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Lipopolysaccharides (LPS) were extracted and purified from the type strain and from a clinical isolate of Branhamella catarrhalis. Chemical analysis revealed the presence of glucose, galactose, and glucosamine in different molar proportions in the LPS from these two isolates, whereas there was no difference between the two isolates in the ratios of ketodeoxyoctonate, phosphate, and the fatty acids C12, 3-OH-C12, and 3-OH-C11 present. Heptose or 3-OH-C14 was not detectable in either preparation. LPS from both strains appeared semirough according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, presenting a core polysaccharide plus one repeating unit. Immunoblotting, passive hemolysis, and hemolysis inhibition assays using anti-LPS antibodies from immunized rabbits demonstrated cross-reactivity between the LPS preparations; however, antigenic dissimilarities were also found, suggesting that more than one serotype may exist. The lipid A isolated from the two LPS was serologically identical and exhibited cross-reactivity with lipid A of members of the family Enterobacteriaceae. The B. catarrhalis LPS were biologically active, causing lethality in D-galactosamine-sensitized C57/BL6 mice and inducing Limulus amoebocyte lysate gelation.

A retrospective survey of non-typhoid Salmonella bacteraemia in the period 1984 to 1988 was carried out by the five departments of clinical microbiology in Greater Copenhagen. A total of 168 patients were identified. A gradual increase was observed from 11 cases in 1984 to 58 cases in 1988. The corresponding incidence per 100,000 inhabitants in Copenhagen rose from 0.9 in 1984 to 5.0 in 1988. During the same period the total registered incidence of human Salmonella infections in Denmark increased from 17.6 to 67.4 per 100,000 inhabitants. The serotype most often isolated from bacteraemic patients was Salmonella dublin followed by Salmonella enteritidis and Salmonella typhimurium. Salmonella dublin demonstrated enhanced invasive and pathogenic properties. Predisposing factors were present in 56% of the patients; the most common was malignant disease. A fatal or complicated course of the bacteraemia was observed more frequently in patients with underlying diseases than in persons who had previously been healthy. A total of 17% of the patients died; one-fifth of these had a ruptured aortic aneurysm. It is concluded that the substantial increase in the number of cases and the often serious course taken by the infection demonstrate a need for increased efforts at prophylaxis.

A case of preterm premature rupture of the membranes (PPROM) in the 31st week of gestation is reported. Initial cultures from the cervix and urine were without pathogenic microorganisms. After 8 days of prophylactic pivampicillin and metronidazol, culture from the cervix showed profuse growth of Group B Streptococci (GBS) and the patient developed symptoms of chorioamnionitis. Cesarean section was performed and the infant presented GBS-septicemia. In spite of continued treatment with pivampicillin, culture from the cervix on day 6 post partum still showed profuse growth of GBS. Prolonged prophylactic per oral administration of broad-spectrum antibiotics after PPROM may not always protect against infectious complications. Literature is reviewed, and it is discussed whether the applied regimen in some cases even may favour the occurrence of serious infections.

Eighty-one episodes of bacteremia and candidemia were recorded in 78 infants in our neonatal intensive care unit during the years 1984 to 1988. The species isolated from monomicrobial episodes were as follows: Enterobacteriaceae 38%, group B streptococci or pneumococci 12%, staphylococci 20%, Candida albicans 9%, and other species 15%. Polymicrobial bacteremia occurred in 6%. Group B streptococci and pneumococci were predominantly isolated from early infections, whereas Enterobacteriaceae, Staphylococcus epidermidis and C. albicans were associated mainly with late septicemia. More than 80% of the episodes occurred in premature infants. During the study period, initial empiric antibiotic therapy consisted of ampicillin plus gentamicin. In spite of the fact that Enterobacteriaceae isolates, often ampicillin resistant, and penicillinase producing staphylococci, were the dominant etiologic agents, choice of this initial therapy did not seem to contribute to mortality. Mortality was most convincingly associated with overwhelming infections caused by group B streptococci and pneumococci.

118 episodes of fungemia occurring at Rigshospitalet, Copenhagen, between 1984 and 1988 were reviewed retrospectively. Underlying diseases in the patients were dominated by malignancies, primarily hematological disorders, and intraabdominal diseases requiring major abdominal surgery. Predisposing factors identified in the patients were ongoing antibacterial chemotherapy (83%), central venous catheters (72%), major abdominal surgery (39%), and neutropenia (32%). 120 fungal strains were isolated, of which 88 (73%) were Candida albicans, 23 strains representing 8 other Candida species were also isolated, as were 9 strains belonging to 7 other fungal genera. There were only 5 strains resistant to 5-fluorocytosine (MICs greater than or equal to 25 mg/l), and no strain was resistant to amphotericin B. Treatment with antifungal agents was given in 78 patients, generally a combination of amphotericin B and 5-fluorocytosine. In 14 patients (all non-hematological) the only treatment was removal of a permanent central venous catheter. The outcome was poor in patients with hematological disorders (mortality 76%), whereas patients with malignant and non-malignant intraabdominal diseases had a mortality of 35%. All patients with a permanent central venous catheter as the only risk factor recovered rapidly after removal of the catheter.

Branhamella catarrhalis isolates from lower respiratory tract specimens collected in 1983/84 (n = 50) and 1988 (n = 30) were examined for beta-lactamase production. The percent of beta-lactamase-producing strains increased from 25% to 63% from 1983/84 to 1988. beta-lactamases from 30 strains could be typed, and of these 28 were of the BRO-1 type and two the BRO-2 type. The two beta-lactamase inhibitors, clavulanic acid and brobactam were very active against beta-lactamase extracts whether of the BRO-1 or the BRO-2 type. Susceptibility to a number of antimicrobial agents or combinations of agents was determined by a plate dilution method (MICs) and by a tablet diffusion method. The penicillin tablet (Rosco Neo-Sensitabs) was found to be useful in discriminating between beta-lactamase-producing and non-beta-lactamase- producing strains of B. catarrhalis.

During the five-year period 1984-1988 we received 192 specimens from 180 patients infected with the human immunodeficiency virus (HIV) for investigation of Legionella infection. The majority of specimens were bronchoalveolar lavage (BAL) fluids (84%), but tracheal suctions and lung tissue from autopsies were also examined. The diagnostic methods used were a direct immunofluorescence assay (DFA) for the detection of Legionella antigen, and culture on buffered charcoal yeast extract (BCYE-alpha) media. All specimens were also examined for the presence of other bacterial lung pathogens, and all BAL specimens additionally for Pneumocystis carinii and mycobacteria. Legionellosis was not found to be common among HIV-infected patients, as only six specimens (3%) from six patients were found positive by DFA, and no specimens were culture-positive for Legionella species. Dual infection with Legionella and P. carinii occurred in two patients. Clinical data of the six patients are presented, and currently used methods for diagnosing legionellosis are discussed.

At all ages, Staphylococcus aureus is the most common microorganism responsible for septic arthritis, but in children below five years of age Haemophilus influenzae is frequently found. In about 1/3 of the cases cultures are negative, primarily because of initiation of antibiotic therapy prior to sampling. A distended joint should be treated as infection until otherwise proved. Today repeated aspirations of the infected joint are recommended as initial treatment, if possible with additional joint lavage, together with systemic antibiotic therapy. In case with no clinical response within 2-4 days, surgical debridement of the joint with synovectomy is of utmost importance. Exercises without weight-bearing are initiated at an early stage and weight-bearing avoided for six weeks.

We report a case of Candida norvegensis invasive disease in an immunosuppressed renal transplant patient on continuous ambulatory peritoneal dialysis. Multiple cultures of peritoneal fluid, blood, and tracheal suction done over a 2-week period were positive for this unusual isolate. Despite treatment with amphotericin B and flucytosine the patient died. This is the first report of C. norvegensis fungemia documented by culture.

We report a case in which a strain of the U.S.A. Centers for Disease Control (CDC) dysgonic fermenter (DF) 3, together with Citrobacter freundii, was isolated from an abscess in a diabetic patient. DF 3 may be easily overlooked due to its fastidious nature, a characteristic shared with two former DF groups now placed in the genus Capnocytophaga. To our knowledge, this is the first European case report of DF 3-associated infection.

The five departments of clinical microbiology in Greater Copenhagen have together carried out a retrospective review of bacteraemia caused by the zoonotic Salmonella serotypes in the period 1984-1988 in the municipalities of Copenhagen and Frederiksberg and in the County of Copenhagen. A gradual increase in frequency was observed from 11 cases in 1984 to 58 cases in 1988. The serotype most commonly isolated was Salmonella dublin followed by Salmonella enteritidis and Salmonella typhimurium. S. dublin was found to be more invasive and more virulent than the other serotypes. Predisposing factors were present in 56% of the patients; the commonest of these was malignant disease. Fatal or complicated course of the disease were observed more frequently in predisposed patients than in persons who had previously been healthy. A total of 17% of the patients died and one fourth of these had ruptured aortic aneurysm probably on account of Salmonella arteritis. 20% developed recurrence of bacteraemia while in the remaining patients the disease ran an uncomplicated course. It is concluded that the marked increase in the number of cases and the serious course taken by the infection demonstrate a definite need for increased prophylactic efforts in the food industry.

Invasive fungal disease with Saccharomycetaceae is very rarely reported and these fungi are usually considered nonpathogenic. We report here 4 cases of positive blood cultures with fungal species belonging to this family. Severe neutropenia, permanent central venous catheter, ongoing antibacterial chemotherapy, and major abdominal surgery were identified as risk factors for fungemia in patients. In the immunocompromised host isolation of such species from sterile fluids cannot be ignored but should be considered an opportunistic infection and treated as such.

The kinetics of cefuroxime in serum and wound fluid were investigated in the period after vascular prosthetic implantation. Cefuroxime was administered as intravenous bolus injections in doses of 0.75 gm (five patients) or 1.5 gm (five patients). The concentration of cefuroxime in wound fluid increased after the injection and reached a maximal level corresponding to serum concentration levels with 1.5 hours. The subsequent elimination of cefuroxime from the wound fluid closely paralleled the elimination from the serum. The wound fluid concentrations were found to be greater than the minimum inhibitory concentration for Staphylococcus aureus and Escherichia coli in 8 and 5 hours, respectively, after injection of 0.75 gm of cefuroxime, and in 11 and 7 hours, respectively, after injection of 1.5 gm of cefuroxime.

The Harlem regional stroke program has been a demonstration model designed to detect, treat, and follow up stroke and hypertension patients through collaboration with a municipal teaching hospital, community practitioners, local service organizations and a major medical school. Many aspects of the stroke program appear suitable for replication at the local or regional levels in varied settings. In particular, the program has demonstrated the need to link community outreach programs, stressing early detection and preventive-care education, with sustained treatment and follow-up programs. The stroke program has also suggested ways in which specialized programs can be incorporated into long-term comprehensive health planning and care facilities at local and regional levels.

By means of crossed immunoelectrophoresis and crossed-line immunoelectrophoresis, antigens were compared from four strains and five strains, respectively, of Flavobacterium meningosepticum DNA hybridization groups I and II. Two reference antisera raised against the pooled antigens of each of the two DNA groups were used. The two reference antigen-antibody systems produced 81 and 51 immunoprecipitates, respectively, and within each group nearly all antigens were common to all strains, giving matching coefficients of 0.98-1.00. Group specific antigens were not detected. When antigens from individual strains from each group were compared to individual strains of the other group, however, 5-15 of the antigens were not cross-reactive, giving matching coefficients of 0.83-0.94. The antigenic relatedness between strains belonging to the two different groups was, therefore, just below the level commonly found for strains belonging to the same species. The results thus confirm the previously published differences detected by a DNA hybridization technique.

Antimicrobial susceptibilities of 52 strains of Flavobacterium meningosepticum were determined by an agar plate dilution technique and by a tablet diffusion method. No useful susceptibility differences between the two main DNA relatedness groups within F. meningosepticum were found. All strains produced beta-lactamase and were resistant to most beta-lactam antibiotics. Ciprofloxacin, rifampicin and clindamycin appear to be the most promising for treating infections caused by F. meningosepticum.

47 staphylococcal reference strains representing 13 species were tested for polymyxin sensitivity using tablet and disc diffusion methods. Corresponding MIC and IC50 values were determined with a plate dilution assay. Coagulase-positive strains were found to be more resistant towards polymyxin, and could thereby be separated from coagulase-negative strains. Furthermore, 100 recently isolated staphylococci from clinical specimens were tested, and again the coagulase-positive strains could be identified by their smaller tablet inhibition zones. We conclude that the polymyxin test is an aid in the fast differentiation of staphylococci in laboratories performing primary sensitivity tests.

116 cystic fibrosis patients were observed, by monthly examinations over an eight-month period, to investigate the importance of non-bacterial respiratory infections (NBI) in exacerbations of the respiratory disease. Sputum was examined for bacteria, and serum investigated for antibody response against virus, mycoplasma and chlamydia and for antibodies against Pseudomonas aeruginosa. During this period each patient had, on an average, 2.9 exacerbations of which 76% were associated with bacteria, most frequently P. aeruginosa (51%), and 20% with NBI, although bacteria were also present in most of these cases. No etiology was established in 18% of the exacerbations. The NBI were caused by respiratory syncytial virus (RSV) (9%), parainfluenza virus (5%), influenza virus (3.6%), adenovirus (2.4%), mycoplasma (0.6%) and chlamydia (0.6%). The incidence of exacerbations was higher in patients with chronic P. aeruginosa infections. RSV infections were more common in patients who developed chronic P. aeruginosa infection during the study period, and RSV infections were frequently associated with a rise of P. aeruginosa antibodies in patients who harboured these bacteria. The important role of NBI as mediators of onset of chronic P. aeruginosa infections in cystic fibrosis patients is suggested.

Previous surveys of stroke populations have offered only cursory information on language disturbance, and, conversely, few surveys of aphasic populations have dealth exclusively with stroke or with acute phenomena. This paper describes aphasia in 850 acute stroke patients consecutively registered by the Harlem Regional Stroke Program, of whom 177 (21%) were aphasic; of these, nine were of Broca's type, 24 were of Wernicke's type, 14 were of anomic, ten were conduction, seven were of "isolation" type, and 107 were "mixed." An unexpected finding was a significant over-representation of men among the nonfluent aphasics. During the following four to 12 weeks, 12% of fluent aphasics died, and 12% remained moderately or severely impaired; among survivors, aphasia improved in 74%, and in 44% it cleared completely. During the same period, 32% of nonfluent aphasics died, and 34% remained moderately or severely impaired; among survivors, aphasia improved in 52%, and in only 13% did it clear completely. In both fluent and nonfluent groups, hemiparesis and/or visual field cut were associated with poor prognosis.

DNA-DNA reassociation studies on 52 strains of Flavobacterium meningosepticum showed two main hybridization groups about 40-55% interrelated and comprising 4 and 48 strains respectively. The larger group could be further divided into four subgroups if differences in thermal stability of the reassociated duplexes were taken into consideration. Of a total of 20 strains isolated from cerebrospinal fluid, 18 were found in one subgroup of 28 strains, which also contained seven blood isolates. The results indicate that genetically defined subgroups within the species might differ with regard to pathogenic significance.

171 episodes of bacteremia and candidemia in 142 patients were recorded during the period 1981-1985 in patients with hematological malignancies. Overall mortality, within 1 week of onset of bacteremia, was 20%. Increased mortality was found in patients with poor disease-prognosis (39%), with granulocytopenia for more than 6 days (30%), and in those developing hypotension (49%). Compared to a similar previous study from 1970-1974, the incidence of bacteremia remained unchanged, but the mortality had decreased by 50%. This result could not be explained by changes in microbial spectrum, in patient groups, or in the initial antibiotic regimen used. An improved general condition of these patients due to better supportive treatment may be the single most important factor in improving the outcome of septicemia.

A total of 55 different antigens were demonstrated in pooled lysate of six Flavobacterium meningosepticum strains by crossed immunoelectrophoresis against homologous rabbit antibody. The six strains used represented the two DNA relatedness groups within this species. All 52 strains of F. meningosepticum investigated were found to cross-react with 53 to 55 of these F. meningosepticum reference antigens. Using this F. meningosepticum reference system, cross-reactions between the F. meningosepticum reference antigen and antigens from representatives of other Flavobacterium taxa were also investigated. A positive correlation was demonstrated between the number of cross-reacting antigens and the taxonomic relatedness of the investigated taxa as determined by other classification methods, including DNA-DNA hybridization. The crossed immunoelectrophoretic techniques were not useful in differentiating the two main DNA groups of F. meningosepticum.

Among 148 strains of Haemophilus influenzae isolated in 1975 from different localities in Denmark, none was found to be resistant to ampicillin. A majority of the strains belonged to biotypes I, II and III and the presence of a partial correlation between biotype and origin was confirmed.

Eleven strains of Cardiobacterium hominis, two strains of Kingella indologenes and six strains of CDC group EF-4 were characterized. Since all three taxa are oxidase-positive, fastidious gram-negative rods with relatively few positive reactions, they may be easily confused in the microbiological laboratory. Common characteristics are acid production from glucose, aerobic growth in semi-solid agar and very slow anaerobic growth. Group EF-4 is distinguished from the other two species by its capacity to produce catalase and its lack of indole production. Cardiobacterium hominis can be distinguished from Kingella indologenes by its ability to ferment mannitol and sorbitol and by differences in enzyme production. The three taxa also differ considerably with regard to morphological and cultural characteristics.

The microorganisms isolated in 1981-1985 from 171 cases of septicemia in patients with hematological malignancies were on the whole the same as those found in 1970-1972. The distribution between species was also quite similar for the two periods except within staphylococci, where the isolation rate of coagulase-negative staphylococci was higher in the latter period while that of Staphylococcus aureus was lower. Of 67 strains of Enterobacteriaceae tested for an aminoglycoside, 6% were found to be resistant, whereas 8% of 48 Enterobacteriaceae strains were found to be cefotaxime resistant. Methicillin- or aminoglycoside resistant S. aureus did not occur.

A three-month survey revealed 29 patients at our hospital with symptoms of acute pulmonary infection, from whom Branhamella catarrhalis was isolated from lower respiratory tract specimens, in 18 cases in pure culture. Approximately 2% of all respiratory tract specimens examined during the period yielded growth of B. catarrhalis. All except one patient suffered from chronic pulmonary disease, notably chronic bronchitis. A phenotypic comparison was made between 55 strains of B. catarrhalis, of which 50 were recent isolates from lower respiratory tract specimens, and 23 Neisseria strains representing Neisseria meningitidis, Neisseria gonorrhoeae, Neisseria cinerea, Neisseria flavescens, Neisseria mucosa, Neisseria pharyngis, and Neisseria lactamica. The morphology of B. catarrhalis colonies is very characteristic, and when the diagnosis is suspected, testing for the ability to hydrolyze tributyrin may confirm it within hours. Ability to produce deoxyribonuclease is another property which differentiates B. catarrhalis from the Neisseria species. Otherwise, the combination of nitrate reduction and failure to produce acid from glucose, maltose, and sucrose establishes the diagnosis.

Fifty-two strains found to belong to Flavobacterium meningosepticum on the basis of DNA-DNA hybridization analyses were characterized and found to be phenotypically homogeneous. All strains were oxidase, catalase, indole, gelatinase and beta-galactosidase positive, and produced acid from glucose, mannose, fructose, maltose and mannitol; nitrate was not reduced. F. meningosepticum could be differentiated from Flavobacterium group IIb by its ability to produce beta-galactosidase, and by the latter taxon's ability to produce a bright yellow pigment in contrast to the weak or non-existing pigmentation of F. meningosepticum. Phenotypic characteristics that could differentiate between the two main DNA relatedness groups of F. meningosepticum were not discovered, wherefore a subdivision of the present species into two species cannot be recommended. Strains from the DNA relatedness groups comprising 19 of 20 CSF isolates were found to be able to grow at 40 degrees C and to produce a weak yellow pigment; in contrast, strains from the three other DNA relatedness groups were unable to grow at 40 degrees C and produced no pigment.

An outbreak of infections due to Flavobacterium meningosepticum type C in a neonatal intensive care unit is described. During a period of two weeks, two infants developed meningitis and a third was colonized in the respiratory tract and had transient bacteremia. The two meningitis patients were treated with clindamycin, rifampicin and cefotaxime systemically, plus rifampicin intraventricularly. Bacteriological eradication was achieved within 48 h, and both infants recovered from the meningitis without apparent neurological sequelae; however, one infant died two months later of unrelated causes. Environmental surveillance cultures failed to demonstrate a reservoir for the epidemic strain, but other Flavobacterium strains were recovered. Two clinically healthy infants were found to be colonized in the nasopharynx with strains that were extremely difficult to differentiate phenotypically from the epidemic strain. Extensive characterization of strains is necessary in order to differentiate between strains and subsequently to determine a certain source of infection.