Showing papers in "Journal of Clinical Microbiology in 1987"

Duration of tick attachment and Borrelia burgdorferi transmission.

Abstract: Nymphal Ixodes dammini transmitted Borrelia burgdorferi to 1 of 14 rodents exposed for 24 h, 5 of 14 rodents exposed for 48 h, and 13 of 14 rodents exposed for greater than or equal to 72 h. Prompt removal of attached ticks is a prudent public health measure, especially in regions where Lyme disease is endemic.

Application of DNA typing methods to epidemiology and taxonomy of Candida species.

Abstract: Methods are described for extraction of DNA from the yeast form of Candida spp., followed by digestion and electrophoresis of DNA fragments. The resulting gel patterns (greater than 100 bands) were used to type Candida isolates. Four intense bands identified, three of which are present in each isolate (6 to 7, 3.7 or 4.2, and 2.5 to 3 kilobases), appear to be DNA encoding the rRNA. The methods proved to be both simple and reproducible. The patterns were shown to be stable through several hundred doublings from multiple single colonies. A survey of isolates showed that, on the basis of similarity of gel patterns, several Candida species could be sorted into mutually exclusive groups, and subgroups could be created. Analyses of this survey suggested the possible epidemiologic and taxonomic applications of these methods. DNA typing methods appear to offer important potential advantages over phenotyping methods. The methods provide a base for further epidemiologic studies and for further development of techniques, such as the use of cloned probes for studies of DNA homology. Images

Urease-positive bacteriuria and obstruction of long-term urinary catheters.

Abstract: Long-term urethral catheterization (greater than or equal to 30 days), a management technique for urinary incontinence, results in polymicrobial bacteriuria. We frequently found urease-producing bacteria: of 1,135 weekly urine specimens from 32 long-term-catheterized patients, 86% had urease-positive bacterial species at greater than or equal to 10(5) CFU/ml. The most common species were Proteus mirabilis and Morganella morganii, each found in over half the specimens. P. mirabilis, but not other urease-positive species, was significantly associated with the 67 obstructions observed in 23 patients. M. morganii had a more complex association and in some way may protect the catheter from obstruction.

Rotavirus isolate WI61 representing a presumptive new human serotype.

Abstract: A virus (strain WI61) representing a presumptive new human serotype was isolated from an 18-month-old child with gastroenteritis admitted to Children's Hospital of Philadelphia in February 1983. The WI61 virus was clearly distinguished by cross-neutralization tests from human rotaviruses of serotypes 1, 2, 3, and 4, human 69M, and representative bovine (NCDV), porcine (OSU), and chicken (Ch2) rotaviruses. Antisera generated in guinea pigs hyperimmunized to WI61 virus displayed a partial cross-reactivity with rotaviruses of human serotypes 1, 2, 3, and 4. By means of studies with reassortant rotaviruses, it was presumptively determined that the WI61 virus cross-reactive antigenic determinants are localized on the vp3 surface polypeptide coded by gene segment 4. The characteristic RNA genome electropherotype of WI61 virus was observed in 5 of 59 cases of infant gastroenteritis detected in 1983 and 1984 but has not been observed in a subsequently at Children's Hospital. Serotype WI61-specific neutralizing antibodies were observed in a majority of sera of normal adults and infants of less than 4 or greater than 12 months of age collected in the Philadelphia area. Median antibody titers to WI61 equaled or exceeded those to rotaviruses of serotype 1 or 3. Each of seven samples of commercial cow's milk exhibited neutralizing antibodies to WI61 virus at a titer greater than or equal to that to serotype 1 or 3 or bovine (strain NCDV) rotavirus. However, WI61 rotavirus did not induce disease or a specific serum-neutralizing antibody response when fed to a caesarean-derived colostrum-deprived newborn calf. WI61 rotavirus caused diarrhea in newborn mice with a 50% diarrhea-inducing dose of 10(7.0) PFU.

Crohn's disease-isolated mycobacteria are identical to Mycobacterium paratuberculosis, as determined by DNA probes that distinguish between mycobacterial species.

Abstract: DNA extracted from an unclassified Crohn's disease-isolated Mycobacterium strain was cloned. The recombinant clones were radiolabeled and hybridized to restriction digests of mycobacterial DNA transferred to nylon membranes. Restriction fragment length polymorphisms (RFLPs) were identified that distinguished between mycobacterial DNA samples. Quantitative estimates of frequencies of DNA base substitution were also obtained. No RFLPs were detected between the DNA of three unclassified Crohn's disease-isolated mycobacteria and Mycobacterium paratuberculosis, although several RFLPs were detected that distinguished between M. paratuberculosis and both M. avium complex serovars 2 and 5. The frequency of DNA base substitution between M. paratuberculosis and M. avium complex serovar 2 was measured as 0.87 (+/- 1.2)%.

High-frequency switching in Candida strains isolated from vaginitis patients.

Abstract: High-frequency switching and strain variability at the site of infection was assessed in 11 patients with acute Candida albicans vaginitis. By cloning cells directly from the site of infection, it was demonstrated that 4 of the 11 isolates contained multiple-switch phenotypes at the site of infection and that 9 of the 11 isolates were in a high-frequency mode of switching (10(-2) to 10(-3)). Isolates could be separated into four general categories of switching repertoires. To demonstrate that multiple phenotypes at the site of a single infection represented the same strain, EcoRI digests of total cell DNA were separated on agarose gels, and Southern hybridization patterns with two cloned midrepeat sequences were compared. Images

Pythium insidiosum sp. nov., the etiologic agent of pythiosis.

Abstract: Pythium insidiosum sp. nov., the etiologic agent of pythiosis, a cosmopolitan disease of horses, cattle, and dogs, is described and illustrated. Images

Aeromonas veronii, a new ornithine decarboxylase-positive species that may cause diarrhea.

Abstract: In 1983, the vernacular name Enteric Group 77 was coined for a group of strains that had been referred to our laboratory as "possible Vibrio cholerae except for gas production." By DNA-DNA hybridization (hydroxyapatite, 32P), 8 of 10 strains of Enteric Group 77 were very highly related to the labeled strain 1169-83 (74 to 100% at 60 degrees C and 75 to 100% at 75 degrees C; percent divergence, 0.0 to 2.5). Type strains of six other Aeromonas species were 45 to 66% related (60 degrees C) to strain 1169-83, but type strains of 27 Vibrio species were only 2 to 6% related. The name Aeromonas veronii is proposed for the highly related group of nine strains formerly known as Enteric Group 77. The type strain is designated as ATCC 35604 (CDC 1169-83). Strains of A. veronii grew well at 36 degrees C and had positive reactions at this temperature for indole, methyl red, Voges-Proskauer, citrate, lysine and ornithine decarboxylases, DNase, lipase, and motility; the strains had negative reactions for arginine decarboxylase, H2S, urea, and malonate. The following sugars were fermented: D-glucose (acid and gas), cellobiose (seven of nine strains), D-galactose, maltose, D-mannitol, D-mannose, alpha-methyl-D-glucoside (eight of nine strains), salicin, sucrose, and trehalose. The following sugars were not fermented: adonitol, L-arabinose, D-arabitol, dulcitol, erythritol, myo-inositol, lactose, raffinose, L-rhamnose, D-sorbitol, and D-xylose. The positive ornithine decarboxylase reaction differentiates A. veronii from other Aeromonas species. The antibiogram of A. veronii is typical of other Aeromonas strains (resistance to ampicillin and carbenicillin and susceptibility to most other agents). A. veronii strains were isolated from three clinical sources: respiratory secretions of four victims of drowning or near drowning in fresh water (probably not clinically significant); infected wounds of two patients previously exposed to fresh water (unknown clinical significance); and stools from three patients with diarrhea (probably clinically significant).

Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses.

Abstract: An enzyme immunoassay for serotyping human rotaviruses in stools and in cell culture was developed Hyperimmune rabbit antisera to rotaviruses were used as capture antibodies, and rotavirus-neutralizing mouse monoclonal antibodies specific for serotypes 1, 2, 3, and 4 were used as detection reagents Partial purification of monoclonal antibodies and inclusion of skim milk powder in antibody diluents contributed to assay specificity The sensitivity of this assay was greater than that of a direct enzyme immunoassay in which rotaviruses of the appropriate serotype were adsorbed directly to the solid phase When fecal extracts were concentrated threefold, this serotyping enzyme immunoassay was of equal specificity and approached the sensitivity of electron microscopy for rotavirus detection This assay is simple and rapid and is suitable for serotyping the large numbers of isolates obtained from epidemiological studies and vaccine trials

Improved methods for isolation and enumeration of Malassezia furfur from human skin.

Abstract: A medium for the isolation and enumeration of Malassezia furfur is described. Incubation at 34 degrees C yielded geometric mean counts (in CFU per square centimeter) of 2.6 X 10(3) on the forehead, 8.5 X 10(2) on the cheek, and 9.6 X 10(3) on the back. These counts compared favorably with microscopic counts and greatly exceeded those obtained with previously described media.

High incidence of hemolysin production by Enterococcus (Streptococcus) faecalis strains associated with human parenteral infections.

Abstract: Hemolysin production, clumping (pheromone) response, transferability of the hemolytic trait, and drug resistance were examined in 97 clinical isolates of Enterococcus (Streptococcus) faecalis. The isolates were derived from various sources (i.e., urine, pus, vagina, sputum, bile, and blood), and approximately 60% were found to be hemolytic. About 85% of the hemolytic strains exhibited a clumping response, compared with about 49% of the nonhemolytic strains. Over 50% of the hemolytic strains carried transferable hemolysin determinants, and in no case were drug resistance genes linked. The hemolytic strains exhibited multiple drug resistance more frequently than did the nonhemolytic strains. In contrast to the high frequency of hemolysin producers among parenteral isolates, strains derived from fecal specimens of healthy individuals exhibited a low (17%) incidence of hemolysin production.

Selected characteristics of pathogenic and nonpathogenic strains of Bacteroides gingivalis.

Abstract: Strains of Bacteroides gingivalis were compared for the presence of properties associated with pathogenicity. Some strains were infectious in pure culture in an in vivo model (guinea pig), and all but one of these were more collagenolytic than those which failed to cause lesions in guinea pigs. However, other factors seem to be necessary for the induction of an infection in this animal model.

Improved medium for antimicrobial susceptibility testing of Haemophilus influenzae.

Abstract: The need for complex growth media has complicated routine susceptibility testing of Haemophilus influenzae because of antagonism of certain antimicrobial agents by the medium or because of difficulties in interpretation of growth endpoints. Haemophilus test medium (HTM) is a simple, transparent medium for broth- or agar-based tests with H. influenzae. HTM incorporates Mueller-Hinton medium with additions of 15 micrograms of hematin per ml, 15 micrograms of NAD per ml, and 5 mg of yeast extract per ml as growth-promoting additives. Agar or broth microdilution MICs of 10 antimicrobial agents for a collection of 179 H. influenzae isolates determined by using HTM compared favorably with MICs determined by the conventional agar or broth dilution methods recommended by the National Committee for Clinical Laboratory Standards. Disk diffusion tests performed with HTM allowed accurate categorization of susceptible and resistant strains and were easier to interpret than tests performed with Mueller-Hinton chocolate agar. A particular advantage of HTM was the reliability of broth- or agar-based test results with trimethoprim-sulfamethoxazole. The results of the study suggest modification of current National Committee for Clinical Laboratory Standards MIC-interpretive criteria for H. influenzae with amoxicillin-clavulanate, chloramphenicol, and trimethoprim-sulfamethoxazole. Error rate-bounded analysis of MICs and disk diffusion zone sizes also suggest modified zone-interpretive criteria for ampicillin, amoxicillin-clavulanate, chloramphenicol, and tetracycline with HTM or conventional media. Interpretive zone sizes are newly proposed for cefaclor and rifampin disk diffusion tests.

Isolation of immunodominant antigens from sera of patients with systemic candidiasis and characterization of serological response to Candida albicans.

Abstract: Candidal antigens were isolated by affinity chromatography from the sera of patients with disseminated Candida albicans infections. The immunodominant 47-kilodalton (kDa) antigen appeared to be a heat-stable breakdown product of several larger heat-labile components (84 to 92, 74 to 79, and 66 to 72 kDa). It was undetectable in normal sera and sera from four patients with systemic C. parapsilosis, C. tropicalis, and C. krusei infections. Serum samples from 92 patients with proven systemic C. albicans infections were examined by the immunoblot technique. Seventy-four patients had detectable antibody, and 92% of these produced antibody to the 47-kDa antigen. All survivors had major serological responses to this antigen, whereas patients who died had no, minor, or fading responses. Fifty-five of the patients were neutropenic following cytotoxic chemotherapy for malignancies, usually lymphoproliferative disorders (hematological patients). The remainder were surgical or medical patients (nonhematological). Hematological patients differed from nonhematological patients in the range of antigens that were commonly recognized by their immune systems, although antibodies to the 47- and 60-kDa antigens were frequently present in both groups. They also differed in that they produced mainly an immunoglobulin M (IgM) response, failing to seroconvent to IgG. This did not reduce survival rates, which were similar in both groups. It may be responsible, however, for the lower antigen titers that were observed in hematological patients when measured by reverse passive latex agglutination. Images

Rapid identification of Mycobacterium avium complex in culture using DNA probes.

Abstract: A commercial DNA probe procedure for identification of Mycobacterium avium complex isolates was evaluated for accuracy and applicability for use in the clinical laboratory. The test (Gen-Probe Rapid Diagnostic System for Mycobacterium Avium Complex; Gen-Probe Corp., San Diego, Calif.) uses hybridization in solution of two 125I-labeled cDNA probes. One probe is complementary to rRNA from M. avium, and the other is complementary to rRNA from M. intracellulare. Results are expressed as absolute percent hybridization, with values greater than or equal to 10% considered positive. The procedure accurately identified all 134 M. avium complex isolates and concomitantly identified them to species level. There were no false-positives with 66 other mycobacteria, including 22 M. tuberculosis and 18 M. kansasii isolates, or with 8 Nocardia isolates. The mean percent hybridization (+/- the standard deviation) of M. avium probe-positive isolates (94 isolates) was 48.0 +/- 9.9 (range, 11.5 to 72.7); for M. intracellulare (40 isolates), it was 45.7 +/- 8.8 (range, 22.7 to 60.7). Among the 74 non-M. avium complex isolates, the percent hybridization range was 1.0 to 4.2, except for a single value of 9.7 which was less than 5 when the test was repeated. Four M. avium complex isolates reacted positively with both probes on initial testing, and three were confirmed. On repeat testing of subcultures, each reacted with only one probe, suggesting the presence of a mixed culture. The procedure can be completed in as little as 2 h and could easily be performed in most clinical laboratories.

Candida parapsilosis fungemia associated with parenteral nutrition and contaminated blood pressure transducers.

Abstract: During the period September 1983 through May 1985, Candida parapsilosis was isolated from intravascular sites (blood or vascular catheter tips) in 12 patients at a pediatric hospital. Of 205 patients with cultures of any site positive for Candida species, 32 (16%) had cultures positive for C. parapsilosis. In contrast, of 23 patients with intravascular cultures positive for Candida species, 12 (51%) had cultures positive for C. parapsilosis (P less than 0.001, Fisher's exact test). The 12 patients with intravascular cultures positive for C. parapsilosis were more likely to have received central venous nutrition therapy (10 of 12 versus 7 of 23; P less than 0.01, Mantel-Haenzel chi-square test) and had a longer duration of exposure to blood pressure transducers (P less than 0.08, paired t test) than the 23 ward- and age-matched controls. C. parapsilosis was isolated from 11 (32%) of 34 in-use and stored blood pressure transducers. After ethylene oxide sterilization of blood pressure transducers was begun, in-use pressure transducers showed no growth of C. parapsilosis. This study emphasizes the role of C. parapsilosis as a nosocomial pathogen associated with invasive devices and parenteral nutrition; it also emphasizes the importance of adhering to recommended procedures for sterilizing blood pressure transducers.

Fluorescence detection of Cryptosporidium oocysts in human fecal specimens by using monoclonal antibodies.

Abstract: With the discovery that the coccidian parasite Cryptosporidium sp. can cause severe symptoms in humans, implementation of many diagnostic techniques rapidly followed. The infection is self-limiting in patients with normal immune systems but chronic in the immunosuppressed patient. With the eventual development and use of therapeutic agents, it will become very important to find Cryptosporidium sp., even in low numbers, in fecal specimens. Production of a highly specific and sensitive antibody by use of cloning techniques has provided another diagnostic tool. Formalinized positive human fecal specimens (n = 99) and negative specimens (n = 198), of which 115 contained yeastlike fungi and other organisms, were tested in blind trials by use of a monoclonal antibody. Sensitivity was 100% with 3- to 4+ fluorescence on all cryptosporidial oocysts, both in light and heavy infections. The organisms were round and easily visible (4 to 6 micron), showing apple-green to yellow fluorescence against a dark background free of nonspecific fluorescence. Specificity was also 100% with all 99 positive Cryptosporidium sp. specimens exhibiting fluorescence and all 198 negative specimens showing no fluorescence. All positive and negative specimens were previously confirmed by the hot modified acid-fast technique. However, seven specimens previously considered negative by this acid-fast method were positive by the monoclonal antibody technique. These specimens were confirmed as positive, after extensive examination of additional smears prepared by the modified hot acid-fast method revealed rare organisms, emphasizing the increased sensitivity of the monoclonal antibody technique. Since acid-fast stains do not always consistently stain all oocysts, the increased sensitivity of the monoclonal reagent provides an excellent screening method.

Local and systemic antibody response to bovine respiratory syncytial virus infection and reinfection in calves with and without maternal antibodies.

Abstract: Enzyme-linked immunosorbent assays for the detection of immunoglobulin M (IgM), IgA, IgG1, and IgG2 antibodies against bovine respiratory syncytial virus (BRSV) were used to measure antibody responses of calves after experimental or natural infection with BRSV. Serially collected sera, lung lavage samples, nasal and eye secretions, and feces were tested for the presence of these antibodies. Lung lavage fluids and nasal secretions were further examined for the presence of virus. After experimental infection of 3- to 4-week-old, colostrum-deprived (seronegative) calves, the virus was detected from days 3 to 8 post-initial inoculation day (PID). An immune response was first detected 8 to 10 days PID, when BRSV-specific IgM and IgA appeared nearly simultaneously in serum, secretions, and feces. BRSV-specific IgG1 appeared only in serum on days 13 to 17 PID, and IgG2 was first detected in sera from 1 to 3 months PID. Specific IgM and IgA were detectable in the different samples for various periods. In the respiratory and eye secretions, IgA usually remained detectable for long periods, that is, for up to 3.5 months or longer. In lung lavage samples, BRSV-specific IgG1 was only incidentally demonstrated and appeared to be blood derived. The immune response of a 5-month-old calf strongly resembled that of the 3- to 4-week-old calves (feces excepted), indicating that an age effect on the immune response to BRSV is unlikely. After experimental infection of colostrum-fed, seropositive calves, both local and systemic antibody responses were largely or totally suppressed. The degree of suppression seemed to be related to the level of preinoculation virus-specific serum IgG1. Of all isotypes, IgM was least affected. Colostrum-fed animals shed virus in about equal amounts and for the same length of time as colostrum-deprived calves. Clinical signs were mild in both groups. After reinfection, no virus shedding was detected in either colostrum-deprived or colostrum-fed calves. In both groups, a secondary immune response developed, characterized by strong and rapid (from about day 6 PID) mucosal and systemic IgA responses, but reaching higher titers in colostrum-deprived calves. Also, strong mucosal, but not serum, IgM responses were observed, which, however, did not develop faster than those observed after primary infection. Naturally infected calves, showing severe signs of respiratory disease, had various levels of, most likely, maternally derived antibodies on the first day of illness. Mucosal and systemic antibody responses of various heights and durations were observed, but in general these responses were stronger than those observed after experimental infection. The results point to an important role for local IgA, rather for serum IgG1, in the protection against BRSV infection. The capacity to mount a local memory IgA response seems especially important. Priming for such a mucosal memory response is possible even when the primary immune response is severely suppressed because of the presence of material antibodies.

Serotyping and electron microscopy studies of Staphylococcus aureus clinical isolates with monoclonal antibodies to capsular polysaccharide types 5 and 8.

Abstract: Monoclonal antibodies to Staphylococcus aureus capsular polysaccharide types 5 and 8 were prepared and used to serotype 821 clinical isolates of S. aureus from four countries. The capsular polysaccharide-binding sites on the bacterial membrane were examined by transmission and scanning electron microscopy.

Isolation of enterotoxigenic Bacteroides fragilis from humans with diarrhea.

Abstract: Enterotoxigenic Bacteroides fragilis was isolated from stool specimens of 8 of 44 diarrheic individuals (ages, 4 months to 69 years). The individuals had watery diarrhea and intestinal cramping; and infants had hyperthermia, vomiting, and blood in the stools. No recognized enteric pathogens were detected in seven of the eight diarrheic individuals positive for enterotoxigenic B. fragilis. The bacterium produced an enterotoxin detectable in concentrated broth that supported bacterial growth. Fifteen adult rabbits with ligated ceca developed fatal enteric disease following intraileal injection with 5 x 10(9) CFU of enterotoxigenic B. fragilis. Conversely, eight control rabbits injected with nonenterotoxigenic B. fragilis remained clinically normal. As few as 5 x 10(3) CFU of enterotoxigenic B. fragilis caused fatal enteric disease in the rabbit model. Disease in rabbits was characterized by mucoid, often hemorrhagic, diarrhea. The bacterium colonized the caudal small intestine and the colon of the rabbits and caused moderate to severe necrotizing colitis. Enterotoxigenic B. fragilis is widespread in the intestinal tract of diarrheic humans and is enteropathogenic in adult rabbits with ligated ceca. Its possible role in the enteric disease complex merits further study.

Cytotoxic necrotizing factor production by hemolytic strains of Escherichia coli causing extraintestinal infections

Abstract: Two hundred and nineteen strains of Escherichia coli from extraintestinal infections and feces of healthy subjects were examined for hemolysin (Hly) and cytotoxic necrotizing factor (CNF) production and for mannose-resistant hemagglutination. Of 105 strains from extraintestinal infections, 42 (40.0%) were positive for production of both Hly and CNF, and 21 (20.0%) were positive for Hly alone; on the contrary, only 1 Hly- and CNF-positive strain and 2 Hly-positive strains were found among 114 strains from normal stools. CNF production was not found to occur among the nonhemolytic strains, confirming the close association existing between these toxic factors. Hemolytic strains positive for CNF showed mannose-resistant hemagglutination less frequently than did Hly-positive, CNF-negative strains (25.6 versus 82.6%), suggesting the existence of two distinct classes among hemolytic strains of E. coli.

Indicator medium for isolation of Campylobacter pylori.

Abstract: The use of a new indicator culture medium, Belo Horizonte medium, is proposed for better colony recognition and a presumptive identification of Campylobacter pylori. This medium, containing brain heart infusion sheep blood agar, was supplemented with 40 mg of 2,3,5-triphenyltetrazolium chloride per liter in addition to vancomycin, nalidixic acid, and amphotericin B. On Belo Horizonte medium, Campylobacter pylori present unique golden colonies.

Subgroup characteristics of respiratory syncytial virus strains recovered from children with two consecutive infections.

Abstract: Respiratory syncytial virus strains from 13 children who had repeat infections at least 1 year apart were identified as either subgroup A or subgroup B according to reaction patterns with several monoclonal antibodies directed against the large surface glycoprotein (G), fusion protein (F), nucleoprotein (NP), and matrix protein (M). The virus strains were characterized by enzyme immunoassay, polyacrylamide gel electrophoresis, and immunofluorescence procedures. During the first infection, 10 children had subgroup A strains and 3 had subgroup B strains. Of the 10 children with subgroup A strains during their first infection, 6 had subgroup B and 4 had subgroup A strains during the second infection. Of the three children with subgroup B strains during their first infection, one had subgroup A and two had subgroup B strains during their second infection. No child experienced unusually severe respiratory tract illnesses during second infections with respiratory syncytial virus. Fourfold or greater rises in serum antibody as determined by enzyme immunoassay were as common after the first infection as after the second infection among the seven children tested. Thus, second infections with strains of either subgroup of respiratory syncytial virus did not potentiate respiratory illness, and infection with subgroup A strains of respiratory syncytial virus provided some protection from a second infection with the homologous, but not the heterologous, subgroup of the virus.

Prevalence and characterization of hippurate-negative Campylobacter jejuni in King County, Washington.

Abstract: A total of 593 strains of thermophilic Campylobacter species were isolated either from humans with diarrhea or from poultry in King County, Washington. Of these strains, 98 (52 hippurate-positive strains and all 46 of the hippurate-negative strains) were selected for further phenotypic characterization and genetic classification. Hippurate hydrolysis, the test typically used to differentiate Campylobacter jejuni and C. coli, did not always correlate with the genetic classification. All hippurate-positive strains were classified as C. jejuni. Of the hippurate-negative strains, 20% were C. jejuni, 78% were C. coli, and 2% were C. laridis. Assuming that the remaining hippurate-positive strains were all C. jejuni, then hippurate-negative C. jejuni represented a small percentage (9 of 556 or 1.6%) of C. jejuni strains but a significant percentage (9 of 46 or 20%) of hippurate-negative strains. This finding suggests that hippurate hydrolysis should not be used as the sole criterion for differentiating thermophilic Campylobacter species, particularly when describing the disease states associated with these organisms.

Characterization of Borrelia burgdorferi strains isolated from Ixodes pacificus ticks in California.

Abstract: In a survey of 1,714 adult Ixodes pacificus ticks collected in northern California, 24 (1.4%) were found to be infected with spirochetes that reacted with an anti-Borrelia burgdorferi polyvalent conjugate in direct immunofluorescence tests. Eleven isolates of B. burgdorferi from these ticks were characterized by monoclonal antibody, polyacrylamide gel electrophoresis, and Western blot (immunoblot) analyses. Ten of the isolates had molecular and antigenic characteristics similar to those of other U.S. isolates. One strain, cloned by limiting-dilution techniques, was different from any previously reported U.S. strain, but similar to reported European strains. The cloned strain, DN127-Cl9-2, did not react with monoclonal antibodies to Osp A and Osp B major proteins found in most of the U.S. strains. It exhibited an abundant protein with an apparent molecular weight of 25,000.

Serogroup F, a new capsule serogroup of Pasteurella multocida.

Abstract: Four capsule serogroups (A, B, D, and E) have been described by using passive hemagglutination tests. Serogroups A and D predominant in pasteurelloses of avian species. A new capsule serogroup of Pasteurella multocida has been isolated from turkeys in Arkansas, California, Indiana, Iowa, Missouri, New Jersey, and Virginia. Strains belonging to the new serogroup were somatic serotype 1, 3, 7, or 12, and they varied in virulence for mice and poults. Antisera made in rabbits passively protected mice against challenge with the same serogroup regardless of somatic serotype.

Reactivity of monoclonal antibodies to Rickettsia rickettsii with spotted fever and typhus group rickettsiae.

Abstract: Analysis of 15 spotted fever group (SFG) and 2 typhus group strains of rickettsiae with a panel of monoclonal antibodies revealed a number of shared and unique epitopes of the 120- and 155-kilodalton surface proteins. All of the SFG strains but neither of the typhus group strains reacted with antibody to the lipopolysaccharidelike antigen of Rickettsia rickettsii; possibly the lipopolysaccharidelike antigen is the common antigen which defines the SFG. North Carolina and Montana strains of R. rickettsii known to differ slightly in virulence for guinea pigs differed in at least one epitope of the 120-kilodalton protein. Images

Nonvalue of sputum culture in the management of lower respiratory tract infections.

Abstract: Establishment of the microbiological etiology of bacterial pneumonia by sputum culture is confounded by both lack of recovery of fastidious pathogens and contamination of specimens with oropharyngeal flora. We reviewed the clinical records from 249 patients over a 3-month period for evidence of pneumonia. Gram staining and cultures were performed on 381 specimens isolated from this population of patients. Recovery of respiratory tract pathogens was accomplished with 354 specimens from 226 patients; 27 specimens yielded normal flora in culture but were smear positive. An additional 256 specimens submitted to our microbiology laboratory did not meet smear criteria for purulence nor did they yield respiratory tract pathogens in culture. A total of 637 specimens submitted to the microbiology laboratory were evaluated for sputum purulence by the criteria of Bartlett. Of the total 354 specimens which were positive in culture for a pathogen, 182 (52%) were submitted from 150 patients with no objective evidence of pneumonia. The majority of specimens obtained from patients without pneumonia were nonpurulent. However, 71 of 182 culture-positive specimens obtained from 50 patients without pneumonia were purulent. Approximately half of these patients (31 of 50) had other pulmonary or upper respiratory tract pathology which could account for the sputum purulence. Among the 172 culture-positive specimens from 76 patients with pneumonia, only 100 (58%) were acceptable by smear criteria. An additional 23 patients provided expectorated purulent sputum from which no respiratory tract pathogen could be isolated. Of these 23, 7 had pneumonia. We conclude that sputum culture and Gram staining are neither specific nor sensitive as diagnostic tools. Objective criteria for purulence of Gram-stained specimens must be applied before their inoculation into culture media. Specimens should be sought only from patients with objective evidence of pneumonia.

Clinical features and therapeutic interventions in 17 cases of Bacillus bacteremia in an immunosuppressed patient population.

Abstract: We retrospectively examined episodes of Bacillus bacteremia at a hospital with a large proportion of immunosuppressed patients Seventeen episodes in 95 years met our case definition: two of two bottles of one blood culture or one of two bottles of two or more separately obtained blood cultures drawn on the same date During the same period, there were 59 additional episodes in which a single blood culture had only one of two bottles positive for Bacillus species Only 2 of 59 such episodes resulted in recurrent bacteremia (3%), as compared with 5 of 17 episodes meeting our case definition (29%) (P = 0004) In four of five episodes complicated by recurrent bacteremia and in which appropriate antibiotics were used, a Hickman-Broviac catheter was in place and was not removed We suggest that our case definition permits the differentiation of infection from contamination based on outcome and that patients with Bacillus bacteremia have chronic venous catheters removed as well as receive antibiotic treatment

Comparison of nasopharyngeal aspirate and nasopharyngeal swab specimens for respiratory syncytial virus diagnosis by cell culture, indirect immunofluorescence assay, and enzyme-linked immunosorbent assay.

Abstract: Paired nasopharyngeal aspirate (NPA) and nasopharyngeal swab (NPS) specimens obtained from each of 32 hospitalized infants with X-ray-confirmed pneumonia (91%) or bronchiolitis were tested for respiratory syncytial virus (RSV) infection by virus culture, the indirect immunofluorescent-antibody (IFA) technique, enzyme-linked immunosorbent assay (ELISA; Ortho Diagnostic Systems, Inc.), and spot hybridization with a human genomic probe to quantitate cellular DNA. RSV was isolated in cell cultures from 72% (23 of 32) of patients by using NPA specimens compared with 47% (15 of 32) by using NPS specimens. With tissue culture positivity as the reference test, the sensitivities of the ELISA on NPA and NPS specimens were found to be 69% (16 of 23) and 61% (14 of 23), respectively, with a specificity and a positive predictive value from both sites of 100%. The sensitivities of the IFA technique compared with the cell culture on NPA and NPS specimens were 61% (14 of 23) and 52% (12 of 23) with specificities of 89 and 78% and positive predictive values of 96 and 92%, respectively. Despite the recovery of significantly more cells (as shown by detection of more cellular DNA by using NPA specimens), virus was detected by the IFA technique or ELISA at similar frequencies in paired specimens. However, virus was recovered more often from NPA than NPS specimens by cell culture, and ELISA optical density readings and the number of RSV-positive fluorescing cells were greater for NPA specimens. NPA specimen collection was less traumatic for the patient, was an easier procedure for the physician to perform, and provided a superior laboratory specimen for RSV diagnosis than the NPS technique.