bluuebunny/crossref_metadata_2025 huggingface dataset:
Semiquantitative culture results and pathogenic significance of obligate anaerobes in peritonsillar abscesses
Authors: Jokipii A M , Jokipii L , Sipilä P , Jokinen K
Published: January 2020

We studied the bacteria in consecutive peritonsillar abscesses using semiquantitation of the primary culture findings and correlated the results to clinical parameters. Puncture-aspirated pus from 42 abscesses yielded 133 isolates. Group A streptococci were isolated 10 times and, unlike other bacteria, were isolated 4 times in pure culture; other beta-hemolytic streptococci were found in 8 abscesses, and anaerobes were found in 28. The infections were polymicrobial, with two to seven bacteria in 83%. Anaerobes were more abundant than nonanaerobes; members of the genera Streptococcus, Bacteroides, Peptostreptococcus, and Fusobacterium were the most important quantitatively, considering both frequency and abundance. In patients with ongoing antibiotic treatment, nonanaerobes (but not anaerobes) were less abundant than in untreated patients. The abundance of obligate anaerobes (specifically cocci and gram-positive rods) correlated to the severity of illness as defined by fever and short duration before hospitalization. With other groups of bacteria, no such correlation was found. The correlation was not explained by a difference between the antibiotic-treated and the untreated patients. The results indicate the value of the semiquantitation of culture data and the frequency and pathogenic significance of obligate anaerobes in peritonsillar abscesses.
Biochemical and ultrastructural study of Blastocystis hominis
Authors: Zierdt C H , Donnolley C T , Muller J , Constantopoulos G
Published: January 2020

This study was prompted by the paradox of strong presence of mitochondria in an anaerobic protozoan, recently reclassified from the yeasts. Stemming from publication in 1911 to 1912, Blastocystis hominis has been generally accepted as a harmless intestinal yeast of humans, with short standardized textbook (parasitology) descriptions, even to the present day. Reports since 1967 have changed the classification of B. hominis from yeast to protozoan (Sarcodina), and this has been followed by interest in B. hominis-caused disease, resulting in documentation of disease in humans and other primates. In this study of B. hominis, the basic ultrastructure of the mitochondria was shown by thin-section electron microscopy to be identical to that of an archetypical mitochondrion. There were hundreds of them in large B. hominis cells (100 to 200 microns in diameter). Mitochondria were confined to a peripheral ring of cytoplasm bounded by the outer cell membrane (there is no cell wall) and the membrane of the large, spherical, organelle-free central body that constitutes 75% of the cell's volume. Mitochondria tended to surround the cell's usual two to four nuclei. Rhodamine 123 stained the mitochondria selectively, visualized by fluorescence microscopy. The cell was devoid of cytochromes. Addition of 0.1% cytochrome c to the growth medium increased utilization of glucose by 34% and that of lactate by 17%. Furthermore, it markedly increased the number of mitochondrion-filled cells. At higher concentrations, cytochrome c inhibited the growth of the cells. Despite the presence of large numbers of mitochondria, activities of the mitochondrial enzymes pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase complex, isocitrate dehydrogenase, glutamate dehydrogenase, and cytochrome c oxidase were absent. Thus, the function of the mitochondria in B. hominis remains unknown. Considerable activities of aspartate aminotransferase and alanine aminotransferase were found. Aldolase activity was prominent. Pyruvate decarboxylase was present. Diaphorase and lactate dehydrogenase were detectable but in suspect quantities. Other missing enzymes were gamma glutamyl transpeptidase, alkaline phosphatase (a lysosomal marker), and creatine kinase isoenzymes.
Production of lipase by clinical isolates of Pseudomonas cepacia
Authors: Lonon M K , Woods D E , Straus D C
Published: January 2020

Ten clinical isolates of Pseudomonas cepacia from the sputum of cystic fibrosis patients were examined for the ability to produce lipase. Lipase substrates used included egg yolk agar, four different polyoxyethylene sorbitans (Tweens), and p-nitrophenylphosphorylcholine, a chromogenic substrate used to assay for phospholipase C. Lipase activity was detected in the filtrates of organisms grown to the exponential phase in either tryptose minimal medium or chemically defined medium. Lipase activity increased in the filtrates if the cultures were allowed to proceed into the stationary phase. None of the isolates produced phospholipase C. Lipase activity on Tween 20 ranged from 41.6 X 10(-3) to 640.0 X 10(-3) U/micrograms of protein. The activity was similar or slightly lower when Tween 40, 60, or 80 was used as the substrate. There was no correlation between lipase activity on Tween and that demonstrated on egg yolk agar. Lipase activity increased as pH increased from 7.0 to 9.0. Boiling for 5 min resulted in 66% loss of enzyme activity. The remaining activity continued to decrease with increasing boiling time. The enzyme was purified by gel filtration on Sephadex G-200, and the resultant preparation, when subjected to polyacrylamide gel electrophoresis, resulted in a single protein band (molecular weight, approximately 25,000) from which lipase activity could be eluted. The purified lipase was not cytotoxic to HeLa cells, nor was it toxic when injected intravenously into mice.
Pneumococcosuria in adults
Authors: Nguyen V Q , Penn R L
Published: January 2020

Pneumococcal bacteriuria is usually felt to indicate systemic pneumococcal infection, especially in children. To determine the frequency and significance of pneumococcosuria in adults, we reviewed the Shreveport Veterans Administration Medical Center microbiology laboratory log books for 1982 through 1985. During these 4 years, more patients had pneumococcal bacteriuria than bacteremia (38 versus 33), but only 2 patients had both. The medical records from 31 bacteriuric patients with 35 positive urine specimens were available for review. The collection technique was reliable for 23 (66%) urine specimens; the type of collection technique used was unknown for 12 (34%). The urine pH was less than or equal to 6 in 24 (68%) specimens, and the specific gravity was greater than 1.020 in 12 specimens (34%). A total of 24 specimens grew only Streptococcus pneumoniae, and 11 yielded mixed growth. All 31 patients were men, 25 (81%) were 60 years or older, and 13 (42%) had underlying genitourinary disorders. A total of 7 patients had genitourinary symptoms, 7 had fever, 5 had leukocytosis, and 12 had pyuria; however, these symptoms and signs were frequently accounted for by factors other than pneumococcosuria. Pneumococcosuria was an unexpected finding in all but two (6%) bacteremic patients. These were the only patients with systemic pneumococcal infections, and both died despite appropriate antibiotics. Among the 20 surviving patients with follow-up urine samples, pneumococcosuria resolved whether or not they had received antibiotics. Thus, pneumococcosuria in our patient population was frequently not accompanied by systemic pneumococcal infection and by itself did not require antibiotic therapy.
Effect of treatment of shell vial cell cultures with dimethyl sulfoxide and dexamethasone for detection of cytomegalovirus
Authors: Espy M J , Wold A D , Ilstrup D M , Smith T F
Published: January 2020

Urine specimens submitted for the diagnosis of cytomegalovirus infection were inoculated into shell vials that had been pretreated with a combination of dimethyl sulfoxide (DMSO) and dexamethasone (DEX). The results were compared with those for inoculated shell vials which had received no drug treatment. Of 664 specimens, 100 (15%) were positive for cytomegalovirus. Of the 100 strains of cytomegalovirus, 88 (88%) were detected in both DMSO-DEX-treated and untreated shell vials. Of the remaining 12 positive specimens, 6 were detected with untreated shell vials exclusively and 6 were detected with DMSO-DEX-treated shell vials alone (not significant by the sign test). The median number of fluorescent foci was not significantly higher in DMSO-DEX-treated shell vials compared with that in untreated cultures (Wilcoxon signed-rank test; P = 0.1). DMSO-DEX-treated monolayers did not enhance the sensitivity detection of cytomegalovirus in shell vial cell cultures.
New rat model of Pneumocystis carinii infection
Authors: Bartlett M S , Fishman J A , Queener S F , Durkin M M , Jay M A , Smith J W
Published: January 2020

Rats free of latent Pneumocystis carinii organisms were immunosuppressed with adrenal corticosteroids and transtracheally injected with P. carinii. These animals subsequently developed P. carinii pneumonia. Infection was accomplished by using organisms from infected rat lung or from culture. Diffuse infection was produced with no significant differences in the numbers of organisms found in various lobes of the lungs. Infections progressed over time so that by 6 weeks postinoculation all animals were heavily infected. Infection by transtracheal injection has three advantages over current models. First, transtracheal injection provides a reliable model which is not dependent on naturally occurring latent Pneumocystis infection. Second, transtracheal injection allows the perpetuation of specific Pneumocystis strains. Third, transtracheal injection is a more rapid and economical means of producing severe Pneumocystis pneumonia.
Detection of respiratory syncytial virus antigen in nasopharyngeal secretions by Abbott Diagnostics enzyme immunoassay
Authors: Masters H B , Bate B J , Wren C , Lauer B A
Published: January 2020

We compared a rapid respiratory syncytial virus (RSV) antigen enzyme immunoassay (EIA) (Abbott Diagnostics, North Chicago, Ill.) with virus culture and with the indirect fluorescent-antibody test (FAT) by using nasopharyngeal washings from children with suspected RSV pneumonia or bronchiolitis. Fresh washings were used in all three tests. Specimens were inoculated into HEp-2 cells and human embryonic lung fibroblasts and observed for cytopathic effect. Cells in the centrifuged sediments of the nasal washes were examined for typical cytoplasmic fluorescence of RSV by FAT. The EIA cutoff was an optical density (OD) at 492 nm that was greater than the mean OD of the negative controls plus 0.1. An OD within +20% of the cutoff was considered borderline, and these specimens were retested. Of 289 specimens, 118 (41%) were positive by culture, 150 (52%) were positive by FAT, and 154 (53%) were positive by EIA. Eight borderline EIAs were all negative when the specimens were retested after storage at -70 degrees C. Of 17 specimens positive by EIA but negative by culture and FAT, 9 were blocked in a competitive EIA, indicating that they were true-positives and that the culture and FAT were falsely negative. The sensitivity, specificity, and predictive value (positive) of the EIA versus culture, FAT, or blocking assay were 90, 94, and 95%, respectively. We conclude that the Abbott RSV antigen EIA is highly sensitive and specific.
Simultaneous infections with different serogroups of Legionella pneumophila investigated by routine methods and Fourier transform infrared spectroscopy
Authors: Horbach I , Naumann D , Fehrenbach F J
Published: January 2020

Simultaneous infections with different Legionella spp. have rarely been described in the literature. We now report on seven sporadic cases of legionellosis of which three were simultaneous infections caused by multiple Legionella pneumophila serogroups. Four different legionellae were involved. L. pneumophila serogroup 1, two different types of L. pneumophila serogroup 4, and L. pneumophila serogroup 10 have been identified simultaneously from a lung tissue specimen of one patient. Specimens from two other patients each revealed two different legionellae of serogroups 1 and 4. The existence of different L. pneumophila serogroups in simultaneous infections has not only been documented by identifying the incriminated Legionella spp. by classical methods. In addition, preliminary results of Legionella spp. identification with the novel physical procedure of Fourier transform infrared spectroscopy have been presented to evaluate its possible applicability for routine diagnostic procedures.
Staphylococcus aureus antigens reactive with milk immunoglobulin G of naturally infected dairy cows
Authors: Adams D S , McDonald J S , Hancock D , McGuire T C
Published: January 2020

A 14- to 26-kilodalton fraction of Staphylococcus aureus exoproteins isolated by molecular sieve chromatography and electroelution from polyacrylamide gels was shown to specifically react with antibodies in milk of naturally infected dairy cows. Silver staining of the antigen preparation electrophoresed in polyacrylamide gels showed the strongest reactivity in the 24- to 26-kilodalton region with lesser staining at lower apparent molecular sizes. An enzyme-linked immunosorbent assay was developed to differentiate infected from uninfected cows for diagnostic purposes. Samples from S. aureus-infected cows reacted in the assay, and samples from uninfected cows did not. There was no correlation between numbers of somatic cells in the samples and reactivity to the antigens. Samples from cows infected with coagulase-negative staphylococci did not react with the antigens. It was found, however, that some samples from uninfected cows that were recently postpartum or producing low amounts of milk contained antibodies which bound the antigens. This was believed to be due to transport from blood to the mammary gland of antibodies which were generated by previous intramammary infections or infections at other sites.
Rapid screening method with a cell multisizer for inhibitors of human immunodeficiency virus-induced cell fusion in vitro
Authors: Nakashima H , Tanabe A , Tochikura T S , Yamamoto N
Published: January 2020

Cocultivation of MOLT-4 and MOLT-4/HIVHTLV-IIIB cells induces syncytium formation very efficiently and is an appropriate model for evaluation of various substances which might inhibit human immunodeficiency virus (HIV)-induced multinucleated giant cell formation in vitro. We attempted here to quantify the grade of the syncytium formation by using a cell multisizer. The size distribution pattern of the cocultivated cells in the presence of glycyrrhizin sulfate, polysaccharide Krestin, dextran sulfate, ribofuranan sulfate, and lentinan sulfate was indistinguishable from that of cocultured cells grown in the absence of inhibitors. However, the pattern of cocultured cells without an inhibitor was quite different from that of MOLT-4 or MOLT-4/HIVHTLV-IIIB cells alone. Moreover, the size distribution pattern of cocultured cells after treatment with two nucleoside analogs, 3'-azido-2',3'-dideoxythymidine and 2',3'-didehydro-2',3'-dideoxythymidine, which were known to inhibit cell-free but not cell-to-cell infection, was similar to that of cocultured cells without an inhibitor. These data are well correlated with the fusion index which was reported previously. Application of the cell multisizer is very quantitative for evaluating the syncytium formation induced by HIV and providing a simple and rapid screening for anti-HIV substances, especially for virus-induced cell fusion.
Prospective study of microbial colonization of the nose and skin and infection of the vascular access site in hemodialysis patients
Authors: Kaplowitz L G , Comstock J A , Landwehr D M , Dalton H P , Mayhall C G
Published: January 2020

We conducted a prospective study of nasal and skin floras in 71 patients receiving chronic hemodialysis. We wished to determine whether a sterile skin preparation technique was more effective than a clean technique in removing microorganisms from the skin of the vascular access site. We also examined the effect of administration of antibiotics and status of patient hygiene on microbial flora. The presence of Staphylococcus aureus in the nose had a low predictive value for the simultaneous presence of the microorganism on the skin. The status of skin colonization can be accurately assessed only by culture of the skin. Sterile technique was no more effective at removing microorganisms from skin than was clean technique. Antibiotics significantly affected nasal flora but not skin flora. S. aureus was significantly more likely to remain on the skin after application of an antiseptic in patients with poor hygiene than in patients with good hygiene (P = 0.002). Patients with poor hygiene also had a significantly higher concentration of S. aureus on the skin of the vascular access site after application of antiseptic than patients with good hygiene (P = 0.005). We found no evidence to support a change from clean to sterile technique for skin preparation, but improvement in personal hygiene may be an effective strategy for prevention of vascular access infections.
Serial propagation of porcine group C rotavirus (pararotavirus) in a continuous cell line and characterization of the passaged virus
Authors: Saif L J , Terrett L A , Miller K L , Cross R F
Published: January 2020

The Cowden strain of porcine group C rotavirus (pararotavirus) was adapted to serial passage in a continuous monkey kidney cell line (MA104). Key factors in its successful adaptation included use of virus passaged in primary porcine kidney cells as the initial inoculum, use of roller tubes, and addition of pancreatin to the maintenance medium. A cell culture immunofluorescence test was used to quantitate the virus at each passage level, since a possible cytopathic effect was obscured by the effects of pancreatin. The virus titers dropped after initial passage into MA104 cells but increased thereafter, with peak titers evident after 16 passages (10(7) immunofluorescence U/ml). Immune electron microscopy and genome electropherotyping were used to identify group C rotavirus particles and confirm group C rotavirus double-stranded RNA gel migration patterns, respectively, from infected cell culture supernatants. The electropherotype of the cell culture-propagated group C rotavirus was identical to that of the gut virulent virus from which it was derived. The cell culture-passaged group C rotavirus also retained its infectivity for gnotobiotic pigs. No group A rotavirus was detected in the intestinal contents of the pigs or in cell culture fluids from group C rotavirus-inoculated monolayers with the two former techniques or the cell culture immunofluorescence test. This is the first verified report of serial propagation of a non-group A rotavirus in a continuous cell line.
Multiply high-level-aminoglycoside-resistant enterococci isolated from patients in a university hospital
Authors: Nachamkin I , Axelrod P , Talbot G H , Fischer S H , Wennersten C B , Moellering R C , MacGregor R R
Published: January 2020

Enterococci isolated from different body sites were tested for high-level gentamicin resistance. A total of 259 enterococcal isolates were screened for resistance (MIC, greater than 2,000 micrograms/ml) by a broth-tube method. Thirty-nine (15.1%) were found to exhibit resistance and were confirmed by agar screening (1,000 micrograms/ml) and agar dilution MIC determinations. The majority of isolates also showed high-level resistance to kanamycin and streptomycin. The remaining isolates showed high-level resistance to gentamicin and kanamycin but not streptomycin. Synergy testing of several isolates confirmed the correlation between lack of synergy and high-level resistance. A retrospective clinical review was performed. Most patients had a source of definite or likely infection (79%). Serious infections such as endocarditis or meningitis were not observed during the course of this study. Retrospective clinical data suggest that in cases not involving endocarditis or meningitis, neither infection refractory to therapy nor relapse of infection is a common sequela of infection with gentamicin-resistant enterococci in hospitalized patients.
DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages
Authors: Newland J W , Neill R J
Published: January 2020

A set of DNA probes has been developed to study the genes for Shiga-like toxins (SLT) and the bacteriophage from which these toxin genes were isolated. Under stringent conditions of hybridization (80 to 90% homology), these probes detect strains containing (i) SLT I-related genes, (ii) SLT II-related genes, (iii) phage sequences from the SLT I-converting phage H19A/933J, and (iv) phage sequences from the SLT II-converting phage 933W. Strain characterization by hybridization with the toxin gene probes was as accurate as methods that used toxin-specific antibody to determine toxin synthesis. Screening of different gram-negative bacteria with the toxin probes revealed that only two species carry sequences related to the SLT genes, Escherichia coli and Shigella dysenteriae 1. These results indicated that the lower levels of toxin activity observed in shigellae other than S. dysenteriae 1 are due to a gene(s) that is genetically distinct from that which encodes Shiga toxin. Analysis of enterotoxigenic, enteroinvasive, enteropathogenic, and enterohemorrhagic E. coli indicated that SLT genes are found primarily in the enterohemorrhagic E. coli strain group. Use of both the toxin and the phage probes has identified a variety of genotypic combinations of phage and toxin sequences which differ from those observed for the original toxin-converting phage isolates, for E. coli O157:H7 strain 933, and for E. coli O26:H11 strain H19.
Distribution of a monoclonal antibody-recognized protective protein immunogen on the outer membranes of Pasteurella multocida rabbit isolates
Authors: Lu Y S , Gerrity L W , Afendis S J , Watkins L , Pakes S P
Published: January 2020

The distribution of a monoclonal antibody (MAb)-recognized protective protein immunogen on the outer membrane of 153 Pasteurella multocida rabbit isolates was determined by dot blot (DB) analysis. MAb 1608 reacted with 36 (24%) of the 153 clinical isolates. The DB-positive clinical isolates expressed capsular antigens A, D, and nontypable and somatic antigens 2, 3, 10, 12, 15, and nontypable. Western blot (immunoblot) analysis with adsorbed and eluted MAb 1608 confirmed that the antigenic determinant identified was located on the cell surface. With MAb 1608 as a probe for antibody-accessible radioimmunoassay, 31 of 36 DB-positive P. multocida rabbit isolates were shown to have surface-exposed and antibody-accessible antigenic determinants, while 44 of 44 DB-negative isolates were negative by antibody-accessible radioimmunoassay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed DB-negative P. multocida isolates both with (6 of 13, 46%) and without (7 of 13, 54%) the 37.5-kilodalton protein. This study establishes that the protective antigenic determinant of the 37.5-kilodalton outer membrane protein is present in 24% of rabbit clinical isolates tested and is detectable in P. multocida strains distributed among the major somatic types (3, 10, 12, and 15) and the capsular types (A and D) commonly isolated from rabbits in North America.
Rapid detection and identification of pathogenic mycobacteria by combining radiometric and nucleic acid probe methods
Authors: Ellner P D , Kiehn T E , Cammarata R , Hosmer M
Published: January 2020

The combination of radiometric methodology (BACTEC 12B) and probe technology for recovery and identification of mycobacteria was studied in two large hospital laboratories. The sediment from vials with positive growth indices was tested with DNA probes specific for Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium intracellulare. The sensitivity of the radiometric method and the specificity of the probes resulted in a marked reduction in the time to the final report. Biochemical testing could be eliminated on isolates giving a positive reaction with one of the probes. Some 176 isolates of M. tuberculosis, 110 of M. avium, and 5 of M. intracellulare were recovered. Two-thirds of these isolates were detected and identified within 2 weeks of inoculation and the remainder was detected by 4 weeks, a reduction of 5 to 7 weeks to the final report.
Clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli infections detected by DNA hybridization
Authors: Taylor D N , Echeverria P , Sethabutr O , Pitarangsi C , Leksomboon U , Blacklow N R , Rowe B , Gross R , Cross J
Published: January 2020

To determine the clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli (EIEC) infections, we investigated 410 children with diarrhea and 410 control children without diarrhea who were seen at Children's Hospital, Bangkok, Thailand, from January to June 1985. Shigella spp. were isolated from 96 (23%) and EIEC were isolated from 17 (4%) of 410 children with diarrhea and from 12 (3%) and 6 (1%) of 410 control children, respectively. The isolation rates of both pathogens increased with age and peaked in children 3 to 5 years old from whom Shigella spp. were isolated from 38% and EIEC were isolated from 9%. Shigella spp. were isolated from 52% and EIEC were isolated from 7% of 91 children with bloody diarrhea and from 15 and 3% of 319 children with nonbloody diarrhea. Fifteen (65%) of 23 EIEC were lactose positive, and all belonged to recognized EIEC serotypes. Among children with diarrhea, the stool blots of 76% of 17 children infected with EIEC, 45% of 96 children infected with Shigella spp., and 1% of 297 culture-negative children hybridized with the 17-kilobase EcoRI digestion fragment of pRM17, a recombinant plasmid containing DNA derived from the 140-megadalton Shigella flexneri plasmid. Although EIEC colonies can be reliably detected by DNA hybridization, detection by stool blot is less sensitive. Shigella spp. and EIEC are important causes of endemic diarrhea among children greater than 1 year old in Thailand.
Problems with rapid agglutination methods for identification of Staphylococcus aureus when Staphylococcus saprophyticus is being tested
Authors: Gregson D B , Low D E , Skulnick M , Simor A E
Published: January 2020

Six rapid agglutination tests for identification of Staphylococcus aureus were evaluated by using 62 strains of S. aureus, 63 strains of S. saprophyticus, and 67 strains of other coagulase-negative staphylococci. S. saprophyticus was responsible for 19 of 26 false-positive results and 20 uninterpretable reactions. Thus, urinary staphylococcal isolates that are positive by rapid agglutination tests may require other confirmatory tests for the identification of possible S. saprophyticus.
Legionella jordanis isolated from a patient with fatal pneumonia
Authors: Thacker W L , Wilkinson H W , Benson R F , Edberg S C , Brenner D J
Published: January 2020

A Legionella-like organism was isolated from an open-lung biopsy of a patient with fatal pneumonia. The isolate was shown by growth, physiologic, serologic, and genetic characteristics to belong to the species Legionella jordanis, which had previously been isolated only from the environment.
Serratia plymuthica osteomyelitis following a motorcycle accident
Authors: Zbinden R , Blass R
Published: January 2020

Serratia plymuthica was isolated from the sinus tract and from the infected bone in a case of chronic osteomyelitis following an accident of a motorcyclist. This is the second case in which S. plymuthica was a significant pathogen. Previously, a case of S. plymuthica sepsis associated with infection of a central venous catheter was described.
Rapid and sensitive viral culture method for human immunodeficiency virus type 1
Authors: Jackson J B , Coombs R W , Sannerud K , Rhame F S , Balfour H H
Published: January 2020

Peripheral blood mononuclear cells from 142 consecutive patients with antibodies to human immunodeficiency virus type 1 (HIV-1) were cultured for HIV-1. All 72 patients with symptoms of HIV-1 infection were culture positive, as were 69 of 70 asymptomatic patients. Of the 142 patients, 132 (93%) were culture positive within 10 days after initiation of the culture.
Antibody response to Campylobacter coli in children during intestinal infection and carriage
Authors: Martin P M , Mathiot J , Ipero J , Georges A J , Georges-Courbot M C
Published: January 2020

In the Central African Republic, etiological studies of diarrhea have shown that Campylobacter coli accounts for almost 40% of Campylobacter enteric isolations. This prompted us to investigate the antibody response to C. coli infection in children. As expected from the literature on Campylobacter jejuni infections, our results show that both infection and carriage elicited antibodies against glycine-extracted membrane antigens, flagella, and cholera toxin. The human antibody response to C. coli resembles the response to C. jejuni, and this similarity will allow comparative studies on larger numbers of infections, both symptomatic and asymptomatic. Anti-cholera toxin antibodies were directed against both the A and B subunits.
Spurious oxacillin resistance in Staphylococcus aureus because of defective oxacillin disks
Authors: Boyce J M , Lonks J R , Medeiros A A , Papa E F , Campbell S
Published: January 2020

Clinical isolates of Staphylococcus aureus wee inappropriately categorized as intermediate or resistant to oxacillin on the basis of tests with two lots of oxacillin disks. The potency of one lot was tested and found to be below accepted limits. Routine quality control tests failed to detect the defective disks.
Simplified culture procedure for large-scale screening for Chlamydia trachomatis infections
Authors: Lees M I , Newnan D M , Garland S M
Published: January 2020

A method that uses a 48-well tissue culture cluster tray system for the isolation of Chlamydia trachomatis is described. The cluster tray system was as sensitive (100%) as and more time efficient than the conventional cover slip method, thereby being considerably cost saving. With both culture methods, the prevalence rates of genital carriage of C. trachomatis in women attending clinics for legal abortion and for cervical dysplasia were 5% (31 of 641 patients) and 2% (3 of 148 patients), respectively.
Isolation and propagation of enteric adenoviruses in HEp-2 cells
Authors: Perron-Henry D M , Herrmann J E , Blacklow N R
Published: January 2020

Eighty-two stool samples from children with gastroenteritis in Canada, England, and Thailand which had been shown to contain adenovirus antigen (by a group-specific enzyme-linked immunosorbent assay) or adenovirus particles (by electron microscopy) or both, were tested for primary isolation of enteric adenoviruses in HEp-2 and Graham 293 cells. Graham 293 cells are known to support the replication of enteric adenovirus types (Ad40 and Ad41) on primary isolation, whereas HEp-2 cells reportedly do not. Of the 82 adenovirus isolates, 73 could be typed as Ad40 or Ad41 by type-specific monoclonal antibody enzyme-linked immunosorbent assay and by analysis of SmaI endonuclease digests. Of these 73, 30 (41%) could be isolated in HEp-2 cells, which included 43% (9/21) of those typed as Ad40 and 40% (21/52) of those typed as Ad41. On the basis of these results, the growth characteristics of adenoviruses in HEp-2 cell cultures, commonly used to distinguish enteric from nonenteric adenovirus types, are not valid for either diagnosis or epidemiological studies. For the samples studied here, use of these nondefinitive criteria would result in underestimation of the incidence of enteric adenoviruses in viral gastroenteritis.
Clonal origin, restricted natural distribution, and conservation of virulence factors in isolates of enterotoxigenic Escherichia coli serogroup O126
Authors: Yam W C , Lung M L , Ng M H
Published: January 2020

Enterotoxigenic Escherichia coli serogroup O126 isolates have been isolated in Hong Kong since 1982 from sporadic cases of infantile diarrhea and from one outbreak in a neonatal ward. A 64-megadalton plasmid encoding colonization factor antigen I and heat-stable enterotoxin was identified in all 23 isolates. Enterotoxigenic E. coli strains producing heat-stable enterotoxin from different regions of Southeast Asia were collected and compared by biotyping, antibiotic resistance patterns, and plasmid profiles. Restriction endonuclease digestion of plasmids and subsequent Southern blot analysis with the heat-stable enterotoxin gene probe of representative strains showed a unique plasmid was harbored by all heat-stable enterotoxin-producing O126 strains tested. These results are consistent with conservative inheritance of enterotoxin plasmids within enterotoxigenic E. coli strains over a 2-year period in Hong Kong.
Comparison of immunofluorescence, enzyme immunoassay, and Western blot (immunoblot) methods for detection of antibody to human T-cell leukemia virus type I
Authors: Gallo D , Hoffman M N , Cossen C K , Diggs J L , Hurst J W , Penning L M
Published: January 2020

A total of 3,349 serum samples were screened by the immunofluorescence (IF) method for antibody to human T-cell leukemia virus type I (HTLV-I). Only 9 of 2,409 specimens from selected individuals, blood bank donors, patients with encephalitis-meningitis, and human immunodeficiency virus antibody-positive homosexual or bisexual men were reactive by IF. In addition, 940 serum samples from intravenous drug abusers were tested by IF and also by an HTLV-I enzyme immunoassay (EIA) method. Of these, 222 (24%) were positive for both HTLV-I and HTLV-II antigens by IF, and 191 of these 222 were also reactive in the HTLV-I EIA. Of the 31 IF-positive, EIA-negative serum samples, 20 exhibited optical density readings greater than or equal to 70% of the positive cutoff in the EIA, and 29 samples reacted with 1 or more bands in the Western blot (immunoblot) test. An additional 10 specimens that were EIA negative reacted only with HTLV-I by IF. Differences in staining morphology and in reactions on HTLV-I and HTLV-II antigens before and after absorption of the serum specimens with HTLV-I and HTLV-II-infected cell pellets revealed six distinct serological patterns by IF. These results indicate that infections by HTLV-I or by another closely related retrovirus(es) occur in California. Further studies utilizing statistically valid sampling methods are needed to estimate true prevalence rates among various groups. IF and Western blot tests should supplement the EIA method to maximize sensitivity and specificity of test procedures.
Performance and reliability of five commercial enzyme-linked immunosorbent assay kits in screening for anti-human immunodeficiency virus antibody in high-risk subjects
Authors: Ozanne G , Fauvel M
Published: January 2020

Anti-human immunodeficiency virus enzyme-linked immunosorbent assay kits marketed by Electro-Nucleonics Inc. (ENI), Genetic Systems Corp. (GSC), Organon Teknika Inc. (OTI), Ortho Diagnostic Systems Inc. (ODSI), and Wellcome Diagnostics (WD) were evaluated by using 289 randomly selected serum samples from a high-risk population and 53 serum samples likely to produce false-positive results. The radioimmunoprecipitation assay was used as the reference test. Sensitivities ranged from 96.51% (ODSI, WD) to 97.67% (ENI, GSC, OTI). Sera showing antibodies to viral glycoproteins only produced the false-negative results. Specificities ranged from 99.6% (ENI, GSC, ODSI, OTI) to 100% (WD). False-positive results were obtained with sera from patients with autoimmune disease or Epstein-Barr virus infection. Only results from GSC and OTI kits were distributed in two compact clusters well segregated on either side of the cutoff point. ODSI and GSC kits had the best intralot reproducibility. The GSC kit had the best interlot reproducibility. Cutoff values for ODSI and GSC kits were the least variable. Intraplate repeatability was good for all kits. Sample localization was not an important source of variability. Our results do not point out one outstanding kit among the five evaluated. However, the GSC kit showed the best overall results.
Development of neutralizing antibodies and group A common antibodies against natural infections with human rotavirus
Authors: Zheng B J , Han S X , Yan Y K , Liang X R , Ma G Z , Yang Y , Ng M H
Published: January 2020

We determined the levels of group A common and neutralizing antibodies against human rotavirus in paired serum specimens obtained from 38 infants within 12 days of the onset of diarrhea. Thirty of the infants excreted rotavirus in stools, and eight did not. Nine patients (30%) with rotavirus diarrhea and seven patients (88%) with diarrhea due to other causes had detectable levels (greater than or equal to 1: 80) of immunoglobulin (IgG) common antibodies in acute-phase sera. All the patients with rotavirus diarrhea showed at least fourfold rises in titers of IgG or IgM common antibodies or both, while only two control patients showed significant rises in either IgG or IgM common antibodies in their convalescent-phase sera. Of the 19 patients excreting "short" electropherotypes of rotavirus, 18 showed at least fourfold rises in titers of neutralizing antibodies against serotype 2 human rotavirus but not against serotype 1, 3, or 4. Nine of the ten patients excreting "long" electropherotypes showed significant rises in neutralizing antibodies against serotype 3, and the other patient showed a significant rise in neutralizing antibodies against serotype 1. One patient excreted long and short electropherotypes simultaneously, and he also showed a significant rise in neutralizing antibodies against serotype 2 and 3 viruses. The control patients with diarrhea did not show significant changes in titers of antibodies against any of the serotypes. These results demonstrated that the neutralizing antibody response within 2 weeks after clinical onset is specific for the infecting serotype of rotavirus.
Rotavirus survival on human hands and transfer of infectious virus to animate and nonporous inanimate surfaces
Authors: Ansari S A , Sattar S A , Springthorpe V S , Wells G A , Tostowaryk W
Published: January 2020

We tested the survival of the Wa strain of human rotavirus on the hands of volunteers and also studied infectious virus transfer between animate and inanimate (stainless steel disks) surfaces. The virus was diluted in a 10% suspension of feces, and 10 microliters (1 X 10(3) to 4 X 10(4) PFU) was placed on each of the four fingerpads of the left hand. One milliliter of 20% tryptose phosphate broth in Earle balanced salt solution was used for virus elution from each fingerpad, and the hands were disinfected with 70% ethanol before they were washed with an antiseptic soap and water. At 20, 60, and 260 min after inoculation, approximately 57, 43, and 7%, respectively, of the input infectious virus could be recovered. For virus transfer, the inoculum (2 X 10(4) to 8 X 10(4) PFU) was allowed to dry, and the donor surface was kept in contact with the recipient surface for 10 s at a pressure of approximately 1 kg/cm2. At 20 and 60 min after virus inoculation, 16.1 and 1.8%, respectively, of the input infectious virus could be transferred from the contaminated hand to a clean disk; when a clean hand was pressed against a contaminated disk, virus transfer was 16.8 and 1.6%, respectively. Contact between a contaminated and a clean hand 20 and 60 min after virus inoculation resulted in the transfer of 6.6 and 2.8%, respectively, of the input infectious virus. These findings indicate the potential vehicular role for human hands in the spread of rotaviral infections.
Detection of nephropathia epidemica (Puumala virus)-specific immunoglobulin M by enzyme-linked immunosorbent assay
Authors: Niklasson B , Kjelsson T
Published: January 2020

Nephropathia epidemica (NE), a less severe form of hemorrhagic fever with renal syndrome, is caused by Puumala virus (PUU). This communication reports the development of a mu-capture enzyme-linked immunosorbent assay (ELISA) for detection of specific immunoglobulin M (IgM) antibodies to PUU virus in human sera. Acute- and early-convalescent-phase sera (collected 1 to 41 days after disease onset) from 29 Swedish patients with clinical NE were tested for PUU virus-specific IgG and IgM antibodies by the indirect immunofluorescence test and ELISA, respectively. Late-convalescent-phase serum was also collected from 18 of these patients 3 to 24 months postinfection and assayed. The IgM ELISA values were strongly positive in sera collected during the first 2 months; at 3 to 9 months, they were negative or in the lower range of significance, and at 24 months all sera were negative. Paired sera from NE patients often fail to show seroconversion or a significant titer rise when tested by indirect immunofluorescence. Since all acute- and early-convalescent-phase sera were positive by IgM ELISA, this test could become an important tool for early diagnosis of acute human NE infections.
rRNA gene restriction patterns of Haemophilus influenzae biogroup aegyptius strains associated with Brazilian purpuric fever
Authors: Irino K , Grimont F , Casin I , Grimont P A
Published: January 2020

The rRNA gene restriction patterns of 92 isolates of Haemophilus influenzae biogroup aegyptius, associated with conjunctivitis or Brazilian purpuric fever in the State of São Paulo, Brazil, were studied with 16 + 23S rRNA from Escherichia coli as a probe. All strains were classified into 15 patterns. Isolates from Brazilian purpuric fever cases were seen only in patterns 3 (most frequently) and 4 (rarely), whereas isolates from conjunctivitis were found in all 15 patterns. The study demonstrated that rRNA from E. coli can serve as a probe for molecular epidemiology.
Particle agglutination assays for rapid detection of fibronectin, fibrinogen, and collagen receptors on Staphylococcus aureus
Authors: Naidu A S , Paulsson M , Wadström T
Published: January 2020

Latex beads (0.8-micron diameter; Difco Laboratories) were coated with fibronectin, fibrinogen, collagen type I, or denatured collagen (gelatin) and evaluated in a particle agglutination assay (PAA) for the rapid detection of fibronectin, fibrinogen, or collagen binding to Staphylococcus aureus. These assays were compared with a commercial test for detecting the binding of fibrinogen and immunoglobulin G (Staphaurex). Bacterial cells (approximately 10(10) cells per ml) suspended in 0.02 M potassium phosphate buffer (pH 6.8) caused the clumping of standard fibronectin, collagen, gelatin, and fibrinogen latex suspensions within 2 min on glass slides. The test results were scored semiquantitatively from strongly positive ( ) to weakly positive (+) and negative (-) reactions. The negative PAA reactions corresponded to a median value of 11.5% relative to the binding of 125I-labeled protein to strain Cowan 1, indicating the high sensitivity of the test. The reactions with fibronectin and fibrinogen latex suspensions and with Staphaurex were optimal for cells grown on tryptic soy and brain heart infusion broth media. Blood agar was optimal for reactions with collagen and gelatin latex suspensions. Media containing high salts (mannitol salt agar and staphylococcus medium 110) enhanced the tendency of cells to autoaggregate. These assays were also clinically evaluated on 187 S. aureus isolates. The PAA reagents were stable, and the assays were highly specific, sensitive, and reproducible, thus making PAA suitable for the rapid screening of the binding of various bacterial pathogens to serum and connective-tissue proteins.
Aeromonas schubertii, a new mannitol-negative species found in human clinical specimens
Authors: Hickman-Brenner F W , Fanning G R , Arduino M J , Brenner D J , Farmer J J
Published: January 2020

In 1983 the vernacular name Enteric Group 501 was coined for a group of strains that had been referred to our laboratory as "possible Vibrio damsela that does not require NaCl for growth." By DNA-DNA hybridization (hydroxyapatite method, 32P, 60 and 75 degrees C), six strains of Enteric Group 501 were closely related to the labeled strain 2446-81 (70 to 95% at 60 degrees C and 71 to 93% at 75 degrees C; 0 to 1% divergence). Type strains of all Aeromonas species and reference strains of six other Aeromonas DNA hybridization groups were 26 to 42% related (60 degrees C) to strain 2446-81, but type strains of 27 Vibrio and Photobacterium species, including V. damsela, were 0 to 1% (75 degrees C) related. We propose the name Aeromonas schubertii for the highly related group of seven strains formerly known as Enteric Group 501. The type strain is designated as ATCC 43700 (CDC 2446-81). Strains of A. schubertii grew well at 36 degrees C and had positive reactions at this temperature for methyl red, Voges-Proskauer (1% NaCl, Coblentz method), lysine decarboxylase, arginine dihydrolase, motility, lipase, DNase, nitrate reduction to nitrite, oxidase, and growth in nutrient broth with 0 and 1% NaCl. There was no growth in 6% NaCl or on thiosulfate-citrate-bile salts-sucrose agar. The following sugars were fermented: D-glucose, D-galactose, maltose, D-mannose, and trehalose. The following sugars were not fermented: adonitol, L-arabinose, D-arabitol, cellobiose, dulcitol, erythritol, myo-inositol, lactose, D-mannitol, melibiose, alpha-CH3-D-glucoside, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose, and D-xylose. Esculin was not hydrolyzed, and the string test was negative. The mannitol-negative reaction differtiates A. schubertii from other Aeromonas species. The antibiogram of this organism is typical of other Aeromonas strains (resistance to ampicillin and carbenicillin and susceptibility to most other agents). A. schubertii strains have been isolated from abscesses (two strains), wound (one), skin (one), pleural fluid (one), and blood (two). The two blood isolates suggest clinical significance typical of other Aeromonas species , but further information is needed on this group.
Cost containment of formalin-preserved stool specimens for ova and parasites from outpatients
Authors: Peters C S , Hernandez L , Sheffield N , Chittom-Swiatlo A L , Kocka F E
Published: January 2020

Three individual formalinized stool specimens from each of 123 patients were pooled and examined for ova and parasites. Results obtained from the pooled specimens were compared with those obtained with the three individual specimens used to make the pooled specimens. Of 123 sets examined, 92 were negative and 31 were positive for ova and parasites. The pooled specimens were positive and all individual specimens were negative eight times, whereas the pooled specimens were negative and the individual specimens were positive twice. The data indicated that the pooled system is a useful and economical method of screening for ova and parasites.
Rapid separation of Pneumocystis carinii from lung lavage fluids
Authors: Pesanti E L , McCarron B A
Published: January 2020

A simple, rapid procedure to separate Pneumocystis carinii obtained by lavage of lungs of steroid-treated rats from rat leukocytes is described. Commercially available monoclonal antibody to rat common leukocyte determinants is used to sediment the leukocytes, resulting in supernatant fluid containing P. carinii cells virtually free of intact rat cells.
Salt-supplemented medium for testing methicillin-resistant staphylococci with newer beta-lactams
Authors: Jorgensen J H , Redding J S , Maher L A , Ramirez P E
Published: January 2020

The addition of 2% NaCl to cation-supplemented Mueller-Hinton broth (CSMHB) was evaluated for microdilution testing of the susceptibility of staphylococci to five cephalosporins, imipenem, amoxicillin-clavulanate, and ticarcillin-clavulanate. With Staphylococcus aureus, NaCl improved the recognition of methicillin (oxacillin) resistance to cefamandole, imipenem, or ticarcillin-clavulanate. Resistance to amoxicillin-clavulanate was readily determined, irrespective of the presence of added salt. The addition of 2% NaCl to CSMHB did not significantly improve detection of resistance to any of the beta-lactams among coagulase-negative staphylococci. Since the addition of NaCl did not have significant adverse effects on tests with coagulase-negative staphylococci, the routine addition of 2% NaCl to oxacillin or methicillin tests with staphylococci may be justifiable on the basis of convenience or standardization. However, addition of NaCl to susceptibility tests of other beta-lactams does not consistently improve recognition of resistance among staphylococci and thus cannot be recommended for routine use.