bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of Fasciola hepatica tegument antigens
Authors: Hillyer G V
Published: January 2020

Fasciola hepatica tegument antigens were isolated from intact worms in the cold by using Nonidet P-40. Proof of the tegumental nature of the antigens was shown by the peroxidase-antiperoxidase immunocytochemical technique at the light microscope level. The potential of F. hepatica tegument antigens for the immunodiagnosis of rabbit and human fascioliasis was shown by Ouchterlony immunodiffusion, although cross-reactivity was evident in one of six serum samples from patients infected with Schistosoma mansoni. A genus-specific Fasciola antigen was found in F. hepatica tegument. Finally, F. hepatica tegument contained antigens which protected mice from challenge infection with S. mansoni.
Source of Pseudomonas in osteomyelitis of heels
Authors: Goldstein E J , Ahonkhai V I , Cristofaro R L , Pringle G F , Sierra M F
Published: January 2020

Pseudomonas aeruginosa is the most common cause of osteomyelitis following puncture wounds of the feet of children. The source of the initial inoculum is unknown. Only one strain of P. aeruginosa was cultured from paired samples of the heel or corresponding shoe's surface or both obtained from 100 children. Neither the skin of the heel nor the shoe appears to be the source of the initial inoculum.
Improvements in the passive immune hemolysis test for assaying enterotoxigenic Escherichia coli
Authors: de Castro A F , Serafim M B , Gomes J A , Gatti M S
Published: January 2020

The sensitivity of the passive immune hemolysis test for the detection of heat-labile enterotoxin from Escherichia coli was increased when the test was carried out with Veronal-buffered saline plus Ca2+ and Mg2+ as diluent, instead of phosphate-buffered saline. The performance of the test was further improved by using stationary cultures to which mitomycin C had been added at the end of the lag phase.
Cryptococcus neoformans: pitfalls in diagnosis through evaluation of gram-stained smears of purulent exudates
Authors: Bottone E J
Published: January 2020

The recognition of Cryptococcus neoformans in Gram-stained smears of purulent exudates may be hampered by the presence of the large gelatinous capsule which apparently prevents definitive staining of the yeast-like cells. In such stained preparations, C. neoformans may appear either as round cells with gram-positive granular inclusions impressed upon a pale lavender cytoplasmic background or as gram-negative lipoid bodies.
Rifampin as a Selective Agent for Isolation of Oral Spirochetes
Authors: Leschine S. B. , Canale-Parola E.
Published: January 2020

Spirochetes indigenous to the healthy gingival crevice of the human mouth were isolated directly from colonies in agar medium containing rifampin as a selective agent.
Epidemiological usefulness of changes in hemolytic activity of Vibrio cholerae biotype El Tor during the seventh pandemic
Authors: Barrett T J , Blake P A
Published: January 2020

Hemolytic Vibrio cholerae biotype El Tor strains were isolated in the United States in 1973 and 1978 after they had supposedly disappeared worldwide during the 1960s and 1970s. We decided to examine the change in prevalence of hemolytic El Tor strains since the beginning of the seventh pandemic and evaluate the usefulness of hemolytic activity as an epidemiological marker. A total of 48 isolates of V. cholerae biotype El Tor isolated in the Eastern Hemisphere between 1960 and 1979, along with 1 Texas (1973) and 38 Louisiana (1978) isolates, were tested for hemolytic activity by each of four methods. One method (utilizing heart infusion broth with 1% glycerol) was slightly superior for detecting hemolytic activity. Titers obtained with this method ranged from less than 2 to 1,024. Of 13 (76.9%) strains from the earliest part of the current pandemic, 10 were hemolytic, compared with 1 of 26 (3.8%) strains isolated in the period from 1966 to 1979 in the Eastern Hemisphere, indicating that nonhemolytic El Tor strains have replaced the hemolytic variety there. In contrast, all 38 Louisiana isolates and the Texas isolate were strongly hemolytic. Hemolytic activity was concluded to be a useful epidemiological marker.
Quantitative method for enumeration of enterotoxigenic Escherichia coli
Authors: Calderon R L , Levin M A
Published: January 2020

A rapid method was developed to quantify toxigenic Escherichia coli, using a membrane filter procedure. After filtration of samples, the membrane filter was first incubated on a medium selective for E. coli (24 h, 44 degrees C) and then transferred to tryptic soy agar (3%; 6 h, 37 degrees C). To assay for labile toxin-producing colonies, the filter was then transferred to a monolayer of Y-1 cells, the E. coli colonies were marked on the bottom of the petri dish, and the filter was removed after 15 min. The monolayer was observed for a positive rounding effect after a 15- to 24-h incubation. The method has an upper limit of detecting 30 toxigenic colonies per plate and can detect as few as one toxigenic colony per plate. A preliminary screening for these enterotoxigenic strains in polluted waters and known positive fecal samples was performed, and positive results were obtained with fecal samples only.
Variable assimilation of carbon compounds by Candida albicans
Authors: Syverson R E
Published: January 2020

A total of 215 typical strains of Candida albicans were studied for their ability to assimilate 11 carbon compounds. All isolates assimilated lactic acid, ribitol, succinic acid, methyl alpha-D-glucopyranoside. None of the isolates assimilated cellobiose and salicin; 1.9% of the isolates assimilated L-arabinose. Citric acid, glycerol, and L-sorbose were assimilated by greater than 97% of the isolates, whereas melezitose was assimilated by 81% of the isolates. Assimilation results depended on duration of incubation, temperature, and methodology.
Evaluation of a finger-stick specimen collection method for seroepidemiology of antibody to hepatitis A virus
Authors: McCaustland K A , Bond W W , Bradley D W , Favero M S , Petersen N J , Maynard J E
Published: January 2020

A finger-stick swab method of collecting blood specimens was shown to compare favorably with the conventional venipuncture method in serological determinations of antibody to hepatitis A virus by radioimmunoassay.
Effects of antibodies, trypsin, and trypsin inhibitors on susceptibility of neonates to rotavirus infection
Authors: McLean B S , Holmes I H
Published: January 2020

Levels of antirotaviral secretory immunoglobulin A were measured by enzyme-linked immunosorbent assay in colostrum and milk samples collected daily for the first 5 days postpartum from 49 mothers breast-feeding their infants. The trypsin-inhibitory capacity of these lacteal secretion samples was assessed by their ability to inhibit the hydrolysis of alpha-N-benzoyl-DL-arginine-p-nitroanilide by trypsin. Stools passed by these breast-fed infants and by an additional 43 bottle-fed infants were pooled by individual and examined by electron microscopy for rotavirus. Stool trypsin levels were estimated with the gelatin hydrolysis test. Breast-fed infants were significantly less likely to become infected with rotavirus and showed significantly lower stool tryptic activity than did bottle-fed infants. Breast-fed infants who did not excrete rotavirus over the 5-day period received milk of significantly higher antirotaviral secretory immunoglobulin A or trypsin-inhibitory capacity or both than breast-fed infants who were infected with rotavirus. A case of probable maternal rotavirus infection during pregnancy, producing greatly elevated lacteal antirotaviral secretory immunoglobulin A levels lasting for 2 years, was detected. Results of this study suggest that both antibodies and trypsin inhibitors in human milk can be associated with the protection of neonates against rotavirus infection in the first 5 days of life.
Otitis media caused by beta-lactamase-producing Branhamella (Neisseria) catarrhalis
Authors: Lee W S , Fordham T , Alban J
Published: January 2020

A beta-lactamase-producing Branhamella catarrhalis was isolated in pure culture from the right middle ear aspiration of an otitis media in a 3-month-old girl. The patient responded well to cefamandole treatment.
Relative efficacy of embryonated eggs and cell culture for isolation of contemporary influenza viruses
Authors: Monto A S , Maassab H F , Bryan E R
Published: January 2020

The efficiency of embryonated eggs and primary monkey kidney cell culture for isolation of contemporary influenza viruses was compared over three seasons by parallel inoculation of clinical materials. For the type A (H1N1) strains of 1977-1978 and for type B strains eggs were very inefficient, and nearly all viruses were isolated in cell culture alone. For the type A (H1N1) strains of 1978-1979 and for type A (H3N2) strains, isolations were more frequently made in eggs, but even here eggs were not as efficient for primary isolation as were cell cultures. It is concluded that primary monkey kidney cell cultures can be used as the single system for primary isolation of current type A and B influenza viruses.
Naturally acquired rabies in an eastern wood rat (Neotoma floridana)
Authors: Dowda H , DiSalvo A F , Redden S
Published: January 2020

Rabies in an eastern wood rat was detected by fluorescent antibody testing and mouse inoculation. The results were independently confirmed.
Characterization of antibody activity in oligoclonal immunoglobulin G synthesized within the central nervous system in a patient with tuberculous meningitis
Authors: Kinnman J , Link H , Frydén A
Published: January 2020

Thin-layer polyacrylamide gel isoelectric focusing of cerebrospinal fluid (CSF) and serum obtained from one patient 48 and 65 days after the onset of tuberculous meningitis revealed five oligoclonal immunoglobulin zones in CSF without any counterpart in serum, indicating local immunoglobulin production. Subsequent immunofixation with specific antisera revealed that three of the zones consisted of immunoglobulin G lambda present simultaneously. Immunofixation with Mycobacterium tuberculosis and bacillus Calmette-Guérin (BCG) as antigens and autoradiography revealed zones of specific antibodies in the CSF which, regarding mobility, corresponded to oligoclonal and polyclonal CSF immunoglobulin G zones. No antibody activity was detectable in the corresponding serum, indicating that the antibodies present in CSF were synthesized within the central nervous system. In seven control patients (three with multiple sclerosis, four with chronic inflammatory central nervous system diseases of unknown cause) with oligoclonal CSF immunoglobulin, no evidence for local production of antibodies against M. tuberculosis or BCG was detectable. Immunofixation with M. tuberculosis or BCG as antigens and autoradiography may prove to be a useful diagnostic complement to conventional techniques in patients with suspected tuberculous meningitis.
Properties of binding of Escherichia coli endotoxin to various matrices
Authors: Maitra S K , Yoshikawa T T , Guze L B , Schotz M C
Published: January 2020

Binding of Escherichia coli O127:B8 endotoxin to a variety of resins and column materials was investigated by measuring the beta-hydroxy myristic acid content (a major component of the lipid A moiety) of endotoxin after hydrolysis by selected ion-monitoring gas chromatography-mass spectrometry. More than 80% of the endotoxin was bound to hydroxylapatite, polystyrene, Dowex 1-X2, and charcoal. The binding of endotoxin to these materials was markedly reduced by the addition of normal or delipidated serum. Phenyl- and octyl-Sepharose bound 56 and 50% of the endotoxin from saline solutions, respectively. Their percent binding was increased significantly in 1 M ammonium sulfate solutions, indicating hydrophobic interactions between endotoxin and phenyl- and octyl-Sepharose. Only 5% of the endotoxin was bound to plastic polymer PSI-HAP-100 beads, and no binding was observed with concanavalin A- and heparin-Sepharose. Study of the in vitro binding of endotoxin to the above material in the presence of serum suggests that the use of these materials in removing circulating endotoxin in vivo is limited.
Differential identification of Mycobacterium fortuitum and Mycobacterium chelonei
Authors: David H L , Traore I , Feuillet A
Published: January 2020

Mycobacterium fortuitum and Mycobacterium chelonei are distinguished unambiguously by the combined use of five test characters: nitrate reductase, beta-glucosidase, acid production from fructose, penicillinase, and trehalase. Typically, M. fortuitum was nitrate reductase positive, beta-glucosidase positive; M. chelonei was nitrate reductase negative, beta-glucosidase negative, penicillinase positive, and trehalase positive and did not produce acid from fructose.
Methodological variation in antibiotic synergy tests against enterococci
Authors: Ryan R W , Kwasnik I , Tilton R C
Published: January 2020

Thirty-two human isolates of enterococci were tested for antibiotic synergy by using penicillin and one of six aminoglycosides. Three methods were used: synergy screen, microdilution checkerboard, and time-kill curves. The synergy screen accurately predicted synergy for gentamicin-penicillin combinations, and this synergy was later confirmed by time-kill curves. The microdilution checkerboard method suffered from inherent variation, and agreement with time-kill curves ranged from 92% (twofold reduction in minimum inhibitory concentration) to 4.2% (fourfold reduction in minimum inhibitory concentration). We suggest that enterococci be screened for synergy (i.e., presence or absence of high-level resistance) by using the criterion of growth or no growth in the presence of 2,000 microgram of an aminoglycoside per ml. The microdilution checkerboard test for synergy is not recommended.
Characterization of Vibrio cholerae protease activities with peptide digest analysis
Authors: Schneider D R , Sigel S P , Parker C D
Published: January 2020

A simple method for the analysis of microbial proteases is described that was used to characterize the proteolytic activities of various Vibrio cholerae isolates. This method utilized the unique peptides generated from the degradation of a standard protein by proteases of various specificities. These peptides were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The unique patterns of peptides seen in gels can be used to type proteases according to their relative specificities. Culture supernatants of V. cholerae isolates from a variety of environmental and human sources were analyzed for the presence of a protease previously isolated and characterized in this laboratory from V. cholerae strain CA401. Supernatants from most isolates showing dimethyl casein proteolytic activity exhibited the presence of enzymes similar to the CA401 protease in their peptide digest patterns against bovine serum albumin and in their immunological reactivities. The probable widespread presence of this virulence-associated protease in V. cholerae isolates is discussed.
Detection of bacteriuria: manual screening test and early examination of agar plates
Authors: Murray P R , Niles A C
Published: January 2020

We assessed two procedures for the initial processing of urine specimens: (i) manual screening of urine samples by inoculation into tryptic soy broth and incubation for 3 to 6 h, and (ii) examination of agar plates inoculated with urine and incubated for 4 to 6 h. Of the urine samples with greater than 10(5) colony-forming units per ml, 73.6 and 90.1% were detected with the broth cultures after 3 and 6 h of incubation, respectively. The organisms not detected at 6 h were Corynebacterium spp., Lactobacillus spp., and one of six isolates of yeasts. When inoculated plates were examined for growth, 53.3 and 80.0% of the specimens with greater than 10(5) colony-forming units per ml were detected after 4 and 6 h of incubation, respectively. Of the cultures with significant growth of Enterobacteriaceae or Pseudomonas spp., 95% were detected on the plates after 6 h of incubation.
Carbohydrate fingerprints of streptococcal cells
Authors: Pritchard D G , Coligan J E , Speed S E , Gray B M
Published: January 2020

The carbohydrates of whole cells of group A, B, C, D, F, and G streptococci were analyzed with a highly sensitive gas chromatographic procedure. Characteristic chromatographic fingerprints were obtained for each group of streptococci grown in broth cultures or as single colonies on a blood agar plate. Rhamnose, glucose, and N-acetylglucosamine were major components of all the groups. Groups A, C, and F contained very little galactose, and groups A and B showed almost a complete absence of N-acetylgalactosamine. Chromatograms obtained for group B streptococci were distinguished by the presence of 1,4-anhydroglucitol.
Further studies on antigen variation in Staphylococcus aureus
Authors: Pereira A T
Published: January 2020

The plating of successive Staphylococcus aureus subcultures of daily transfers proved that discontinuous variation resembling a genetic mutation and selective outgrowth of the variant are responsible for antigen variation. Every subculture of S. aureus, when repeatedly transferred, contained a mixture of cells with original antigen 17 (or 13) and final antigen 1 (or 3) that are relevant for research, serological diagnosis, and epidemiological study of staphylococcal diseases.
K99 antigen-positive enterotoxigenic Escherichia coli from piglets with diarrhea in Sweden
Authors: Smyth C J , Olsson E , Moncalvo C , Söderlind O , Orskov F , Orskov I
Published: January 2020

K88 antigen-negative enterotoxigenic Escherichia coli and non-enterotoxigenic strains isolated from piglets with diarrhea were examined for K99 antigen by agglutination tests after growth on Minca-IsoVitaleX (BBL Microbiology Systems, Cockeysville, Md.) agar medium. Of 64 K88-negative enterotoxigenic strains from as many piglets, 17 were found to be K99 positive. Of these, 10 were Swedish and 7 were of Norwegian origin. All 17 produced heat-stable enterotoxin detectable in the infant mouse assay, but only 2 gave positive ligated loop tests in 3- to 7-week-old piglets. Ligated loop tests in 5- to 12-day-old piglets were positive for each of the 15 K99-positive strains tested. Each of the Swedish K99-positive isolates was from a piglet of lesser than or equal to 7 days of age. One piglet harboring a K99-positive strain also harbored an O141:K88 enterotoxigenic strain producing only heat-stable enterotoxin. Five of the Swedish piglets yielding K99-positive isolates were from dams vaccinated wih a multicomponent bacterial vaccine containing K88 antigen. The K99 strains were O:K:H serotyped. The O serogroups represented were O8, O9, O64, O101, and O140. None of the 101 non-enterotoxigenic porcine isolates, representing 42 serogroups and non-O-groupable and rough strains, was found to be K99 positive. The findings indicate that so-called class 2 or atypical porcine enterotoxigenic E. coli should be routinely examined for the presence of K99 antigen.
Hydrolytic enzymes of Moraxella bovis
Authors: Frank S K , Gerber J D
Published: January 2020

Certain strains of Moraxella bovis produce tissue-damaging enzymes which may initiate or potentiate infectious bovine keratoconjunctivitis. Thirteen reference strains of this species were characterized physiologically and screened for production of various enzymes by some conventional biochemical tests and the API ZYM system (Analytab Products, Plainview, N.Y.). All 13 strains were hemolytic. All hydrolyzed Tween 80 and Tween 85 and displayed C4 esterase, C8 esterase-lipase, and C14 lipase activities. All produced phosphoamidase and phosphatase. All were able to hydrolyze casein and gelatin. All produced leucine and valine aminopeptidases and fibrinolysin. Twelve produced hyaluronidase or were agarolytic. Three hydrolyzed chondroitin sulfate. Nine strains autoagglutinated. Five produced catalase, and two produced cystine aminopeptidase.
Automated immunofluorescence test for toxoplasmosis
Authors: Gordon M A , Duncan R A , Kingsley L C
Published: January 2020

A soluble-antigen fluorescent-antibody technique for serodiagnosis of toxoplasmosis, employing a special fluorometer (FIAX; International Diagnostic Technology, Santa Clara, Calif.), was evaluated in comparison with a standard indirect fluorescent-antibody method which uses a fluorescence microscope. Titers obtained by the new method on 324 serum samples correlated very well with those of the conventional procedure. There was good intralaboratory and interlaboratory agreement in runs on selected samples. The FIAX method is simple, rapid, and reliable and has the advantage of objectivity.
Enzyme-linked immunosorbent assay for detection of pertussis immunoglobulin A in nasopharyngeal secretions as an indicator of recent infection
Authors: Goodman Y E , Wort A J , Jackson F L
Published: January 2020

An enzyme-linked immunosorbent assay was developed for detection of immunoglobulin A (IgA) antibody to Bordetella pertussis (PsIgA) in nasopharyngeal secretions as an indicator of recent infection. Secretion specimens submitted for pertussis culture were examined for PsIgA by this technique. Of 348 specimens tested, B. pertussis was cultured from 57, and PsIgA was detected in 8 culture-positive and 40 culture-negative specimens. The average time between onset of symptoms and specimen collection for the culture-positive, PsIgA-negative specimens was 10 days; for the culture-positive, PsIgA-positive specimens, 15 days; and for the culture-negative, PsIgA-positive specimens, 36 days. Examination of paired samples available from several culture-proven cases demonstrated conversion from a negative PsIgA in the early sample to a positive PsIgA in the follow-up sample. Our results indicate that PsIgA is produced during natural human infection and does not arise as a result of parenteral vaccination. PsIgA usually appears in the nasopharyngeal secretions during the second or third week of illness and persists for at least 3 months. The detection of PsIgA in secretions may be a valuable diagnostic aid in culture-negative patients with pertussis.
Technical aspects of the Staphylococcus aureus teichoic acid antibody assay: gel diffusion and counterimmunoelecrophoretic assays, antigen preparation, antigen selection, concentration effects, and cross-reactions with other organisms
Authors: Sheagren J N , Menes B L , Han D P , Sanders J L , Sheagren M A
Published: January 2020

Because variable results are being reported from laboratories performing the teichoic acid antibody assay in patients with serious infections due to Staphylococcus aureus, we have thoroughly reviewed all technical aspects of the test. This paper reports on the importance of agar and antigen preparation in standardizing results of the assay and reducing the prevalence of false-negative and -positive tests. Once standardized, the counterimmunoelectrophoretic method is as accurate as the gel diffusion method for both initial screening and generating titers; practically, however, unless numerous tests are to be performed, the gel diffusion technique will suffice for most purposes. The cell wall of the Lafferty strain of S. aureus was used as the standard antigen in assays for serum antibodies. We studied whether it is an appropriate antigen and found that antibody titers obtained with the Lafferty strain antigen were, in three patients with endocarditis, the same as those obtained with antigens from the individual blood stream isolates. We have also confirmed that pooled human gamma globulin can be used, by back titration against newly prepared lots of antigen, to select optimal antigen concentration and is as good as more specific, higher titer serum specimens for that purpose. Finally, cell wall antigens from Staphylococcus epidermidis and a variety of streptococci may react with normal human sera, but such antigens are distinct by immunoprecipitation from those from S. aureus.
Analysis of parameters affecting the hemagglutination activity of Escherichia coli possessing colonization factor antigens: improved medium for observing erythrocyte agglutination
Authors: Sedlock D M , Bartus H F , Zajac I , Actor P
Published: January 2020

The hemagglutination (HA) activity of two strains of Escherichia coli, each possessing different colonization factor antigens (CFA), was examined under different test conditions. The effects of ionic strength, temperature, pH, cations, and reaction surface on erythrocyte (RBC) agglutination were analyzed. Strain H-10407 (CFA/I) caused the agglutination of human, bovine, and chicken RBC, whereas strain CL-9699 (CFA/II) agglutinated only bovine and chicken RBC. The HA activity of both strains increased with decreasing ionic strength, pH, and temperature, the effects of temperature being negligible at low ionic strength. When accounting for ionic strength, the presence of Ca2+, Mg2+, Fe2+, or Fe3+ ions did not increase the HA activity of these bacteria. Optimum conditions for HA of reactive RBC by bacteria included low ionic strength (less than 50 mM) and slightly acidic pH (6.0 to 7.0). Use of a low-ionic-strength medium permitted application of microtitration methods to visualize the HA reactions. Storage of RBC in low-ionic-strength medium did not change their HA properties, and the use of this medium proved superior to saline in overcoming HA variation observed with different preparations of RBC.
Differentiation and characterization of Klebsiella pneumoniae strains by pyrolysis-gas-liquid chromatography-mass spectrometry
Authors: Abbey L E , Highsmith A K , Moran T F , Reiner E J
Published: January 2020

Nine coded duplicate strains of capsular nontypable Klebsiella pneumoniae were analyzed by pyrolysis-gas-liquid chromatography-mass spectrometry. All duplicate strains were correctly matched, and individual strains, including seven nontypable strains, were clearly distinguishable from one another. The addition of mass spectrometry to the analysis has aided the process of identification and has provided chemical structural information on K. pneumoniae strains. Application of this technique to the identification of some disease outbreaks or nosocomial problems could be of epidemiological importance, especially when conventional methods do not identify the epidemic strain.
Evaluation of an automated, computerized system (automicrobic system) for Enterobacteriaceae identification
Authors: Burdash N M , Teti G , West M E , Bannister E R , Manos J P
Published: January 2020

The automated and computerized AutoMicrobic system (AMS; Vitek Systems, Inc., subsidiary of McDonnell Douglas, Hazelwood, Mo.) was evaluated as a means of identifying the Enterobacteriaceae. The Micro-ID system (General Diagnostics, Morris Plains, N.J.) and, when necessary, conventional tubed media were used for comparison. Identification by AMS and Micro-ID differed in only 12 of 1,528 isolates (0.8%). Disagreements occurred primarily with Enterobacter spp. Precision testing of the AMS showed only 1 of 72 tests (1.4%) deviating from the expected. The AMS was found to be an accurate and precise method for the identification of Enterobacteriaceae.
Clinical evaluation of the AutoMicrobic system Yeast Biochemical Card for rapid identification of medically important yeasts
Authors: Oblack D L , Rhodes J C , Martin W J
Published: January 2020

The autoMicrobic system Yeast Biochemical Card (Vitek Systems, Inc., subsidiary of McDonnell Douglas, Hazelwood, Mo.), a fully automated and computer-assisted method for identifying seven yeast genera based on 26 biochemical reactions, was compared with the API 20C (Analytab Products, Plainview, N.Y) yeast identification system, using 253 clinical yeast isolates. There was an 84% correlation between the Yeast Biochemical Card and API 20C systems based on biochemical tests alone and a 96% correlation when morphology was combined with the biochemical profile. Of 22 species examined, 14 were definitively identified within 24 h, using only the biochemical tests; the remaining 8 species were presumptively identified, using biochemical tests within 24 h, with definitive identification being completed in 48 to 72 h when morphological characteristics were ascertained. The Yeast Biochemical Card system was both a facile and, with concomitant morphology, an accurate system for performing yeast identification.
Human infective dose determinations for oral poliovirus type 1 vaccine in infants
Authors: Minor T E , Allen C I , Tsiatis A A , Nelson D B , D'Alessio D J
Published: January 2020

The 50, 10, and 1% human infective doses of poliovirus type 1 vaccine administered orally to 32 infants were estimated to be 72, 39, and 20 tissue culture infective doses, respectively.
Quantitation of dengue precipitating antibody by inhibition countercurrent immunoelectrophoresis
Authors: Churdboonchart V , Bhamarapravati N , Harisdangkul V , Futrakul P , Chiengsong R
Published: January 2020

The inhibition countercurrent immunoelectrophoresis test was employed to detect dengue virus antibody in patients' sera. Anti-dengue type 2 titers determined by inhibition countercurrent immunoelectrophoresis correlated well with hemagglutination inhibition titers. In secondary cases, more than fourfold increases in precipitating antibodies were observed. The control sera were negative except for sera from a few patients with systemic lupus erythematosus, which showed low titers. Simultaneous detection of dengue virus antigen and antibody in sera collected during the acute phase could confirm at least 90% of cases. This method is recommended as a routine technique to quantitate antibody in sera from suspected cases of dengue hemorrhagic fever.
Identification of Pasteurella multocida and Pasteurella haemolytica by API 20E, Minitek, and Oxi/Ferm systems
Authors: Collins M T , Weaver N , Ellis R P
Published: January 2020

Fifty serotyped isolates each of Pasteurella multocida and Pasteurella haemolytica were tested on the API 20E strip (Analytab Products, Plainview, N.Y.), the Oxi/Ferm tube (Roche Diagnostics, Nutley, N.J.), and the Minitek system (BBL Microbiology Systems, Cockeysville, Md.). None of the rapid test systems reliable identified these organisms. With the API system, discrepancies between expected and actual results for the oxidase test and nitrate test frequently resulted in misidentification or no identification. The Minitek system misidentified 68% of the P. haemolytica isolates. The Minitek identification of Pasteurella depends on 100% positive xylose reactions, whereas only 56% of the P. haemolytica strains were positive for xylose fermentation. The Oxy/Ferm system, instead of giving a definitive identification, in most instances merely placed Pasteurella in a category of similar organisms.
Antibiotic-resistant group JK bacteria in hospitals
Authors: Gill V J , Manning C , Lamson M , Woltering P , Pizzo P A
Published: January 2020

The organisms designated as Center for Disease Control group JK are gram-positive rods that have previously been described as causing serious infection in compromised hosts. Four years of hospital experience with this group of organisms in Clinical Center patients was reviewed. Studies were also undertaken on specific wards to determine frequency of occurrence and distribution patterns. Inguinal cultures taken on two wards showed that 30 to 35% of patients were colonized with group JK and that newly admitted patients may already be colonized at the time of admission. Colonization was shown to persist for weeks and sometimes months. Isolates obtained throughout the hospital were predominantly from cancer patients, particularly in wounds, abscesses, and drainage sites. Most blood isolates were from granulocytopenic patients with hematological malignancies.
In vitro antibiotic removal and bacterial recovery from blood with an antibiotic removal device
Authors: Lindsey N J , Riely P E
Published: January 2020

The antibiotic removal device manufactured by Marion Laboratories (Kansas City., Mo.) is intended for treatment, before culture, of blood specimens from hospital patients being treated with antibiotics. Measurement of 13 antibiotics showed that the antibiotic removal device removed amikacin, ampicillin, carbenicillin, cefazolin, cephalothin, chloramphenicol, gentamicin, nafcillin, tetracycline, tobramycin, and vancomycin and reduced cefoxitin and ticarcillin to extremely low levels. Three combinations of antibiotics were similarly removed or reduced. Five species of anaerobic bacteria, one yeast species, and six species of facultative or aerobic bacteria were used to challenge the possibility that the antibiotic removal device would trap or inhibit microorganisms. All were recovered from the device in the same numbers as were inoculated.
Distribution of
Legionella pneumophila
Antibody Among Primate Species
Authors: Helmke R. J. , Kalter S. S. , Heberling R. L.
Published: January 2020

Sera representing 16 different primate species were surveyed by indirect immunofluorescence for evidence of antibody to
Legionella pneumophila.
The presence of antibody in Old and New World monkeys and in apes supports previous observations of the ubiquity of
Legionella pneumophila
.
Yersinia pseudotuberculosis infection: study of an epizootic in squirrel monkeys
Authors: Buhles W C , Vanderlip J E , Russell S W , Alexander N L
Published: January 2020

An epizootic of an acutely fatal enteric disease in a colony of squirrel monkeys (Saimiri sciureus) was attributed to infection by Yersinia pseudotuberculosis serotype III. Of a total adult population of 96 animals at risk, there were six fatal cases of yersiniosis. Serological evaluation of the colony just after the outbreak ended revealed that 22 of 60 monkeys tested (37%) had significant antibody to Y. pseudotuberculosis (microagglutination titer of greater than or equal to 1:80) but did not have clinical disease. The outstanding pathological lesions noted in dying monkeys were acute, purulent, necrotic and focal enteritis primarily affecting the jejunum and ileum and focal hepatic necrosis and abscessation. Y. pseudotuberculosis was isolated from the organs of two of the dying monkeys. Using cold enrichment techniques, Yersinia was also isolated from the feces of two apparently healthy monkeys (both seropositive), from the spleen of a monkey dying of other causes, and from the colon contents of a stillborn squirrel monkey baby. All isolates had the same biotype and serotype. An episode of abortions was associated both temporally and spatially with the fatal cases of yersiniosis, and Y. pseudotuberculosis was cultured from the uterus of two of the dying monkeys, suggesting that yersinia infection may be associated with abortion, as well as with enteric infection, in these animals.
Production of Hemagglutinating Antigens of La Crosse Virus by Polyethylene Glycol Precipitation
Authors: Rabson Michael , Beaty Barry J. , Hildreth Stephen W. , Shope Robert E.
Published: January 2020

Hemagglutination titers of La Crosse virus antigens prepared in BHK-21 suspension cell cultures were substantially increased by precipitating the antigens with polyethylene glycol.
Unique temperature-sensitive nutritional requirements of bacteremic Escherichia coli isolates
Authors: Welch W D , Spurgeon L , Kitts D , Moyed H S , Thrupp L D
Published: January 2020

Of 50 strains of Escherichia coli isolated from blood cultures of bacteremic patients, 14 (28%) were unable to grow on minimal medium at 42 degrees C, compared to only 2 of 50 nonbacteremic strains. In 7 of the 14 bacteremic strains, growth at 42 degrees C was restored by adding nicotinic acid. These unique temperature-sensitive auxotrophic patterns warrant evaluation as a marker correlating with clinical pathogenicity in E. coli.
Multiple-antigen slide test for detection of immunoglobulin M antibodies in newborn and infant sera by immunofluorescence
Authors: Gallo D , Riggs J L , Schachter J , Emmons R W
Published: January 2020

A microimmunofluorescence test was evaluated for use in measuring immunoglobulin M (IgM) antibodies in infant sera to five of the agents implicated in congenital and neonatal disease. Pen point dots of Toxoplasma gondii, cytomegalovirus, rubella virus, herpes simplex virus, and chlamydial cell culture antigens were applied to each circle of eight-circle printed slides. These multiple-antigen slides greatly facilitated the screening of 607 sera from infants and 117 sera from mothers for the presence of IgM antibody to these agents. Forty sera could be examined microscopically in approximately 30 min. All sera reacting with one or more antigens were tested for rheumatoid factor by the latex method, absorbed with glutaraldehyde-cross-linked human IgG, and retested for the presence of IgM antibody. IgM antibody to cytomegalovirus was demonstrated in sera from four newborns, but IgM antibody to rubella virus could not be detected until 21 days after birth, although rubella virus was isolated from sera from five younger infants. This may indicate that rubella IgM levels in many congenitally infected newborns are too low to be measured by the immunofluorescence method. Five percent of the sera from infants in this study possessed demonstrable IgM antibody to one of the antigens.
Use of rapid auxanographic procedures for recognition of an atypical Candida
Authors: Baker J G , Salkin I F , Pincus D H , D'Amato R F
Published: January 2020

An atypical Candida which can cause diagnostic problems in clinical laboratories has recently been characterized. Assimilation patterns of 29 clinical isolates of an atypical Candida were obtained by the API 30C (Analytab Products, Plainview, N.Y.)., Uni-Yeast-Tek (Flow Laboratories, Inc., Rockville, Md.), and dye pour-plate auxanographic methods. The low frequency of assimilation of cellobiose, sucrose, and melezitose noted in all of these procedures permitted the early recognition of the atypical character of the isolates.
Serotyping of Ureaplasma urealyticum by immunoperoxidase assay
Authors: Quinn P A , Arshoff L U , Li H C
Published: January 2020

The immunoperoxidase method was applied to the identification of Urea-plasma urealyticum serotypes. The assay used highly diluted antisera and could be run directly on primary plate isolates. It was ideal for detecting and identifying mixed serotypes because stained and unstained colonies could be visualized simultaneously by conventional light microscopy. Antisera run against eight serotypes revealed one-way cross-reactions between serotypes 3 and 5 and antiserum to 2, and between serotype 4 and antiserum to 8, at dilutions of less than 1:150. This cross-reactivity could be diluted out at the optimal antiserum dilution for the immunoperoxidase assay, but not for the growth inhibition assay or immunofluorescence test. The immunoperoxidase assay therefore proved ideal for serotyping U. urealyticum.
Improved method for cholera diagnosis
Authors: Rogol M , Sechter I , Gerichter C B
Published: January 2020

The use of the oxidase test is proposed to discover sparse colonies of cholera vibrios on a plate of Pril nutrient agar seeded directly with the suspected stool sample or after enrichment. This method also enables the detection of other oxidase-positive potential pathogens, such as non-agglutinating vibrios, Aeromonas hydrophila, Pseudomonas spp., and other nonfermentative organisms.
Isolation of Legionella spp. from environmental water samples by low-pH treatment and use of a selective medium
Authors: Bopp C A , Sumner J W , Morris G K , Wells J G
Published: January 2020

A selective medium was developed and used successfully to isolate Legionella pneumophila and Legionella-like organisms from environmental specimens previously positive by animal inoculation methods. This medium consists of charcoal-yeast extract agar to which have been added cephalothin (4 micrograms/ml), colistin (16 micrograms/ml), vancomycin (0.5 microgram/ml), and cycloheximide (80 micrograms/ml). Pretreating of the environmental water samples with an acid buffer (pH 2.2), followed by plating on the selective medium, improved the rate of recovery of both Legionella and Legionella-like organisms relative to that with direct plating on selective media.
Detection of antibodies to Legionella pneumophila in immune guinea pig serum by solid-phase immunofluorescence
Authors: Vogel F R , Klein T W , Specter S C , Hitchings M , Friedman H
Published: January 2020

A semiautomated solid-phase immunofluorescence apparatus (FIAX; International Diagnostic Technology, Santa Clara, Calif.) was utilized to develop a rapid method for detection of antibody to Legionella pneumophila. The sera from guinea pigs immunized with a mixture of killed L. pneumophila and Freund complete adjuvant displayed markedly enhanced antibody activity as measured by FIAX when compared with that obtained from adjuvant-injected or unimmunized animals. A correlation was observed between FIAX net fluorescence units and microagglutination titers of serum samples obtained from immunized animals. Within-run and between-run coefficients of variation performed on selected immune serum samples were low. These results demonstrated that the FIAX method could readily and reproducibly detect Legionella-specific antibodies in the sera of actively immunized animals and suggest the possibility of a broader application of FIAX in the serological detection of exposure to L. pneumophila antigen.
Sensitive plaque neutralization assay for parainfluenza virus types 1, 2, and 3 and respiratory syncytial virus
Authors: Hawthorne J D , Albrecht P
Published: January 2020

A sensitive plaque neutralization assay for parainfluenza virus types 1, 2, and 3 and respiratory syncytial virus was developed in Vero and MA 104 cell cultures. The tests were performed in semimicrotoiter trays containing 24 wells, 16 mm in diameter. Parainfluenza virus type 1 formed plaques in Vero and MA 104 cells only when trypsin was added to the overlay medium. Plaquing of parainfluenza virus type 1 was more sensitive and technically reproducible in MA 104 cells than in Vero cells. Parainfluenza virus types 2 and 3 and respiratory syncytial virus readily formed plaques in Vero cells. Plaques with all viruses were necrotic in character, except for plaques produced by parainfluenza virus type 3, which appeared red due to an increased uptake of neutral red by infected cells. Different conditions for plaquing of the four viruses had to be used to obtain plaques of suitable size. Antibody titers of commercially prepared guinea pig typing sera were 5- to 50-fold higher by the plaque neutralization test than by complement fixation. The addition of guinea pig immunoglobulin G antiglobulin to the serum-virus mixtures enhanced the conventional neutralization test 5- to 10-fold. The sensitivity and specificity of the plaque neutralization test was also determined with sera of marmosets experimentally infected with parainfluenza virus types 1 and 3. The generally low postinfection titers could be enhanced, on the average, 40-fold by using human immunoglobulin G antiglobulin in the neutralization test. A low degree of cross-reactivity was shown between parainfluenza virus types 1 and 3 both in the conventional neutralization test and in the anti-immunoglobulin enhanced neutralization test.
Investigation of enzyme immunoassay time courses: development of rapid assay systems
Authors: Yolken R H , Leister F J
Published: January 2020

An enzyme immunoassay (EIA) consists of a series of antigen-antibody reactions which result in the binding of an enzyme-labeled antibody to a solid phase. The performance time of an EIA determination is thus largely dependent upon the time required for the antigen-antibody reactions. In an attempt to develop a rapid EIA system, we investigated the time course of an EIA system for the measurement of Haemophilus influenzae type b polysaccharide. We found that, although the use of short incubations led to a decrease in sensitivity, an assay system utilizing 10-min incubation periods was still capable of detecting antigen at a concentration of 1 ng/ml. Important factors in the sensitivity of EIAs with short incubation times were the performance of the reaction at 37 degrees C and the incubation of the solid phase with constant agitation. Utilizing these techniques, we developed an EIA system for the measurement of H. influenzae type b polysaccharide which could be completed in less than 30 min. This system was sufficiently sensitive to detect H. influenzae polysaccharide in the cerebrospinal fluids of nine patients with proven H. influenzae meningitis. Thus, EIA systems utilizing short incubation times might be useful for the rapid detection of infectious antigens in body fluids.
Association of Aeromonas sobria with human infection
Authors: Daily O P , Joseph S W , Coolbaugh J C , Walker R I , Merrell B R , Rollins D M , Seidler R J , Colwell R R , Lissner C R
Published: January 2020

Fifteen Aeromonas isolates from various human infections and nine isolates from polluted water were identified as either Aeromonas hydrophila or Aeromonas sobria and examined for cytotoxigenicity, enterotoxigenicity, adherence to epithelial cells, and other virulence-associated factors, including proteases, lipases, elastases, and hemolysins. Two groups of organisms (I and II) were distinguishable based on differences in median lethal doses in mice and cytotoxicity for Y-1 adrenal cells. Group I clinical and environmental strains had median lethal doses of less than 10(7) colony-forming units, were cytotoxic, frequently possessed several virulence-associated factors, and had lysine decarboxylase-positive or Voges-Proskauer-positive phenotypes or both. Piliation of Aeromonas was associated strongly with ability to adhere to human buccal cells, and these characteristics were associated with group I strains. Group II clinical and environmental strains had median lethal doses of greater than or equal to 10(7) colony-forming units, were not cytotoxic, and usually were lysine decarboxylase negative or Voges-Proskauer negative or both. Clinical strains in group II exhibited enterotoxigenicity, which was not detected in group II environmental strains. A sobria was more frequently associated with human infections; 13 of the 15 clinical strains were A. sobria, and 2 were A. hydrophila. On the other hand, the majority of the environmental strains (seven of nine) were A. hydrophila.
Fluorescent-antibody detection of Legionella dumoffii in a fatal case of pneumonia
Authors: Harris P P , Aufdemorte T , Ewing E P , Johnson J E , Tio F O
Published: January 2020

Legionella dumoffii organisms were detected in the lung tissue from a patient with a fatal case of pneumonia, the second reported to date. A fluorescent-antibody conjugate specific for the Tex-KL organism isolated in 1979 revealed many L, dumoffii organisms in lung tissue obtained postmortem from this patient.
Plaque Assay for Ebola Virus
Authors: Moe James B. , Lambert Rhonda D. , Lupton Harold W.
Published: January 2020

A plaque assay for Ebola virus is reported. The procedure has real potential for future research, although it is less sensitive than indirect fluorescent-antibody and mouse inoculation tests.
Gentamicin and amikacin disk susceptibility tests with Pseudomonas aeruginosa: definition of minimal inhibitory concentration correlates for susceptible and resistant categories
Authors: Barry A L , Thornsberry C , Jones R N
Published: January 2020

With currently recommended disk susceptibility tests, minimal inhibitory concentration correlates for amikacin were greater than 16 micrograms/ml for resistance and less than or equal to 12 micrograms/ml (not less than or equal to 8 micrograms/ml) for susceptibility. For gentamicin, they were greater than 8 micrograms/ml for resistance and less than or equal to 6 micrograms/ml (not less than or equal to 4 micrograms/ml) for susceptibility. Minor discrepancies between disk tests and minimal inhibitory concentration determinations will occur if only doubling dilutions of drug are used for measuring minimal inhibitory concentration values.