bluuebunny/crossref_metadata_2025 huggingface dataset:
Aeromonas hydrophila typing scheme based on patterns of agglutination with erythrocytes and yeast cells
Authors: Adams D , Atkinson H M , Woods W H
Published: January 2020

An agglutination typing scheme has been developed for strains of Aeromonas hydrophila. Primary agglutination typing is based on testing agar-grown A. hydrophila cells with human, horse, rat, and guinea pig erythrocytes and Saccharomyces cerevisiae cells. Further subdivision of primary groups is based firstly on whether yeast cell agglutination is inhibited by a D-mannose polymer, yeast mannan, and secondly on patterns of inhibition of hemagglutination by yeast mannan and the monomeric sugars L-fucose, D-galactose, and D-mannose. A total of 320 isolates were tested, and these were divisible into 39 distinct types on the basis of this scheme. Application of this typing scheme in the future to isolates of A. hydrophila known to be associated with human infection may enable correlations to be made between particular agglutination types and human pathogenicity.
Comparison of Passive Hemagglutination with Turkey Erythrocyte Assay, Enzyme-Linked Immunosorbent Assay, and Counterimmunoelectrophoresis Assay for Serological Evaluation of Tetanus Immunity
Authors: Pitzurra L. , Bistoni F. , Pitzurra M. , Bastianini L. , Perito S. , Vecchiarelli A. , Marconi P.
Published: January 2020

Antibody titers to tetanus toxin in human sera were assayed by passive hemagglutination with turkey erythrocytes, enzyme-linked immunosorbent assay, and counterimmunoelectrophoresis. The first two of these tests were shown to be the most sensitive for antibody detection, having the same range of sensitivity and reproducibility. The antibody levels determined by these assays were up to 400-fold higher than those determined by counterimmunoelectrophoresis. The turkey erythrocyte hemagglutination assay requires only 40 min, whereas the immunosorbent assay method requires 24 h. These results suggest that the hemagglutination assay is the more appropriate method for rapid and sensitive determination of tetanus antibody levels.
Production in guinea pigs of antibodies to cross-reactive antigenic determinants of human enteroviruses
Authors: Hasegawa A , Inouye S
Published: January 2020

We studied the type specificity of the antisera which were produced in guinea pigs by immunization with heat-disrupted virions of human enteroviruses (echovirus types 7 and 11 and coxsackievirus type B5). When guinea pigs were immunized with heated virions (H particles), the antibodies produced reacted type specifically by complement fixation with the homologous H particles. By contrast, when guinea pigs were immunized with disrupted virions prepared by heating in the presence of sodium dodecyl sulfate, antibodies were produced which reacted broadly. Heterotypic complement-fixing antibody responses of humans to enterovirus infections are discussed.
Comparison of reticulate and elementary body antigens in detection of antibodies against Chlamydia trachomatis by an enzyme-linked immunosorbent assay
Authors: Jones R B , Bruins S C , Newhall W J
Published: January 2020

Reticulate bodies from a type C and elementary bodies from a type L2 strain of Chlamydia trachomatis were isolated and used as antigens in an enzyme-linked immunosorbent assay (ELISA). Results obtained for human sera with these two antigens used in the ELISA were compared with each other and with results obtained for the same sera by the micro-immunofluorescence test. Negative control populations included cloistered nuns and children with respiratory infections. Populations at risk for chlamydial infection consisted of 42 men with nongonococcal urethritis attending a sexually transmitted diseases clinic and 42 college women who had contact with men with nongonococcal urethritis. ELISAs done with the two antigens were equivalent to each other and to the micro-immunofluorescence test in the ability to predict the presence or absence of infection. None of the tests had high predictive values for the men with urethritis. However, the negative predictive value of both the micro-immunofluorescence test and the elementary body ELISA was 0.92 for the college women. Such serological tests may be of value in screening selected populations for subclinical infections with C. trachomatis.
Chloramphenicol and penicillin resistance in pneumococci isolated from blood and cerebrospinal fluid: a prevalence study in metropolitan Denver
Authors: Istre G R , Humphreys J T , Albrecht K D , Thornsberry C , Swenson J M , Hopkins R S
Published: January 2020

From January through October 1981, we screened blood and cerebrospinal fluid pneumococcal isolates from 101 patients in the Denver, Colorado area. Isolates from seven patients (6.9%) showed relative resistance to penicillin, with minimal inhibitory concentrations ranging from 0.12 to 1.0 microgram/ml. Two isolates (2.0%) were resistant to chloramphenicol, both with a minimal inhibitory concentration of 16 micrograms/ml. One of these was multiply resistant (to penicillin, chloramphenicol, and tetracycline). All isolates were susceptible to erythromycin, sulfamethoxazole-trimethoprim, and rifampin. On the basis of penicillin susceptibilities performed by participating hospitals on the isolates from 215 patients in the Denver area (101 included in the survey, 114 not included), we estimated the rate of relative resistance to penicillin to be approximately 4.3%. Compared with resistance rates reported in a previous study in Denver, these penicillin and chloramphenicol resistance rates may represent a trend of increasing resistance to these antibiotics in the Denver area. We recommend screening all isolates from invasive pneumococcal infections for penicillin and chloramphenicol susceptibility.
Cross-reactivity of Haemophilus somnus antibody in agglutination and complement fixation tests and in the enzyme-linked immunosorbent assay
Authors: Canto J , Biberstein E L , Schulte T A , Behymer D
Published: January 2020

The specificity and sensitivity of agglutination, complement fixation, and enzyme-linked immunosorbent assay (ELISA) procedures in the detection of antibodies to Haemophilus somnus was investigated. H. somnus rabbit immune sera were found to agglutinate Pasteurella multocida, Staphylococcus aureus, and Haemophilus agni and, in some instances, also Pasteurella haemolytica, Salmonella dublin, Streptococcus agalactiae, and Corynebacterium pyogenes. In complement fixation tests with saline extracts as antigens, only H. agni reacted with H. somnus antisera to any significant degree. In ELISA tests with sonicated or heat-extracted antigens, cross-reactions were seen with the two Pasteurella spp. and with H. agni. When whole cells and saline extracts were used as antigens in ELISAs, only H. agni showed any cross-reactivity. The greatest specificity in distinguishing homologous from heterologous reactions was achieved by ELISA with saline extracts as antigens. Escherichia coli and Brucella abortus antigens failed to react with H. somnus antibody in any of the tests. A rabbit serum containing antibody to bovine type isolates of P. multocida, P. haemolytica, S. aureus, S. agalactiae, S. dublin, C. pyogenes, and E. coli gave no positive reaction in ELISA tests with saline extract of H. somnus as antigen. It is concluded that such saline extract, which appears to consist largely of H. somnus common antigen, has the potential of being a useful diagnostic reagent in the study by ELISA of antibody response to H. somnus.
Characterization of the hemolytic activity of Staphylococcus aureus strains associated with toxic shock syndrome
Authors: Chow A W , Gribble M J , Bartlett K H
Published: January 2020

The hemolytic activity of 32 vaginal isolates of Staphylococcus aureus from patients with typical toxic shock syndrome (TSS) was contrasted with that of 50 vaginal isolates from patients without TSS, using a standardized inoculum (10(5) CFU) on 5% sheep blood agar after 48 h of incubation under 30% CO2. Additionally, 7 nongenital isolates from patients with nonmenstrual TSS and 57 strains of nongenital control isolates were included for comparison. Vaginal TSS strains were significantly less hemolytic than non-TSS S. aureus strains of either genital (P less than 0.001) or nongenital (P less than 0.01) origin. Vaginal TSS S. aureus strains were also less hemolytic than were nongenital TSS S. aureus strains (P less than 0.02). This reduced hemolytic activity of genital TSS S. aureus strains may provide a useful marker for screening and further delineation of toxigenic S. aureus associated with menstrually related TSS.
Distinctive biochemical features of Salmonella dublin isolated in California
Authors: Fierer J , Fleming W
Published: January 2020

We examined 34 strains of Salmonella dublin that were isolated in California between 1978 and 1982. All were of a characteristic biotype; they did not grow on Simmons citrate or acetate and did not ferment arabinose. Their apparent inability to use citrate as the only carbon source was due to a nutritional requirement for nicotinic acid. Because S. dublin strains are of a characteristic biotype, are host adapted to bovines, and are unusually virulent for humans, we suggest that S. dublin be considered a separate species of the genus Salmonella. It is important that clinical laboratories recognize and differentiate this organism from less pathogenic salmonellae so that they can alert clinicians to the presence of this invasive microorganism.
Evaluation of a commercial counterimmunoelectrophoresis kit for detection of Staphylococcus aureus teichoic acid antibodies
Authors: West T E , Burdash N M , Boehm A M , West M E
Published: January 2020

A commercial kit from Diagnostica, Inc., Miami Fla., was studied for its ability to detect antibodies to the teichoic acids of Staphylococcus aureus. A comparative study of the Diagnostica counterimmunoelectrophoresis (CIE) system and our gel double-diffusion method was undertaken with 156 serum samples from 142 patients. Included were 25 cases of staphylococcal and non-staphylococcal endocarditis, 30 cases of S. aureus bacteremia, 19 cases of nonbacteremic S. aureus infection, 39 cases of hospitalized patients without a staphylococcal infection, and 29 normal controls. Agreement between methodologies was attained in 138 (88.5%) of the 156 samples tested and in 127 (89.4%) of the 142 patients. Of 13 patients with culture-proven S. aureus endocarditis, significant antibody titers were found in all patients (100%) by CIE and in 12 patients (92.3%) by double diffusion. No significant titers were found in normal sera by CIE, but four sera were positive by double diffusion. Of 80 sera from patients with no evidence of S. aureus infection, 4 (5.0%) were positive by CIE and 7 (8.8%) were positive by double diffusion. The Diagnostica CIE kit appears to provide a suitable means for the detection of deep-seated S. aureus infections.
Detection by a staphylococcal coagglutination test of heat-labile enterotoxin-producing enterotoxigenic Escherichia coli
Authors: Honda T , Samakoses R , Sornchai C , Takeda Y , Miwatani T
Published: January 2020

For detection of heat-labile enterotoxin-producing enterotoxigenic Escherichia coli, the staphylococcal coagglutination test reported by Brill et al. (J. Clin. Microbiol. 9:49-55, 1979) was modified to give better results. Staphylococcal cells were sensitized with anti-heat-labile enterotoxin antiserum and suspended in phosphate-buffered saline containing 0.5% bovine serum albumin, 0.05% Tween 80, 0.01% gelatin, and 0.02% NaN3. The test strain was cultured in 0.25 ml of Biken broth no. 2 in a test tube (12 by 100 mm) stood at an inclination of about 10 degrees to the horizontal. After incubation for 5 h at 37 degrees C, the cells were collected by centrifugation at 2,500 rpm for 15 min and suspended in 50 microliters of polymyxin B solution (20,000 IU/ml). The suspension was then incubated for 1 h at 37 degrees C and centrifuged at 2,500 rpm for 15 min, and 10 microliters of the supernatant was used for the test on a slide. The results of the modified test correlated completely with those obtained by the Biken test. The modifications of the staphylococcal coagglutination test described here allow for detection of heat-labile enterotoxin-producing enterotoxigenic E. coli within 6 to 7 h after inoculation of a test strain.
Rapid GM1-enzyme-linked immunosorbent assay with visual reading for identification of Escherichia coli heat-labile enterotoxin
Authors: Svennerholm A M , Wiklund G
Published: January 2020

A modified, quicker and simpler GM1-enzyme-linked immunosorbent assay procedure for detection of Escherichia coli heat-labile enterotoxin (LT) has been developed with the intent that it should be useful in less well-equipped laboratories. The method, which makes use of stable reagents including commercially available horseradish peroxidase immunoglobulin conjugate, can detect LT in overnight cultures within 1 working day (8 h), and the tests can be read with the naked eye. This GM1-horseradish peroxidase-enzyme-linked immunosorbent assay shows excellent quantitative and qualitative correlation with the conventional GM1-enzyme-linked immunosorbent assay. When 100 human E. coli strains were analyzed blindly and in parallel by the two methods, LT production was identified in 50 out of 50 LT-positive strains and in 0 out of 50 LT-negative strains by either method.
Broadly reactive immunofluorescence test for measurement of immunoglobulin M and G antibodies to Ureaplasma urealyticum in infant and adult sera
Authors: Gallo D , Dupuis K W , Schmidt N J , Kenny G E
Published: January 2020

The indirect immunofluorescence test was used to measure immunoglobulin M (IgM) and IgG antibodies to acetone-fixed Ureaplasma urealyticum organisms in sera from 128 adults with genital infections and from 713 symptomatic newborns and babies 1 day to 18 months old. Thirty-four percent of the adults had demonstrable IgG antibody to ureaplasma. IgM antibody was detected in 2 of the adult sera and in 17 of the infant sera. These babies were divided into two distinct groups. Ten of the infants presented at birth with various physical findings, whereas the onset of symptoms for the other 7 occurred 3 to 13 weeks after birth, and the major clinical finding in 6 of the 7 was respiratory distress. The results of this study suggested that U. urealyticum infection may be associated with fetal damage and infant pneumonia, and if this is substantiated, the indirect immunofluorescence test employing acetone-fixed antigen to measure IgM antibody to U. urealyticum may be an important diagnostic tool.
Limitations of available tests for diagnosis of infectious mononucleosis
Authors: Fleisher G R , Collins M , Fager S
Published: January 2020

Among 500 students seen at a university health service for illnesses resembling infectious mononucleosis (IM), the diagnosis of IM was established in 124 (25%) on the basis of the initial presence or subsequent emergence of the spectrum of Epstein-Barr virus (EBV) antibodies characteristic of a primary EBV infection. Of these 124 patients, 113 had an EBV-specific antibody pattern in the initial serum indicative of current primary infection; however, 11 (9%) had no detectable immunoglobulin G antibodies to EBV-specific antigens in their first serum. The sensitivity of this panel of EBV antibody assays was 91% and the specificity was 100%. Initial sera had detectable heterophil antibodies for 107 (86%) of the 124 students with IM and for 2 with other illnesses. Among our patients, the Monospot (Ortho Diagnostics Inc.) test had a sensitivity of 86% and a specificity of 99%. Reliance on hematological criteria (lymphocyte count greater than or equal to 50% and atypical lymphocyte count greater than or equal to 10%) gave a sensitivity of only 39%, but a specificity of 99%. Students with IM who showed a delayed emergence of the spectrum of EBV-specific antibodies characteristic of an acute infection were compared with control patients who had such antibodies at the time of their initial visit to the health service. They were found to have a briefer duration of illness (P greater than 0.05), lower leukocyte (P less than 0.005), lymphocyte (P less than 0.005), and atypical lymphocyte (P less than 0.05) counts, and a less frequent occurrence of heterophil antibodies (P less than 0.05).
Comparison of serological reactions of typed Fusobacterium nucleatum strains with those of isolates from humans, canines, and a Macaca mulatta monkey
Authors: Vincent J W , Falkler W A , Craig J A
Published: January 2020

In the present study, we compared typed F. nucleatum strains with isolates from various human periodontal diseases, canines, and a Macaca mulatta monkey. All isolates displayed biochemical reactions similar to those of the typed strains in the API 20A system (Analytab Products, Plainville, N.Y.). The human and monkey isolates displayed both type I and type II colonial morphologies on crystal violet erythromycin agar, whereas the canine isolates displayed only type I. Antigen preparations of the human isolates, typed strains, and two of the monkey strains shared lines of identity in immunodiffusion testing. An enzyme-linked immunosorbent assay in which rabbit anti-F. nucleatum and human sera were used revealed that the human and monkey isolates, when used as antigens, allowed the detection of similar degrees of antibody activity; however, no activity was observed with the canine isolates. None of the canine isolates displayed the characteristic hemagglutination of sheep erythrocytes demonstrated by the human and monkey strains. These results suggest that F. nucleatum isolates from various human periodontal diseases share antigenic determinants. Also, differences were observed in F. nucleatum isolates from canines when compared with human and monkey strains. These findings are important when considering the immunopathological mechanisms involved in human periodontal diseases and when comparing animal model systems with naturally occurring disease in humans.
Enzyme-linked immunosorbent assay for antibodies against teichoic acid in patients with staphylococcal infections
Authors: Granström M , Julander I G , Hedström S A , Möllby R
Published: January 2020

A highly purified teichoic acid preparation was used in an enzyme-linked immunosorbent assay to measure the specific immunoglobulin G (IgG) and IgM response in staphylococcal disease. Antibody determination in a normal population, showing a difference of up to 20-fold in the mean IgG titers between the youngest children and adults, was used to establish age-correlated upper normal values. IgM antibodies were found to be of little diagnostic value since their response was often low or absent. Increased IgG titers were found in 24 of 27 (89%) patients with endocarditis, in 11 of 14 (79%) with complicated septicemia, and in 10 of 20 (50%) with uncomplicated septicemia with serum samples drawn between days 7 and 30 of disease. With paired samples, the numbers of patients with increased IgG titers were 17 of 17, 3 of 4, and 6 of 7, respectively, in the same patient groups. Increased IgG titers were less often demonstrated in patients with chronic osteomyelitis (7 of 22). The enzyme-linked immunosorbent assay for teichoic acid antibodies was found to be a sensitive and specific method for diagnosing staphylococcal endocarditis and septicemia. For optimal results, both the substantial age-correlated variation in normal titers and the importance of adequately spaced samples should be considered.
Principles, problems, and strategies in the use of antigenic mixtures for the enzyme-linked immunosorbent assay
Authors: Kenny G E , Dunsmoor C L
Published: January 2020

Competition between proteins and other macromolecules for adsorption sites on plastic was studied with the enzyme-linked immunosorbent assay (ELISA) to determine effects of the use of antigenic mixtures or extracts of organisms on assays of antibodies and antigens by ELISA. A comparison of a number of different polystyrene microplates with bovine albumin and human immunoglobulin G (IgG) as antigens showed two major classes of plates; those which adsorbed albumin poorly and those which adsorbed albumin well. IgG adsorbed well on all plates, but plates which adsorbed albumin best also gave significant background levels of nonspecific binding of conjugate. When mixtures of IgG and bovine serum albumin were used as coating antigens, significant competition was observed; the component present at 1% or less in the mixture was essentially undetectable unless excessive amounts of conjugate were used. The important factor was the ratio of competitor to antigen, not the absolute amount. Other proteins (ovalbumin, rabbit albumin, human albumin, and gelatin) were equally effective competitors for adsorption sites on plastic. Nonionic detergents (Tween 20, and Triton X-100) were strong competitors even at 10:1 competitor-to-antigen ratios. In antigen capture assays, normal serum components blocked attachment of antigen-specific IgG, but this competition could be lessened to a degree by the use of strongly binding polystyrene plates. In indirect ELISA for measurement of serum antibody, the use of antigenic mixtures gave significantly lower antibody titers when the desired antigen was less than 1% of the total protein coated. Therefore, the use of mixed or crude antigens in ELISA presents significant problems concerning the sensitivity and specificity of tests.
Detection of Chlamydia trachomatis inclusions in Mccoy cell cultures with fluorescein-conjugated monoclonal antibodies
Authors: Stamm W E , Tam M , Koester M , Cles L
Published: January 2020

We compared two methods for identification of Chlamydia trachomatis inclusions in McCoy cell monolayers: conventional iodine staining and immunofluorescence staining with monoclonal antibodies against the species-specific major outer membrane protein antigen of C. trachomatis. Among 878 urethral and cervical specimens tested in parallel, the immunofluorescence method detected eightfold more inclusions per monolayer, identified a higher proportion of positive specimens on first passage (98 versus 62% by iodine staining; P less than 0.01), and improved overall sensitivity (98% of total positive specimens detected versus 84% by iodine staining; P less than 0.01). Improved sensitivity was most evident in specimens with low numbers of inclusions. Compared with conventional iodine staining, immunofluorescence staining with monoclonal antibodies improves sensitivity and offers more rapid detection of chlamydial inclusions in cell culture.
Cold-reactive immunoglobulin M antilymphocyte antibodies directed against B cells in Thai children with dengue hemorrhagic fever
Authors: Gilbreath M J , Pavanand K , MacDermott R P , Ussery M , Burke D S , Nimmannitya S , Tulyayon S
Published: January 2020

Antilymphocyte antibodies were found in 51 of 83 serum specimens from Thai children with dengue hemorrhagic fever (DHF). The lymphocytotoxic activity was complement dependent, and cytotoxicity was detected in the 19S immunoglobulin M-associated serum fractions at a temperature optimum of 15 degrees C. Sera with lymphocytotoxic activity were cytotoxic to autologous as well as allogeneic lymphocytes from patients and healthy adult donors and were directed primarily against B cells, with some T cell cross-reactivity. This study suggests that infection with DHF induces predominately cold-reactive antilymphocyte antibodies in DHF patients that could potentially interact with peripheral blood cells of patients and modulate the humoral immune responses of patients during infection.
Effect of dietary regimen on rotavirus-Escherichia coli weanling diarrhea of piglets
Authors: Lecce J G , Clare D A , Balsbaugh R K , Collier D N
Published: January 2020

Previously, we induced weanling diarrhea in piglets by infecting them with rotavirus followed by hemolytic enteropathogenic Escherichia coli. We postulated that rotavirus, by damaging the epithelium of the small intestines, produced an enteroenvironment which favored the selection and growth of enteropathogenic E. coli. Furthermore, diet might affect the enteroenvironment and influence the course of the disease. To test this, newly weaned 3-week-old piglets were assigned to one of four dietary regimens and infected with rotavirus followed 24 h later with enteropathogenic E. coli. The course of the disease was followed by monitoring the severity of diarrhea and the fecal shedding of rotavirus and enteropathogenic E. coli in these dually infected piglets. The dietary regimen designed to tax the digestive and absorptive capacities of the piglets (high nutrient intake fed three times a day) produced the most prolonged diarrhea, colonization of the gut by hemolytic enteropathogenic E. coli, and persistent shedding of rotavirus (P less than 0.01). The same nutrient intake divided into 24 equal increments and fed hourly produced a less severe response (P less than 0.01). The least severe response was seen in piglets fed one-third the nutrient intake either hourly or three times a day (P less than 0.01). We conclude that dietary regimen plays an important role in rotavirus-E. coli-induced weanling diarrhea.
Outbreak of food poisoning caused by lactose-fermenting Salmonella tuebingen
Authors: Dube S D
Published: January 2020

An outbreak of food poisoning occurred in a cafeteria and involved 102 student nurses. A lactose-fermenting strain of Salmonella tuebingen was isolated. The source was traced to the chicken consumed. Production of H2S was not evident on triple sugar iron but was detected on lysine iron agar. Therefore, the present report emphasizes the importance of bismuth sulfite agar and lysine iron agar for routine use in the isolation of salmonellae from stool.
Rapid Determination of Log
10
50% Lethal Doses or 50% Infective Doses
Authors: Kuehne Ralph W.
Published: January 2020

A reference table is presented for rapid determination of 50% lethal doses or 50% infective doses after administering 10-fold serial dilutions of toxins, viruses, sera, etc., to six animals per dilution.
Efficacy of enzyme-linked immunosorbent assay in serodiagnosis of aspergillosis
Authors: Mishra S K , Falkenberg S , Masihi K N
Published: January 2020

Sera from 43 cases of aspergillosis, 73 cases of other bronchopulmonary diseases, and 50 healthy persons were examined for the presence of antibodies to Aspergillus fumigatus. Enzyme-linked immunosorbent assay proved to be more efficacious than the immunodiffusion test and counterimmunoelectrophoresis in the cases of allergic bronchopulmonary and invasive aspergillosis.
Identification of Mycobacterium fortuitum and Mycobacterium chelonei
Authors: Lévy-Frébault V , Daffé M , Goh K S , Lanéelle M A , Asselineau C , David H L
Published: January 2020

The study of 52 strains of rapidly growing mycobacteria showed that Mycobacterium fortuitum and M. chelonei were clearly distinguished by the aid of seven key tests (nitrate reductase, iron uptake, beta-glucosidase, penicillinase, growth on fructose, resistance to pipemidic acid, and resistance to capreomycin) and by analysis of their respective mycolic acids. However, the subdivision of these species into M. fortuitum var. fortuitum and M. fortuitum var. peregrinum and M. chelonei subsp. chelonei and M. chelonei subsp. abscessus was not satisfactorily accomplished.
Fibrinogenolytic and fibrinolytic activity in oral microorganisms
Authors: Wikström M B , Dahlén G , Linde A
Published: January 2020

Samples were taken from blood accumulated in dental alveoli after surgical removal of mandibular third molars, from subgingival plaque of teeth with advanced periodontal destructions, from teeth with infected necrotic pulps, and from subjects suffering from angular cheilitis. Of the microorganisms subcultured from these samples, 116 strains were assayed for enzymes degrading fibrinogen and fibrin. Enzymes degrading fibrinogen were assayed with the thin-layer enzyme assay cultivation technique. This assay involves the cultivation of microorganisms on culture agars applied over fibrinogen-coated polystyrene surfaces. Enzymes degrading fibrin were assayed with both a plate assay and a tube assay, in which fibrin was mixed with a microbial culture medium. Microorganisms degrading fibrinogen or fibrin or both were isolated from all sampling sites. Activity was mainly detected in strains of Actinomyces, Bacteroides, Fusobacterium, Peptococcus, Propionibacterium, and Staphylococcus aureus. Most Fusobacterium strains degraded fibrinogen only. Enzymes degrading fibrinogen as well as enzymes degrading fibrin via activation of plasminogen were revealed in strains of Clostridium, S. aureus, and Streptococcus pyogenes. It was generally found that fibrinogen was degraded by more strains than was fibrin, which indicates that different proteases may be involved.
Heptakis(2,6-O-dimethyl)beta-cyclodextrin: a novel growth stimulant for Bordetella pertussis phase I
Authors: Imaizumi A , Suzuki Y , Ono S , Sato H , Sato Y
Published: January 2020

The effect of cyclodextrins on the growth of Bordetella pertussis Tohama phase I in synthetic medium was evaluated. The addition of cyclodextrins, especially heptakis(2,6-O-dimethyl)beta-cyclodextrin (Me beta CD), to a complete synthetic medium such as Stainer-Scholte medium gave the same number of individual colonies and growth rates as those on Bordet-Gengou medium. Furthermore, with the addition of Me beta CD, growth inhibition by fatty acids such as oleic or palmitic acid was overcome and normal cell growth was observed. This modified Stainer-Scholte medium, designated as cyclodextrin solid medium (CSM), supported excellent growth of 20 lyophilized clinical isolates. Serotypes of the organisms after 10 passages on this CSM plate were not changed. These results suggest that Me beta CD is a significant growth stimulant and CSM is one of the most suitable synthetic media for culture of B. pertussis phase I.
Evaluation of the AutoSCAN-3 and Sceptor systems for Enterobacteriaceae identification
Authors: Woolfrey B F , Lally R T , Quall C O
Published: January 2020

To evaluate the accuracy and cost effectiveness of the AutoSCAN-3 (Micro-Scan Systems of America, Sacramento, Calif.) and Sceptor (BBL Microbiology Systems, Cockeysville, Md.) systems for identification of members of the Enterobacteriaceae, we performed parallel tests on 678 stock cultures of well-characterized clinical isolates of Enterobacteriaceae. Automated results by AutoSCAN-3 correctly identified 95.1% at the genus level and 94.9% at the species level. However, 15 of 42 Shigella isolates were misidentified as members of other genera. In contrast to the automated results, visual interpretation of panels produced 97.9% agreement at the genus level, missing only three Shigella isolates. Sceptor correctly identified 96.8% at the genus level and 93.4% at the species level. Of 42 Shigella isolates, 3 were missed and were designated as Salmonella spp. Although all Salmonella spp. were correctly identified, six other isolates were misidentified as Salmonella spp. Test costs were found to be comparable for each system, with the cost per test increasing markedly with fewer than 10 to 15 tests performed per day.
Evaluation of two serological tests for Trichomonas vaginalis infection
Authors: Mathews H M , Healy G R
Published: January 2020

Trichomonas vaginalis is a widely prevalent, sexually transmitted protozoan infecting both males and females. Despite its prevalence, little is known about its contribution to the morbidity rates for urogenital-tract infections. Currently accepted diagnostic methods are limited to the demonstration of the organism in fresh material, identification in stained material, or in vitro cultivation of organisms from the urogenital tract. We have evaluated the indirect hemagglutination test and the gel diffusion test for efficacy in detecting antibodies in serum samples drawn from two population groups. Sera from patients attending a vaginitis clinic had a seropositivity rate of 69% by indirect hemagglutination and 34% by gel diffusion. Seropositivity rates among culture-positive patients were 78% with indirect hemagglutination and 43% with gel diffusion. A group of normal female hospital employees showed seropositivity rates of 30% by indirect hemagglutination and 3% by gel diffusion. Absorption of reactive sera with Trichomonas antigens reduced or abolished the serological reactivity, confirming the specificity of the test. Serological methods can provide a rapid, sensitive, and economical tool to study the epidemiology of this common protozoan infection.
Detection and quantitation of simulated anaerobic bacteremia by centrifugation and filtration
Authors: Lamberg R E , Schell R F , LeFrock J L
Published: January 2020

Fresh human whole blood was inoculated with various anaerobic bacteria or with combinations of anaerobic, facultatively anaerobic, and aerobic microorganisms (3 to 28 microorganisms per ml). The seeded blood was then layered on a reduced Ficoll-Hypaque gradient (density, 1.093 g/ml) and centrifuged (400 X g) for 30 min at ambient temperature. The entire gradient (plasma, leukocytes, and Ficoll-Hypaque) was removed and filtered anaerobically through a 0.45-micron membrane filter. The filters were then placed on reduced chocolate Mueller-Hinton agar plates and incubated at 35 degrees C in humidified atmosphere containing 85% N2, 10% H, and 5% CO2 or in air containing 5% CO2. No statistically significant differences were detected between the numbers of microorganisms recovered (alone or in combination) by filtration and by direct culturing of the original inoculum. All organisms were detected within 30 h after filtration. This technique has excellent sensitivity.
Prospective study of gram-stained stool smears in diagnosis of Clostridium difficile colitis
Authors: Shanholtzer C J , Peterson L R , Olson M N , Gerding D N
Published: January 2020

Gram stains of stools from patients with diarrhea and control patients with no diarrhea were examined for a predominance of gram-positive rods and the presence of polymorphonuclear leukocytes. Results were compared with those from lower gastrointestinal endoscopy for pseudomembranes. Clostridium difficile culturing, and C. difficile toxin assay. The Gram stain was moderately difficult to interpret and was not useful in diagnosing diarrheal disease associated with C. difficile.
Factors influencing the reactivity of Legionella antigens in immunofluorescence tests
Authors: Benson R F , Malcolm G B , Pine L , Harrell W K
Published: January 2020

We examined several factors for their effects on the serological reactivity of Legionella antigens used for direct or indirect fluorescent-antibody tests. These factors included media, methods of killing, strain differences, and the nature of the reactivity with diverse human sera. The maximum serological reactivities were obtained with charcoal-yeast extract agar; the relative antigenicity of cells grown on a chemically defined medium could be fourfold less than those grown on the charcoal-yeast extract agar. Cells grown at 25 degrees C showed only small antigenic differences from those grown at 35 degrees C but had better morphological and staining characteristics. Cells killed by 1% Formalin or 37% Formalin vapors showed a 20% less relative antigenicity than those killed by heat, but their cell walls stained more clearly and they had fewer aberrations. As tested with several human sera, cells of Philadelphia 1 showed great variation in relative antigenicity with changes in media or methods of preparation; Bellingham 1 was quite stable under these same conditions. The data suggest that Bellingham 1 had serogroup 1-specific antigens, reactive with human sera, which were not present in Philadelphia 1.
Age-Specific Prevalence of Antibody to Rotavirus,
Escherichia coli
Heat-Labile Enterotoxin, Norwalk Virus, and Hepatitis A Virus in-a Rural Community in Thailand
Authors: Echeverria Peter , Burke Donald S. , Blacklow Neil R. , Cukor George , Charoenkul Chanin , Yanggratoke Samreong
Published: January 2020

A serological survey in rural Thailand demonstrated that inhabitants acquired antibody to rotavirus between the ages of 6 months and 6 years, to Norwalk virus between the ages of 4 and 5 years, and to hepatitis A between the ages of 6 and 35 years. Antibody of
Escherichia coli
heat-labile toxin was most prevalent between 1 and 4 years and 18 and 25 years of age.
Group VE-1 Septicemia
Authors: Berger Stephen A. , Siegman-Igra Yardena , Stadler Jona , Campus Anne
Published: January 2020

A severely immunocompromised patient developed septicemia while under treatment for pancreatitis. An organism isolated from three separate blood cultures was identified as CDC group VE-1. The biochemical characteristics, taxonomy, and antibiotic sensitivity of this organism are presented.
Endocarditis caused by M6
Authors: Simor A E , Salit I E
Published: January 2020

M6 is a Moraxella-like organism which is usually of pharyngeal origin and is generally nonpathogenic. Endocarditis caused by M6 occurred in a 31-year-old female with known mitral valve prolapse. The infection quickly responded to ampicillin, but valve replacement was required because of progressive left ventricular failure.
Neisseria lactamica septicemia in an immunocompromised patient
Authors: Schifman R B , Ryan K J
Published: January 2020

Neisseria lactamica was isolated from the blood of a 7-year-old girl who was immunosuppressed from chemotherapy for acute lymphocytic leukemia. She was receiving trimethoprim-sulfamethoxazole prophylactically. The isolate was resistant to trimethoprim-sulfamethoxazole and sensitive to penicillin. The patient responded to intravenous penicillin therapy. The organism did not produce immunoglobulin A1 protease.
Corrected Version Specificity of the Latex Test for Cryptococcal Antigen: a Rapid, Simple Method for Eliminating Interference Factors
Authors: Stockman Leslie , Roberts Glenn D.
Published: January 2020

An enzymatic method involving a protease (pronase) for the elimination of interference factors in the latex test for cryptococcal antigen was developed and compared with dithiothreitol treatment. The two were equivalent in their ability to remove interference factors; however, the enzymatic method generally yielded higher titers. The method is simple, requires only 20 min, and makes the latex test for cryptococcal antigen specific.
Polyacrylamide gel electrophoresis of isoenzymes from Entamoeba species
Authors: Mathews H M , Moss D M , Healy G R , Visvesvara G S
Published: January 2020

In this preliminary report, we describe a polyacrylamide gel electrophoresis technique for the resolution of isoenzyme patterns of four isolates of Entamoeba histolytica and one isolate of Entamoeba coli. Our findings were similar to previous findings for three enzyme systems: maleic enzyme (malate dehydrogenase [EC 1.1.1.40]), hexokinase (EC 2.7.1.1), and phosphoglucomutase (EC 2.7.5.1). We found preliminary evidence that glucosephosphate isomerase (EC 5.3.1.9) may also differentiate invasive amoebae from noninvasive amoebae, when the isoenzymes are separated by polyacrylamide gel electrophoresis, whereas this differentiation is not evident with starch-gel electrophoresis. We used an Rf system to relate isoenzyme band mobility to the migration distance of a standard E. histolytica strain (HK-9). The numerical identification of isoenzyme bands can simplify the grouping of isolates into zymodemes.
New, sensitive rocket immunoelectrophoretic assay for measurement of the reaction between endotoxin and Limulus amoebocyte lysate
Authors: Baek L
Published: January 2020

To study the active proteins which participate in the reaction of Limulus amoebocyte lysate (LAL) with lipopolysaccharide, antibody was raised in rabbits against LAL. When LAL was run against rabbit antiserum in crossed immunoelectrophoresis, a complex precipitin pattern appeared. Profound changes took place after reaction of LAL with lipopolysaccharide. The most distinct change was the complete disappearance of the cathodic migrating protein coagulogen, because the antigenicity of coagulogen was lost. Based on this observation, a new rocket immunoelectrophoretic method was developed to detect the disappearance of coagulogen after reaction of LAL with lipopolysaccharide. This assay method was used on clinical specimens (cerebrospinal fluid, plasma, ascites, and urine). It was used as a qualitative test, when a single sample is tested, or as a quantitative assay, when a number of sample dilutions were tested. The new method showed a higher degree of accuracy and sensitivity in comparison with the tube test and it can be used for both research and diagnostic purposes.
Rapid detection by a coagglutination test of heat-labile enterotoxin in cell lysates from blood agar-grown Escherichia coli
Authors: Rönnberg B , Wadström T
Published: January 2020

Colonies of Escherichia coli from blood agar plates were suspended and lysed in saline containing polymyxin B and a detergent (Triton X-100). The lysates were assayed for heat-labile enterotoxin (LT) by a coagglutination test (coa-test). The coa-test reagent consisted of Formalin-treated and heat-killed cells of Staphylococcus aureus, strain Cowan 1, sensitized with a high-titer rabbit anti-LT serum. Purified LT was detected in the coa-test at the nanogram level (2 to 5 ng), whereas larger amounts of cholera toxin (50 ng) were required to give a positive test. The coa-test was compared with the CHO cell test for detection of LT among E. coli strains isolated from human and animal stool cultures. The results of the coa-test and the CHO cell test correlated with 63 of 67 strains of human origin. Six of nine animal strains, defined as LT positive by the CHO cell test, gave positive results in the coa-test. We conclude that the coa-test is probably accurate and sensitive enough to be used in routine diagnosis of LT-producing E. coli strains isolated from human stool cultures. No special laboratory equipment is required, which makes the coa-test suited for diagnosis of enterotoxigenic E. coli in small hospital laboratories and in developing countries.
Controlled evaluation of hypertonic sucrose medium at a 1:5 ratio of blood to broth for detection of bacteremia and fungemia in supplemented peptone broth
Authors: Reimer L G , Reller L B , Mirrett S , Wang W L , Cox R L
Published: January 2020

The value of hypertonic media in the detection of bacteremia and fungemia is controversial, since prior clinical trials have yielded conflicting results with different media. Earlier, we showed that the addition of 10% sucrose to supplemented peptone broth at a 1:10 ratio of blood to broth yielded better recovery of Staphylococcus epidermidis, the Enterobacteriaceae, Pseudomonas aeruginosa, and yeasts. To evaluate the effect of 10% sucrose on blood cultured at a 1:5 ratio, we compared the yield and speed of detection of clinically important microorganisms from adult patients in 5,839 blood samples cultured in supplemented peptone broth with 0.03% sodium polyanetholesulfonate with and without 10% sucrose. The atmosphere of incubation (open venting units), 1:5 ratio of blood to broth, and methods of processing were the same for both bottles. Recovery of facultative gram-positive (P less than 0.02) and gram-negative (P less than 0.02) bacteria was improved, but the recovery of anaerobic gram-negative bacteria was both reduced (P less than 0.01) and delayed (P less than 0.02) by sucrose. The total yield of microorganisms including fungi, however, was increased with sucrose. The effect of sucrose on blood cultures appears to depend on the ratio of blood to broth as well as on the medium used and strains of microorganisms encountered.
Rapid identification of group D streptococci with the API 20S system
Authors: Jorgensen J H , Crawford S A , Alexander G A
Published: January 2020

The API 20S system (Analytab Products) was evaluated as a means of identifying 209 isolates of Lancefield group D streptococci to the species level. Results were compared with those from a conventional tube biochemical identification system and from serological grouping. Use of the latest 20S computerized data base allowed species identification of 97% (203 of 209) of test isolates after a 4-h incubation period and 99% (208 of 209) of test isolates if supplemental overnight biochemical tests were used to clarify the identity of five Streptococcus bovis-variant isolates. The API 20S system appears to be a convenient and accurate method for rapid, same-day species identification of group D streptococci.
Human Rotavirus Detection by Agglutination of Antibody-Coated Erythrocytes
Authors: Sanekata Takeshi , Okada Hidechika
Published: January 2020

We sensitized sheep erythrocytes (SRBC) with antibodies against human rotavirus strain Wa (SRBC-antiWa) and antibodies against calf rotavirus strain NCDV (SRBC-antiNCDV). These were readily agglutinated in the presence of homologous antigens, i.e., human rotavirus and calf rotavirus. By the hemagglutination of SRBC-antiWa and SRBC-antiNCDV (reverse passive hemagglutination [RPHA]), titration of rotavirus in extracts from feces of children suffering from diarrhea (61 specimens) was carried out. We found that the ratio of titers determined with SRBC-antiWa and SRBC-antiNCDV varied remarkably from specimen to specimen. This indicated that the antigenic determinants on human rotavirus in patients feces cross-react with antibodies against NCDV to varying extents. To express the cross-reactivity of human rotavirus with antibodies to NCDV, we propose a Wa/NCDV rotavirus index which can be calculated from the RPHA titer with SRBC-antiWa and SRBC-antiNCDV as follows: Wa/NCDV rotavirus index = (antiWa-RPHA titer of specimen/antiWa-RPHA titer of NCDV)/(antiNCDV-RPHA titer of specimen/antiNCDV-RPHA titer of NCDV).
Enhanced primary isolation of Legionella pneumophila from clinical specimens by low-pH treatment
Authors: Buesching W J , Brust R A , Ayers L W
Published: January 2020

During the 12-month study period, 919 clinical specimens were submitted to the laboratory for culture of Legionella pneumophila. All specimens were plated onto nonselective and semiselective media both directly and after treatment with a KCl-HCl solution, pH 2.2. Of the 51 specimens (5.5%) positive for L. pneumophila and Legionella-like organisms, 20 were recovered by acid pretreatment only, 5 were recovered by direct plating only, and 26 were recovered by both acid pretreatment and direct inoculation. These data demonstrated that acid pretreatment substantially improved the recovery of L. pneumophila and Legionella-like organisms from the clinical specimens examined.
Use of a Novobiocin-Containing Medium for Isolation of
Staphylococcus saprophyticus
from Urine
Authors: Latham Robert H. , Grootes-Reuvecamp Grada A. , Zeleznik Dolores , Stamm Walter E.
Published: January 2020

The use of a novobiocin-containing medium provided little benefit over observable quantitative growth on blood agar in detecting
Staphylococcus saprophyticus
in urine cultures.
Simultaneous determination of arabinitol and mannose by gas-liquid chromatography in experimental candidiasis
Authors: de Repentigny L , Kuykendall R J , Reiss E
Published: January 2020

A method is described for the simultaneous quantitation of D-arabinitol and D-mannose in serum by gas-liquid chromatography as an aid for the diagnosis of disseminated candidiasis. Both variables were observed as per-O-acetylated aldononitrile derivatives in each chromatographic run of sera from immunosuppressed rabbits experimentally infected with Candida albicans 3181A.
Isolation of Pseudomonas putrefaciens in intra-abdominal sepsis
Authors: Marne C , Pallarés R , Sitges-Serra A
Published: January 2020

We report the isolation of Pseudomonas putrefaciens from an intra-abdominal abscess in a patient with colonic carcinoma and from bile in two patients with biliary tract disease. In all three cases, P. putrefaciens was isolated in mixed culture with enteric bacteria.