bluuebunny/crossref_metadata_2025 huggingface dataset:
Urea-hydrolyzing Vibrio parahaemolyticus associated with acute gastroenteritis
Authors: Oberhofer T R , Podgore J K
Published: January 2020

A case of gastroenteritis caused by a urea-hydrolyzing strain of Vibrio parahaemolyticus is presented. Urea-hydrolyzing strains of Vibrio parahaemolyticus have rarely been reported and have not been described previously as a cause of gastroenteritis in the United States. With the exception of urea hydrolysis and the methyl red test, the isolate had all the characteristics of V. parahaemolyticus. The need to screen suspicious non-lactose-fermenting colonies from stool specimens with the oxidase test is emphasized.
Acute Pyelonephritis Caused by
Staphylococcus xylosus
Authors: Tselenis-Kotsowilis Angelina D. , Koliomichalis Maria P. , Papavassiliou John T.
Published: January 2020

Staphylococcus xylosus
was recovered from the urine of a patient with pyelonephritis. Antibodies against the teichoic acid of the microorganism were demonstrated in the patient's serum by the agar gel diffusion technique.
Comparison of the complement fixation test and the indirect hemolysis test for cattle vaccinated and infected with Brucella abortus
Authors: Sutherland S S , Le Cras D V , Evans R J
Published: January 2020

The complement fixation test (CFT) and indirect hemolysis test (IHLT) were applied to sera collected from 60 cattle challenged with Brucella abortus 544. Of the 60 cattle, 48 were vaccinated with either B. abortus 19 or B. abortus 45/20 as calves or as adults. The remaining 12 cattle were not vaccinated. Of the 27 sera from cattle found to be infected, 9 showed aberrant reactions to the CFT. The advantages of the IHLT for these cattle were as follows. After challenge, the titers to the IHLT became positive earlier than or at the same time as the titers to the CFT, they persisted longer than the titers to the CFT, and they failed to show prozone reactions, which are a problem with the CFT. An additional advantage was that before challenge and after vaccination with strain 19, the titers to the IHLT rose later and declined earlier than the titers to the CFT. We concluded that the CFT used in conjunction with the IHLT improves the detection of infected cattle.
Enzyme-linked immunosorbent assay for detection of immunoglobulin M antibody against the Reiter treponeme flagellum in syphilis
Authors: Pedersen N S , Petersen C S , Axelsen N H
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin M (IgM) antibodies against the periplasmatic flagellum of the Reiter treponeme is described. IgM in the test samples was bound to anti-IgM-coated microtest plates, and flagellum-specific IgM antibody was subsequently detected by incubation with a purified flagellum preparation and monospecific anti-flagellum conjugate. Rheumatoid factor, antinuclear antibodies, or flagellum-specific IgG did not interfere. The specificity of the ELISA for IgM antibodies was 99.5% for sera from 200 blood donors and 98.6% for 147 patient sera that gave false-positive reactions in other syphilis serological tests. The sensitivity was 88.5% for sera from 87 patients with first-time primary syphilis, 93.5% for sera from 62 patients with first-time secondary syphilis, 21.4% for sera from 42 patients who were reinfected, and 0% for sera from 13 patients with late syphilis. Of the sera from 153 patients with treated syphilis, 7.2% had IgM antibodies, and sera from patients with primary or secondary syphilis generally had no IgM antibodies 6 months after treatment. The finding of IgM antibodies indicates that patients should receive antisyphilis treatment if they have not been treated recently, but a negative result does not exclude the possibility of active syphilis. The method may prove useful for the diagnosis of congenital syphilis in newborns.
Family outbreak of yersiniosis
Authors: Martin T , Kasian G F , Stead S
Published: January 2020

Yersinia enterocolitica biotype 1, serotype O:21 was isolated from feces or rectal washings of three members of one family in northwestern Saskatchewan. The three isolates gave positive pathogenicity tests in guinea pigs with cultures grown at 22 degrees C as inoculum. All three cases showed clinical symptoms consistent with yersiniosis. All three cases had symptoms of diarrhea and abdominal pain, and two cases had recorded fever. In two cases, appendicitis was initially suspect. One case with ileitis and peritonitis was fatal. The environmental source of the infection was not found, but river water, milk, and person-to-person spread are discussed as possible sources of the infections. The need for microbiology laboratories to culture stool specimens specifically for Y. enterocolitica, using cold-enrichment techniques is emphasized. This family outbreak of yersiniosis provides further evidence that certain biotype 1 strains of Y. enterocolitica are pathogenic.
Isolation of Two
Brucella abortus
Biotypes from Tissues of a Naturally Infected Cow
Authors: Jones Robert L. , Deyoe Billy L. , Meyer Margaret E. , Buening Gerald M. , Fales William H.
Published: January 2020

Brucella abortus
biotypes 1 and 2 were recovered from various tissues of a naturally infected cow. This result indicates a need for biotyping multiple isolates of
B. abortus
from several affected animals in a herd before the biotype of the infecting strain can be definitively established.
Comparative study of selective media for isolation of Legionella pneumophila from potable water
Authors: Edelstein P H
Published: January 2020

A total of 100 water samples, 95% of which were taken from hospital potable water fixtures, were cultured on three different media used for the isolation of Legionella pneumophila. The media used were buffered charcoal-yeast extract medium (BCYE alpha medium), BCYE alpha medium with antimicrobial agents (BMPA alpha medium), and BCYE alpha medium with antimicrobial agents, glycine, and differential dyes (MWY medium). An acid wash procedure was also used for specimens plated on BCYE alpha and BMPA alpha media. A total of 24 samples were culture positive for L. pneumophila by one or more techniques. MWY medium detected 92% of positive cultures, BCYE alpha medium with the acid wash detected 83% of positive cultures, BMPA alpha medium detected 79% of positive cultures. BCYE alpha medium detected 71% of positive cultures, and BMPA alpha medium with the acid wash detected 62% of positive cultures. MWY medium was the best medium for isolating L. pneumophila from potable water specimens and can probably be depended upon as the sole medium for this type of testing.
Quantitation of pathogenic potential of Staphylococcus aureus
Authors: Zierdt C H , MacLowry J D , Robertson E A
Published: January 2020

Numerical estimates of the pathogenicity of Staphylococcus aureus strains were made for phage-typed strains from a relative incidence of significant to nonsignificant isolates from hospital patients. For a specific phage-patterned strain, the number of isolates from significant (wounds, abscesses, blood, etc.) sites was divided by the number of isolates from nonsignificant (respiratory tract, body surfaces, etc.) sites. This value, multiplied by 100, was the index of infection potential (IIP). IIP values for the S. aureus strains studied ranged from a low of 8 to a high of 50. The average IIP for all phage-patterned strains that occurred 50 or more times was 20. There was an inverse relationship between length of the phage pattern (number of the 26 typing phages that lysed the strain) and pathogenicity. Those strains with shorter phage patterns had higher IIP values and were more pathogenic. Strains lysed by one phage had an average IIP of 27, whereas those lysed by 18 phages had an average IIP of 14.
Comparative evaluation of media for isolation of ureaplasma urealyticum and genital mycoplasma species
Authors: Leland D S , Lapworth M A , Jones R B , French M L
Published: January 2020

Two media systems were compared for isolation of Ureaplasma urealyticum and genital Mycoplasma sp. System 1 (S-1) consisted of arginine agar and an arginine biphasic medium for isolation of Mycoplasma sp. and urea agar and urea broth for isolation of U. urealyticum. System 2 (S-2) utilized Boston broth, which is a urea-containing broth, and A7 agar, both of which support the growth of both species. Urine samples, some freshly collected and some known-positive frozen samples, were used as inocula for the two media systems. With S-1, U. urealyticum was recovered in 68% of U. urealyticum-positive cultures: 58% were detected in urea broth, and 60% were identified on urea agar. When the S-2 system was used for culture of the same samples. U. urealyticum was recovered in 98% of the cultures, with 94% detected in Boston broth and 92% identified on A7 agar. Mycoplasma sp. was recovered in S-1 and S-2 in 97 and 100% of Mycoplasma sp.-positive cultures, respectively. The S-1 arginine broth gave positive results for 89% of the cultures, and the arginine agar was positive for 97% of the cultures. The S-2 Boston broth and A7 agar gave positive results for 92 and 97% of the cultures, respectively. For the isolation of U. urealyticum, colony counts were higher, growth was seen sooner, and colonies were larger when the S-2 media system was used. Overall, the cost per test of the S-2 system was less both in technologist time and in reagent costs.
Detection of a Rotavirus-Like Agent Associated with Diarrhea in an Infant
Authors: Rodger Suzanne M. , Bishop Ruth F. , Holmes Ian H.
Published: January 2020

A rotavirus-like agent was detected in the feces of a child with diarrhea. Although morphologically indistinguishable from rotavirus, the agent was serologically distinct, and electrophoresis of its nucleic acid also showed that it was dissimilar.
Isolation of human rotavirus subgroups 1 and 2 in cell culture
Authors: Kutsuzawa T , Konno T , Suzuki H , Kapikian A Z , Ebina T , Ishida N
Published: January 2020

One strain of human rotavirus subgroup 1 (KUN) and one strain of subgroup 2 (MO) were isolated with the MA104 cell line, a fetal rhesus monkey kidney cell line. Their subgroup specificities and RNA patterns were identical to those of rotaviruses present in stools before cultivation. Distinct cytopathic effects consisting of obscure cell boundaries, cell fusion, cell rounding, cell detachment, and lytic foci were recognized at passage 3 of MO and passage 6 of KUN. No differences in cytopathic changes were found between the two isolates.
Campylobacter jejuni peritonitis complicating continuous ambulatory peritoneal dialysis
Authors: Pepersack F , D'Haene M , Toussaint C , Schoutens E
Published: January 2020

We report the occurrence of Campylobacter jejuni peritonitis complicating C. jejuni enteritis in a patient treated with continuous ambulatory peritoneal dialysis. Cure followed oral administration of erythromycin and intraperitoneal therapy with gentamicin.
Reduction of Brucella species and Francisella tularensis cross-reacting agglutinins by dithiothreitol
Authors: Behan K A , Klein G C
Published: January 2020

Cross-reaction agglutinin titers to Brucella abortus antigen were found in 42 of 128 tularemia serum specimens, and cross-reaction titers to Francisella tularensis antigen were found in 8 of 34 brucellosis serum specimens. The cross-reaction titers were reduced to 10 or less by dithiothreitol, suggesting that the titers are due to immunoglobulin M antibody.
Serum Antibody Response to
Clostridium botulinum
Toxin in Infant Botulism
Authors: Rubin Lorry G. , Dezfulian Manuchehr , Yolken Robert H.
Published: January 2020

A serum antibody response has not been previously demonstrated after infection with
Clostridium botulinum.
We developed an enzyme immunoassay for measuring serum antibody to
C. botulinum
toxins A, B, and E. This assay system detected a specific immunoglobulin G and immunoglobulin M antibody response to
C. botulinum
toxin in two patients with infant botulism.
Bacteremia Due to
Cedecea neteri
sp. nov
Authors: Farmer J. J. , Sheth Neela K. , Hudzinski June A. , Rose Harold D. , Asbury Mary F.
Published: January 2020

Three of four blood cultures from a patient with possible endocarditis were positive for a new species of
Enterobacteriaceae
which is named
Cedecea neteri.
This is the first reported case of bacteremia caused by a strain of
Cedecea.
Cellular fatty acids of Capnocytophaga species
Authors: Dees S B , Karr D E , Hollis D , Moss C W
Published: January 2020

The cellular fatty acid composition of 18 clinical isolates and 4 reference strains of Capnocytophaga species (Capnocytophaga ochracea, Capnocytophaga gingivalis, and Capnocytophaga sputigena) was determined by gas-liquid chromatography. The fatty acid profiles of the 22 cultures were essentially identical and were characterized by major amounts (60% or greater) of a saturated, iso-branched-chain, 15-carbon acid (13-methyl-tetradecanoate) and the presence of two relatively uncommon saturated, iso-branched, 3-hydroxy acids (13-methyl-3-hydroxy-tetradecanoate and 15-methyl-3-hydroxy-hexadecanoate). The presence and relative amounts of these acids distinguish Capnocytophaga spp. from other gliding bacteria.
Analysis of a single reference strain for determination of gentamicin regression line constants and inhibition zone diameter breakpoints in quality control of disk diffusion antibiotic susceptibility testing
Authors: Kronvall G
Published: January 2020

An equation was derived from known formulas to express the size of the inhibition zone diameter in the disk diffusion antibiotic susceptibility test as a function of the disk content of antibiotic. The equation permitted a calculation of regression line constants for the correlation between zone diameter and the minimum inhibitory concentration (MIC) with a single reference strain. Analysis of reference strains Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853, as well as 12 clinical isolates belonging to these species, showed a linearity between zone size squared and the logarithm of disk content in tests with 10-, 30-, and 100-micrograms gentamicin disks. All three species, however, gave regression line constants which were characteristic for the individual bacterial species. Calculations of zone diameter breakpoints corresponding to recommended MIC limits with E. coli and P. aeruginosa reference strains gave an accurate prediction of gentamicin susceptibility. Histogram analysis of 48 strains of Streptococcus faecalis from clinical specimens showed a distribution of zone diameter values which would result in false classification of susceptibility with breakpoints calculated for the other bacterial species studied. Single reference strain analysis of S. faecalis ATCC 29212 (gentamicin MIC, 8 micrograms/ml) permitted the calculation of breakpoints which accurately assigned the strains tested to the intermediate category of susceptibility. Single reference strain analysis offers a quality control method for individual laboratories that allows the determination of inhibition zone diameter breakpoints corresponding to recommended MIC limits with no MIC determinations required.
Detection of antibodies to influenza virus M protein by an enzyme-linked immunosorbent assay
Authors: Khan M W , Bucher D J , Koul A K , Kalish G , Smith H , Kilbourne E D
Published: January 2020

An enzyme-linked immunosorbent assay test system was developed in which purified influenza virus M protein was used for the detection of M antibody in human sera. Antibody levels to influenza A virus M protein were monitored in sera from a vaccine study population by using an enzyme-linked immunosorbent assay technique with purified M protein as the adsorbent antigen. A 10-fold variation in titers of preexisting M antibody was observed in this population of young adults. Increases of anti-M titer of 7- to 24-fold were observed upon immunization with Formalin-inactivated vaccine or after natural infection. The antibody response to M protein was dissociated from the response to the hemagglutinin or neuraminidase antigens. The M antibody response preceded or was coincident with the antibody response to H1 hemagglutinin upon natural exposure to circulating virus.
Achromopeptidase for lysis of anaerobic gram-positive cocci
Authors: Ezaki T , Suzuki S
Published: January 2020

Achromopeptidase, which has potent bacteriolytic activity for most of the gram-positive aerobic bacteria, was for the first time used for the lysis of anaerobic cocci. Most of the lysozyme-resistant gram-positive anaerobic cocci were lysed with this new enzyme. Peptococcus magnus was the only organism tested resistant to achromopeptidase. P. saccharolyticus was quite unusual because it was very sensitive to both achromopeptidase and lysozyme.
Dengue virus replication in a polyploid mosquito cell culture grown in serum-free medium
Authors: Kuno G
Published: January 2020

A subline of a polyploid cell line (TRA-284) derived from a nonbiting mosquito, Toxorhynchites amboinensis, was adapted to a serum-free medium. The sensitivity of the subline (TRA-284-SF) to all serotypes of adapted dengue viruses was generally comparable to that of Aedes albopictus (C6/36), and DEN 3 viruses replicated to higher titers in TRA-28F-SF cells than in C6/36 cells. The subline was found to be useful for isolation of dengue viruses from human serum, since isolation rates were higher in TRA-284-SF cells than in C6/36 cells. The advantages of using a serum-free medium and mosquito cells for virus isolation are discussed.
Rapid method for simultaneous detection of the arginine dihydrolase system and amino acid decarboxylases in microorganisms
Authors: Chen K C , Culbertson N J , Knapp J S , Kenny G E , Holmes K K
Published: January 2020

A specific procedure has been developed for the detection of the first two enzymes involved in the arginine dihydrolase system and the detection of the decarboxylases of arginine, glutamic acid, histidine, lysine, ornithine, phenylalanine, tryptophan, and tyrosine. A loopful of growth of each organism from dihydrolase-decarboxylase induction agar medium (or broth) was washed and incubated separately with 0.2-ml samples of three test media supplemented with different amino acids. Each spent test medium was dansylated, and the dansyl derivatives were separated by two-dimensional thin-layer chromatography on polyamide sheets. The end products (citrulline, ornithine, gamma-amino-n-butyric acid, and amines) produced during incubation were estimated by comparing the fluorescent intensities of end products from the spent test media and of the corresponding parent amino acids from test medium controls after thin-layer chromatography. The method is reproducible, requiring incubation of an organism in three test media for 1 h for simultaneous detection of the first two enzymes involved in the arginine dihydrolase system and of eight amino acid decarboxylases. This method has been successfully applied to gram-positive and gram-negative microorganisms and also to Mycoplasmatales. It could simplify and improve the accuracy of the corresponding biochemical tests performed in clinical laboratories for the identification and differentiation of microorganisms, and it may prove particularly useful for the differentiation of species of Pseudomonas and Mycoplasma.
Development and evaluation of a biochemical scheme for identification of endocervical lactobacilli
Authors: Fagnant J E , Sanders C C , Sanders W E
Published: January 2020

A biochemical scheme for the species identification of endocervical lactobacilli was developed and evaluated with 10 isolates obtained from the American Type Culture Collection (ATCC) and 106 endocervical isolates obtained from women reporting to a local venereal disease clinic and a local hospital clinic. The scheme consisted of two stages. Stage I included six tests and was tested and modified with results obtained with ATCC strains. From the modified stage I, stage II was developed. Tests to be performed in this stage were determined from expected characteristics of lactobacilli. Stage II was also tested with the ATCC strains. Of the 106 endocervical isolates, 78 (74%) were identified with the two-stage scheme as developed with the ATCC strains. Unexpected results were obtained in one or both stages with the other 28 isolates. For 10 isolates, the final species identified were not previously expected to be recovered. A "best-fit" method was used to determine the most likely identification of the remaining 18 isolates. In a few instances, the use of a third stage was necessary to reach an identification. The final identification scheme, although complicated in appearance, generated a species identification with a total of 12 tests with a range of 7 to 10 tests per isolate.
Practical procedure for demonstrating bacterial flagella
Authors: Kodaka H , Armfield A Y , Lombard G L , Dowell V R
Published: January 2020

We developed a simple, rapid method for demonstrating bacterial flagella with Ryu staining solution that gave satisfactory results for numerous motile and nonmotile bacteria. Two major advantages of this method are that the staining solution, ready for use, is stable at ambient temperature indefinitely and that microscopic examination of bacteria in the stained drop preparations can be performed rapidly.
Formalin-killed versus heat-killed Legionella pneumophila serogroup 1 antigen in the indirect immunofluorescence assay for legionellosis
Authors: Wilkinson H W , Brake B J
Published: January 2020

Indirect immunofluorescence assay titers of human sera obtained against the Legionella pneumophila serogroup 1 antigen (Philadelphia 1 strain) killed with 10% Formalin showed a tendency to be lower than those obtained against the reference heat-killed antigen (geometric mean titer of 194 and 370, respectively) when all other variables in the test were held constant. Test results were interpreted the same for 96% of 60 paired sera if the cutoff level used to interpret a positive test result for the formalinized antigen was lowered by one twofold dilution factor.
Inactivation of Penicillins by Thiol Broth
Authors: Murray Patrick R. , Niles Ann C.
Published: January 2020

Thiol broth with sodium polyanetholesulfonate inactivated penicillin G, carbenicillin, nafcillin, oxacillin, and gentamicin, but had no effect on cephalothin, cefoxitin, clindamycin, chloramphenicol, erythromycin, and tetracycline. Only Thiol broth was capable of this inactivation, which was not influenced by the presence of blood.
Identification of virus-specific oligoclonal bands in subacute sclerosing panencephalitis by immunofixation after isoelectric focusing and peroxidase staining
Authors: Mehta P D , Patrick B A , Thormar H
Published: January 2020

We developed a sensitive peroxidase staining procedure to identify oligoclonal band specificity in subacute sclerosing panencephalitis by subjecting serum and cerebrospinal fluid to isoelectric focusing and immunofixation with measles virus. The gel was washed and stained with horseradish peroxidase-conjugated goat antihuman immunoglobulin G. The samples showed dark brown measles-specific oligoclonal bands in the alkaline pH region of the gel, whereas the controls showed no significant staining. This technique may be useful in identifying specific antibody activity against viral and other antigens in oligoclonal bands in cerebrospinal fluid and serum.
Equi factors in the identification of Corynebacterium equi Magnusson
Authors: Prescott J F , Lastra M , Barksdale L
Published: January 2020

The production of equi factor(s) by 173 serologically verified Corynebacterium equi isolates was tested by streaking strains at right angles to a culture of Corynebacterium pseudotuberculosis or Staphylococcus aureus on a cattle blood agar plate. All strains produced equi factor(s). This characteristic was more specific than other tests used on the strains.
New provisional serovar of Shigella boydii
Authors: Gross R J , Thomas L V , Day N P , Cheasty T , Rowe B
Published: January 2020

Ten bacterial strains are described that give biochemical reactions characteristic of the genus Shigella but do not belong to any of the established or provisional serovars. The strains ferment mannitol and are antigenically indistinguishable. They do not possess any of the group antigens of Shigella flexneri, and it is therefore proposed that they be regarded as members of a new provisional serovar of Shigella boydii. Strain E16553 is designated as the test strain for the new serovar.
Disk diffusion testing with polymyxin and amikacin for differentiation of Mycobacterium fortuitum and Mycobacterium chelonei
Authors: Wallace R J , Swenson J M , Silcox V A , Good R C
Published: January 2020

Disk diffusion is one method of susceptibility testing of the Mycobacterium fortuitum complex to antibacterial agents. We utilized disks of polymyxin B (300 U), amikacin, and kanamycin to determine whether they could also be used for species identification when compared with standard biochemical methods. With the polymyxin disk, 100% of 75 M. fortuitum strains produced zones of inhibition, whereas none (0%) of 58 Mycobacterium chelonei subspecies abscessus and chelonei strains had any zone of inhibition. With the amikacin disk, 99% of M. fortuitum biovariant fortuitum had zones of greater than or equal to 30 mm compared with 6% of M. chelonei. The rare M. chelonei-like organisms gave variable results, and 42% of the unnamed "third group" biovariant of M. fortuitum exhibited an unusual but diagnostic pattern of small zone sizes to amikacin and no zone to kanamycin. The kanamycin disk was otherwise not helpful, although it resulted in larger zone sizes for M. chelonei than did amikacin. Thus, disk diffusion susceptibilities which include these disks (especially polymyxin) will provide presumptive evidence of species as well as susceptibility data.
Immunoglobulin M antibody titers in the diagnosis of Legionnaires disease
Authors: Zimmerman S E , French M L , Allen S D , Wilson E , Kohler R B
Published: January 2020

The purpose of this study was to determine whether measurement of immunoglobulin M (IgM) antibodies against Legionella pneumophila serogroup 1 can aid in the diagnosis of Legionnaires disease. On the basis of measurements of antibody levels in 1,942 control sera, we used an IgM titer of 1:256, observed in 2.3% of the controls, as presumptive evidence of Legionnaires disease. Measurement of IgM titers permitted us to presumptively or definitively diagnose Legionnaires disease in 13 of 34 patients earlier than we would have if only IgG titers had been measured. Of the 13 patients, 5 were diagnosed serologically only by IgM antibody determination. IgM titers were presumptively diagnostic in week 1 of clinical symptoms in 4 of the 13 patients. We conclude that conjugates used for antilegionella indirect fluorescent-antibody tests should be capable of detecting IgM antibodies so that the value of serological results in diagnosing and managing Legionnaires disease will be maximized.
Microsticks as solid-phase carriers for enzyme-linked immunosorbent assays
Authors: Shekarchi I C , Sever J L , Ward L A , Madden D L
Published: January 2020

Microsticks are machine-tooled or molded pegs of plastic or stainless steel which were developed as solid-phase carriers for the enzyme-linked immunosorbent assay (ELISA). They consist of a stem, which can be coated with plastic to be used as the reactive surface, and a shaft designed for easy handling and labeling and for positioning the sticks in microtiter wells and transfer plates. Microsticks permit a wide selection of coating materials and afford the user greater control over quality and standardization of the solid-phase surface. Polycarbonate and nitrocellulose coatings were tested in ELISAs for antibodies to measles virus, toxoplasma, and human gamma globulin. The Microsticks were found to give reproducible, sensitive, and specific ELISA results and minimized problems due to lot-to-lot and batch-to-batch variations found with other plastic carriers.
Alternatives to the standardized laboratory branch complement fixation test for detection of antibodies to Coccidioides immitis
Authors: Wood J C , Friedly G , Zartarian M , Aarnaes S , de la Maza L M
Published: January 2020

Antibody titers to Coccidioides immitis, using coccidioidin antigen, were determined by three methods: the standardized Laboratory Branch complement fixation method (LBCF), a modified version of the Viral and Rickettsial Disease Laboratory complement fixation test (VRDL-CF), and a quantitative immunodiffusion test (QID). Of the 133 samples evaluated, 72 were negative by each method and 57 (42 serum samples, 15 cerebrospinal fluid samples) were positive by all three methods. Four additional specimens (1 serum sample, 3 cerebrospinal fluid samples) were positive by QID alone. All positive patients were diagnosed clinically as having pulmonary or extrapulmonary coccidioidomycosis or both. When titers from two methods were compared, the agreement within +/- 1 dilution was VRDL-CF/QID, 88.5%; VRDL-CF/LBCF, 85.2%; and LBCF/QID, 82.0%. The agreement of these methods within +/- 2 dilutions was VRDL-CF/QID, 98.4%; VRDL-CF/LBCF, 96.7%; and LBCF/QID, 93.4%. The VRDL-CF and QID methods are simpler to perform; however, they are yet unrecognized as suitable alternatives to the more cumbersome LBCF. Our data show that they should be considered as options for C. immitis serology.
Detection of fungemia obscured by concomitant bacteremia: in vitro and in vivo studies
Authors: Hockey L J , Fujita N K , Gibson T R , Rotrosen D , Montgomerie J Z , Edwards J E
Published: January 2020

Our recent clinical experience suggested that bacteremia may interfere with the detection of concomitant fungemia when standard blood culture methods are used. To determine the extent to which bacteria may interfere with fungal isolation from blood cultures, an in vitro model simulating blood cultures taken during concomitant fungemia and bacteremia was created. Each of six bacteria (Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus faecalis, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa) was combined with each of three pathogenic yeasts (Candida albicans, Candida tropicalis, and Torulopsis glabrata) in vented blood culture bottles containing enriched brain heart infusion broth and fresh normal human blood. Blood culture bottles were analyzed at 1, 2, and 7 days of incubation. Gram strains and subcultures onto chocolate and MacConkey agars failed to detect fungi in 37.0, 66.7, and 100% of samples, respectively. However, subcultures onto Sabouraud dextrose agar failed in only 13% of the samples (occurring only with P. aeruginosa). In a rabbit model of concomitant fungemia with C. albicans and bacteremia with P. aeruginosa, no yeasts were recovered from blood cultures despite 100% detection of P. aeruginosa. Therefore, the usual microbiological techniques may be inadequate to detect fungemia when concomitant bacteremia is present.
Statistical evaluation of a quality control method for isolation of pathogenic Vibrio species on selected thiosulfate-citrate-bile salts-sucrose agars
Authors: West P A , Russek E , Brayton P R , Colwell R R
Published: January 2020

The recovery of Vibrio cholerae, Vibrio fluvialis, Vibrio parahaemolyticus, and Vibrio vulnificus, employing eight strains of each species, was studied by using four brands of thiosulfate-citrate-bile salts-sucrose (TCBS) agar prepared according to manufacturers' instructions and following a standardized procedure. A standardized broth inoculum of each strain was placed on duplicate plates of each brand of TCBS agar and also on tryptic soy agar (Difco Laboratories) containing 1% (wt/vol) NaCl, the latter serving as the control. Plates were inoculated in a sequence designed to compensate for bias associated with multiplication of the bacteria during the inoculation procedure. Colony counts and quality of growth were recorded after incubation for 18 h at 35 degrees C. The comparison procedure was repeated four times at weekly intervals. Data were analyzed by using an analysis of variance model. The recovery and quality of growth of each species varied significantly on the different brands of TCBS agar. Significant variability was also identified for some components of the inoculation procedure. Modifications of the inoculation procedure are suggested to minimize sources of variance. A simplified statistical procedure, based on the t test, is described for media quality control for laboratories routinely isolating pathogenic Vibrio spp.
Quality Control Limits for the Agar Overlay Disk Diffusion Antimicrobial Susceptibility Test
Authors: Barry Arthur L. , Badal Robert E.
Published: January 2020

Replicate control tests were performed with the agar overlay and Kirby-Bauer techniques, using two sources of Mueller-Hinton agar. Mean zones observed with the two methods differed by 1 to 3 mm. Despite the minor differences between methods, National Committee for Clinical Laboratory Standards quality control limits which were established for the Kirby-Bauer method could also be used for the agar overlay technique.
Effect of incubation conditions on anaerobic susceptibility testing results
Authors: Murray P R , Niles A C
Published: January 2020

We determined the effect of performing antimicrobial susceptibility tests in five different anaerobic incubation systems: GasPak jar, large GasPak jar, evacuated-gassed anaerobic jar, anaerobic chamber, and Bio-Bag. Growth of the anaerobes was equivalent in all five incubation systems. The results of testing 38 anaerobes against 11 antimicrobial agents were comparable for the anaerobic jars and anaerobic chamber. However, discordant results were observed for metronidazole and cefamandole tests when incubated in the Bio-Bag.
Use of Mueller-Hinton agar to determine novobiocin susceptibility of coagulase-negative staphylococci
Authors: Almeida R J , Jorgensen J H
Published: January 2020

A group of 254 isolates of coagulase-negative staphylococci were tested in parallel for novobiocin susceptibility by using P agar and Mueller-Hinton agar. Zones of inhibition of 16 mm or less around a 5-micrograms novobiocin disk on Mueller-Hinton agar indicated novobiocin resistance, as demonstrated by Staphylococcus saprophyticus.
Antigenic Relationships Among Type Strains of
Yersinia enterocolitica
and Those of
Escherichia coli, Salmonella
spp., and
Shigella
spp
Authors: Thomas Linda V. , Gross Roger J. , Cheasty Tom , Shipp Christine R. , Rowe Bernard
Published: January 2020

The antigenic type strains for
Yersinia enterocolitica
antigens O:1 to O:34 were examined and their antigenic relationships with the type strains of
Escherichia coli, Shigella
spp., and
Salmonella
spp. were determined.
Y. enterocolitica
O:5,27 was antigenically identical to
E. coli
O97 and
Y. enterocolitica
O:11 was antigenically identical to
E. coli
O98.
Microenzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G and Immunoglobulin M Antibodies to
Legionella pneumophila
Authors: Elder E. M. , Brown A. , Remington J. S. , Shonnard J. , Naot Y.
Published: January 2020

The microenzyme-linked immunosorbent assay (ELISA) for the detection of immunoglobulin M and G (IgM, IgG) antibodies to
Legionella pneumophila
serogroup 1 antigens was evaluated. IgM antibodies were measured by both double-sandwich and single-sandwich techniques. These assays were compared with the previously standardized indirect immunofluorescence test in four groups of subjects: (i) pneumonia patients with culture-proven Legionnaires disease with serogroup 1 isolates, (ii) pneumonia patients with serogroup 1 organisms detected by direct immunofluorescence testing of respiratory secretions but without culture confirmation, (iii) pneumonia patients with negative culture and direct immunofluorescence tests, and (iv) healthy hospital employees. In addition, the sensitivity and specificity of the IgG ELISA were evaluated with larger groups of controls and Legionnaires disease patients. The ELISA was more sensitive than the indirect immunofluorescence test. However, it detected antibody rises in pneumonia patients without culture or direct immunofluorescence evidence of
L. pneumophila
serogroup 1 infection, thereby suggesting that the specificity of the ELISA was slightly lower than that of the indirect immunofluorescence test. The double-sandwich ELISA was a sensitive method for detecting IgM antibodies and, as previously reported, appeared to be free from interference by rheumatoid factor. IgM anti-
Legionella
antibodies detected by the ELISA appeared earlier and were less persistent than IgG antibodies. In addition, the IgM ELISA was useful in detecting antibodies in necropsy serum samples obtained from patients dying acutely of Legionnaires disease. The data presented show that the ELISA is a reliable method for the detection of specific anti-
Legionella
antibodies.
Evaluation of the MicroScan Urinary Combo Panel and API 20E system for identification of glucose-nonfermenting gram-negative bacilli isolated from clinical veterinary materials
Authors: Mathewson J J , Simpson R B , Brooks F L
Published: January 2020

Many isolates of glucose-nonfermenting gram-negative bacilli (NFB) cultured from clinical veterinary specimens are not identified because of the large number of identification tests required. We evaluated two commercial identification systems to determine if they could accurately identify NFB isolated from animals. Of 182 strains of NFB, the MicroScan Urinary Combo Panel (MicroScan, Inc., Campbell, Calif.) correctly identified 72%, and the API 20E system (Analytab Products, Plainview, N.Y.) correctly identified 74%. Of the 118 strains of the three most common species of NFB isolated from animals, the MicroScan Urinary Combo Panel identified 86% correctly, and the API 20E system identified 92% correctly. The use of either of these systems could improve the accuracy of identification of NFB from clinical veterinary materials.
Evaluation of differential and selective media for isolation of Aeromonas and Plesiomonas spp. from human feces
Authors: von Graevenitz A , Bucher C
Published: January 2020

We studied nine solid and two liquid media for their suitability to select Aeromonas and Plesiomonas spp. from human stools, using artificially contaminated samples as well as 254 samples from outpatients with and without diarrhea. Media with optimal sensitivity and specificity for Aeromonas spp. were alkaline peptone-water, Trypticase soy broth with ampicillin, inositol-brilliant green-bile salts agar, dextrin-fuchsin-sulfite agar, xylose-sodium desoxycholate-citrate agar, and Pril-xylose-ampicillin agar. For Plesiomonas sp., alkaline peptone-water and inositol-brilliant green-bile salts agar were optimal. Four strains of Aeromonas spp. were detected in patient samples with these media.
Intestinal parasites in a remote village in Nepal
Authors: Estevez E G , Levine J A , Warren J
Published: January 2020

Stool samples for parasitological examination were collected in a remote area of western Nepal. Of 40 specimens collected, 36 were positive for parasites as determined by examination of direct wet mounts and trichrome smears. All but one of the positive specimens contained several parasite species, averaging four species per specimen. Four negative specimens were found in infants under 1 year of age. The parasitic burden in this population appeared to be high, and the prevalence of parasitic infection approached 100%.
Epidemiology of rotavirus diarrhea in the Highlands of Papua, New Guinea, in 1979, as revealed by electrophoresis of genome RNA
Authors: Albert M J , Bishop R F , Shann F A
Published: January 2020

Results of gel electrophoresis of rotavirus genome RNA from feces of children in two provinces in Papua, New Guinea, suggest that the epidemiology of rotavirus infection in small communities with a total population of 3,000 may differ from that in urban or closely settled rural areas.
Human Edwardsiellosis traced to ornamental fish
Authors: Vandepitte J , Lemmens P , de Swert L
Published: January 2020

Protracted diarrhea in a 2-month-old Belgian infant was associated with Edwardsiella tarda as the only potential pathogen. The same organism was isolated from a tropical aquarium fish in the home of the patient. The enteropathogenic role of E. tarda and the role of fish as a reservoir of infection are reviewed. Additional observations, volunteer studies, and careful prospective surveys in tropical countries will be needed to produce firm evidence of the enteropathogenic potential of E. tarda.
Single radial hemolysis test for rubella immunity and recent infection
Authors: Neumann P W , Weber J M
Published: January 2020

The single radial hemolysis (SRH) test was compared with the hemagglutination inhibition (HI) test for establishing rubella immune status and diagnosing recent infection. Correlation between mean SRH diameters and HI titers greater than or equal to 1:8 was high (R = 0.99). It is suggested that a level of greater than or equal to 5 IU represents low-level antibody and that greater than or equal to 15 IU is a conservative threshold for designation of immunity. Of 343 sera tested, only 1 false-positive was found by SRH with the 5 IU cutoff level. The SRH test could detect serum antibody levels as low as 2.5 IU, whereas 15 IU was generally the limit of resolution of the HI test. Data from sucrose density gradient fractionation of serum demonstrated that neither rubella-specific immunoglobulin M (IgM) nor early postinfection HI-reactive IgG was detected by SRH. However, diagnostic changes in antibody titer measured by SRH were in general greater than those measured by HI. The SRH test showed diagnostic titer changes in some sera containing specific IgM for which no such changes were detected by the HI test. A 2.5-mm difference in hemolytic zone diameters (a fourfold rise in international units) between acute- and convalescent-phase serum pairs was chosen as being of diagnostic significance. This difference was less than the minimum observed difference of 2.9 mm from 59 serum pairs showing diagnostic changes by HI and far exceeded (greater than 3.6 standard deviations) the within-test and individual variability seen for 95 pregnant women from whom samples were obtained during each trimester.
Enhanced detection of bacteremia with a new BACTEC resin blood culture medium
Authors: Appelbaum P C , Beckwith D G , Dipersio J R , Dyke J W , Salventi J F , Stone L L
Published: January 2020

In this multicenter study, 621 sets of blood culture specimens were drawn from 280 patients who were suspected of being septic and who were receiving antimicrobial therapy. Equal volumes of each specimen were inoculated into BACTEC 6B and 16B media. The 16B medium contained adsorbent and cationic resins for neutralizing the effects of the drugs. Of the 621 sets drawn, there were 72 positive cultures in 16B and 52 positive cultures in 6B. In 23 cases the organism was detected only in the 16B medium, and in 3 cases the organism was detected in 6B only. The remaining 49 positives were detected in both culture bottles. In 13 of these 49 cultures, detection in 16B was made between 1 and 5 days earlier than in 6B, whereas 3 of 49 specimens were detected 1 day earlier in 6B; the remaining 33 cultures became positive at approximately the same time in both media. There were a total of 43 patients with positive cultures in this study. Of these patients, 28 had sepsis detected in both the 16B and 6B media. The 6B medium alone detected an additional three cases of sepsis, and the 16B resin medium alone identified 12 additional cases. Supplementary culturing of samples from patients receiving antimicrobial therapy significantly increased the number of positive cultures and positive patients, as well as significantly shortening the time to positivity in these cultures.
Protease phenotypes of Pseudomonas aeruginosa isolated from patients with cystic fibrosis
Authors: Jagger K S , Bahner D R , Warren R L
Published: January 2020

The protease phenotypes expressed by isolates of Pseudomonas aeruginosa from cystic fibrosis (CF) patients were evaluated. The majority of isolates tested produced elastase (65%) or alkaline protease (64%) or both. The mucoid phenotype expressed by many CF isolates of P. aeruginosa did not absolutely restrict the expression of protease activity, although a higher percentage of nonmucoid isolates was proteolytic. When isolates from CF patients chronically infected with P. aeruginosa were compared to isolates from CF patients colonized with this organism, both groups were found to contain comparable percentages of elastase-producing strains and mucoid strains. However, the group of isolates from colonized patients contained a higher percentage of strains producing alkaline protease and expressing general protease activity. In addition, the group of isolates from chronically infected patients contained more weakly proteolytic isolates than either the group from colonized CF patients or a group of isolates from pediatric patients without CF. These data suggest that protease production may be important in the initial colonization of the respiratory tract of CF patients by P. aeruginosa.
Enzyme-linked immunosorbent assay for Clostridium difficile toxin A
Authors: Lyerly D M , Sullivan N M , Wilkins T D
Published: January 2020

Antibodies against Clostridium difficile toxin A were purified by affinity chromatography from antiserum prepared against crude C. difficile toxin preparations. The affinity-purified antibody preparation was free of detectable amounts of antibodies to other C. difficile antigens, as demonstrated by crossed immunoelectrophoresis, and specifically neutralized the cytotoxicity of toxin A. An indirect enzyme-linked immunosorbent assay (ELISA) was subsequently developed using the antibody preparation for the specific detection of toxin A. The ELISA, which could detect 1 ng (5 ng/ml) of toxin A, was used to quantitate the toxin in the culture supernatant fluids of strains of C. difficile. The ELISA values for toxin A closely correlated with the toxin A and B cytotoxic titers of the supernatant fluids. In addition, toxin A was detected by ELISA in human fecal specimens from persons with antibiotic-associated colitis, demonstrating that this toxin is produced during C. difficile colitis.
Antigen-specific proliferation responses of peripheral blood lymphocytes to Trichomonas vaginalis antigen in patients with Trichomonas vaginitis
Authors: Yano A , Aosai F , Yui K , Kojima S , Kawana T
Published: January 2020

This report describes the development of an assay system which overcomes the difficulty of detecting immune responses of patients with Trichomonas vaginitis by making use of peripheral blood leukocytes obtained from such patients. When peripheral blood leukocytes from the patient were stimulated in microcultures with the soluble antigen extracted from Trichomonas vaginalis, significant degrees of proliferation ensued, as measured by the incorporation of [methyl-3H]thymidine 4 to 5 days after initiation. The antigen-induced proliferation response of peripheral blood leukocytes is specific for T. vaginalis antigen. The T. vaginalis-specific [methyl-3H]thymidine incorporation is mediated by Leu-1-positive cells, namely, T lymphocytes, in the peripheral blood leukocyte population. This assay system should prove useful for the analysis of the immune response to the protozoa in patients with Trichomonas vaginitis.