bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of Campylobacter fetus subsp. jejuni from Bangladeshi children
Authors: Blaser M J , Glass R I , Huq M I , Stoll B , Kibriya G M , Alim A R
Published: January 2020

To determine the prevalence of infection with Campylobacter fetus subsp. jejuni in Bangladesh, culture surveys were conducted among three populations. In Dacca, Campylobacter was isolated from 5.2% of 97 individuals with clinical dysentery and from 12.3% of 204 patients with only diarrhea. This difference may have resulted from a greater representation of young children in the second group. Campylobacter was isolated from 17.7% of the 141 healthy village children aged 1 to 5.5 years and from 38.8% of the 1-year-old children. More infected children (48%) had a history of recent diarrheal illness than did a group of matched controls (20%; P = 0.016). These findings suggest that campylobacter infection is common for Bangladeshi children. However, this organism may not cause diarrheal illness in all instances in which it is isolated.
Comparison of modified New York City medium with Martin-Lewis Medium for recovery of Neisseria gonorrhoeae from clinical specimens
Authors: Granato P A , Paepke J L , Weiner L B
Published: January 2020

A modified formulation of New York City medium was comparatively evaluated with Martin-Lewis medium for the recovery of Neisseria gonorrhoeae from clinical specimens. A total of 240 strains of gonococci were recovered from 1,250 specimens collected from walk-in patients attending a sexually transmitted disease clinic. N. gonorrhoeae was cultivated on both of these media from 182 clinical specimens with an additional 58 gonococcal strains isolated on either of the media. Of these discrepant gonococcal isolates, 27 strains were recovered on only modified New York City medium, whereas the remaining 31 strains were recovered on only Martin-Lewis agar. The differences in these isolation rates were not statistically significant. The overall results showed that modified New York City and Martin-Lewis media were comparable in their ability to grow gonococci from clinical material. Since modified New York City medium is capable of supporting the growth of N. gonorrhoeae, Mycoplasma pneumoniae, and urogenital mycoplasmas and inhibiting the growth of commensal microorganisms, it is possible that it may have considerable application as a multifunctional plating medium within the clinical laboratory.
Evaluation of a radial immunodiffusion test with polysaccharide B antigen for diagnosis of bovine brucellosis
Authors: Jones L M , Berman D T , Moreno E , Deyoe B L , Gilsdorf M J , Huber J D , Nicoletti P
Published: January 2020

A radial immunodiffusion (RID) test employing a polysaccharide antigen (poly B) was compared with tests currently used in the diagnosis of bovine brucellosis. Over 1,000 sera from vaccinated and infected cattle, all of which had been examined bacteriologically, were used to determine the sensitivity and specificity of the RID, card, Rivanol, and complement fixation tests. The RID test identified 90% of the cattle that were shedding Brucella in their milk. Although the complement fixation test was more sensitive, it was less specific than the RID test in cattle vaccinated as adults with Brucella abortus strain 19. A sensitive screening test, such as the card test, in combination with the RID test could be used in diagnostic laboratories, or even in the field, with little additional expense or technical expertise. An additional advantage is that the RID could be applied to sera from adult cattle as early as 2 months after vaccination, when postvaccinal agglutinins and complement-fixing antibodies may still be present. The indirect hemolytic test was used with some of the sera and was found to be a very sensitive test which could be useful in areas of low incidence but would not be practical for large-scale testing in adult-vaccinated herds.
Value of passive immune hemolysis for detection of heat-labile enterotoxin produced by enterotoxigenic Escherichia coli
Authors: Tsukamoto T , Kinoshita Y , Taga S , Takeda Y , Miwatani T
Published: January 2020

The method of passive immune hemolysis of Evans and Evans (Infect. Immun. 16:604-609, 1977) for detection of heat-labile enterotoxin produced by enterotoxigenic Escherichia coli was modified. A total of 373 strains of E. coli were tested by this method using materials obtained by treating the cells with polymyxin B and rabbit antiserum against cholera enterotoxin, purified by affinity gel column coupled with purified cholera enterotoxin, in N-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer (pH 6.7). The results correlated very well with those obtained in an assay with Chinese hamster ovary cells. It is concluded that passive immune hemolysis is useful as a routine clinical method for identifying E. coli strains that produce heat-labile enterotoxin.
Comparison of media and culture techniques for detection of Streptococcus pneumoniae in respiratory secretions
Authors: Wu S C , Trask L M , Phee R E
Published: January 2020

We compared the relative efficacy of three methods for the isolation of Streptococcus pneumoniae in lower respiratory secretions. Based on results from 294 clinical specimens, we found that S. pneumoniae was isolated at a frequency of 65% with 5% sheep blood agar or 5% sheep blood agar containing 5 micrograms of gentamicin per ml, both incubated in 5% CO2. Anaerobic incubation of 5% sheep blood agar enhanced the recovery rate of S. pneumoniae to 93%. The improved efficacy with anaerobic incubation is due to the greater ease of recognition of the larger and more mucoid colonies of S. pneumoniae, and to the suppression of the growth of other oral bacteria present in the respiratory sections.
Immunoglobulin G and immunoglobulin M polar staining of Toxoplasma gondii in the indirect immunofluorescence test
Authors: Franco E L , Sulzer A J , Higby R W , Peralta J M
Published: January 2020

Polar staining (PS) of Toxoplasma gondii in the indirect immunofluorescence test has been considered a nonspecific reaction caused exclusively by certain immunoglobulin M (IgM) antibodies and confined to the anterior end of the parasite. After we observed a patient with positive serology for Chagas' disease who presented an IgG PS reaction, we studied sera from 120 patients with Chagas' disease, 20 sera from patients with Leishmania donovani infection, and 30 sera from patients with Leishmania braziliensis infection. When only those specimens having no detectable anti-Toxoplasma activity were considered, a significantly (P less than 0.01) higher prevalence of IgG PS was found in the Chagas' disease and L. donovani groups than in sera from normal American and Brazilian adults. Those sera also showed higher PS titers (1:64 to 1:1,024) when compared with controls (1:16 to 1:64). IgG PS titers did not decrease after serum treatment with 2-mercaptoethanol. However, the same treatment removed completely IgM PS. IgG PS, but not IgM PS, could be removed by adsorption with Trypanosoma cruzi epimastigotes. IgM PS was found in all samples studied, except in 41 of a group of 43 umbilical cord sera. It was found that the antigen source and the microscopy system can influence the detection of PS. It is proposed that after finding an intense IgG PS reaction, the laboratory should screen such serum also for anti-T. cruzi antibodies which may be undetected in the sample.
Comparison of techniques for recovering murine cytomegalovirus from a macrophage-enriched subpopulation of mice
Authors: Winkler S R , Booss J
Published: January 2020

We previously demonstrated the transmission of murine cytomegalovirus to syngeneic mice and preformed monolayers of mouse embryo fibroblasts with plastic-adherent peritoneal exudate cells from infected mice as a source of macrophages. In the present studies we compared this standard feeder layer method with a reverse feeder layer method in which the adhering peritoneal exudate cells remained attached to plastic and to which were added mouse embryo fibroblasts. Recovery of virus from the adherent peritoneal exudate cells of infected mice was achieved with both methods. Whereas the standard method achieved greater accuracy and usually recovered more virus, the reverse method appeared to recover virus more frequently and required fewer cells to perform the assay. Furthermore, the reverse method demonstrated the survival of murine cytomegalovirus in adhering cells after culture periods of up to 2 weeks.
Clostridium difficile
Toxin as a Confounding Factor in Enterovirus Isolation
Authors: Schmidt Nathalie J. , Ho Helen H. , Dondero Marjorie E.
Published: January 2020

A peculiar cytotoxic effect, occasionally encountered in the course of inoculating cell cultures with fecal specimens for routine enterovirus isolation attempts, was shown to be produced by
Clostridium difficile
toxin.
Preservation of nasopharyngeal smears for fluorescent antibody detection of Bordetella pertussis
Authors: Harris P P , Thomason B , McKinney R M
Published: January 2020

Smears from throat and nasal washings seeded with Bordetella pertussis were either treated with aprotinin (a protease inhibitor) or fixed with 1 or 10% Formalin. These smears were stored at 23, 4, and -70 degrees C. Smears were removed and stained with a fluorescent antibody conjugate for B. pertussis at intervals during 22 days of storage. Results indicate that treatment of smears with 4 or 8 U of aprotinin per ml preserved fluorescent antibody staining qualities of B. pertussis for 22 days; fixation with either concentration of Formalin was unsatisfactory.
Candida antigenemia, as detected by passive hemagglutination inhibition, in patients with disseminated candidiasis or Candida colonization
Authors: Meunier-Carpentier F , Armstrong D
Published: January 2020

A passive hemagglutination inhibition assay was studied by using a hyperimmune serum from rabbits immunized with whole yeast cells (Candida albicans group A). This technique was effective at detecting small amounts of laboratory-prepared mannan or a whole-cell extract of C. albicans. Of 32 patients with documented disseminated candidiasis that were tested, 19 showed evidence of circulating antigen by passive hemagglutination inhibition. Three of these patients showed only partial, rather than complete, inhibition. Among 22 colonized patients, 4 showed partial inhibition, and none of 49 normal controls demonstrated inhibition. All of the sera were tested for antibody by agglutination, immunodiffusion, and passive hemagglutination. This last technique added increased sensitivity, but not specificity, to the standard tests already in use. Fourfold or greater titer rises by passive hemagglutination occurred in fewer than one-third of patients with invasive candidiasis and developed in more than one-half of patients who were colonized and did not require systemic anticandida therapy.
Demonstration of Actinomyces and Arachnia species in cervicovaginal smears by direct staining with species-specific fluorescent-antibody conjugate
Authors: Pine L , Malcolm G B , Curtis E M , Brown J M
Published: January 2020

For direct observation of microaerophilic actinomycetes by fluorescent antibody, a procedure was developed in which pepsin treatment and rhodamine conjugate of normal serum were used to reduce nonspecific staining in cervicovaginal smears. Actinomyces israelii, Actinomyces naeslundii, and Arachnia propionica were observed in cervicovaginal smears from women who did use and who did not use an intrauterine contraceptive device. A. israelii was found more commonly in women with an intrauterine contraceptive device, but no evidence was obtained that the use of an intrauterine contraceptive device influenced the presence of either A. propionica or A. naeslundii.
Septic Arthritis and osteomyelitis caused by an organism of the genus Rhodococcus
Authors: Broughton R A , Wilson H D , Goodman N L , Hedrick J A
Published: January 2020

We describe a previously healthy, immunologically normal young girl who presented painless swelling of fingers, a toe, and one knee. Roentgenograms were consistent with osteomyelitis of the phalanges and knee effusion. Rice bodies (corpora oryzoidea) were identified in viscous fluid obtained from the knee during arthroscopy. Culture of this fluid grew an organism initially believed to be a member of the genus Nocardia but which was later presumptively identified as a member of the genus Rhodococcus. The patient was successfully treated with a combination of erythromycin and amoxicillin for a total of 6 months. Previously reported cases of this unusual infection and the microbiological features of the organism are reviewed. The significance of rice bodies found in joint fluid and the therapy of this infection are discussed.
Detection of typhus antibodies by latex agglutination
Authors: Hechemy K E , Osterman J V , Eisemann C S , Elliott L B , Sasowski S J
Published: January 2020

A latex test for assay of antibodies to endemic and epidemic typhus rickettsiae is simple, group-specific, sensitive, and reproducible. Cross-reactivity within the typhus group was extensive.
Comparison of three techniques for isolation of Rhodococcus (Corynebacterium) equi from contaminated sources
Authors: Barton M D , Hughes K L
Published: January 2020

Inoculation of a liquid medium comprised of Trypticase soy broth (BBL Microbiology Systems), cycloheximide, nalidixic acid, penicillin, and potassium tellurite and subcultured onto M3 medium plus potassium tellurite was highly successful for the isolation of Rhodococcus (Corynebacterium) equi from soil.
Iodometric spot test for detection of beta-lactamase in Haemophilus influenzae
Authors: Lee W S , Komarmy L
Published: January 2020

A simple iodometric spot test for detecting beta-lactamase activity in Haemophilus influenzae was compared with the capillary procedure for detecting beta-lactamase and the Bauer-Kirby disk susceptibility procedure. Isolates were classified similarly by all three procedures.
Reader error in determining minimal inhibitory concentrations with microdilution susceptibility test panels
Authors: Barry A L , Braun L E
Published: January 2020

Microdilution susceptibility test panels were independently read by two different technologists. One or more discrepancies between readers occurred with 5% of the 25,022 recorded minimal inhibitory concentrations, but only 0.4% of the tests varied by greater than or equal to 2 dilution intervals.
Facilitating quality control of the antimicrobial susceptibility test
Authors: Fluornoy D J
Published: January 2020

Standard reference strains of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were suspended in sterile deionized water and tested daily via disk agar diffusion for their antimicrobial susceptibilities. Inocula of E. coli and P. aeruginosa yielded acceptable results for up to 32 days; however, results on S. aureus were unacceptable due to loss of viability of the organism in water. E. coli and P. aeruginosa inocula, in water, can be used daily for quality control of the disk gear diffusion test.
Hippurate hydrolysis by Legionella pneumophila
Authors: Hébert G A
Published: January 2020

Strains of Legionella pneumophila were shown to hydrolyze sodium hippurate in an overnight test system, but strains of L. bozemanii, L. micdadei, L. dumoffii, and some other organisms similar to the legionellae did not. Although only a small number of strains of legionellae other than L. pneumophila have been classified and tested, the results indicate that the hippurate hydrolysis test may prove useful for differentiating among Legionella species.
Correlation of herpes simplex virus antibody titers and specific lymphocyte stimulation in adult blood donors
Authors: Moser H , Behrens F , Ziegelmaier R , Hilfenhaus J , Mauler R
Published: January 2020

Antibody titers to herpes simplex virus type 1 in sera from healthy adult donors were assayed by complement fixation, microneutralization, and an enzyme immunoassay (ELISA). This last test proved to be the most sensitive method for antibody detection. It was estimated that ELISA antibody titers were up to 40-fold higher than neutralizing antibody titers and up to 100-fold higher than complement fixation antibody titers. Due to the higher sensitivity of ELISA, only 3 of 36 blood donors tested in this assay were shown to be seronegative, whereas 6 additional persons of the same group were termed seronegative by the microneutralization assay. Furthermore, four of the latter also did not respond in the complement fixation test. In vitro stimulation of peripheral lymphocytes by using a partially purified herpes simplex virus type 1 particle antigen was achieved for all seropositive blood donors. Only those three donors who were ELISA negative reacted negatively in this stimulation assay. From these results it may be concluded that ELISA is an appropriate method not only for rapid and sensitive antibody determination but also for selecting herpes simplex virus-negative patients.
Microplate enzyme-linked immunosorbent assay for bovine leukemia virus antibody
Authors: Poli G , Balsari A , Toniolo A , Ponti W , Vacirca G
Published: January 2020

A microplate enzyme-linked immunosorbent assay method was developed for the measurement of bovine immunoglobulin G antibody specific to the envelope antigen (glycoprotein 60) of bovine leukemia virus. The test was then performed on 440 serum samples from dairy cows belonging to herds in which bovine leukemia was suspected or which were leukemia free, and the results were compared with those obtained with the gel-diffusion technique.
Analysis of tests used to differentiate Trichophyton rubrum from Trichophyton mentagrophytes
Authors: Sinski J T , Van Avermaete D , Kelley L M
Published: January 2020

Differential identification of Trichophyton rubrum and Trichophyton mentagrophytes was evaluated by using the standard in vitro hair perforation test as a criterion for identification and comparing its effectiveness with that of nine other tests used to differentiate these two taxa. Correlation and multiple regression analysis showed that no single test or combination of these tests was able to make the differentiation as effectively as the standard in vitro hair perforation test. Using relative error analysis, we suggest an identification scheme based upon selective utilization of potato-carrot agar for sporulation and the standard in vitro hair perforation test. Using potato-carrot agar for development of typical microconidia on each of the nonsporulating primary isolates and subsequently performing the standard in vitro hair perforation test only on those isolates that failed to produce spores on potato-carrot medium, we showed that this combination scheme provides more expedient results, without any sacrifice in reliability, than the standard in vitro hair perforation test alone.
Rapid method for identification of group B streptococci in neonatal blood cultures
Authors: Holmes R L , Harada W A
Published: January 2020

A rapid technique used for the identification of Streptococcus agalactiae, Lancefield group B, from the blood cultures of two neonatal infants is reported. The method utilized the Phadebact Streptococcus Test System (Pharmacia Diagnostics, Piscataway, N.J.) and the supernatant from 13- and 14-h blood cultures. Additional studies with simulated neonatal blood cultures revealed that this method was reproducible. Additional studies also revealed that some non-specific agglutination did occur, which could not be eliminated with dithiothreitol, but was visibly reduced by treatment with soluble staphylococcal protein A.
Antimicrobial susceptibility of Ureaplasma urealyticum
Authors: Davis J W , Hanna B A
Published: January 2020

An antimicrobial susceptibility test, a tow-tube broth dilution and disk elution method for Ureaplasma urealyticum, was modified to incorporate some of the standard procedures followed in traditional antimicrobial testing. The susceptibility pattern of the species was reevaluated by determining the effect of various antimicrobial agents on 21 vaginal isolates. All isolates were inhibited by tetracycline congeners (1 to 6 micrograms/ml) and killed by methenamine mandelate (0.6 mg/ml). All but one isolates were inhibited by erythromycin (0.4 to 3 micrograms/ml). Only eight isolates were inhibited by nalidixic acid (1 to 6 micrograms/ml), and seven were inhibited by nitrofurantoin (20 to 60 micrograms/ml), whereas all isolates were resistant to rifampin (1 microgram/ml) and trimethoprim-sulfamethoxazole (5 micrograms/ml). The in vitro technique described can readily be performed on individual patient isolates before the initiation of antimicrobial therapy.
Comparison of selective media for primary isolation of Campylobacter fetus subsp. jejuni
Authors: Patton C M , Mitchell S W , Potter M E , Kaufmann A F
Published: January 2020

Three selective media, Skirrow, Butzler, and a modification of Butzler medium, were compared for the primary isolation of Campylobacter fetus subsp. jejuni. This organism was isolated from 87 of 347 specimens (72 from 240 dogs rectal swabs and 15 from 107 cats rectal swabs). The positive rate for dogs (30%) was twice as high as that for cats (14%). Skirrow and Butzler media were comparable in their isolation of C. fetus subsp. jejuni. A significantly higher rate of positive results was obtained with modified Butzler medium. The best combination of two media was that of modified Butzler and Skirrow media, which detected 98% of the isolates obtained. The percentage of Campylobacter-positive specimens was increased by 9% by holding primary isolation plates 72 h.
Phospholipase D activity of Corynebacterium pseudotuberculosis (Corynebacterium ovis) and Corynebacterium ulcerans, a distinctive marker within the genus Corynebacterium
Authors: Barksdale L , Linder R , Sulea I T , Pollice M
Published: January 2020

A search has been made for corynebacterial phospholipase D, "ovis toxin," a sphingomyelinase (phosphatidylcholine phosphohydrolase, EC 3.1.4.4), among a wide variety of corynebacteria. Phospholipase D activity has been found in strains exhibiting the biochemical properties characteristic of Corynebacterium pseudotuberculosis or of Corynebacterium ulcerans and in no other species of Corynebacterium. Methods for the assay of phospholipase D as a sphingomyelinase and methods for screening for phospholipase D in the presence of Corynebacterium equi on washed sheep blood agar are discussed.
Serodiagnosis of La Crosse virus infections in humans
Authors: Calisher C H , Bailey R E
Published: January 2020

Serological diagnoses of human infections with California serogroup viruses are usually made by means of complement fixation or hemagglutination inhibition tests or both. An analysis of antibody titers in sera from 128 humans with California (La Crosse) virus infections indicated that exclusive use of the complement fixation tests would have detected only 50% of the actual seroconversions, whereas hemagglutination inhibition and neutralization tests alone would have been sufficient to make the diagnosis in 79.3% and 84.8% of the cases, respectively. These and other results presented demonstrate that the complement fixation test is not sufficiently useful as a primary tool for diagnosis of La Crosse virus infections. We suggest the use of the hemagglutination inhibition test for preliminary screening of sera from individuals with suspect infections caused by California serogroup viruses. The neutralization test should then be used or confirmation and subtype identification. If hemagglutination inhibition and neutralization titers are high and stable, the complement fixation test should then be used as a last, but specific, resort.
Dye-containing buffered charcoal-yeast extract medium for differentiation of members of the family Legionellaceae
Authors: Vickers R M , Brown A , Garrity G M
Published: January 2020

The addition of 0.001% bromocresol purple and 0.001% bromothymol blue to buffered charcoal-yeast extract agar allowed differentiation between members of the family Legionellaceae. On this medium, Legionella pneumophila grew as relatively flat, pale green colonies, whereas Tatlockia micdadei (gen. nov., comb. nov., Pittsburgh pneumonia agent) produced blue-gray colonies. Fluoribacter spp. (gen. nov., atypical Legionella-like organisms) developed glistening colonies which were brighter green than those of L. pneumophila.
Simple procedure for measuring neutrophil chemiluminescence
Authors: Prendergast E , Proctor R A
Published: January 2020

A simple method is described whereby measurement of neutrophil chemiluminescence was performed at variable reaction temperatures even though the scintillation counting chamber was refrigerated.
Rapid detection of yeast enzymes by using 4-methylumbelliferyl substrates
Authors: Bobey D G , Ederer G M
Published: January 2020

In a preliminary study with a limited number of isolates, the usefulness of 17 4-methylumbelliferyl substrates for identifying yeast isolates was investigated. Substrates to detect acid phosphatase, glucosidase, and pyrophosphate diesterase showed promise.
Antigenically intermediate human adenovirus strain associated with conjunctivitis
Authors: Hierholzer J C , Rodriguez F H
Published: January 2020

Two strains of an antigenically intermediate human adenovirus were isolated from eye specimens from cases of conjunctivitis. The viruses were doubly neutralized by antisera to adenoviruses 15 and 29 but possessed the hemagglutinins of adenovirus 8 or 9.
Safranin O-stained antigen microagglutination test for detection of brucella antibodies
Authors: Brown S L , Klein G C , McKinney F T , Jones W L
Published: January 2020

A microagglutination test was compared with a standard tube agglutination test for the detection of brucella antibodies. Advantages of the microagglutination test were that it required less time to perform, had a shorter incubation period, and used less antigen.
Enzyme-linked immunosorbent assay for detection of immunoglobulin M antibody to hepatitis B core antigen
Authors: Kryger P , Mathiesen L R , Møller A M , Aldershvile J , Hansson B G , Nielsen J O
Published: January 2020

An enzyme-linked immunosorbent assay for detection of specific immunoglobulin M (IgM) antibodies against the core antigen of the hepatitis B virus (anti-HBc IgM) is described. The interference of IgM rheumatoid factor was evaluated quantitatively. In the anti-HBc IgM test, the rheumatoid factor gave false-positive results when the concentration exceeded 20 IU/ml. The rheumatoid-positive sera were disclosed by a control and retested for anti-HBc IgM after absorption of rheumatoid factor with latex particles aggregated with human IgG. In five of seven selected patients with acute hepatitis B followed to biochemical and clinical recovery, anti-HBc IgM was present transiently until antibodies against hepatitis B surface antigen (anti-HBs) appeared. Two patients had persistent anti-HBc IgM during the follow-up period. Four patients with hepatitis B surface antigenemia and progression to chronic liver disease did not clear their anti-HBc IgM in the period of observation (11 to 24 months). Anti-HBc IgM could not be demonstrated in 223 of 225 Danish blood donors. The two donors found positive for anti-HBc IgM also had anti-HBs. Twenty patients with acute A or non-A non-B hepatitis were negative for anti-HBc IgM. The enzyme-linked immunosorbent assay for anti-HBc IgM described here has a high specificity and sensitivity. The diagnostic relevance needs further evaluation, including quantitation of anti-HBc IgM, but the results presented indicate that anti-HBc IgM may be helpful in differentiating between prior and recent or ongoing hepatitis B infection.
Hemadsorption immunosorbent technique for determination of rubella immunoglobulin M antibody
Authors: van der Logt J T , van Loon A M , van der Veen J
Published: January 2020

A highly specific and sensitive hemadsorption immunosorbent technique for measuring rubella immunoglobulin M (IgM) antibody is described. IgM from human sera was absorbed into anti-human IgM-coated wells in plates and rubella-specific IgM was detected by adding rubella virus hemagglutinin and a small quantity of sheep erythrocytes. Centrifugation of the plates facilitated reading of the test. Specific IgM-positive sera showed hemadsorption, whereas negative sera showed hemagglutination. Rheumatoid factor and rubella-specific IgG antibody did not interfere with the results. The test was clearly more sensitive than the solid-phase immunosorbent technique for detection of rubella IgM antibody by hemagglutination inhibition and at least as sensitive as the hemagglutination inhibition test on IgM fractions from a sucrose density gradient and the indirect immunofluorescence test for IgM antibody with absorbed serum. All of 40 sera from 17 rubella patients taken 4 to 49 days after the onset of rash were positive in the new test, with antibody titers ranging from 2,560 to 81,920 between 4 and 28 days. The test is reliable, practical, and suitable for general diagnostic use.
Enzymatic profile of Legionella pneumophilia
Authors: Müller H E
Published: January 2020

The enzyme activities of four strains of Legionella pneumophilia were investigated by using the API ZYM system (API System S.A., F-38390 Montalieu Vercieu, France) and synthetic substrates. Aminopeptidases were detected specifically against L-alanine, L-arginine, L-aspartic acid, L-cystine, L-glutaminic acid, glycine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-tryptophan, L-tyrosine, and L-valine. Furthermore, the bacteria possesses esterase activity splitting propionate, butyrate, caproate, caprylate, and caprate, but not laurate, myristate, palmitate, and stearate, esters. The enzymes studies were inhibited partially by aprotinin. No inhibition of phosphatase (pH range, 5.4 to 8.5) or of phosphoamidase was observed. Activities of arylsulfatase, chymotrypsin, trypsin, and glycosidases could not be detected.
Amino acid requirements of a Chlamydia trachomatis genital strain in McCoy cell cultures
Authors: Karayiannis P , Hobson D
Published: January 2020

Amino acid requirements of a Chlamydia trachomatis were defined in McCoy cell monolayer cultures, depleted of their nutrient reserves by maintenance in Earle balanced salt solution for at least 2 days, and then treated at infection with 1 microgram of cycloheximide per ml, an inhibitor of host cell protein and deoxyribonucleic acid synthesis. Serum-free Waymouth MB 752/1 medium lacking one amino acid or a group of related amino acids was added to depleted cultures before infection. Eight of the amino acids normally present in MB 752/1 were essential for chlamydial growth, six were advantageous for growth, and five were nonessential.
Evaluation of transport and storage techniques for isolation of Campylobacter fetus subsp. jejuni from turkey cecal specimens
Authors: Luechtefeld N W , Wang W L , Blaser M J , Reller L B
Published: January 2020

Immediate culturing of fecal specimens is not always possible, and appropriate methods for transport and storage of Campylobacter fetus subsp. jejuni specimens have not been fully evaluated. Using nine techniques, we studied the survival of C. fetus subsp. jejuni in cecal specimens from infected turkeys. The organisms survived in specimens held without transport medium for 3 to 15 days (median, 9 days) at 4 degrees C, and 2 to 9 days (median, 4 days) at 25 degrees C. Only 20% of specimens frozen for 24 h at either -20 or -70 degrees C yielded C. fetus subsp. jejuni. Specimens dried on filter paper strips were negative for C. fetus subsp. jejuni within 1.5 h. Cary-Blair medium with decreased agar was the best of the six transport media tested, it enabled recovery of the organism from 100% (3 days) and 71% (7 days) of cecal samples held at 4 degrees C and 94% (3 days) and 85% (7 days) of cecal specimens held at 25 degrees C. In contrast, more than half of all cecal specimens held at 4 or 25 degrees C in Culturettes or buffered glycerol saline were negative by 3 days, and all were negative at 7 days. Results with the other three media studied (Campy-thio, thioglycolate medium, and alkaline peptone water) were intermediate. Overnight incubation of specimens in alkaline peptone water at 37 or 42 degrees C did not enhance recovery of C. fetus subsp. jejuni. Therefore, refrigeration without a transport medium is satisfactory for up to 3 days for recovery of C. fetus subsp. jejuni from specimens, however, we recommend the use of Cary-Blair medium with decreased agar for specimens that must be transported or stored for longer than 3 days and for rectal swabs, to prevent drying.
Comparison of spot esculin hydrolysis with the PathoTec strip test for rapid differentiation of anaerobic bacteria
Authors: Qadri S M , Johnson S , Smith J C , Zubairi S , Gillum R L
Published: January 2020

The ability of several anaerobic bacteria to hydrolyze esculin to esculetin is used by clinical microbiologists and taxonomists in the differentiation and identification of both gram-positive and gram-negative microorganisms. Conventional methods used for determining esculin hydrolysis by anaerobic bacteria require 24 to 48 h for completion. In this paper we evaluate two procedures which yield rapid results. A total of 738 anaerobic bacteria were used in this study. A total of 99% of the esculin-hydrolyzing anaerobic bacteria gave positive results with the spot test in 1 h, whereas the other test method, the PathoTec strip test (General Diagnostics, Morris Plains, N.J.), required 4 h for 96% of the strains tested to yield positive reactions. Both tests showed a 100% specificity when compared with the standard broth test and are easy to perform, accurate, and economical. The spot test is superior to the PathoTec strip test in yielding results more rapidly.
Rapid identification of Enterobacteriaceae with microbial enzyme activity profiles
Authors: Godsey J H , Matteo M R , Shen D , Tolman G , Gohlke J R
Published: January 2020

A total of 539 clinical isolates belonging to 10 species of the Enterobacteriaceae family were identified by enzyme activity profiles within 30 min of test inoculation. Each isolate was grown at 37 degrees C for 18 h on Mueller-Hinton agar and suspended to an optical density of 200 Klett units on 0.85% saline. Enzyme activity profiles were obtained by inoculating 18 fluorogenic substrates with the standardized bacterial suspension and monitoring initial rates of hydrolysis over the first 30 min of analysis. Individual enzyme activity profiles were entered into a coded data bank, and identifications were based on the Bayesian theory of probabilities. At a confidence level of 95%, five species were identified with a greater than 90% efficiency, three species were identified between 83 and 88% efficiency, and two species demonstrated a 72 and 75% efficiency of identification. The enzyme activity profile method of bacterial identification is rapid, easily automated, and reproducible.
Assessment of Haemophilus influenzae type b opsonins by neutrophil chemiluminescence
Authors: Kaplan S L , Umstead C L , Mason E O , Anderson D C , Parke J C , Feigin R D
Published: January 2020

A luminol-enhancement chemiluminescence assay and a radiolabeled uptake assay were developed to assess opsonins for Haemophilus influenzae type b. Opsonins in acute and convalescent sera from 17 children with H. influenzae type b meningitis, along with pooled normal human sera, were evaluated and compared with anti-polyribosephosphate antibody concentrations. Five children had a rise in the chemiluminescence-area under the curve for convalescent compared with acute sera. Patient chemiluminescence--area-under-the-curve values were significantly (P less than 0.05) more likely to exceed 50% of normal human serum values if sera contained greater than or equal to 0.1 microgram of anti-polyribosephosphate antibody per ml. Magnesium ethylene glycol tetraacetic acid chelation and heat inactivation of patient and normal human sera significantly (P less than 0.05) reduced chemiluminescence--area-under-the-curve activity. Thus, complement appears to contribute significantly to the opsonization of H. influenzae type b in sera of children. Two of nine children had increases in opsonins as assayed by 3H-labeled H. influenzae type b uptake. After natural systemic H. influenzae type b infection, young children are unable to respond acutely with an increase in anti-polyribosephosphate antibody or serum opsonic activity.
Quantitative nephelometric determination of Haemophilus influenzae antigen in body fluids
Authors: Caceci T , Brook I , Daniel A
Published: January 2020

Nephelometry, an immunological technique widely used for the quantification of blood proteins, was adapted to provide a quantitative method of detecting Haemophilus influenzae capsular antigen in body fluids. Using specific antiserum directed against H. influenzae capsular antigen, samples of serum, cerebrospinal fluid, urine, and joint fluid from 38 cases of H. influenzae infections were analyzed. The results were compared for reliability to counterimmunoelectrophoresis, a widely used diagnostic tool. The nephelometric technique has the same advantages of speed and specificity as counterimmunoelectrophoresis and provides the clinician and researcher with a quantitative method that is as reliable as the qualitative counterimmunoelectrophoresis procedure. The method allowed directly quantitative readouts on patient specimens, with no necessity for serial dilutions or densitometric readings.
Distribution of serotypes of Nocardia asteroides from animal, human, and environmental sources
Authors: Pier A C , Fichtner R E
Published: January 2020

The antigenic types of 129 isolates of Nocardia asteroides from diverse clinical, environmental, and geographic origins were determined. The majority of the isolates studied were of bovine (56) or human (44) origin; 11 were derived from six species of animals other than cattle, and 10 were isolated from environmental sources; the source of 8 strains could not be determined. Testing culture filtrate antigens against four standard reference sera in a gel diffusion precipitin test established the antigenic type of 95.3% of the isolates. After excluding strains that weighted the data because of common infection, the distribution of serotypes was examined according to the origin of the isolate. Type I was the most frequently encountered serotype (31.9%); types III (15.0%) and IV (20.4%) were also observed frequently, as was the antigenic mixture III + IV (14.2%). There was an apparent difference in frequency of type III and IV antigens among isolates of bovine and human origin; type III made up 20.0% of the bovine isolates and 13.6% of the human isolates, whereas type IV constituted 10.0% of bovine and 27.3% of human isolates.
Hemagglutinin-specific enzyme-linked immunosorbent assay for antibodies to influenza A and B viruses
Authors: Murphy B R , Phelan M A , Nelson D L , Yarchoan R , Tierney E L , Alling D W , Chanock R M
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies present in human serum or nasal washes directed against influenza A or B hemagglutinin glycoproteins. The assay was modified to measure the immunoglobulin isotype specificity of the anti-hemagglutinin response in serum and nasal secretions. In the postinfection sera anti-hemagglutinin of the immunoglobulin G isotype was predominant, whereas in nasal secretions the antibody was predominantly immunoglobulin A. The antibody response detected by the ELISA manifested hemagglutinin subgroup specificity. In addition, there was a good correlation between the ELISA antibody titer and the hemagglutination-inhibition or neutralizing antibody titer. The ELISA was more sensitive than the hemagglutination-inhibition assay, and the range of antibody titers measurable by ELISA in human serum was from less than 1:20 for children who had never experienced influenza infection to 1:400,000 for adults convalescing from a secondary infection. With more sensitive tests to detect antibody to the influenza hemagglutinin it should be possible to determine the relative contribution of local and systemic immunity to resistance to influenza virus infection.
Characteristics and biotypes of Pasteurella multocida isolated from humans
Authors: Oberhofer T R
Published: January 2020

Fifty-two isolates of Pasteurella (48 strains of Pasteurella multocida and 4 strains of atypical Pasteurella) were identified by conventional and commercial test systems. All strains fermented glucose, sucrose, and fructose in purple broth base (Difco Laboratories) with bromocresol purple as indicator, although the atypical Pasteurella produced fermentation reactions that were barely perceptible. Eleven different biotypes were identified by fermentation reactions in maltose, mannitol, xylose, sorbitol, and trehalose media. There was a correlation of biotypes to cat bites, with 61% of cat bite isolates falling into biotype A and B. A correlation of biotype and dog bite isolates was not seen. The choice of medium used for fermentation tests was critical as evidenced by the inability of the organisms to grow in a second commercially purchased preparation of purple broth base. The reliability of commercial test systems in identifying Pasteurella was 81% for Oxi/Ferm (Roche Diagnostics, Div. Hoffmann-La Roche, Inc., Nutley, N.J.), 68% for API (Analytab Products, Plainview, N.Y.), and 11% for Minitek (BBL Microbiology Systems, Cockeysville, MD.).
Evaluation of 36 Minitek tests and a new approach for identification of nonfermenters
Authors: Yabuuchi E , Yamanaka K , Ohyama A
Published: January 2020

Thirty-six Minitek (BBL Microbiology Systems, Cockeysville, Md.) tests were evaluated with 625 Kansai Medical University stock strains of 22 species and one group of nonfermentative gram-negative rods able to grow on ordinary peptone media. Among the 36 tests, 15 were selected because a clear-cut reaction was shown by all 625 Kansai Medical University strains. Of these 15 tests, 12 were further selected for routine use because they were regarded as useful for the identification of nonfermenters. The 12 tests were arranged into the following four groups: (i) lysine-arginine-ornithine, (ii) urea-ortho-nitrophenyl-beta-D-galactopyranoside-dextrose aerobic, (iii) maltose-xylose-starch, and (iv) esculin-nitrate reduction-indole. A new profile system for four digits, the Minitek Y-Y (Yabuuchi and Yamanaka) system, consisting of 64 numbers which represent each single species and 11 numbers which give two to four species, is herein proposed. The system was designed primarily for a less expensive identification of gram-negative rods already confirmed in a butt of either triple sugar iron or Kligler iron agar for their lack of ability to ferment dextrose. Among the 539 clinical isolates obtained from 3 hospitals, 511 strains identifiable by classical methods were also identified by the Minitek Y-Y system.
Use of guinea pig embryo cell cultures for isolation and propagation of group A coxsackieviruses
Authors: Landry M L , Madore H P , Fong C K , Hsiung G D
Published: January 2020

The isolation of group A coxsackieviruses from clinical specimens generally requires the use of suckling mice. By using guinea pig embryo cells, the following coxsackieviruses were isolated from throat swabs and stool samples obtained from patients with a variety of illnesses: two of type A2, one each of types A6 and A8, and four of type 10. Distinct cytopathic effects were produced in 3 to 5 days in the guinea pig embryo cells inoculated with the clinical specimens. In addition, a number of prototype group A coxsackieviruses, including types 2--6, 8, 10, and 12, were readily propagated in guinea pig embryo cell cultures. Thus, guinea pig embryo cells appeared to be a sensitive alternative cell culture system for the isolation and propagation of certain types of group A coxsackieviruses.
Simple Method for Detecting
Bacteroides
spp. Bacteriocin Production
Authors: Riley Thomas V. , Mee Brian J.
Published: January 2020

Bacteroides
isolates were grown anaerobically on a 0.22-μm membrane filter on an agar plate for 48 h. Cultures of an indicator strain were grown as a lawn on the agar after removal of the filter, and bacteriocin sensitivity was detected by zones of inhibition.
Capsular typing of klebsiellae by coagglutination and latex agglutination
Authors: Onokodi J K , Wauters G
Published: January 2020

A simple method for capsular serotype determination of Klebsiella is described, using staphylococci or latex particles coated by specific antibodies and the supernatant of a bacterial suspension in saline as antigen. These methods may be at least as reliable as other methods and easier to perform and less expensive than other methods.
Co-trimoxazole susceptibility tests improved with separate trimethoprim and sulfamethoxazole disks
Authors: Amyes S G
Published: January 2020

It is impossible to test accurately bacterial susceptibility to the trimethoprim-sulfamethoxazole combination co-trimoxazole with a single combined susceptibility disk. However, a variety of factors still affect the result even when separate trimethoprim and sulfamethoxazole disks are used. Experiments with separate disks showed that the optimum conditions for testing the susceptibilities of enterobacteria to these drugs were to flood-seed an agar plate with an inoculum of 10(4) to 10(5) organisms per ml, take off the excess liquid, and place a disk of 1 microgram of trimethoprim and another of 50 micrograms of sulfamethoxazole on the surface of the agar with their centers exactly 25 mm apart. This method not only allowed the determination of resistance but also distinguished synergy.
Immunoglobulin M antibodies to hepatitis B core antigen: evaluation of enzyme immunoassay for diagnosis of hepatitis B virus infection
Authors: Roggendorf M , Deinhardt F , Frösner G G , Scheid R , Bayerl B , Zachoval R
Published: January 2020

In acute and chronic hepatitis B, antibodies of the immunoglobulin M (IgM) class against the hepatitis B core antigen (anti-HBc IgM) have been demonstrated. For the determination of anti-HBc IgM, a sensitive enzyme immunoassay with anti-mu-coated flat-bottomed microtiter plates is described and evaluated. The specificity of the anti-HBc IgM test system was proven by pretreatment of presumed anti-HBc IgM-positive samples with anti-mu to block anti-HBc IgM. The test system was highly sensitive. In the acute stage of hepatitis B, anti-HBc IgM could be demonstrated in serum dilutions up to 10(-7) (mean titer, 10(-5)), and in sera from patients with chronic hepatitis B, the mean titer was 10(-3). In a study of unselected patients whose sera were sent at irregular intervals for testing, anti-HBc IgM persisted in a high percentage (52%) for at least 13 to 18 months after onset of illness despite the fact that these patients eliminated hepatitis B surface antigen (HBsAg) and produced antibodies to HBsAg (anti-HBs). By using the anti-HBc IgM test as an additional aid in the diagnosis of acute HBsAg-negative hepatitis, the hepatitis B etiology could be established in 13 of 42 patients (31.4%). Investigations of the prevalence of anti-HBc IgM in different groups of patients with chronic hepatitis B infection showed 89.4% anti-HBc IgM-positive results in patients with chronic active hepatitis B, 60% in patients with HBsAg-negative chronic active hepatitis, 58.2% in patients with primary liver carcinoma and markers of hepatitis B infections, and 34.9% in healthy carriers of HBsAg.