laboratory animals, researchers, and the environment.


Subject(s)
Bioterrorism , Medical Waste Disposal , Occupational Health , Safety , Security Measures , Animals , Animals, Laboratory , Animals, Wild , Humans , Laboratories , Medical Laboratory Personnel , Organizational Policy , Poliovirus/pathogenicity , Prions , Risk Assessment , Toxins, Biological/adverse effects
3.
J Clin Microbiol ; 39(11): 4160-2, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11682548

ABSTRACT

Ten strains of Lautropia mirabilis (ATCC 51599(T) and nine phenotypically similar clinical isolates) were examined for cellular fatty acid (CFA) composition to evaluate their chemical relatedness to known bacterial species and groups. The CFAs were liberated from whole cells by base hydrolysis, methylated, and analyzed by gas-liquid chromatography. CFA profiles were generated by using a commericial software package (MIDI, Newark, Del.). All strains tested had an identical CFA profile characterized by major amounts of 16:1omega7c (41%) and 16:0 (44%); smaller amounts (1 to 4%) of 3-OH-10:0, 12:0, 14:0, 15:0, and 18:1 omega7c; trace amounts (<1%) of 10:0, 18:2 and 18:0; and no cyclopropane acids. This profile was similar to the CFA profiles of Acidovorax delafieldii, Comamonas terrigena, and strains of an unclassified Centers for Disease Control group designated weak oxidizer group 1. CFA analysis, when supplemented by phenotypic characterization, is useful for the identification of L. mirabilis isolates.


Subject(s)
Bacterial Typing Techniques , Betaproteobacteria/classification , Fatty Acids/analysis , Gram-Negative Bacterial Infections/microbiology , Betaproteobacteria/chemistry , Betaproteobacteria/genetics , Gram-Negative Bacterial Infections/genetics , Humans
4.
Int J Syst Evol Microbiol ; 51(Pt 3): 1151-1160, 2001 May.
Article in English | MEDLINE | ID: mdl-11411684

ABSTRACT

Seven strains of Legionella-like amoebal pathogens (LLAPs) were characterized on the basis of their cultural and staining characteristics, biochemical reactions, serology, cellular fatty acids (CFAs), isoprenoid quinone composition, total DNA relatedness, analysis of 16S rRNA and macrophage infectivity potentiator (mip) gene sequence analyses. All seven strains exhibited limited growth on buffered charcoal yeast extract alpha (BCYE) agar, required cysteine for growth and contained branched-chain CFAs and quinones typical of Legionella species. The bacilli were Gram-negative and catalase-positive. There were varying degrees of serological cross-reactions between these LLAP strains and other previously described Legionella species. Results from the various tests revealed that four LLAP strains represent three unusual new species of Legionella: Legionella drozanskii sp. nov., type strain LLAP-1T; Legionella rowbothamii sp. nov., type strain LLAP-6T; and Legionella fallonii sp. nov., type strain LLAP-10T. Three other LLAP strains, designated LLAP-7FL, LLAP-7NF and LLAP-9, were shown to be members of the species Legionella lytica. The deductions made from the phenetic characteristics of these bacteria were consistent with the phylogenetic relationships inferred from 16S rRNA and mip gene sequence analyses. This study is the first to speciate LLAP strains on the basis of data including quantitative DNA hybridization.


Subject(s)
Acanthamoeba/microbiology , Legionella/classification , Phylogeny , Acanthamoeba/isolation & purification , Animals , DNA, Ribosomal/genetics , Genotype , Legionella/genetics , Legionella/isolation & purification , Molecular Sequence Data , Poland , RNA, Ribosomal, 16S/genetics , Soil
5.
J Clin Microbiol ; 39(5): 1819-26, 2001 May.
Article in English | MEDLINE | ID: mdl-11325997

ABSTRACT

CDC weak oxidizer group 2 (WO-2) consists of nine phenotypically similar human clinical isolates received by the Centers for Disease Control and Prevention between 1989 and 1998. Four of the isolates were from blood, three were from sputum, and one each was from bronchial fluid and maxillary sinus. All are aerobic nonfermentative, motile gram-negative rods with one to eight polar flagella per cell. All grew at 25 and 35 degrees C and were positive for catalase, urease (usually delayed 3 to 7 days), citrate, alkalinization of litmus milk, oxidization of glycerol (weakly), and growth on MacConkey agar and in nutrient broth without NaCl. All except one strain were oxidase positive with the Kovács method, and all except one isolate weakly oxidized D-glucose. All were negative for oxidation of D-xylose, D-mannitol, lactose, sucrose, maltose, and 20 other carbohydrates, esculin hydrolysis, indole production, arginine dihydrolase, and lysine and ornithine decarboxylase. Only two of nine isolates reduced nitrate. Broth microdilution susceptibilities were determined for all strains against 13 antimicrobial agents. Most of the strains were resistant to ampicillin, extended-spectrum cephalosporins, and aminoglycosides, including gentamicin, tobramycin, and amikacin, but they varied in their susceptibility to fluoroquinolones. High-performance liquid chromatographic and mass spectrometric analyses of the WO-2 group identified ubiquinone-8 as the major quinone component. The percent G+C of the WO-2 strains ranged from 65.2 to 70.7% (thermal denaturation method). All shared a common cellular fatty acid (CFA) profile, which was characterized by relatively large amounts (7 to 22%) of 16:1omega7c, 16:0, 17:0cyc, 18:1omega7c, and 19:0cyc(11-12); small amounts (1 to 3%) of 12:0 and 14:0; and eight hydroxy acids, 2-OH-12:0 (4%), 2-OH-14:0 (trace), 3-OH-14:0 (12%), 2-OH-16:1 (1%), 2-OH-16:0 (3%), 3-OH-16:0 (4%), 2-OH-18:1 (2%), and 2-OH-19:0cyc (3%). This profile is similar to the CFA profile of Pandoraea, a recently described genus associated with respiratory infections in cystic fibrosis patients (T. Coenye et al., Int. J. Syst. Evol. Microbiol., 50:887-899, 2000). Sequencing of the 16S rRNA gene (1,300 bp) for all nine strains indicated a high level (> or =98.8%) of homogeneity with Pandoraea spp. type strains. DNA-DNA hybridization analysis (hydroxyapatite method; 70 degrees C) confirmed the identity of WO-2 with the genus Pandoraea and assigned three strains to Pandoraea apista and three to Pandoraea pnomenusa, and identified three additional new genomospecies containing one strain each (ATCC BAA-108, ATCC BAA-109, ATCC BAA-110). This study also shows that Pandoraea isolates may be encountered in blood cultures from patients without cystic fibrosis.


Subject(s)
Betaproteobacteria/classification , Gram-Negative Bacterial Infections/microbiology , Aged , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Betaproteobacteria/chemistry , Betaproteobacteria/drug effects , Betaproteobacteria/genetics , Child, Preschool , Fatty Acids/analysis , Female , Genes, rRNA , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Sequence Data , Oxidation-Reduction , Phenotype , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
6.
Am J Trop Med Hyg ; 63(5-6): 249-54, 2000.
Article in English | MEDLINE | ID: mdl-11421372

ABSTRACT

As part of an investigation of a 1995 outbreak of leptospirosis in Nicaragua, a cross-sectional serologic survey was conducted in the town of El Sauce. Of 566 persons, 85 (15%) were positive for IgM anti-Leptospira antibodies, indicating recent leptospirosis infection. Asymptomatic leptospirosis infection was common, with only 25 (29.4%) of the 85 seropositive inhabitants reporting a febrile illness in the 2 months before the survey. Multivariable analysis revealed that having an indoor water source remained independently protective against leptospirosis. Gathering wood was independently associated with infection. These findings suggest that asymptomatic infection with Leptospira is common in endemic areas of Leptospira transmission. Improvement in water sanitation and behavioral modifications to reduce environmental exposure may reduce the risk of leptospirosis in endemic regions.


Subject(s)
Antibodies, Bacterial/blood , Disease Outbreaks , Leptospira/immunology , Leptospira/isolation & purification , Leptospirosis/epidemiology , Adolescent , Adult , Age Distribution , Aged , Child , Child, Preschool , Cross-Sectional Studies , Enzyme-Linked Immunosorbent Assay , Female , Humans , Leptospirosis/pathology , Leptospirosis/prevention & control , Male , Middle Aged , Nicaragua/epidemiology , Risk Factors , Surveys and Questionnaires
7.
Am J Trop Med Hyg ; 61(3): 399-404, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10497979

ABSTRACT

Leptospirosis has rarely been reported in Puerto Rico, although in the period from 1948 to 1952, 208 cases of leptospirosis and an island-wide seroprevalence of antibody to Leptospira of 14% were documented. In Puerto Rico in October 1996, following rainfall and a period of flooding generated by Hurricane Hortense, serum specimens of 4 patients with suspected dengue fever that were negative for dengue tested positive for Leptospira-specific IgM antibodies in a dipstick assay. Subsequently, we used an island-wide dengue laboratory-based surveillance system to determine the increase in leptospirosis after hurricane-generated floods. All anti-dengue IgM-negative patients (n = 142) with disease onset from August 8 to October 6, 1996 from prehurricane and posthurricane groups were investigated for leptospirosis. Laboratory-confirmed leptospirosis cases were defined as microscopic agglutination test titers > or = 1 :400 to 1 or more serovars, or positive immunohistochemistry in autopsy tissues. Four (6%) of 72 prehurricane and 17 (24%) of 70 posthurricane patients had laboratory-confirmed cases of leptospirosis (relative risk [RR] = 4.4, 95% confidence interval [CI] = 1.6-12.4). The mean age of case-patients was 34 years (range = 13-64). Eighteen (86%) of 21 confirmed case-patients were males, including one patient who died (31 years old). Patients were located in 18 (38%) of 48 municipalities that submitted serum samples. Clinical features significantly associated with leptospirosis were eye pain (RR = 1.5, 95% CI = 1.3-1.9), joint pain (RR = 1.4, 95% CI = 1.1-1.6), diarrhea (RR = 1.7, 95% CI = 1.2-2.5), and jaundice (RR = 3.3, 95% CI = 1.5-7.2). This study demonstrates the utility of a dengue laboratory-based surveillance system for the detection of an increase of leptospirosis, which most likely would have gone unrecognized. Leptospirosis is treatable with antibacterial agents; knowledge of this diagnosis may significantly reduce morbidity and mortality.


Subject(s)
Dengue/epidemiology , Disasters , Leptospirosis/epidemiology , Adolescent , Adult , Antibodies, Bacterial/blood , Antibodies, Viral/blood , Dengue/virology , Dengue Virus/immunology , Female , Humans , Immunoglobulin M/blood , Leptospira interrogans/classification , Leptospira interrogans/immunology , Leptospira interrogans/isolation & purification , Leptospirosis/microbiology , Male , Middle Aged , Population Surveillance , Puerto Rico/epidemiology
8.
J Zoo Wildl Med ; 30(1): 111-8, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10367652

ABSTRACT

Cutaneous fungal infections occurred in four captive brown tree snakes (Boiga irregularis). The ventral scales were most commonly affected, and lesions began as areas of erythema and edema with vesicle formation, followed by development of caseous brown plaques. Lesions usually started where ventral scales overlapped and spread rapidly. All snakes died within 14 days after clinical signs were first noted. The deaths of three of the snakes were directly attributable to the cutaneous disease; the other snake died from renal failure and visceral gout, most likely induced by gentamicin therapy. Histologically, lesions consisted of epidermal hyperplasia and hyperkeratosis, with foci of epidermal necrosis, intraepidermal vesicle formation, and subacute inflammation of the underlying dermis. These lesions were associated with bacteria and numerous septate, branched fungal hyphae within the epidermis and overlying serocelluar crusts. Hyphae that penetrated through the superficial surface of the epidermis often formed terminal arthroconidia. The same species of fungus was isolated in pure culture from the skin of three snakes, but fungal cultures were not performed on samples from the fourth snake. The fungus has been identified as the Chrysosporium anamorph of Nannizziopsis vriesii based on its formation of solitary dermatophytelike aleurioconidia and alternate and fission arthroconidia. The source of the fungus in this outbreak was not determined; however, the warm, moist conditions under which the snakes were housed likely predisposed them to opportunistic cutaneous fungal infections.


Subject(s)
Chrysosporium/isolation & purification , Dermatomycoses/veterinary , Snakes , Animals , Dermatomycoses/microbiology , Dermatomycoses/pathology , Fatal Outcome , Female , Male , Skin/microbiology , Skin/pathology
9.
Int J Syst Bacteriol ; 49 Pt 2: 839-58, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10319510

ABSTRACT

DNA relatedness was determined among 303 strains of Leptospira and Leptonema. Included in the analysis were reference strains from 228 well-characterized and recognized serovars. The study included 268 serovars from 29 named and one or more unnamed serogroups. The strains clustered into 17 DNA hybridization groups, representing 12 previously described species (292 strains) and five new genomospecies (11 strains). The largest groups included Leptospira interrogans (91 strains from 82 serovars), Leptospira santarosai (65 strains from 59 serovars), Leptospira borgpetersenii (49 strains from 43 serovars), Leptospira kirschneri (29 strains from 26 serovars) and Leptospira noguchii (20 strains from 20 serovars). The new genomospecies include Leptospira genomospecies 1 (two strains, serovars pinagchang and sichuan), Leptospira genomospecies 2 (six strains, serovars lushui, manhao 3, manzhuang, nanding, mengla and yunnan), Leptospira genomospecies 3 (one strain, serovar holland), Leptospira genomospecies 4 (one strain, serovar hualin) and Leptospira genomospecies 5 (one strain, serovar saopaulo). With the exception of Ballum, all serogroups with greater than one serovar studied were genetically heterogeneous. Phenotypic tests, including optimal growth temperature, lipase activity and growth inhibition by copper sulfate or 2,6-diaminopurine, were of little use in differentiating DNA relatedness groups. The name Leptospira alexanderi sp. nov. is proposed for Leptospira genomospecies 2 (type strain L 60T = ATCC 700520T, serovar manhao 3).


Subject(s)
DNA, Bacterial/genetics , Leptospira/classification , Leptospiraceae/classification , Leptospirosis/microbiology , Animals , Base Composition , DNA, Bacterial/chemistry , Humans , Leptospira/genetics , Leptospiraceae/genetics , Phenotype , Serotyping , Species Specificity
10.
Clin Infect Dis ; 27(5): 1238-40, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9827276

ABSTRACT

We describe a toddler from Connecticut who developed purulent conjunctivitis, fever, and a morbilliform rash. Blood cultures were positive for Haemophilus influenzae biogroup aegyptius; further investigation was performed to assess the possibility that the illness was consistent with Brazilian purpuric fever, which, to our knowledge, has not been reported in the United States. This isolate shared morphological and some biochemical characteristics with previously studied H. influenzae biogroup aegyptius strains but differed according to slide agglutination testing, plasmid characterization, and ribotyping. Blood and tissue samples obtained during his hospitalization were also positive for Epstein-Barr virus. The child died 8 days after hospitalization. Fifty other cases of invasive H. influenzae infection were identified by active surveillance studies. Of the 49 viable surveillance isolates, 10 were biotype III (two of which had the same ribotype as the strain from our case.


Subject(s)
Haemophilus Infections/complications , Haemophilus influenzae/classification , Herpesviridae Infections/complications , Herpesvirus 4, Human , Purpura/complications , Bacteremia/microbiology , Bacterial Typing Techniques , Fatal Outcome , Haemophilus Infections/microbiology , Haemophilus influenzae/isolation & purification , Herpesviridae Infections/virology , Herpesvirus 4, Human/isolation & purification , Humans , Infant , Male
11.
J Infect Dis ; 178(5): 1457-63, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9780268

ABSTRACT

In October 1995, epidemic "hemorrhagic fever," without jaundice or renal manifestations, was reported in rural Nicaragua following heavy flooding; 2259 residents were evaluated for nonmalarial febrile illnesses (cumulative incidence, 6.1%) and 15 (0.7%) died with pulmonary hemorrhage. A case-control study found that case-patients were more likely than controls to have ever walked in creeks (matched odds ratio [MOR], 15.0; 95% confidence interval [CI], 1.7-132.3), have household rodents (MOR, 10.4; 95% CI, 1.1-97.1), or own dogs with titers >/=400 to Leptospira species (MOR, 23.4; 95% CI, 3.6-infinity). Twenty-six of 51 case-patients had serologic or postmortem evidence of acute leptospirosis. Leptospira species were isolated from case-patients and potential animal reservoirs. This leptospirosis epidemic likely resulted from exposure to flood waters contaminated by urine from infected animals, particularly dogs. Leptospirosis should be included in the differential diagnosis for nonmalarial febrile illness, particularly during periods of flooding or when pulmonary hemorrhage occurs.


Subject(s)
Hemorrhage/complications , Leptospirosis/epidemiology , Lung Diseases/complications , Adolescent , Adult , Animals , Case-Control Studies , Cattle , Child , Child, Preschool , Disasters , Disease Outbreaks , Disease Vectors , Dogs , Hemorrhage/microbiology , Horses , Humans , Incidence , Infant , Leptospira/classification , Leptospira/isolation & purification , Leptospirosis/complications , Leptospirosis/microbiology , Lung Diseases/microbiology , Nicaragua/epidemiology , Rodentia , Swine , Water Microbiology
12.
Syst Appl Microbiol ; 21(1): 89-96, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9741113

ABSTRACT

Seven new Leptospira isolates from rats, a buffalo, and contaminated media showed either reactive serology against more than 1 serogroup or no reactive serology against a reference panel of 22 serovars in the microscopic agglutination test (MAT). Because of these inconclusive results, the 16S rDNA sequences of these isolates were determined and found to resemble that of the type strain of Leptospira inadai (L. inadai), serovar lyme strain 10, which is considered to be nonpathogenic for humans. Comparative analyses of other Leptospira 16S rDNA sequences from databases revealed a L. inadai-specific signature sequence, against which an amplification primer was designed. This primer when used in conjunction with an universal primer enabled the trial of a rapid PCR protocol in which fluorescence emissions due to binding of SYBR Green I dye to PCR products were continuously monitored during rapid thermal cycling. A melting curve acquired immediately after PCR was used to distinguish the intended product. The thermal cycling and continuous monitoring of fluorescence emission were accomplished by the LightCycler; the whole procedure of 30 PCR cycles and melting curve acquisition required only 20 minutes. The primer achieved the required specificity, as the intended PCR product resulted only from 6 confirmed L. inadai reference strains and 7 field isolates that had been verified as L. inadai by the 16S rDNA sequencing, but not from 16 reference strains of Leptospira belonging to 7 other genospecies. Furthermore, these experiments showed that the PCR protocol was robust because target DNA of different conditions, which were extracted by either 1 of the 4 methods used, could be detected.


Subject(s)
Leptospira/classification , Leptospira/isolation & purification , Organic Chemicals , Polymerase Chain Reaction/methods , Animals , Base Sequence , Benzothiazoles , DNA Primers , DNA, Bacterial/analysis , DNA, Ribosomal/analysis , Diamines , Fluorescence , Fluorescent Dyes/metabolism , Molecular Sequence Data , Quinolines , RNA, Ribosomal, 16S/genetics , Rats , Sequence Analysis, DNA , Species Specificity , rRNA Operon
13.
J Clin Microbiol ; 36(6): 1674-8, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9620398

ABSTRACT

Between 1983 and 1994, 13 phenotypically similar unidentified clinical isolates were received by the Special Bacteriology Reference Laboratory, Centers for Disease Control and Prevention (CDC). Sources included blood (four strains), lung (three strains), knee fluid and duodenal tissue (one strain each), bone, and lymph node tissue (two strains each). All were aerobic glucose-oxidizing, slender, long, curved gram-negative rods that utilized xylose, sucrose, and maltose; did not grow on MacConkey agar in 1 to 2 days; were oxidase positive; hydrolyzed esculin; and grew on Campylobacter selective medium. All were negative for urease, indole, nitrate reduction, and gelatin hydrolysis. All were motile by means of a single polar flagellum with a noticeably short wavelength; however, motility was sometimes difficult to demonstrate. The cellular fatty acid compositions of these strains, as analyzed by gas-liquid chromatography, were unique, characterized by relatively large amounts of 16:1omega7c, 16:0, and 18:1omega7c with smaller amounts of 12:0, 3-OH-12:1, 14:0, 15:0, 18:0, Br-19:1, and 19:0cyc11-12. High-performance liquid chromatography and mass spectrometry of the quinone extracts of three representative strains showed ubiquinone-10 as the major component. Based on the breakpoints for the family Enterobacteriaceae, all the strains were susceptible in vitro to aminoglycosides, sulfamethoxazole-trimethoprim, and chloramphenicol but were resistant to most beta-lactams except imipenem. The MICs of amoxicillin-clavulanate and ciprofloxacin for these strains clustered around the breakpoints, which makes it difficult to predict the strains' response in vivo to these agents. This group has been designated CDC oxidizer group 3 (O-3).


Subject(s)
Gram-Negative Bacteria/classification , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacterial Infections/microbiology , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Benzoquinones/analysis , Centers for Disease Control and Prevention, U.S. , Coloring Agents , Fatty Acids/analysis , Female , Glucose/metabolism , Gram-Negative Bacteria/chemistry , Gram-Negative Bacteria/physiology , Humans , Infant , Male , Microbial Sensitivity Tests , Middle Aged , Phenotype , United States
14.
J Clin Microbiol ; 35(2): 504-7, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9003628

ABSTRACT

We present two cases of rapidly progressing, fatal pneumonia caused by Bacillus cereus. These cases are interesting in that B. cereus, even from blood or sputum specimens, may often be considered a contaminant and receive inadequate attention. Also of interest was the fact that the two patients resided in the same area of the state, were welders by trade, and became ill within a few days of each other, yet there was no epidemiologic link between them.


Subject(s)
Bacillaceae Infections/microbiology , Bacillus cereus/isolation & purification , Bacteremia/microbiology , Pneumonia, Bacterial/microbiology , Adult , Humans , Lung/microbiology , Male , Middle Aged , Sputum/microbiology
15.
Infect Immun ; 63(9): 3484-90, 1995 Sep.
Article in English | MEDLINE | ID: mdl-7642281

ABSTRACT

Capnocytophaga canimorsus is a gram-negative rod that causes opportunistic infections resulting in bacteremia, septicemia, meningitis, and death in immunocompromised, splenectomized, and alcoholic individuals. Infections caused by a related species, Capnocytophaga cynodegmi, remain localized at the site of the wound where the organism is introduced. Both organisms are part of the normal canine oral flora and are introduced through puncture wounds via dog bites. We found that both C. canimorsus and C. cynodegmi attach, are phagocytized, and multiply intracellularly in J774 mouse macrophage cells. After 48 h of infection by C. canimorsus, large sections of the macrophage cell layer were observed to detach and lyse, while the monolayer infected with C. cynodegmi demonstrated no cytotoxic effects. Tissue culture supernatants from the C. canimorsus-infected J774 cells filtered through a 0.22-micron-pore membrane produced a similar effect on fresh monolayers, while filtrates from C. cynodegmi and uninfected controls produced no effect. No endotoxin release was observed in these supernatants. We conclude that the cytotoxic phenotype of C. canimorsus is the likely result of a toxin produced by this organism.


Subject(s)
Capnocytophaga/pathogenicity , Macrophages/microbiology , Animals , Bacterial Toxins/biosynthesis , Capnocytophaga/growth & development , Cell Line , Humans , Mice
16.
Infect Immun ; 63(6): 2317-22, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7768615

ABSTRACT

Brazilian purpuric fever (BPF) is a fulminant pediatric disease characterized by fever, with rapid progression to purpura, hypotensive shock, and death. All known BPF cases have been caused by three clones of Haemophilus influenzae biogroup aegyptius and have occurred in either Brazil or Australia. Using an immortalized line of human vascular endothelial cells, we developed an in vitro assay that identifies all known BPF-causing H. influenzae biogroup aegyptius strains (R. S. Weyant, F. D. Quinn, E. A. Utt, M. Worley, V. G. George, F. J. Candal, and E. W. Ades, J. Infect. Dis. 169:430-433, 1994). With multiplicities of infection (MOIs) as low as one bacterium per 1,000 tissue culture cells, BPF-associated strains produce a unique cytotoxic effect in which the tissue culture cells detach and aggregate in large floating masses after 48 h of incubation. In this study, using a BPF-associated strain and a non-BPF-associated control, we demonstrated that strains which produce the cytotoxic phenotype were able to replicate intracellularly whereas non-BPF-associated strains, with MOIs of > or = 1,000 did not replicate and did not produce the phenotype. We also showed that this phenotype is not caused by the activity of an endotoxin or the release of some other compound from the bacterial cell, since neither gamma irradiation-killed whole BPF clone bacteria nor bacterial cell fractions at MOIs of > 1,000 produced the cytotoxic effect. Furthermore, bacteria in numbers equal to MOIs of > 1,000 treated with chloramphenicol did not produce the cytotoxic phenotype, suggesting a requirement for bacterial protein synthesis. In addition, viable bacteria separated from the tissue culture monolayer by a 0.2-micron-pore-size membrane also failed to produce the phenotype. The ability of the bacterium to invade, replicate, and produce the phenotype appears to be primarily parasite directed since phagocytosis, pinocytosis, and eukaryotic protein synthesis inhibitors, including cycloheximide, cytochalasin D, and methylamine, had no effect on the ability of the bacterium to invade and cause a cytotoxic response. Understanding the basic mechanisms involved in this tissue-destructive process should enhance our knowledge of the general pathogenesis of BPF.


Subject(s)
Endothelium, Vascular/microbiology , Fever/etiology , Haemophilus influenzae/pathogenicity , Purpura/etiology , Bacterial Adhesion , Cell Line , Endothelium, Vascular/ultrastructure , Gentamicins/pharmacology , Humans , Virulence
17.
J Clin Microbiol ; 33(1): 1-7, 1995 Jan.
Article in English | MEDLINE | ID: mdl-7699023

ABSTRACT

CDC nonoxidizer group 2 (NO-2) currently consists of 15 gram-negative, rod-shaped, oxidase-negative, asaccharolytic, brown soluble pigment-producing strains isolated from blood cultures, usually from young adults. On the basis of their cellular fatty acid profiles, NO-2 strains formed a single group that was identical with the profile of Bordetella avium. 16S rRNA sequencing of one NO-2 strain and the type strains of B. pertussis, B. parapertussis, B. bronchiseptica, and B. avium showed a high degree of homology (> or = 98% over 1,525 bases). The NO-2 guanine-plus-cytosine content (61.5 to 62.3 mol%) and major ubiquinone analysis (ubiquinone-8) results were both consistent with those for the genus Bordetella. DNA relatedness studies (hydroxyapatite method) confirmed a close relatedness between NO-2 and Bordetella species and demonstrated that NO-2 strains were a single new species. The name B. holmesii sp. nov. is proposed for CDC group NO-2.


Subject(s)
Bordetella Infections/microbiology , Bordetella/classification , Sepsis/microbiology , Adolescent , Adult , Bacterial Typing Techniques , Bordetella/chemistry , Bordetella/genetics , Child , DNA, Ribosomal/genetics , Fatty Acids/analysis , Female , Humans , Male , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Ubiquinone/analysis
18.
J Clin Microbiol ; 33(1): 76-8, 1995 Jan.
Article in English | MEDLINE | ID: mdl-7699070

ABSTRACT

Four slightly yellow-pigmented, alpha-hemolytic, gram-negative coccobacilli, three from wound specimens and one from multiple blood cultures of a patient with endocarditis, were identified as Neisseria elongata subsp. glycolytica on the basis of their overall biochemical and genetic similarities to this subspecies. These strains resembled N. elongata in their guanine-plus-cytosine contents (55.6 to 57.1 mol%) and in their overall cellular fatty acid profiles, which are characterized by large amounts of 16:0, 16:1 omega 7c, and 18:1 omega 7c fatty acids. Their identities were confirmed by species-level DNA relatedness (hydroxyapatite method) to the type strains of all three N. elongata subspecies. The biochemical profiles and cultural characteristics of these strains resembled those of the type strain of N. elongata subsp. glycolytica except for the production of a weak yellow growth pigment and alpha-hemolysis on sheep blood agar. They differed from N elongata subsp. elongata by the production of catalase, by the production of alpha-hemolysis on sheep blood agar, and by acid production from D-glucose. They differed from N. elongata subsp. nitroreducens by the production of catalase and an inability to reduce nitrate. These studies suggest a pathogenic potential for N. elongata subsp. glycolytica, usually considered to be a transient colonizer in humans.


Subject(s)
Blood/microbiology , Endocarditis/microbiology , Gram-Negative Bacterial Infections/microbiology , Neisseria/isolation & purification , Wounds and Injuries/microbiology , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/chemistry , Fatty Acids/analysis , Humans , Male , Middle Aged , Neisseria/chemistry , Neisseria/classification
20.
J Clin Microbiol ; 32(8): 1918-22, 1994 Aug.
Article in English | MEDLINE | ID: mdl-7989543

ABSTRACT

Thirty strains of fermentative coryneform-like bacteria designated CDC fermentative coryneform group 3 and coryneform group 5 were compared biochemically by cellular fatty acid analysis and by DNA relatedness with the type strain of Dermabacter hominis, ATCC 49369. DNA from 22 strains of both CDC groups showed 69 to 96% relatedness (hydroxyapatite method) to labeled DNA from ATCC 49369 and to DNA from CDC group 3 strain G4964, and the strains are considered to belong to D. hominis. The remaining eight strains were genetically but not phenotypically differentiable from D. hominis. They were genetically heterogeneous, but hybridization results indicated that they probably belong to the genus Dermabacter. Thirteen of the 22 D. hominis strains and all 8 of the other Dermabacter strains had been isolated from blood, which indicates the pathogenic potential of this species and genus.


Subject(s)
Actinomycetales/classification , Actinomycetales/pathogenicity , Actinomycetales/enzymology , Actinomycetales Infections/microbiology , Adult , Aged , Bacterial Typing Techniques , Centers for Disease Control and Prevention, U.S. , DNA, Bacterial/genetics , Fatty Acids/analysis , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Nucleic Acid Hybridization , United States
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