bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogenesis of herpes simplex labialis: excretion of virus in the oral cavity
Authors: Spruance S L
Published: January 2020

Excretion of herpes simplex virus (HSV) in the oral cavity was studied in eight human subjects with a history of herpes labialis. Serial intraoral specimens were obtained by gargling broth and examined for virus by centrifugal inoculation of primary human amnion cells. Forty-seven of 637 specimens (7.4%) contained HSV. The majority of isolates (62%) were found in clusters, and the rate of excretion was significantly increased during the common cold (21%) and after oral trauma (17%) (P = 0.001 and 0.04, respectively). Oral HSV excretion often occurred in parallel with episodes of herpes labialis but could not be attributed to viral contamination from a labial lesion. Each patient excreted only one strain of HSV type 1 as determined by restriction endonuclease analysis with KpnI and BamHI. Unexpectedly, prodromal symptoms of herpes labialis were commonly not followed by development of a lesion (false prodrome). False prodromes were associated with a high rate of oral HSV excretion (60%). Intraoral ulcers on the gingivae and hard palate were frequently associated with oral HSV excretion (31%) and are the most likely source of HSV in the oral cavity.
Measurement of levels of aminoglycosides and vancomycin in serum in the presence of new beta-lactam antibiotics
Authors: Calderwood S B , Tung C C , Jackson E
Published: January 2020

We evaluated the effect of new beta-lactam antibiotics (azlocillin, mezlocillin, piperacillin, cefotaxime, moxalactam, and cefoperazone) on assays for aminoglycosides and vancomycin. These antibiotics produced no interference in an immunoassay for gentamicin and tobramycin. The new penicillins produced no interference in a bioassay of amikacin and vancomycin with penicillinase incorporated into the assay agar. Bioassay in the presence of the cephalosporins required predigestion with cephalosporinase. We describe a method for accurate bioassay in the presence of the available cephalosporins, with the exception of moxalactam.
Age-related antibodies to rotavirus in New Zealand rabbits
Authors: DiGiacomo R F , Thouless M E
Published: January 2020

The prevalence of antibodies to rotavirus was determined in 199 New Zealand rabbits from a commercial rabbitry by an indirect immunofluorescent test. Virtually all rabbits less than 1 month old and greater than 2 months old had antibodies, whereas only 25% of rabbits 1 to 2 months old had antibodies.
Disk diffusion antimicrobial susceptibility testing of members of the family Legionellaceae including erythromycin-resistant variants of Legionella micdadei
Authors: Dowling J N , McDevitt D A , Pasculle A W
Published: January 2020

Disk diffusion antimicrobial susceptibility testing of members of the family Legionellaceae was accomplished on buffered charcoal yeast extract agar by allowing the bacteria to grow for 6 h before placement of the disks, followed by an additional 42-h incubation period before the inhibitory zones were measured. This system was standardized by comparing the zone sizes with the MICs for 20 antimicrobial agents of nine bacterial strains in five Legionella species and of 19 laboratory-derived, erythromycin-resistant variants of Legionella micdadei. A high, linear correlation between zone size and MIC was found for erythromycin, trimethoprim, penicillin, ampicillin, carbenicillin, cephalothin, cefamandole, cefoxitin, moxalactam, chloramphenicol, vancomycin, and clindamycin. Disk susceptibility testing could be employed to screen Legionella isolates for resistance to any of these antimicrobial agents, of which only erythromycin is known to be efficacious in the treatment of legionellosis. With selected antibiotics, disk susceptibility patterns also appeared to accurately identify to the species level the legionellae. The range of the MICs of the legionellae for rifampin and the aminoglycosides was too small to determine whether the correlation of zone size with MIC was linear. However, laboratory-derived, high-level rifampin-resistant variants of L. micdadei demonstrated no inhibition zone around the rifampin disk, indicating that disk susceptibility testing would likely identify a rifampin-resistant clinical isolate. Of the antimicrobial agents tested, the only agents for which disk susceptibility testing was definitely not possible on buffered charcoal yeast extract agar were oxacillin, the tetracyclines, and the sulfonamides.
Agglutinating antibody titers to members of the family Legionellaceae in cystic fibrosis patients as a result of cross-reacting antibodies to Pseudomonas aeruginosa
Authors: Collins M T , McDonald J , Høiby N , Aalund O
Published: January 2020

The objective of this study was to evaluate the prevalence and significance of antibody titers to organisms in the family Legionellaceae in 128 serum samples collected from cystic fibrosis patients at routine examinations. Antibody titers were determined for 10 antigenic types of Legionellaceae; Legionella pneumophila serogroups 1 to 6, Fluoribacter (Legionella) bozemanae, Fluoribacter (Legionella) dumoffii, Fluoribacter (Legionella) gormanii, and Tatlockia (Legionella) micdadei. The method of antibody titer determination was the microagglutination test. Elevated titers (greater than or equal to 1:64) to one or more antigens were found in 41.3% of cystic fibrosis patients but in only 9.7% of 103 normal control subjects (P less than 0.01). Titers to 8 of the 10 antigens were directly correlated with the number of Pseudomonas aeruginosa precipitating antibodies in patient sera, as determined by crossed immunoelectrophoresis (correlation coefficients, greater than or equal to 0.74). Cross-reactions between P. aeruginosa and L. pneumophila were substantiated by crossed immunoelectrophoresis of hyperimmune rabbit serum as well as patient sera against P. aeruginosa and Legionellaceae antigens. Monospecific antibody to the "common antigen" of P. aeruginosa was used to demonstrate the presence of this antigen in L. pneumophila. The presence of cross-reacting antibodies in cystic fibrosis patients chronically infected with P. aeruginosa emphasizes the need for cautious interpretation of antibody titers to members of the family Legionellaceae.
Isoprenoid quinone content and cellular fatty acid composition of Campylobacter species
Authors: Moss C W , Kai A , Lambert M A , Patton C
Published: January 2020

A total of 36 strains of Campylobacter species were examined for isoprenoid quinones and cellular fatty acids. The isoprenoid quinone content was determined by reverse-phase high-pressure liquid chromatography, and the fatty acids were determined by capillary gas-liquid chromatography. All Campylobacter species contained menaquinone-6 (2-methyl-3-farnesyl-farnesyl-1,4-naphthoquinone) and a methyl-substituted menaquinone-6 (2,[5 or 8]-dimethyl-3-farnesyl-farnesyl-1,4-napthoquinone) as the major isoprenoid quinones. The latter menaquinone has not been reported in other bacteria and may prove to be a useful chemical marker of Campylobacter species. Campylobacter jejuni and most Campylobacter coli were distinguished from other campylobacteria by the presence of a C19 cyclopropane acid, and Campylobacter sputorum subsp. mucosalis differed from other species by the presence of lauric acid.
Rapid, inexpensive method for specific detection of microbial beta-lactamases by detection of fluorescent end products
Authors: Chen K C , Knapp J S , Holmes K K
Published: January 2020

A rapid method was developed for specific detection of microbial beta-lactamases which uses ampicillin and cephalexin as substrates. The end products (open beta-lactam ring forms) generated after separately incubating either substrate with beta-lactamase-producing organisms initially were separated from the unhydrolyzed substrates by high-voltage electrophoresis at pH 2.1. The end products of both antibiotics were highly fluorescent and could be analyzed visually and semiquantitatively under a long-wave UV lamp. Application of 5 microliters of the same incubation mixture onto filter paper without subsequent electrophoretic separation also resulted in development of fluorescence after brief heating at 120 degrees C for 5 min. This spot test differentiates penicillinase activity from cephalosporinase activity and distinguishes between beta-lactamase and acylase activities, since the end products of acylase [the common side chain, D(-)-alpha-aminophenylacetic acid, and the intact beta-lactam nuclei, 6-aminopenicillanic acid and 7-aminodeacetoxycephalosporanic acid] are not fluorescent. This method was relatively rapid, inexpensive, and more sensitive than the chromogenic cephalosporin (nitrocefin) method when 21 strains of 7 gram-positive species and 77 strains of 29 gram-negative species of bacteria were tested.
Effects of manganese on the growth and morphology of Ureaplasma urealyticum
Authors: Robertson J A , Chen M H
Published: January 2020

All of the 14 serotype standard strains of Ureaplasma urealyticum were inhibited to varying degrees by manganese. A 1 mM concentration of this cation either stopped growth or reduced the rate of growth in liquid medium. The presence of manganese also altered colonial morphology and cellular ultrastructure. Inhibition was dose dependent and strain specific. This differential response allowed the serotype strains to be divided into two broad biotypes. For the first biotype (serotypes 1, 3, 6, and 14), inhibition of growth in broth was temporary. For the second biotype (serotypes 2, 4, 5, 7, 8, 9, 10, 11, and 12), inhibition was permanent. Serotype 13 gave an intermediate response and was not classified. The effect of manganese could be at least partially blocked by magnesium but not by calcium, cobalt, copper, iron, potassium, sodium, or zinc. The concentration of magnesium yielding the maximum blocking effect was directly related to manganese sensitivity. Wild-type isolates of ureaplasma and Mycoplasma hominis also showed a differential response to manganese. Laboratory-adapted strains representing species of the genus Mycoplasma (M. hominis, M. fermentans, and M. pneumoniae) were inhibited by 5 but not by 1 mM manganese. The latter concentration inhibited the growth of Acholeplasma laidlawii and Staphylococcus aureus, and 5 mM manganese had no effect on Escherichia coli.
Isolation of Serratia ficaria from human clinical specimens
Authors: Brouillard J A , Hansen W , Compere A
Published: January 2020

Serratia ficaria was isolated from sputum in a 62-year-old man and from tracheal aspiration fluid in a 62-year-old woman with acute respiratory distress. These strains are the second and third isolations of S. ficaria from human sources. The case reports are presented, together with the laboratory findings and the biochemical activities of the isolates.
Exoantigen test for differentiation of Exophiala jeanselmei and Wangiella dermatitidis isolates from other dematiaceous fungi
Authors: Espinel-Ingroff A , Shadomy S , Kerkering T M , Shadomy H J
Published: January 2020

Concentrated (25X) exoantigens of 105 isolates of pathogenic and saprophytic dematiaceous fungi and 3 isolates of Sporothrix schenckii were analyzed by the microimmunodiffusion method. The reagents used were nonadsorbed and adsorbed sera produced in New Zealand rabbits. One set of rabbits was immunized with soluble antigens of a 1-month-old culture of Exophiala jeanselmei (ATCC 34123), and the other set was immunized with soluble antigens from a culture of Wangiella dermatitidis (ATCC 28869). The reference antigens were 25X-concentrated exoantigens of the above cultures. This exoantigen test permitted the differentiation of E. jeanselmei and W. dermatitidis from one another as well as from other Exophiala species, Fonsecaea species, Phialophora species, Cladosporium species, Rhinocladiella species, and Sporothrix schenckii by presence or absence of lines of identity or of partial identity, or lines of nonidentity. Using adsorbed serum eliminated the problems with cross-reactivity seen with nonadsorbed serum. Thus, with an adsorbed serum as the reagent, it was possible to presumptively differentiate E. jeanselmei and W. dermatitidis from one another and from other dematiaceous fungi.
Revised pyocin typing method for Pseudomonas aeruginosa
Authors: Fyfe J A , Harris G , Govan J R
Published: January 2020

In the Gillies and Govan method of pyocin typing for Pseudomonas aeruginosa a cross-streaking technique was used, and 105 main types and 25 subtypes were identified by the patterns of inhibition observed on 13 indicator strains. Disadvantages of the technique included the need to remove test strain growth before application of the indicator strains, the 48-h period needed to obtain a result, and the inability to reliably type mucoid P. aeruginosa. Recent studies have enabled us to overcome these disadvantages and significantly improve the speed and application of pyocin typing. Our revised technique utilizes the same 13 indicator strains which are already used internationally. Test strains were rapidly applied to the surface of agar plates with a multiple inoculator. After incubation for 6 h and exposure to chloroform, the indicator strains were applied in agar overlays without prior removal of the test strain growth. After 18 h of incubation, the pyocin type was recognized by inhibition of particular indicator strains. Additionally, the activity of particulate (R and F) and nonparticulate (S) pyocins could be distinguished on the basis of inhibition zone size, which thus allowed further discrimination. The revised technique allows typing within 24 h, increases the number of identifiable types, and can be used to type mucoid strains.
Detection of encapsulation in Staphylococcus aureus by use of antiserum agar
Authors: West T E , Apicella M A
Published: January 2020

We examined an antiserum agar method to study its reliability in screening Staphylococcus aureus strains for capsule production. The encapsulated S. aureus Smith diffuse strain was compared with its nonencapsulated variant, Smith compact, in CCY medium containing 0.5% NaCl and 5.0% Smith diffuse rabbit antiserum. A halo was visible surrounding colonies of the Smith diffuse strain but not the Smith compact strain. On this same medium, the protein A-producing Cowan I strain possessed a halo that was visible on photographs. Single high-salt medium is known to inhibit protein A production, halo formation by the strains was also compared in 7.5% NaCl medium. The halo surrounding the Cowan I strain was not present when the salt content of the medium was increased. In contrast, the halo surrounding the Smith diffuse strain persisted in the 7.5% NaCl medium. By use of this medium, the antiserum agar technique may be valuable for the identification of encapsulated staphylococci without appreciable interference from protein A.
Lipopolysaccharide subtypes of Haemophilus influenzae type b from an outbreak of invasive disease
Authors: Inzana T J , Pichichero M E
Published: January 2020

Thirty isolates of Haemophilus influenzae type b were obtained during an outbreak of invasive H. influenzae type b disease and were classified by the electrophoretic profile of their lipopolysaccharide (LPS). The LPS was extracted by a rapid micromethod and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. The isolates could be divided into 1 of 14 subtypes based on the profile of two to four bands. No subtype was predominant. However, all isolates obtained from duplicate sites of the same individual were of the same subtype. Isolates obtained from two patients (6 weeks apart) who attended the same day-care center differed in LPS subtype but were identical in their major outer membrane protein electrophoretic profile. Nasopharyngeal cultures were obtained from healthy children, their immediate families, and employees of the day-care center. Of 13 H. influenzae isolates examined from these contacts, only 1 was type b, which was obtained from a day-care worker and had the same LPS subtype and major outer membrane protein electrophoretic profile as one of the disease isolates. The remaining nasopharyngeal isolates were untypable, and most, but not all, were different in LPS pattern. Thus, LPS subtyping of H. influenzae type b may be useful in examining the predominance or transmission of a strain during an outbreak and may distinguish some strains not differentiated by outer membrane protein pattern.
DNA characterization of the spirochete that causes Lyme disease
Authors: Schmid G P , Steigerwalt A G , Johnson S E , Barbour A G , Steere A C , Robinson I M , Brenner D J
Published: January 2020

Lyme disease, a tick-borne disease long recognized in Europe but only recently recognized in the United States, was shown in 1982-1983 to be caused by a spirochete, the Lyme disease spirochete. Whether one or more species of the spirochete exists is unknown, as is its taxonomic status. To answer these questions, we determined (i) the DNA base (guanidine-plus-cytosine) content for five strains; (ii) the DNA relatedness of 10 strains from Europe or the United States (isolated from ticks, humans, and a mouse) by DNA hybridization (hydroxyapatite assay at 50 and 65 degrees C); and (iii) the DNA relatedness to other pathogenic spirochetes. The guanine-plus-cytosine content of the Lyme disease spirochete strains was 27.5 to 29.0 mol%, most similar to those of Borrelia hermsii (30.6 mol%) and Treponema hyodysenteriae (25.6 mol%) among the other spirochetes tested. DNA hybridization studies with 32P-labeled DNA from Lyme disease spirochete strain TLO-005, a human blood isolate, revealed divergence (unpaired bases) within related nucleotide sequences of only 0.0 to 1.0% for all nine Lyme disease spirochete strains tested for relatedness to TLO-005. Relatedness values of seven strains to TLO-005 were 58 to 98% (mean, 71%) in 50 degrees C reactions and 50 to 93% (mean, 69%) in 65 degrees C reactions. Two other strains, from which very low yields of DNA were obtained, showed less relatedness (36 to 50 degrees C, 38 to 47% at 65 degrees C). These were nonetheless considered to belong to the same species because of the low amount of divergence in the sequences related to TLO-005 and the absence of decreased relatedness in reactions done at 65 degrees Celsius compared with those done at 50 degrees Celsius. DNA from strain TLO-005 showed relatedness of 1% to DNAs of two leptospires and 16% relatedness to DNA from T. hyodysenteriae. B. hermsii DNA was 30 to 40% related to three Lyme disease spirochete strains in 50 degrees Celsius reactions. Divergence in these reactions was 16.5 to 18.5%, and relatedness in 65 degree Celsius reactions was 8 to 10%. On the bases of phenotypic similarity, guanine-plus-cytosine content, and DNA relatedness to B. hermsii, we conclude the Lyme disease spirochete is single previously undescribed species which belongs in the genus Borrelia.
Specific enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus
Authors: Tabatabai L B , Deyoe B L
Published: January 2020

Six soluble antigens prepared from Brucella abortus were compared with a salt-extractable protein (CSP) antigen in an enzyme-linked immunosorbent assay for the detection of antibody to B. abortus in cattle sera. Of seven preparations tested, antigens from B. abortus soluble antigen (prepared from an autoclaved cell suspension) and CSP were stable on frozen storage. Enzyme-linked immunosorbent assay with CSP antigen under optimal conditions was from 100- to 700-fold more sensitive than the standard agglutination, card, Rivanol precipitation-plate agglutination, and the complement fixation tests in detecting immunoglobulin G antibody. From a practical point of view, however, using the most stringent criteria for determining an "upper negative" value, the enzyme-linked immunosorbent assay with CSP was at least 12-fold more sensitive than the standard agglutination test and any of the other serological tests. Furthermore, the enzyme-linked immunosorbent assay with CSP was specific for antibody to B. abortus.
Enzyme-linked immunosorbent assay typing of California serogroup viruses isolated in Canada
Authors: Artsob H , Spence L P , Th'ng C
Published: January 2020

A procedure was developed to type California serogroup viruses by an antibody-capture, enzyme-linked immunosorbent assay. Seven California serogroup members from North America were distinguished, including snowshoe hare, La Crosse, California encephalitis, San Angelo, Jamestown Canyon, Keystone, and trivittatus. Extensive cross-reactions were observed between Jamestown Canyon and the closely related South River strain. The enzyme-linked immunosorbent assay method was successfully applied to the typing of 77 California serogroup viruses isolated in Canada, including 61 snowshoe hare, 13 Jamestown Canyon, and 3 trivittatus topotypes.
Usefulness of plasmid profiles for differentiation of Shigella isolates in Bangladesh
Authors: Tacket C O , Shahid N , Huq M I , Alim A R , Cohen M L
Published: January 2020

We studied the plasmid profiles of 136 Shigella isolates in Bangladesh to determine whether plasmid profiles could be used for differentiation of strains for epidemiological studies. Many different plasmid patterns were observed within each species, indicating that many genetically different strains of Shigella are responsible for illness in Bangladesh.
Human leukocytic pyrogen test for detection of pyrogenic material in growth hormone produced by recombinant Escherichia coli
Authors: Dinarello C A , O'Connor J V , LoPreste G , Swift R L
Published: January 2020

Human growth hormone is biosynthetically produced in recombinant strains of Escherichia coli as methionyl human growth hormone (met-hGH). When purified from the bacterial culture, met-hGH is biologically active in established assays for growth hormone. Therefore, a phase I trial of met-hGH was carried out in healthy human adults; during the first trial, however, signs, symptoms, and clinical laboratory tests characteristic of an acute-phase response to pyrogenic agents was observed. Prior testing of the met-hGH preparation used in the phase I trial did not reveal evidence of toxicity, and the U.S. Pharmacopeial Convention rabbit pyrogen test, as well as the Limulus amoebocyte lysate (LAL) test, had not detected significant levels of exogenous pyrogens or endotoxin. In addition, standard inhibition studies with added endotoxin showed no inhibition by the LAL test. When this preparation of met-hGH was incubated with human blood mononuclear cells, leukocytic pyrogen (LP) was released into the supernatant medium, suggesting that the preparation contained pyrogenic material. Various lots of met-hGH based on different purification and formulating methods were tested by the human LP assay for contaminating pyrogens. The results of these tests aided in the identification of procedures for met-hGH preparations which did not induce LP in vitro. Thus, subsequent lots of met-hGH which had passed the LP test were used in repeat clinical studies, and no inflammatory or pyrogenic reactions were observed. When the LP test was used, experiments revealed that the original lot of met-hGH was contaminated with endotoxin which had not been detected in the LAL or rabbit pyrogen tests. Lyophilization in glycine-phosphate buffer had resulted in a 10- to 20-fold reduction of endotoxin reactivity in the LAL test and the U.S. Pharmacopeial Convention rabbit pyrogen test. These data provide a probable explanation for the negative result from the LAL and rabbit pyrogen test in the initial lot of met-hGH which induced acute-phase reactions. In addition, these studies demonstrate that the release of LP from human cells is a reliable indicator of the presence of materials that are pyrogenic for humans.
Evaluation of new blood culture processing systems
Authors: Moody J A , Fasching C E , Shanholtzer C J , Gerding D N , Peterson L R
Published: January 2020

The Antimicrobial Removal Device (ARD; Marion Scientific) was evaluated in vitro with simulated blood culture samples in fresh blood and clinically with samples from potentially septic patients to test its ability to remove antimicrobial agents and recover bacteria from blood culture specimens containing these drugs. In simulated specimens, the ARD was evaluated for adverse affects on microorganisms as well as compared with lysis-centrifugation (Isolator; Du Pont Co.), biphasic brain heart infusion bottles, and tryptic soy broth bottles for antimicrobial inactivation and organism recovery. There was no adverse effect of the ARD on organisms during a 4-h test period. The ARD was the only system to actually inactivate antimicrobial agents and removed greater than 99.2% of all antimicrobial agents tested from spiked and clinical specimens. Overall, with simulated blood culture specimens, the ARD recovered 90% of bacteria spiked into fresh blood containing antimicrobial agents, Isolator recovered 73%, biphasic brain heart infusion bottles recovered 31%, and tryptic soy broth bottles recovered 24%. In the clinical study, 43 of 86 clinically significant isolates were recovered only by ARD-assisted processing, 6 were recovered only by conventional processing, and 37 were recovered by both methods (the advantage of ARD processing over conventional processing in the clinical study was significant at P less than 0.001). Both clinical and simulated specimens demonstrated the ARD-associated blood culture processing to be the most efficient method for the isolation of microorganisms from specimens containing antimicrobial agents.
Enzyme-linked immunosorbent assay for detection of Clostridium botulinum type A and type B toxins in stool samples of infants with botulism
Authors: Dezfulian M , Hatheway C L , Yolken R H , Bartlett J G
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for Clostridium botulinum type A and type B toxins was assessed for diagnostic accuracy in cases of infant botulism. This test was positive in all 22 cases confirmed by the conventional tests, which included the mouse lethality assay and stool culture. Stool specimens from five cases were positive by culture, but the mouse lethality bioassay was either negative or toxicity was judged nonspecific since it could not be neutralized by specific antitoxin. The positive ELISA results in these specimens suggested that this assay may be more reliable, in some cases, than the mouse bioassay. Of the 21 fecal specimens from suspected foodborne cases, 2 contained botulinal toxin demonstrable by the mouse assay and the ELISA. With regard to specificity, 35 fecal specimens from infants and 19 from suspected foodborne cases which were negative in the bioassay for botulinal toxins A and B were also negative in the ELISA. Only two fecal specimens with negative bioassay gave positive ELISA readings, providing a specificity rate of 96%. These results suggest that the ELISA may serve as a useful screening test to detect C. botulinum toxin in clinical specimens.
Evaluation of the Staph-Ident and STAPHase systems for identification of staphylococci from bovine intramammary infections
Authors: Watts J L , Pankey J W , Nickerson S C
Published: January 2020

The Staph-Ident and STAPHase systems (Analytab Products, Plainview, N.Y.) were compared with conventional methods for identification of staphylococci isolated from bovine intramammary infections. Adjunct testing by colony morphology, pigmentation, and biochemical tests was conducted to resolve discrepant identifications. The initial accuracies of the conventional scheme and Staph-Ident were 92.1 and 89.2%, respectively. Staphylococcus hyicus subsp. chromogenes could not be identified by means of the Staph-Ident test, but the addition of pigment production as a key character permitted identification of most strains. The final accuracy of the Staph-Ident was 94.3%. The STAPHase system was as accurate as the conventional tube coagulase method. The Staph-Ident and STAPHase systems are acceptable alternatives to conventional methods for identification of staphylococcal species isolated from bovine intramammary infections.
Anomalous antibody responses in viral infection: specific stimulation or polyclonal activation?
Authors: Cremer N E , Devlin V L , Riggs J L , Hagens S J
Published: January 2020

Eighteen paired serum samples submitted for serodiagnosis of current infection showed anomalous antibody results by complement fixation test when tested with a battery of agents (viruses, Mycoplasma pneumoniae, and chlamydia) selected for testing on the basis of the symptoms of the patient. Seventeen serum pairs showed a fourfold or greater rise in titer of antibody to two agents in the battery, and one showed only a twofold rise in titer of antibody to the identified causative agent but an eightfold rise in titer of antibody to a heterologous agent. The 18 serum pairs were tested for IgM antibody to the two involved agents to determine whether IgM antibody tests would better distinguish the probable cause of the current infection. The serum pairs were separated into three groups based on their IgM responses. Group I consisted of six serum pairs with IgM antibody to both agents, four pairs of which showed a fourfold or greater rise in titer of IgM antibody to both agents, and two of which showed a rise in titer of IgM antibody to only one of the two agents. Group II consisted of 10 serum pairs with IgM antibody to one of the two agents, 7 pairs of which showed a fourfold or greater rise in titer of IgM antibody to the agent. Group III consisted of two serum pairs with no IgM antibody to either agent. Results show that determination of presence or absence of IgM antibody per se or demonstration of a fourfold or greater rise in specific IgM antibody titer does not always help in distinguishing the causative agent in current infections.
Rapid antimicrobial susceptibility testing of isolates from blood cultures by direct inoculation and early reading of disk diffusion tests
Authors: Coyle M B , McGonagle L A , Plorde J J , Clausen C R , Schoenknecht F D
Published: January 2020

Disk diffusion tests, inoculated directly from positive blood cultures, were evaluated for accuracy of reading zone diameters after 4- and 6-h and overnight incubation. In comparisons with results from standard disk diffusion tests, the 4-h results were in agreement for 83% of tests with gram-positive organisms and 64% of tests with gram-negative organisms. When minor discrepancies were ignored, the 4-h readings were in agreement for 98% of the tests with gram-positive organisms and 95% of the tests with gram-negative organisms. After 6 h of incubation, 91% of the tests with gram-positive organisms and 86% of the tests with gram-negative organisms agreed with standard results. The agreement was 99% for tests with both gram-positive and gram-negative organisms when minor discrepancies were excluded. Very major discrepancies occurred in two tests (0.1%) with gram-positive organisms and were not observed in tests with gram-negative organisms. The frequencies of major discrepancies were 3.5% after 4 h, 0.6% after 6 h, and 0.7% after overnight incubation. Ampicillin and cephalothin tests with Escherichia coli and Klebsiella spp. accounted for 81% of the major discrepancies in tests with gram-negative organisms. Oxacillin tests accounted for more than half of the major discrepancies in tests with staphylococci. The results of this study, which did not include the newer antibiotics, indicate that direct susceptibility tests from blood cultures read after 6 h of incubation are more reliable than 4-h results and produce less than 1% major errors in comparisons with standard susceptibility tests.
Scanning electron microscopy of bacteria adherent to intravascular catheters
Authors: Franson T R , Sheth N K , Rose H D , Sohnle P G
Published: January 2020

Scanning electron microscopy was used to assess the morphological features of coagulase-negative staphylococci adherent to polyvinylchloride intravascular catheter specimens. Clinical specimens were obtained by using patient catheters from which coagulase-negative staphylococci (greater than or equal to 15 colonies per catheter) grew on semiquantitative blood agar roll cultures. In vitro specimens were prepared by a previously published technique in which sterile polyvinylchloride catheters were immersed in 10(6) CFU of coagulase-negative staphylococci per ml suspended in phosphate-buffered saline. Unused sterile polyvinylchloride catheters were also examined. Scanning electron microscopy of unused sterile polyvinylchloride catheters demonstrated multiple linear surface irregularities. Scanning electron microscopy of infected patient catheters showed a diffuse amorphous material covering the entire surface and the presence of bacteria which appeared anchored to that surface by several different means. These included a slime layer, "foot" processes, and lodgement in surface irregularities. Scanning electron microscopy of in vitro specimens demonstrated no background surface coating, but it did show attachment of cocci to the surface by the same mechanisms as described for clinical specimens. These observations of similar means of attachment in clinical and in vitro specimens suggest that intrinsic catheter surface properties, bacterial surface features, and perhaps coating with host substances may all play a role in bacterial attachment to intravascular catheters. More sophisticated analysis of these interactions may clarify mechanisms of pathogenesis.
Frequency-pulsed electron capture gas-liquid chromatographic analysis of metabolites produced by Clostridium difficile in broth enriched with amino acids
Authors: Brooks J B , Nunez-Montiel O L , Wycoff B J , Moss C W
Published: January 2020

Clostridium difficile strain CDC A-567 was cultured in Trypticase (BBL Microbiology Systems)-yeast-salt broth supplemented with 0.2% L-leucine, L-norleucine, L-isoleucine, L-tyrosine, or L-tryptophan. Four extractions were done on the spent medium, three at pH 2 and one at pH 10, using CHCL3 or ether. Derivatizations were done with trichloroethanol, heptafluorobutyric anhydride, and heptafluorobutyric anhydride-ethanol. All samples were analyzed with frequency-pulsed electron capture gas-liquid chromatography. A dedicated computer was used to assist in data analysis. C. difficile produced both short-chain and aromatic acids in Trypticase-yeast-salt broth; hydroxy acids were also detected. p-Cresol, indoleacetic acid, 4-methylthio-2-hydroxybutyric acid, and some unidentified alcohols were observed. The basic chloroform extraction contained cadaverine and putrescine. Leucine, norleucine, and isoleucine influenced the production of C5 and C6 acids and alcohols. L-Tyrosine underwent successive degradation to produce p-cresol and aromatic acids as final products. Tryptophan increased the production of indoleacetic, indolepropionic, and indolebutyric acids. Isocaproic acid was produced in relatively high concentrations regardless of medium substitution. The consistent production of iC6 under various substrate conditions indicates that the production of this compound might be consistent enough in vitro to form the basis of a rapid test for detection of C. difficile in stool specimens by frequency-pulsed electron capture gas-liquid chromatography.
Evaluation of a rapid latex agglutination test for identification of group D streptococci
Authors: Vanzo S J , Washington J A
Published: January 2020

The SeroSTAT latex agglutination test (Scott Laboratories, Inc., Fiskesville, R.I.) for identification of group D streptococci was compared with bile-esculin agar and Lancefield grouping by using 110 clinical isolates of group D streptococci (S. faecalis, 74; S. bovis, 24; S. durans, 3; S. faecium, 9) and 65 viridans streptococci (S. anginosus-constellatus, 2; Streptococcus MG, 15; S. sanguis II, 14; S. mitis, 7; S. mutans, 5; S. salivarius, 8; S. sanguis I, 8; S. acidominimus, 1; S. morbillorum, 2; S. pneumoniae, 3). All strains of group D streptococci were bile-esculin positive. SeroSTAT reactions were falsely positive with 2 strains of S. sanguisII and 1 strain of Streptococcus MG (4.6% of all viridans streptococci tested) and falsely negative with 10 S. bovis, 8 S. faecalis, 1 S. durans, and 6 S. faecium strains (22.7% of all group D streptococci tested). When considered with colonial morphology and hemolytic reaction, SeroSTAT is a rapid (60-s) and useful test for recognition of group D streptococci.
Efficacy of an enzyme immunoassay with uncentrifuged first-voided urine for detection of gonorrhea in males
Authors: Rudrik J T , Waller J M , Britt E M
Published: January 2020

An enzyme immunoassay (Gonozyme; Abbott Laboratories, North Chicago, Ill.) for detection of Neisseria gonorrhoeae antigens was used to screen 184 urethral or uncentrifuged first-voided urine or both specimens from males and 78 cervical specimens. When compared with culture, the sensitivity and specificity of Gonozyme for cervical and urethral specimens were comparable to those in published reports. The sensitivity and specificity for urine specimens were 91.6 and 97.9%, respectively.
Differential detection of cholera enterotoxin and Escherichia coli heat-labile enterotoxin by enzyme-linked immunosorbent assays with antibodies specific to the two toxins
Authors: Honda T , Sato M , Miwatani T
Published: January 2020

Enzyme-linked immunosorbent assays (ELISAs) with antibodies specific to either cholera enterotoxin (CT) of Vibrio cholerae or heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli were developed to detect LT and CT, respectively. With these ELISA systems, LT and CT could be detected only with the respective specific antibody. Both antibody ELISA and ganglioside ELISA were used for differential detection of LT and CT, but the former method seemed to be more specific. By this ELISA, as little as 0.1 ng of purified LT or CT could be detected per ml. The type of toxins in fluids in intestinal loops of experimental animals challenged with living cells of either V. cholerae or LT-producing E. coli was identified correctly by this ELISA. These results suggest that the ELISA systems reported here could be used to detect and differentiate CT and LT in unknown samples; it could also be used for assaying toxins in stool specimens for the diagnosis of diarrhea due to V. cholerae or LT-producing E. coli directly, without or before bacterial isolation.
Comparison of radioimmunoassay and enzyme-linked immunosorbent assay in measurement of antibodies to Neisseria meningitidis group A capsular polysaccharide
Authors: Beuvery E C , Kayhty M H , Leussink A B , Kanhai V
Published: January 2020

Antibodies to meningococcal group A polysaccharide were determined by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) in serum samples from 16 adults vaccinated with bivalent meningococcal group A and C polysaccharide vaccine. The specific antibody levels in the serum samples were expressed as micrograms of antibody protein per milliliter of serum. For RIA the polysaccharide was radiolabeled extrinsically with 125I. Both native polysaccharide and polysaccharide labeled with 127I were used in ELISA. Because these antigens gave similar results, it can be concluded that the introduction of tyramine and iodine by the labeling procedure did not alter the antigenic activity of the polysaccharide. The reproducibility of RIA was clearly better than that of ELISA. The antibody levels detected by the methods were equal, which means that ELISA can be used satisfactorily to measure antibodies to meningococcal group A polysaccharide quantitatively. Some discrepant results were found due to an underestimation of immunoglobulin M antibodies in ELISA. This was shown by a correlation test in which a weakly significant negative correlation was found between the immunoglobulin M antibody level/immunoglobulin G antibody level ratio and the RIA antibody level/ELISA antibody level ratio.
Diagnostic probability matrix for identification of slowly growing mycobacteria in clinical laboratories
Authors: Wayne L G , Krichevsky M I , Portyrata D , Jackson C K
Published: January 2020

A probability matrix is presented for identification of slowly growing mycobacteria that are likely to be encountered in clinical laboratories. The matrix includes 23 features that are useful for identifying members of 14 species or species complexes. The computer program identifies strains as a function of the ID (identification) score, which measures the discrimination among possible alternative identifications, and the R (ratio) score, which measures the degree of fit to the most likely taxa. It is not necessary to employ all 23 tests when initiating an identification; the program will suggest additional tests to perform when a partial data set fails to yield a definitive identification. Two independent sets of cultures comprising a total of 1,212 strains were used to test the matrix. Correct diagnoses were based on clustering behavior in numerical taxonomic analysis with larger numbers of features. The probable efficiencies with the two sets were 94.2 and 83.4%, respectively, and the accuracy of the definitive identifications for both sets exceeded 95%. A discussion is presented of situations when it may be appropriate to override an R score that has caused the rejection of an identification and to thereby enhance the efficiency.
Evaluation of the rapid NFT system for identification of gram-negative, nonfermenting rods
Authors: Appelbaum P C , Leathers D J
Published: January 2020

This study evaluated the ability of the Rapid NFT system (API System SA, Montalieu-Vercieu, France) to accurately identify 262 clinically isolated, gram-negative, nonfermentative rods without additional tests. Identifications were classified as correct; low discrimination, with a spectrum of two or more possibilities (additional tests necessary for accurate identification); and incorrect. Correct identification rates were analyzed in two categories: (i) correct to species or biotype for all organism groups except Alcaligenes faecalis-odorans, Moraxella, Pseudomonas testosteroni-alcaligenes-pseudoalcaligenes, and Acinetobacter calcoaceticus biotype haemolyticus-alcaligenes (in this category, the latter four genus-biotype group identifications were taken as correct) and (ii) correct to species or biotype in all cases, including the above four groups. In category i, 87.4% of the strains were correctly identified, with 4.2% low discrimination and 8.4% incorrect. When the criteria of category ii were used, 71.8% of the strains were correctly identified, with 19.9% low discrimination. The Rapid NFT system provided excellent species identification of Pseudomonas and Flavobacterium spp., Bordetella bronchiseptica, and Achromobacter xylosoxidans strains. Within Acinetobacter calcoaceticus, differentiation between biotypes anitratus and lwoffi was satisfactory, but the system did not differentiate between biotypes haemolyticus and alcaligenes. Species resolution within the genera Moraxella and Alcaligenes was incomplete. All Alcaligenes faecalis strains were misidentified and accounted for 50% of misidentifications with the Rapid NFT system; however, these results may reflect taxonomic differences rather than true misidentifications. The Rapid NFT system is easy to inoculate and interpret and represents a worthwhile advance in the identification of gram-negative, nonfermentative rods.
Comparison of strains of gram-negative, anaerobic, agar-corroding rods isolated from soft tissue infections in cats and dogs with type strains of Bacteroides gracilis, Wolinella recta, Wolinella succinogenes, and Campylobacter concisus
Authors: Love D N , Jones R F , Bailey M , Calverley A
Published: January 2020

A total of 64 strains of gram-negative, asaccharolytic, anaerobic, agar-corroding, rod-shaped bacteria from soft-tissue infections of cats and dogs were compared with other agar-corroding, anaerobic organisms isolated from human periodontal pockets (Wolinella recta ATCC 33238T), bovine rumens (Wolinella succinogenes ATCC 29543T), and gingival crevices of humans (Bacteroides gracilis ATCC 33236T and Bacteroides ureolyticus NCTC 10941T). Campylobacter concisus ATCC 33237T (from human gingival crevices) which did not corrode agar but which biochemically resembled organisms in this group was also included in this study. Although the type strains of W. recta, W. succinogenes, and B. gracilis resembled the animal strains phenotypically and in DNA base ratios, none had bacterial protein patterns (as indicated by isoelectric focusing) identical with the animal strains studied. The animal strains could be divided into motile and nonmotile groups. The motile animal strains were similar biochemically but could be divided into three groups by isoelectric focusing of bacterial proteins. Some had cell wall ultrastructural features identical with W. recta; others had the smooth walls of conventional gram-negative organisms. One group of nonmotile animal strains closely resembled B. gracilis phenotypically, and they had the cell wall ultrastructure of conventional gram-negative bacteria as described previously (4). The other nonmotile group had cell wall ultrastructure like that of W. recta.
Haemophilus influenzae biotype VIII
Authors: Sottnek F O , Albritton W L
Published: January 2020

Six Haemophilus influenzae strains could not be classified as biotypes I through VII. The strains were indole, urease, and ornithine decarboxylase negative. We propose that they be classified as biotype VIII, a previously unreported biotype.
Chocolate agar, a differential medium for gram-positive cocci
Authors: Gunn B A
Published: January 2020

Reactions incurred on chocolate agar by gram-positive cocci were correlated with species identity. Darkening and clearing of the medium was usually associated with the species Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus simulans, and Streptococcus faecalis. Yellowing of chocolate agar was associated with alpha-hemolytic species of Streptococcus. The study demonstrated that reactions occurring on chocolate agar are useful in identifying gram-positive cocci.
Comparison of microdilution and agar dilution procedures for testing antibiotic susceptibility of Neisseria gonorrhoeae
Authors: Shapiro M A , Heifetz C L , Sesnie J C
Published: January 2020

Studies were run in parallel to compare the broth microdilution method and the chocolate agar dilution method for testing antibiotic susceptibility of Neisseria gonorrhoeae. Six clinically relevant drugs were tested against 23 clinical isolates of N. gonorrhoeae, including several penicillinase-producing, as well as multiply resistant, strains. Results showed that the MIC obtained by the two methods were not significantly different. The microdilution method appears to be a more sensitive system for discriminating penicillinase activity. The microdilution system is a more expedient method for screening new antibacterial agents and is more readily adaptable to new automated equipment.
Sporadic occurrence of zoonotic swine influenza virus infections
Authors: Dacso C C , Couch R B , Six H R , Young J F , Quarles J M , Kasel J A
Published: January 2020

Two infections by swine influenza virus, antigenically similar to A/New Jersey/76 (H1N1) virus, were detected during community epidemics with other influenza viruses. The swinelike viruses were obtained during virological surveillance of acute respiratory illnesses, and the clinical symptoms of these two patients were similar to those caused by other respiratory viruses. Both patients reported contact with swine a few days before onset of illness, but in one case it was brief. Serological studies suggested that one patient may have transmitted the virus to his roommate, but spread into the community was not indicated.
Rapid presumptive recognition of diarrhea-associated adenoviruses
Authors: Brandt C D , Rodriguez W J , Kim H W , Arrobio J O , Jeffries B C , Parrott R H
Published: January 2020

The quantity of adenoviruses in a diarrhea stool provided a strong presumptive indication of the presence or absence of an adenovirus from subgenus F or G (proposed species 40 or 41). These adenoviruses were found in the stools of 91% of 56 acutely ill diarrhea patients with one or more than one adenovirus particle per min of direct electron microscopic viewing, as compared with 40% of 20 acutely ill diarrhea patients with less than one detected adenovirus particle per min of viewing.