bluuebunny/crossref_metadata_2025 huggingface dataset:
Cellular fatty acid and soluble protein composition of Actinobacillus actinomycetemcomitans and related organisms
Authors: Calhoon D A , Mayberry W R , Slots J
Published: January 2020

The cellular fatty acid and protein content of twenty-five representative strains of Actinobacillus actinomycetecomitans isolated from juvenile and adult periodontitis patients was compared to that of 15 reference strains of oral and nonoral Actinobacillus species and Haemophilus aphrophilus. Trimethylsilyl derivatives of the fatty acid methyl esters were analyzed by gas-liquid chromatography. The predominant fatty acids of all 40 strains examined were 14:0, 3-OH 14:0, 16 delta, and 16:0. Actinobacillus seminis (ATCC 15768) was unlike the other strains examined because of a greater amount of 14:0 detected. The soluble protein analysis using polyacrylamide gel electrophoresis revealed that A. actinomycetemcomitans, H. aphrophilus, and nonoral Actinobacillus species possessed distinct protein profiles attesting to the validity of separating these organisms into different species. Established biotypes of A. actinomycetemcomitans could not be differentiated on the basis of fatty acid or protein profiles.
Plasmid analyses in clinical isolates of Bacteroides fragilis and other Bacteroides species
Authors: Wallace B L , Bradley J E , Rogolsky M
Published: January 2020

Plasmid analyses were performed on Bacteroides strains isolated from clinical specimens. Of 32 Bacteroides strains, 8 were found to contain plasmids. Seven of these eight strains were B. fragilis, and the other one was B. distasonis. Three of these eight strains harbored only a 3.0-megadalton plasmid. Two strains had only a 2.0-megadalton plasmid, and one had 2.0-, 3.0-megadalton plasmid. Of the remaining two strains, one had 2.0-, 3.0-, and 5.0-megadalton plasmids, and the other had 3.0- and 5.0-megadalton plasmids. Beta-Lactamase was produced by 93% of the clinical isolates. Seven of the eight plasmid-carrying strains were cadmium resistant, five were zinc resistant, four were mercury resistant, and two expressed a brick-red fluorescence under ultraviolet light. None of these traits could be associated with a plasmid after performing either curing experiments or genetic transfer experiments by cell-to-cell contact.
Comparison of a slide blood culture system with a supplemented peptone broth culture method
Authors: Bryan L E
Published: January 2020

A slide blood culture system (Roche Diagnostics, Div. Hoffman-La Roche, Inc., Montreal, Canada; Roche BCB) was compared with a supplemented peptone broth Vacutainer method (Becton, Dickinson & Co., Rutherford, N.J.) on blood samples taken from the same 1,209 patients. Significantly more clinically important isolates were isolates with the Roche BCB system, and technical processing time was reduced. However, significantly more contaminants were isolated with the Roche BCB system, and it could not readily be adapted for anaerobic culture. Contamination was reduced by careful tightening of the slide to the bottle top to prevent any leakage. Overall, the BCB system is a satisfactory method for aerobic blood culture, markedly reducing technical processing time relative to most other blood culture methods.
BrucELISA: an enzyme-antibody immunoassay for detection of Brucella abortus antibodies in milk: correlation with the Brucella ring test and with shedding of viable organisms
Authors: Boraker D K , Stinebring W R , Kunkel J R
Published: January 2020

An indirect enzyme-antibody immunosorbent assay (BrucELISA) is described for the detection of antibody to Brucella abortus in cow's milk. Three series of milk samples were obtained from an adult-vaccinated dairy herd infected with B. abortus. The BrucELISA system was used as a screening test for individual milks diluted 1:200 (BE 200 test), for undiluted bulk milks, and to determine antibody titer (BrucELISA titration assay). The BrucELISA results correlated highly with positive Brucella ring test reactions and culture positivity, eliminated false-positive Brucella ring test reactions, detected antibody in some samples which were Brucella ring test negative, and distinguished between vaccinated and infected animals. BrucELISA titration assay titers of greater than 1:800 were correlated with shedding, or were prognostic for animals which eventually became shedders. Binding of the enzyme-antibody conjugate to bovine immunoglobulin in the absence of rabbit anti-bovine immunoglobulin occurred with culture-positive or -negative milks showing titers of greater than 1:1,600 (the beta effect); the effect was also of predictive value in identifying eventual shedders. The BrucELISA system is a sensitive, specific, and inexpensive method for screening large numbers of individual or bulk milk samples for the presence of antibody to B. abortus.
Evaluation of the Repliscan II System for identification of Enterobacteriaceae
Authors: Woolfrey B F , Fox J M , Quall C O
Published: January 2020

In a precious report (Woolfrey et al., J Clin. Microbiol. 13:58-61, 1981), we indicated that the Repliscan system did not reliably identify Enterobacteriaceae. Recent improvements in the system prompted us to evaluate Repliscan II (Cathra International, Inc., St. Paul, Minn.) by using representative isolates of the population sample previously used to test the system. Isolates (692) representing eight genera were identified in parallel by the Repliscan II and API 20E (Analytab Products, Plainview, N.Y.) systems. Isolates given different identifications by the two systems were assigned reference identifications by using classical microbiological methods. Repliscan II identified 95.2% correctly, 4.0% incorrectly, and 0.7% as unknown. API 20E identified 99.4% correctly, 0.6% incorrectly, and none as unknown. Repliscan II correctly identified Salmonella and Shigella spp. to the genus level and isolates of six other genera to the species level as follows: Salmonella spp., 100%; Shigella spp., 97.7%; Escherichia spp., 95.2%; Citrobacter spp., 82.1%; Enterobacter spp., 85.2%; Klebsiella spp., 98.6%; Proteus spp., 97.2%; and Serratia spp., 97.9%. These findings indicate that Repliscan II is a significantly improved system and provides acceptable identification of Enterobacteriaceae.
Cost analysis of the Auto Microbic system urine identification card
Authors: Hamoudi A C , Lin S
Published: January 2020

The AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.) is a fully automated, computerized instrument. One of the most thoroughly studied aspects of the system is its ability to identify and quantify of nine most common urinary tract pathogens. The major advantages of the AutoMicrobic system are that the results of urine cultures are available in a fraction of the time required by conventional methods and that samples can be processed with fewer man-hours. Although the specificity, sensitivity, and reliability of the system have been amply described in the literature, only one study combines these aspects with a cost analysis (D. P. Nicholson and J. A. Koepke, J. Clin. Microbiol. 10:823-833, 1979). Because one of the major considerations that microbiology laboratory directors must face is the cost-effectiveness of instruments, we studied the relative cost of the system as compared with that of conventional methods. Our findings indicated that the instrument provided valuable savings in technologist and turnaround time. These advantages were most easily realized in processing positive urine specimens. For negative urine specimens, the savings in technologist time were cancelled by the cost of the consumable supplies, which greatly exceeded the cost of supplies for conventional methods. This disadvantage might be ameliorated by efficient and effective screening methods to exclude those urine specimens that are most probably negative.
Enzyme Immunoassays for Measurement of Cytomegalovirus Immunoglobulin M Antibody
Authors: Yolken Robert H. , Leister Flora J.
Published: January 2020

The diagnosis of congenital cytomegalovirus (CMV) infection is often accomplished by the detection of circulating antibody directed against CMV. We devised a method for measuring CMV-specific immunoglobulin M (IgM) based on the isolation of IgM antibody by reaction with a solid phase coated with antihuman IgM. The determination of IgM antibody specific for CMV was accomplished by the subsequent addition of CMV or control antigen and enzyme-labeled CMV antibody (solid phase-IgM method). We compared the sensitivity and specificity of this method with those of a conventional form of solid-phase enzyme immunoassay in which CMV antigen is bound to the solid phase (solid phase-antigen method). Both assay systems were capable of detecting CMV-specific IgM antibody in the sera of 10 babies with documented CMV infection and in those of the mothers of 4 of these babies. The solid phase-IgM method yielded negative results in all 66 sera available from babies who did not have congenital CMV infection. On the other hand, the solid phase-antigen system yielded false-positive results in 12 (18%) of these sera. In addition, the solid phase-antigen system yielded false-positive results in 8 of 12 sera obtained from patients with demonstrable rheumatoid factor. However, the solid phase-IgM system yielded negative results for the rheumatoid sera, provided that appropriate control reactions were performed. The solid phase-IgM system is thus a specific and sensitive method for the determination of CMV IgM antibody.
Improved streptococcal grouping antisera containing polyethylene glycol
Authors: George J R , Ashworth H , Facklam R R , Harrell W K , Palmer D F
Published: January 2020

Antisera to streptococcal groups A through G containing 4% polyethylene glycol 6000 were prepared and evaluated. Seventy strains of homologous and heterologous beta- and non-beta-hemolytic streptococci were included in the evaluation. Homologous reactions were determined against extracts prepared by four extraction methods: hot hydrochloric acid (Lancefield) extraction, autoclave extraction, hot formamide extraction, and nitrous acid extraction. Enhancement of the precipitin reaction in the presence of polyethylene glycol 6000 permitted a fourfold dilution of all antisera for use in the capillary precipitin test. At this dilution, the potency of the antisera exceeded requirements established for these reagents.
Correlation of penicillin minimum inhibitory concentrations and penicillin zone edge appearance with staphylococcal beta-lactamase production
Authors: Gill V J , Manning C B , Ingalls C M
Published: January 2020

Production of staphylococcal beta-lactamase was shown to be correlated with penicillin G minimum inhibitory concentrations (MICs) of greater than 0.05 microgram/ml for 97% of the Staphylococcus aureus and 99% of the Staphylococcus epidermidis strains tested. However, it is important to note that of the isolates for which MICs were equal to or less than 0.05 micro/ml, a significant percentage (16% of S. aureus and 5% of S. epidermidis) were beta-lactamase producers. Thus, lack of beta-lactamase production, which implies susceptibility to penicillin, cannot be presumed solely on the basis of low MICs. Beta-lactamase production can be easily predicted from disk diffusion susceptibility tests by observing the appearance of the penicillin inhibition zone edge. A sharply demarcated edge was correlated with beta-lactamase production for 100% of the S. aureus and 93% of the S. epidermidis strains tested. The presence of this type of zone edge when a penicillin zone measures in the intermediate or susceptible range indicates that the isolate should be checked for beta-lactamase production.
Comparison between a modified counterimmunoelectrophoresis test and the indirect fluorescent-antibody test for detection of antibodies to rabies virus in human sera
Authors: Díaz A M , Myers D M
Published: January 2020

The correlation between the modified counterimmunoelectrophoresis test and the indirect fluorescent-antibody test for detecting rabies virus antibodies in human sera was studied, using as a reference the standard mouse neutralization test. Higher sensitivity was observed with the counterimmunoelectrophoresis technique than with the indirect fluorescent-antibody method. The counterimmunoelectrophoresis test provides a practical quantitative alternative to the indirect fluorescent-antibody method.
Misleading Results from Bone Marrows Cultured for Mycobacteria
Authors: Dawson David J. , Blacklock Zeta M.
Published: January 2020

Re-evaluation of a previous study dealing with atypical mycobacteria grown from specimens of bone marrow provided strong evidence that the isolates were of no clinical significance. Bone marrow culture should not be undertaken as a diagnostic test for pulmonary atypical mycobacteriosis.
Hemolytic reaction of clinical and environmental strains of Vibrio vulnificus
Authors: Johnson D E , Calia F M
Published: January 2020

All Vibrio vulnificus strains tested (four clinical isolates and eight environmental isolates) hemolyzed human erythrocytes. In contrast to findings with Vibrio parahaemolyticus, in which hemolytic activity correlates with isolation from clinical specimens, results from the present study suggest that hemolysis is not usefull in differentiating V. vulnificus strains.
Single radial immune hemolysis test for detection of Escherichia coli thermolabile enterotoxin
Authors: Serafim M B , de Castro A F , Leonardo M B , Monteiro A R
Published: January 2020

A single radial immune hemolysis test for the detection of thermolabile enterotoxin has been developed for routine purposes. Stationary cultures from enterotoxigenic Escherichia coli in Casamino Acids-yeast extract medium may be used for the detection of this enterotoxin, and under the conditions of the experiment, the single radial immune hemolysis test was as sensitive as the passive immune hemolysis test. The results obtained in the single radial immune hemolysis test agreed entirely with those obtained in the passive immune hemolysis test, and no false-positive reactions were obtained when cholera antitoxin diluted 1:80 was used. The assay is easy to perform, inexpensive, and specially designed for less-equipped laboratories.
Rapid and direct staphylocoagulase assay that uses a chromogenic substrate for identification of Staphylococcus aureus
Authors: Engels W , Kamps M A , van Boven C P
Published: January 2020

A rapid and direct assay of staphylocoagulase that uses chromogenic substrate Chromozym TH was developed for the routine identification of Staphylococcus aureus. False-positive and false-negative reactions were eliminated by measuring the activity of staphylocoagulase-prothrombin complex directly. The method was compared with three clotting assays. The results show that the new assay is specific, quantitative, and easily automated. If offers clinical laboratories a more reliable assay of staphylocoagulase and is a valuable aid for the rapid identification of S. aureus.
Production of antiserum to respiratory syncytial virus polypeptides: application in enzyme-linked immunosorbent assay
Authors: Hornsleth A , Grauballe P C , Friis B , Genner J , Pedersen I R
Published: January 2020

By use of crossed immunoelectrophoresis techniques, respiratory syncytial (RS) virus-specific precipitates were produced between RS virus cellular antigen [solubilized in tris(hydroxymethyl)aminomethane-glycine buffer, pH 9] and antiserum raised in rabbits against semipurified RS virus. When these precipitates were employed as antigens for further immunizations in rabbits, antibodies (anti-RSV-precip.I) were produced which reacted with only one RS virus antigen when tested by the crossed immunoelectrophoresis technique. Precipitates obtained between RS virus cellular antigen (labeled with L-[35S]methionine) and anti-RSV-precip.I were examined by polyacrylamide gel electrophoresis, which showed that anti-RSV-precip.I precipitated RS virus polypeptides of molecular weights 28,000 to 84,000. Anti-RSV-precip.I was employed as capture antibodies in the enzyme-linked immunosorbent assay, in which RS virus cellular antigen was used as the second layer. Determination of human RS virus immunoglobulin G antibodies by this enzyme-linked immunosorbent assay technique showed a high degree of sensitivity, specificity, and reproducibility.
Detection of respiratory syncytial virus in nasopharyngeal secretions by inhibition of enzyme-linked immunosorbent assay
Authors: Hornsleth A , Brenøe E , Friis B , Knudsen F U , Uldall P
Published: January 2020

An enzyme-linked immunosorbent assay developed for the demonstration of respiratory syncytial (RS) virus immunoglobulin G antibodies was used for the detection of RS virus in specimens of nasopharyngeal secretions (NPS) obtained from children with acute respiratory disease. Samples of NPS were incubated with a fixed amount of standard serum (human serum antibodies to RS virus) before being added to the enzyme-linked immunosorbent assay test plate. A decrease in the optical density value determined for this standard serum was seen with majority of NPS specimens from which RS virus had been isolated in tissue culture. The reliability and the specificity of this inhibiton test were supported by experiments with purified RS virus and by tests with NPS specimens containing other respiratory viruses.
Bacteriology of acute obstructive suppurative cholangitis of the aged
Authors: Shimada K , Noro T , Inamatsu T , Urayama K , Adachi K
Published: January 2020

Bacteriological examination was performed on bile from 23 patients with acute obstructive suppurative cholangitis. Of 23 bile cultures, 15 yielded aerobic and anaerobic bacteria and 8 yielded aerobic bacteria only. No specimen grew anaerobic bacteria only. A total of 20 cultures yielded a polymicrobial flora, and 3 cultures grew one single aerobic bacterium. Escherichia coli, Klebsiella sp., enterococci, Bacteroids fragilis, and Clostridium perfringens were the predominant bacterial flora in bile of these patients. Bacteremia was documented in 10 of the 12 patients cultured. All bacteremias involved aerobic bacteria and were polymicrobial in three patients. The frequent presence of anaerobes in bile of patients with acute obstructive suppurative cholangitis suggests that antimicrobial therapy should provide adequate coverage for anaerobic bacteria and enteric organisms.
Mannose in body fluids as an indicator of invasive candidiasis
Authors: Monson T P , Wilkinson K P
Published: January 2020

Using gas-liquid chromatography, we measured five mannose in the serum of six nondiabetic patients with autopsy-proven invasive candidiasis. In all patients serum mannose concentrations were higher than mannose levels found in serum from normal adults and children or from patients with catheter-associated candidemia, mucosal candidiasis, and other mycoses. Spinal fluid from two patients with Candida meningitis showed increased free mannose as compared to seven non-inflammatory spinal fluid samples. However, free mannose in the serum of poorly controlled diabetics (blood glucose of greater than or equal to 300 mg/dl) did overlap concentrations in patients with invasive candidiasis. In vitro culture of Candida albicans demonstrated increasing concentrations of mannose associated with growth of the organism. We conclude that physical and chemical assay for mannose in body fluids may be a useful technique to assist in the diagnosis in invasive candidiasis.
Variations in properties of Nocardia asteroides resulting from growth in the cell wall-deficient state
Authors: Beaman B L , Bourgeois L
Published: January 2020

Several revertants possessing a cell wall were obtained from L-forms of Nocardia asteroides 10905. These L-form revertants differed from the parent in respect to colonial and cellular morphology, pigmentation, metabolic, capacities, cell wall structure, and mycolic acid composition. These data suggest a possible mechanism for the taxonomic heterogeneity and phenotypic diversity observed among N. asteroides strains.
Modified colony indirect epifluorescence test for serotyping Ureaplasma urealyticum and an adaptation to detect common antigenic specificity
Authors: Stemke G W , Robertson J A
Published: January 2020

A microtiter method for efficiently serotyping colonies of Ureaplasma urealyticum by immunofluorescence is described. Prior detergent treatment allowed identification of common group determinants.
Evaluation of detection of positive blood cultures by pH changes
Authors: King G S , Niles A C , Murray P R
Published: January 2020

We observed that the pH of positive blood culture broths was significantly lower than that of negative broths. However, significantly pH changes were generally not observed for subculture-positive, macroscopically negative blood culture broths. Therefore, although the pH of most positive blood culture broths was reduced significantly, this change was generally not useful for the early detection of positive cultures.
Enteric group 15 (Enterobacteriaceae) associated with pneumonia
Authors: Bae B H , Sureka S B , Ajamy J A
Published: January 2020

A gram-negative, oxidase-negative, fermentative rod belonging to enteric group 15 of Enterobacteriaceae was isolated in mixed culture from two patients with pneumonia. Both were elderly patients with chronic heart disease.
Atypical toxin variant of Clostridium botulinum type B associated with infant botulism
Authors: Hatheway C L , McCroskey L M , Lombard G L , Dowell V R
Published: January 2020

An atypical toxin variant of Clostridium botulinum (strain 657) was isolated from the feces of a 6-week-old female infant whose symptoms and clinical history were consistent with infant botulism. Toxin detected in the feces and the toxin produced by isolates from the feces and from two rectal swabs could be neutralized by type B botulinal antitoxin only at very high ratios of of antitoxin to toxin in the neutralization mixture. One international unit of type B antitoxin neutralized only about 10 lethal doses of 657 toxin as compared with approximately 10,000 lethal doses of conventional type B toxin from the Beans strain. Antitoxin prepared against 657 toxin was 10 times more effective against the conventional toxin than against the homologous toxin. Toxoid-antitoxin-binding studies indicate that both 657 toxin and type B toxin are heterogeneous and that both toxins may contain the same molecular variants, but that the proportions of the variants are different in each.
Cellular fatty acid composition of organisms frequently associated with human infections resulting from dog bites: Pasteurella multocida and groups of EF-4, IIj, M-5, and DF-2
Authors: Dees S B , Powell J , Moss C W , Hollis D G , Weaver R E
Published: January 2020

The cellular fatty acid composition of Pasteurella multocida and four unclassified groups of gram-negative bacteria (EF-4, M-5, IIj, and DF-2) which are frequently associated with human dog-bite infections was determined by gas-liquid chromatography. Strains of P. multocida were readily distinguished from the unclassified groups by the presence of 3-hydroxy myristic acid (3-OH- 14:0). Groups M-5 and EF-4 were characterized by the presence of 3-hydroxy lauric (3-OH-12:0) acid. Only group EF-4 organisms contained 2-OH-16:0, a-17:0, and 17:0 cyclopropane acids. Groups IIj and DF-2 differed from the other groups by the presence of large amounts of a branched-chain 15-carbon acid (i-15:0); they differed from each other by the presence of i-2-OH-15:0 and i-17:1 acids in IIj, which were absent in DF-2. The data indicate that gas-liquid chromatographic analysis for cellular fatty acids provides an additional test for rapid differentiation of these gram-negative organisms associated with dog-bite infections. Similarities observed in the fatty acid compositions of Flavobacterium, IIj, and DF-2 suggest that these two unclassified groups may be additional species of Flavobacterium.
Evidence against the practicality and cost-effectiveness of a gram-positive coccal selective plate for routine urine cultures
Authors: Bale M J , Matsen J M
Published: January 2020

A total of 899 urine cultures were evaluated to assess the need for and cost-effectiveness of using a gram-positive coccal selective plate in the initial plating of urine cultures. Of these cultures, 437 were examined retrospectively and 462 were examined prospectively. Urines were quantitatively plated to three media: sheep blood sugar, MacConkey agar, and phenyl ethyl alcohol agar. Of all urine samples in both studies, 52% yielded no growth on any of the three media. Of all 899 urine cultures, there were only 5 cultures (less than 1%) in which a significant count of a gram-positive organism was detected only on the phenyl ethyl alcohol agar plate and not recoverable on the sheep blood agar plate. In each of these five instances, the need for the use of the selective plate occurred when a Proteus mirabilis strain swarmed over an enterococcus. The inclusion of a selective gram-positive coccal medium for initial plating of urine cultures is unnecessary and not cost-effective. When Proteus swarms on sheep blood agar, a sweep should be made with an inoculating loop from the sheep blood agar and streaked to phenyl ethyl alcohol agar or a similar gram-positive coccal selective medium.
Comparison of liquid growth media for Legionella pneumophila
Authors: Saito A , Rolfe R D , Edelstein P H , Finegold S M
Published: January 2020

Ten liquid media were compared under standard conditions for their ability to support the growth of Legionella pneumophila. Modified gonococcal-ferric cysteine broth (without sodium chloride) supplemented with 1% yeast extract yielded the best overall growth of the one strain of L. pneumophila examined. Growth rates were independent of pH changes which occurred during incubation. The growth rates of 10 different strains of L.pneumophila were compared in this medium. There appeared to be little difference in the growth rates of strains passaged frequently or infrequently, or between environmental and clinical isolates. Moderate aeration resulted in a faster growth rate and in approximately a 1 log10 higher final cell concentration as compared to a static broth culture. These experiments demonstrate that there are moderate to marked differences among the various media described in the literature and that no liquid medium yet developed supports rapid growth of L. pneumophila incubated without shaking.
Comparison of three collection-preservation methods for detection of intestinal parasites
Authors: Price D L
Published: January 2020

A study was designed to compare the effectiveness and time efficiency of three collection-preservation methods individually and grouped to form systems. Employees and members of their families of an agricultural grower in Hillsborough County, Fla., provided all fecal specimens used in the study. The 160 individuals participating were members of 64 families and represented a cross section of all job activity levels including management. Specimens were collected in Merthiolate/iodine/Formalin, Formalin (10% neutral buffered solution), and polyvinyl alcohol fixative. Parasites were found in the specimens from 39 (24.4%) of the 160 individuals participating, which represented 29.7% of the 64 families. No single method was effective in recovering all of the parasites found. When the methods were grouped to form diagnostic systems, the Merthiolate/iodine/Formalin system appeared to be more effective for parasite recovery as well as more time efficient than the Formalin/polyvinyl alcohol fixation system.
Comparison of the quantitative direct plating method and the BACTEC procedure for rapid diagnosis of Haemophilus influenzae bacteremia in children
Authors: La Scolea L J , Dryja D , Neter E
Published: January 2020

The efficacy and reproducibility of the quantitative direct plating (QDP) method and the semi-automated BACTEC radiometric system (BBL Microbiology Systems, Cockeysville, Md.) were analyzed for the rapid diagnosis of Haemophilus influenzae bacteremia on the basis of 41 positive cultures from 35 patients. The QDP method detected 61% and BACTEC only 19% of the positive cultures within the first 12 h. Similarly, the QDP procedure yielded growth of H. influenzae in 56% of the cultures before a positive growth index reading was obtained with the BACTEC method. The observation that the quantitative procedure recovered H. influenzae in 88% of all positive cultures, using only 1 ml or less of blood, is attributed to the fact that 68% of the cultures had counts in excess of 100 colony-forming units per ml. The reproducibility of the QDP method was documented by the fact that duplicate blood cultures taken within a few hours of each other yielded comparable results on the number of bacteria at low (1 to 100/ml), moderate (100 to 1,000/ml), and high (greater than 1,000/ml) levels of bacteremia. We concluded that the QDP method is a valuable, simple, and inexpensive supplementary technique to the semi-automated BACTEC procedure for the rapid diagnosis of H.influenzae bacteremia in children.
Comparative recovery of fungi from biphasic and conventional blood culture media
Authors: Kiehn T E , Capitolo C , Mayo J B , Armstrong D
Published: January 2020

A brain heart infusion broth and agar biphasic bottle was compared with a vented broth brain heart infusion bottle for the recovery of fungi from blood. A total of 40 fungi, all yeasts, were recovered from 5,000 blood cultures. The biphasic bottle slightly increased the overall recovery of six species of yeasts. In addition, yeasts were first detected more often in the biphasic bottle (73%) than in the vented broth bottle (38%). A routine early (6- to 24-h) or late (5-day) subculture of macroscopically negative cultures may not be required for yeast isolation when a biphasic medium is used. Of the yeasts initially detected in the biphasic medium, 83% were seen to be growing on the agar slant. Only four were detected from a 24-h subculture, a Candida glabrata of questionable clinical significance, was recovered after our routine blood culture period of 7 days; however, other fungi, not recovered in this study, require extended incubation periods.
Lethal Meningoencephalitis and Septicemia Caused by
Haemophilus influenzae
Type f in an Adult with Multiple Myeloma
Authors: Wagener William C. , Myerowitz Richard L. , Dulabon George M.
Published: January 2020

A patient with chemotherapy-treated multiple myeloma developed overwhelming sepsis and meningoencephalitis with
Haemophilus influenzae
type f. Typable
H. influenzae
other than type b has only rarely been reported as a cause of serious disease in adults. The patient's immunosuppressed status presumably predisposed her to this unusual infection.
Toxigenic
Clostridium perfringens
from a Parvovirus-Infected Dog
Authors: Tilton R. C. , Van Kruiningen H. J. , Kwasnik I. , Ryan R. W.
Published: January 2020

A strain of
Clostridium perfringens
, type A, has been isolated from the intestine of a dog which died from parvovirus infection. This
Clostridium
strain produces a toxin which can be detected by counterimmunoelectrophoresis, using
C. difficile
antitoxin, and produces cytotoxicity in WI-38 cell culture. Cytopathology can be blocked by
C. difficile
antitoxin. Its role in canine parvovirus infection is unknown.
Klebsiella ozaenae
Septicemia Associated with Hansen's Disease
Authors: Murray Katherine A. , Clements Bruce H. , Keas Stephen E.
Published: January 2020

Two cases of
Klebsiella ozaenae
septicemia from the National Hansen's Disease Center, Carville, La., are discussed: one fatal and one nonfatal. Although both patients had nasal complications of Hansen's disease (leprosy), the organism was grown initially from spinal fluid, blood, and wound cultures. This report confirms the potential pathogenicity of the
K. ozaenae
species and its widening disease spectrum.
Typing of
Haemophilus influenzae
by Coagglutination and Conventional Slide Agglutination
Authors: Shively Roxanne G. , Shigel Janet T. , Peterson Ellena M. , De La Maza Luis M.
Published: January 2020

Coagglutination was compared with conventional slide agglutination for the typing of 297 clinical isolates of
Haemophilus
sp. A 100% correlation was found with the
H. influenzae
type b isolates. Coagglutination showed no false-positive reactions with the nontypable strains of
H. influenzae
and
H. parainfluenzae
isolates; however, conventional slide agglutination exhibited many false-positive and non-interpretable reactions.
Improved In Vitro Excystation Procedure for
Giardia lamblia
Cysts
Authors: Rice Eugene W. , Schaefer Frank W.
Published: January 2020

Giardia lamblia
cysts obtained from human symptomatic and asymptomatic donors were excysted in vitro. Excystation averaged 87% for cysts from symptomatic donors and 70% for cysts from asymptomatic donors.
Quantitation of Clostridium botulinum organisms and toxin in the feces of an infant with botulism
Authors: Paton J C , Lawrence A J , Manson J I
Published: January 2020

A 4-month-old boy presented with symptoms and signs characteristic of infant botulism. Examination of feces revealed Clostridium botulinum type B and type B toxin. The numbers of C. botulinum and the amount of toxin in feces were measured throughout the 4-week period in hospital. The maximum numbers and amounts were detected in a fecal specimen collected 16 days after admission: this contained 8.4 X 10(6) C. botulinum type B colony-forming units and 61,440 mouse 100% lethal doses of type B toxin per g (wet weight) of feces. This latter figure is the highest fecal toxin titer reported yet for a case of infant botulism. By day 16, however, substantial improvement in the patient's clinical condition had occurred. This suggests that initiation of recovery from infant botulism is not necessarily preceded by a reduction in the numbers of C. botulinum organisms and the quantity of toxin in the gut.
Presence of hydrogen peroxide in media used for cultivation of Neisseria gonorrhoeae
Authors: Norrod E P , Morse S A
Published: January 2020

Defined complex media used for cultivation of Neisseria gonorrhoeae were tested for the presence of H2O2 by both a spectrophotometric and a polarographic assay. H2O2 (35 to 165 microM) was present in all media tested. In the defined media, H2O2 was generated by the interaction of cysteine with other amino acids. The addition of the chelator 8-hydroxyquinoline prevented formation of detectable H2O2, suggesting that metal ions were necessary. The persistence of H2O2 varied greatly among different media. Medium components which affected the presence of H2O2 were pyruvate, oxalacetate, and sodium sulfite. Sodium sulfite also generated superoxide radical. In liquid medium containing H2O2, the endogenous gonococcal catalase present in an inoculum of about 2 X 10(7) colony-forming units/ml destroyed detectable H2O2. The long lag phase which resulted from a 10-fold lower inoculum could not be shortened by the addition of exogenous catalase. Small amounts of residual H2O2 in agar plates of complex medium affected the viability of gonococci which had been suspended in buffer and incubated for 60 min at 37 degrees C. Addition of pyruvate or catalase increased viable counts in medium containing H2O2.
Impaired response of lymphocytes from non-insulin-dependent diabetics to staphage lysate and tetanus antigen
Authors: Casey J , Sturm C
Published: January 2020

We examined the blastogenic responses of lymphocytes to Staphylococcus aureus antigen in 49 uninfected non-insulin-dependent diabetic subjects and in 56 uninfected nondiabetic subjects. We found a significantly decreased response to this antigen in the diabetic patients (P less than 0.05). Diabetics whose blood sugar levels were less than 150 mg/dl showed significantly decreased responses, as did diabetics whose blood sugar levels were greater than 200 mg/dl. The use of fetal calf serum in place of autologous plasma did not correct the defect. In a smaller number of patients, we observed a decreased response to tetanus toxoid, which was significant (P less than 0.05). The response to phytohemagglutinin was not significantly different from normal controls (P greater than 0.4). These results indicated that non-insulin-dependent diabetics, as well as insulin-dependent diabetics (previously reported), show decreased responses to S. aureus antigen and to tetanus toxoid. This defect apparently is not related to blood glucose levels or other serum factors.
Medium for selective isolation and presumptive identification of the Bacteroides fragilis group
Authors: Lyznicki J M , Busch E L , Blazevic D J
Published: January 2020

We developed and evaluated a new medium (FRAG agar) for the selective isolation and presumptive identification of the Bacteroides fragilis group. This medium contains 1% D-glucuronic acid as a fermentable carbon source, a reduced peptone content, gentamicin, and 20% bile. Presumptive identification of the B. fragilis group was based on growth, fermentation, and typical colony morphology. A total of 75 stock culture isolates of the B. fragilis group grew well on this medium, and 69 showed evidence of fermentation. Of 90 other anaerobes, none grew well or fermented glucuronic acid. In a clinical trial of 100 specimens sent for anaerobic culture, FRAG agar inhibited 71 of 71 anaerobes not belonging to the B. fragilis group, as well as 104 of 110 facultative organisms. A total of 33 isolates of the B. fragilis group were recovered on the selective medium, whereas only 23 were recovered by routine methods. Of 23 cultures positive for the B. fragilis group on routine plates, 22 were positive on FRAG agar.
Selective differential human blood bilayer media for isolation of Gardnerella (Haemophilus) vaginalis
Authors: Totten P A , Amsel R , Hale J , Piot P , Holmes K K
Published: January 2020

New selective and differential human blood bilayer agar media with Tween 80 (HBT medium) or without Tween 80 (HB medium), developed for the isolation of Gardnerella (Haemophilus) vaginalis, permitted significantly higher G. vaginalis isolation rates than have been obtained for other media used for this purpose. HB medium consists of a basal layer of Columbia agar base containing colistin and naladixic acid with added amphotericin B and an overlayer of the same composition plus 5% human blood. HBT agar also contains Proteose Peptone No. 3 (Difco Laboratories) and Tween 80 in the basal layer and the overlayer. Both Tween 80 and the bilayer composition enhanced G. vaginalis production of human blood hemolysis, permitting detection of this organism even in the presence of heavy growth of other vaginal flora. The use of HB or HBT medium thus permitted the demonstration that G. vaginalis was present in vaginal fluid from a large percentage (up to 68%) of normal women. However, the concentration of G. vaginalis was found by semiquantitative analysis to be significantly higher in vaginal fluid from women with nonspecific vaginitis than in fluid from normal women.
Detection of rotavirus antibody by inhibition of reverse passive hemagglutination
Authors: Sanekata T , Yoshida Y , Oda K , Okada H
Published: January 2020

A reverse passive hemagglutination inhibition (RPHI) test was developed for detecting rotavirus antibody in patients' sera. Sheep erythrocytes coated with guinea pig antibody against Nebraska calf diarrhea virus (NCDV) were readily hemagglutinated by NCDV in a reaction called reverse passive hemagglutination (RPHA). Inhibition of this RPHA reaction was used to detect the presence of rotavirus antibody in patients' sera which cross-reacted with NCDV. The sensitivity of RPHI was consistently at least 10 times greater than that of the complement fixation test for detection of rotavirus antibody in patients' sera. Since immunoglobulin M (IgM) antibody activity detected by RPHI was destroyed by pretreatment of serum with dithiothreitol (DDT), whereas DTT-resistant antibodies such as IgG remained unchanged, the titer of IgM antibody could be distinguished from that of DTT-resistant antibody by comparing the results of two RPHI tests performed with and without DTT pretreatment. From the IgM antibody response pattern, primary and secondary rotavirus infections could be distinguished.
Visualization of Histoplasma capsulatum in bone marrow with Prussian blue iron stain
Authors: Caldwell C W , Taylor H
Published: January 2020

Bone marrow specimens from patients with disseminated histoplasmosis revealed yeast cells that stained positively with Prussian blue.
Bluetongue virus: comparative evaluation of enzyme-linked immunosorbent assay, immunodiffusion, and serum neutralization for detection of viral antibodies
Authors: Poli G , Stott J , Liu Y S , Manning J S
Published: January 2020

Comparative studies on the detection of bovine serum immunoglobulin G antibodies to bluetongue virus with an enzyme-linked immunosorbent assay, an immunodiffusion method, and a serum neutralization assay demonstrated complete concordance between the enzyme-linked immunosorbent assay and the serum neutralization assay results. However, the immunodiffusion method failed to detect bluetongue virus antibody in a substantial number of sera found to possess bluetongue virus immunoglobulin G with the enzyme-linked immunosorbent assay.
Comparison of Enteric-Tek with API 20E and conventional methods for identification of Enterobacteriaceae
Authors: Bruckner D A , Clark V , Martin W J
Published: January 2020

Enteric-Tek (Flow Laboratories, Inc., Roslyn, N.Y.) is a new system designed for the identification of Enterobacteriaceae and consists of a round, multicompartmented plastic plate with 11 peripheral wells and 1 center well incorporating 14 biochemical reactions. Only one reagent (Kovacs reagent) is required to complete the biochemical test results for generation of a five-digit number for computer code identification. The identification accuracy of the Enteric-Tek system was compared with those obtained by conventional methods and API 20E. The Enteric-Tek system was found to be not only a convenient and simple method for rapid identification of Enterobacteriaceae, but also to be highly reliable, giving excellent identification performance as compared to API 20E and conventional methods.
New medium for differentiation of Cryptococcus neoformans serotype pairs
Authors: Salkin I F , Hurd N J
Published: January 2020

A new medium effectively differentiates B and C serotypes of Cryptococcus neoformans from A and D serotypes on the basis of resistance to low concentrations of cycloheximide and assimilation of glycine as the sole carbon source.
Blood-lysing solution nontoxic to pathogenic bacteria
Authors: Zierdt C H
Published: January 2020

A blood-lysing solution is described that is noninjurious to fastidious microbial pathogens. Lysed blood is filtered, and the filter with collected microbes is cultured.
Identification of Gardnerella (Haemophilus) vaginalis
Authors: Piot P , Van Dyck E , Totten P A , Holmes K K
Published: January 2020

Different tests for the identification of Gardnerella (Haemophilus) vaginalis and for its differentiation from catalase-negative unclassified coryneforms from the vagina were evaluated on over 200 bacterial strains, with special emphasis on optimal test conditions. A presumptive identification of G. vaginalis in the clinical laboratory can be made on the basis of colonial morphology, clear beta-hemolysis with diffuse edges on human blood bilayer-Tween agar, a negative catalase test, and typical cell morphology in the Gram stain. This procedure will correctly identify 90 to 98% of suspect colonies of G. vaginalis with human blood bilayer-Tween agar as primary isolation medium. Useful additional reactions for the confirmation of G. vaginalis include positive hippurate and starch hydrolysis, positive alpha-glucosidase but negative beta-glucosidase tests, the production of acid from glucose and maltose but not from mannitol, and susceptibility to disks containing metronidazole, nitrofurantoin, sulfonamides, and bile.
Accuracy of a rapid carbohydrate oxidation microtube method for identification of nonfermentative gram-negative bacilli
Authors: Qadri S M , Carr D L , Narayanan R , Ottman J
Published: January 2020

A rapid carbohydrate oxidation microtube system (Carr Microbiologicals, Wichita, Kans.), designed for detecting the saccharolytic activity of gram-negative, nonfermenting bacilli, was evaluated and compared with the conventional oxidation-fermentation method. The oxidation of glucose, maltose, lactose, and xylose was tested with 430 strains of Pseudomonas, Acinetobacter, Achromobacter, Alcaligenes, Moraxella, Flavobacterium, and Bordetella species. More than 95% of the isolates tested gave correct oxidation reactions within 4 h in the rapid carbohydrate oxidation microtubes, whereas oxidation-fermentation media required 24 h to achieve the same sensitivity. The microtube system was found to be simple, accurate, rapid, and economical.
Biochemical characteristics of enterotoxigenic Aeromonas spp
Authors: Burke V , Robinson J , Atkinson H M , Gracey M
Published: January 2020

Biotypes of Aeromonas spp. correlated well with enterotoxin production in a study of 174 strains. Using biochemical characteristics determined by conventional methods and multitest systems, we correctly classified 93% of the strains with regard to enterotoxin production. Most of the enterotoxigenic strains were Voges-Proskauer (VP) positive and did not hydrolyze arabinose, but VP-positive strains which hydrolyzed arabinose were mainly non-enterotoxigenic. Aeromonas punctata subsp. caviae, which is VP negative and does not oxidize gluconate or produce gas from glucose, was non-enterotoxigenic. Although the number of other VP-negative strains was small, most were enterotoxigenic. Discrimination was improved so that 97% of the strains were correctly classified if the hemolysin assay was used either for all strains or for only the VP-positive, arabinose-positive and VP-negative, non-A. punctata subsp. caviae strains. Because the proposed classification system does not require facilities for carrying out in vivo assays such as suckling mouse or ileal loop methods, the identification of enterotoxigenic Aeromonas strains should be possible in diagnostic laboratories and will facilitate epidemiological studies of the role of these organisms in acute diarrhea.
Comparison of atmospheres of incubation for primary isolation of Campylobacter fetus subsp. jejuni from animal specimens: 5% oxygen versus candle jar
Authors: Luechtefeld N W , Reller L B , Blaser M J , Wang W L
Published: January 2020

An atmosphere with reduced oxygen tension is required for the primary isolation of Campylobacter fetus subsp. jejuni. Therefore, we compared use of the conventional atmosphere of 5% oxygen and 8% carbon dioxide with use of a candle jar (17% oxygen and 3% carbon dioxide) for primary isolation of C. fetus subsp. jejuni from 263 positive canine, cattle, and turkey fecal or cecal specimens. At an incubation temperature of 42 degrees C, the atmosphere with 5% oxygen resulted in more Campylobacter colonies per plate (P less than 0.005) and consistently larger Campylobacter colonies (P less than 0.005) than did the candle jar, whereas the growth of interfering flora was similar. Overall, 96% of the 263 specimens were positive for C. fetus subsp. jejuni with 5% oxygen, and 90% were positive with the candle jar (P less than 0.02). More striking differences in isolation rates were seen when both the temperature and the atmosphere were varied: 5% oxygen at 42 degrees C enabled recovery of 93% of the isolates from 70 positive specimens, versus 46% recovery with the candle jar at 37 degrees C. Results with 5% oxygen at 37 degrees C were intermediate. The addition of FBP supplement (0.25% each of ferrous sulfate, sodium metabisulfite, and sodium pyruvate) to Campy-BAP selective medium made no improvement over unsupplemented medium at 42 degrees C (whether in 5% oxygen or in the candle jar), but there was significant improvement over unsupplemented medium when both media were incubated at 37 degrees in the candle jar.