bluuebunny/crossref_metadata_2025 huggingface dataset:
Presence of an acid-labile alpha-interferon in sera from fetuses and children with congenital rubella
Authors: Lebon P , Daffos F , Checoury A , Grangeot-Keros L , Forestier F , Toublanc J E
Published: January 2020

In congenital rubella an acid-labile alpha-interferon was present in sera collected from fetuses between weeks 21 and 29 of gestation and from children with active congenital rubella. This interferon was different from the interferon detected in normal amniotic fluid and was not found in sera from uninfected fetuses or from children with postnatally acquired rubella. The fetal interferon is of interest as a complementary marker to confirm the virus contamination of the fetuses during maternal rubella. The role of the prolonged synthesis of this interferon in congenital rubella disease and its immune defects are discussed.
Intracellular multiplication of leprosy-derived mycobacteria in Schwann cells of dorsal root ganglion cultures
Authors: Mukherjee R , Antia N H
Published: January 2020

Organized nerve cultures of dorsal root ganglia from neonatal mice were infected with Mycobacterium leprae, the causative agent of leprosy. A significant multiplication of the acid-fast bacilli was observed within the Schwann cell component of the culture. The growth of these bacilli was sensitive to antileprosy drugs and was not observed directly in bacteriological media. These organisms were brightly stained with the monoclonal antibody to phenolic glycolipid-I, a M. leprae-specific marker. The antigenic, pathogenic, and biochemical characteristics of this mycobacterium are under investigation.
Diagnosis of spirochetal meningitis by enzyme-linked immunosorbent assay and indirect immunofluorescence assay in serum and cerebrospinal fluid
Authors: Stiernstedt G T , Granström M , Hederstedt B , Sköldenberg B
Published: January 2020

The antibody response against a spirochetal strain isolated from Swedish Ixodes ricinus ticks was determined by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay of cerebrospinal fluid (CSF) and serum specimens from 45 patients with chronic meningitis. Samples of CSF, serum, or both from patients with various infections of the central nervous system, multiple sclerosis, syphilis, or infectious mononucleosis and from healthy individuals served as control samples. Probable spirochetal etiology could be demonstrated for 41 of 45 (91%) patients with clinical symptoms of chronic meningitis. Approximately 25% of the patients had significantly elevated titers of antibody to the spirochete in CSF but not in serum. The highest diagnostic sensitivity, 91%, was demonstrated by measurement of CSF antibodies and calculation of a spirochetal CSF titer index, which is the ratio of (ELISA titer in CSF/ELISA titer in serum) to (albumin in CSF/albumin in serum) and which also considers the degree of blood-CSF barrier damage. The highest specificity, 98%, was obtained by calculation of a CSF titer index. Patients with short duration of disease were especially prone to be antibody negative in serum but positive in CSF. Significant rise in serum antibody titers was seldom demonstrated in patients treated with antibiotics. It is concluded that measurement of CSF antibodies, especially by ELISA, is a highly sensitive and specific method for the immunological diagnosis of spirochetal meningitis.
Aminopeptidase activity in arginine-utilizing Mycoplasma spp
Authors: Ball H J , Neill S D , Reid R L
Published: January 2020

In an examination of arginine-utilizing Mycoplasma species, simple fluorogenic and chromogenic tests were used to demonstrate high levels of arginine aminopeptidase activity in unwashed, unconcentrated broth and agar cultures in all strains of 5 species and low levels of activity in all strains of 14 species. Four species contained strains which exhibited either high or low levels of activity.
Rapid method for detection of adherent bacteria on Foley urinary catheters
Authors: Ladd T I , Schmiel D , Nickel J C , Costerton J W
Published: January 2020

An accurate, rapid, and inexpensive method was developed for detecting and enumerating bacteria adherent to Foley urinary catheters based on malachite green staining of acridine orange-prestained specimens. This method has proven to be quick and reliable and will find application in quantitative studies of biomaterial-related sepsis.
Evaluation of three broth disk methods for testing the susceptibility of anaerobic bacteria to imipenem
Authors: Shungu D L , Weinberg E , Cerami A T
Published: January 2020

Imipenem is a member of a new class of highly potent beta-lactam antibiotics, carbapenems, with an antibacterial spectrum that includes nearly all currently known aerobic and anaerobic bacterial species of clinical significance. Although relatively stable in most standard laboratory media used for antimicrobial susceptibility testing, imipenem undergoes rapid inactivation in thioglycolate broth, a recommended medium for susceptibility testing of anaerobic bacteria by the broth disk method. In the current study, a panel of 36 anaerobic bacteria consisting of 28 clinical isolates and eight quality control strains was used to determine the suitability and accuracy of the broth disk methods with brain heart infusion, Schaedler, and anaerobic broths, in comparison to the reference agar dilution method, for the anaerobic susceptibility testing of imipenem. To achieve single test concentrations of approximately 8, 16, and 64 micrograms/ml for imipenem, cefoxitin, and piperacillin, respectively, which correspond to the MIC breakpoints of the test drugs, four 10-microgram imipenem disks, three 30-microgram cefoxitin disks, and three 100-microgram piperacillin disks were used in 5 ml of broth. The correlation between the reference agar dilution method and each of the three broth disk elution procedures evaluated was excellent, for imipenem (100% agreement) and somewhat less so for cefoxitin and piperacillin. Therefore, brain heart infusion, Schaedler, and anaerobic broths, but not thioglycolate broth, are suitable for anaerobic susceptibility testing of imipenem by the disk elution method.
Ciprofloxacin disk susceptibility tests: interpretive zone size standards for 5-microgram disks
Authors: Barry A L , Fass R J , Anhalt J P , Neu H C , Thornsberry C , Tilton R C , Painter B G , Washington J A
Published: January 2020

Evaluations of 5-microgram ciprofloxacin disk diffusion susceptibility tests were performed independently by seven different investigators. The results of the separate tests were combined to increase the number of resistant strains in the challenge set of microorganisms. Based on data with 2,652 isolates, the following interpretive breakpoints are tentatively proposed for use in ongoing clinical trials of ciprofloxacin: less than or equal to 15 mm, resistant (MIC greater than 2.0 micrograms/ml); 16 to 20 mm, intermediate (1.0 less than MIC less than or equal to 2.0 micrograms/ml); and greater than or equal to 21 mm, susceptible (MIC less than or equal to 1.0 micrograms/ml). Disk tests with Streptococcus spp. and with Pseudomonas maltophilia were not reliable; other microorganisms were accurately categorized by the disk diffusion test.
Antigenic distinctiveness of Mobiluncus curtisii and Mobiluncus mulieris
Authors: Roberts M C , Baron E J , Finegold S M , Kenny G E
Published: January 2020

A total of 26 Mobiluncus strains (17 M. curtisii and 9 M. mulieris strains) were compared serologically by double immunodiffusion and immunoblotting against antisera prepared against representative isolates of each species. All strains from the same species were strongly reactive with homologous antisera but generally weakly reactive with antisera to the heterologous Mobiluncus spp. The antisera did not react with strains of the unrelated genera Campylobacter, Succinivibrio, Wolinella, Actinomyces, Anaerobiospirillum, and Anaerovibrio.
Evaluation of the Wayson variation of a methylene blue staining procedure for the detection of microorganisms in cerebrospinal fluid
Authors: Daly J A , Gooch W M , Matsen J M
Published: January 2020

Meningitis of bacterial origin is a severe infection that must receive immediate attention and prompt treatment. We evaluated a basic fuchsin-methylene blue, ethyl alcohol-phenol staining procedure (Wayson stain) and compared it with the Gram stain procedure for evaluation of cerebrospinal fluid. All smears were prepared within 30 min of receiving the specimen and examined without knowledge of the results of the companion smear or culture. Of 546 specimens entered into the study, 84 were culture positive. Based on the culture results, the sensitivity and specificity of Wayson stain were 90 and 98%, respectively, compared with 73% sensitivity and 99% specificity by Gram stain. We observed that Wayson staining is a particularly sensitive method for screening clinical specimens that contain proteinaceous background material. The Wayson staining procedure is a simple and sensitive technique for early detection of meningitis.
Biochemical and genetic characteristics of atypical Campylobacter fetus subsp. fetus strains isolated from humans in the United States
Authors: Edmonds P , Patton C M , Barrett T J , Morris G K , Steigerwalt A G , Brenner D J
Published: January 2020

During a 2-year period, 14 biochemically atypical Campylobacter fetus subsp. fetus-like strains were received by the Campylobacter Reference Laboratory at the Centers for Disease Control. Sources of the isolates were blood, nine strains; stools, two strains; amniotic fluid, one strain; and abscesses, two strains. Atypical phenotypic characteristics exhibited by one or more strains were growth at 42 degrees C, 10 strains; no H2S by lead acetate paper, 3 strains; resistance to a 30-micrograms cephalothin disk, 2 strains; and nonmotility, 1 strain. By DNA-DNA hybridization, all 14 isolates and the type strain of C. fetus subsp. fetus (ATCC 27374) were 94 to 100% related in reassociation reactions at 50 degrees C, with 0.0 to 0.5% divergence, and were 86 to 100% related in reassociation reactions at 65 degrees C. Thus, all of these atypical strains were C. fetus subsp. fetus. MICs of 11 antimicrobial agents for these 14 strains were variable. All strains were susceptible to chloramphenicol, erythromycin, gentamicin, and tetracycline, and most were susceptible to ampicillin, clindamycin, and penicillin. Eleven strains were resistant to cephalothin (MIC greater than or equal to 16 micrograms/ml), nine were resistant to rifampin (MIC greater than or equal to 8 micrograms/ml), and all were resistant to nalidixic acid (MIC greater than 32 micrograms/ml) and vancomycin (MIC greater than 32 micrograms/ml). One can expect to see biochemical variability in C. fetus subsp. fetus strains and to encounter such strains from a variety of human sources, the most important of which appears to be blood.
Improved selective medium for the isolation of lipase-positive Clostridium botulinum from feces of human infants
Authors: Mills D C , Midura T F , Arnon S S
Published: January 2020

Isolation of lipase-positive Clostridium botulinum from fecal specimens establishes the diagnosis of infant botulism, contributes to the diagnosis of food-borne botulism, and is most easily accomplished by use of selective media. Modification of an available selective medium, C. botulinum inhibitory medium (CBI), enabled more rapid isolation of C. botulinum. The modified medium (botulinum selective medium [BSM] contained (per liter) 25 g of dehydrated heart infusion broth, 20 g of agar, 30 ml of egg yolk suspension, 250 mg of cycloserine, 76 mg of sulfamethoxazole, 4 mg of trimethoprim, and 100 IU of thymidine phosphorylase at pH 7.4. The two media were compared by using 15 fresh fecal specimens from infant botulism patients (10 type A and 5 type B) and a C. botulinum isolate that had been obtained from an infant botulism patient and that was mixed into a fresh stool specimen from a healthy human infant. In comparison to CBI, BSM always provided better suppression of the nonbotulinum fecal flora and earlier emergence of lipase-positive colonies. Diagnosis of infant botulism was accomplished sooner with BSM than with CBI because isolation of lipase-positive C. botulinum was easier.
Confirmation of Legionella pneumophila cultures with a fluorescein-labeled monoclonal antibody
Authors: Tenover F C , Carlson L , Goldstein L , Sturge J , Plorde J J
Published: January 2020

We compared a fluorescein-labeled monoclonal antibody directed against an outer membrane protein of Legionella pneumophila (Genetic Systems Corp. [GSC], Seattle, Wash.) with a similarly labeled polyclonal reagent (L. pneumophila serogroups 1 to 6, poly; BioDx, Inc., Denville, N.J.) for the confirmation of L. pneumophila isolates grown in culture. Duplicate suspensions of 52 organisms, including 21 L. pneumophila and 8 non-L. pneumophila species of legionella, were placed on individual glass slides, fixed, and stained with both reagents, and the results were compared. Both antisera correctly identified all L. pneumophila serogroups 1 to 6, but only the GSC reagent produced definitive staining of the L. pneumophila isolates of serogroups 7, 8, and 9. Additionally, the GSC reagent produced more uniform staining patterns around the legionella bacilli and displayed little background fluorescence when compared with the BioDx reagent.
Evaluation of a rapid identification method for Neisseria spp
Authors: Germer J J , Washington J A
Published: January 2020

A comparison was made of carbohydrate degradation reactions of Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria lactamica, Neisseria mucosa, Neisseria sicca, Neisseria subflava, and Branhamella catarrhalis in a rapid (0.5- to 1-h) identification micromethod (RIM-N Kit; Austin Biological Laboratories, Inc., Austin, Tex.) and in a serum-free agar slanted medium (72 h). Reactions after 1 h in the RIM-N system agreed completely with those after 72 h in the conventional system.
Evaluation of the phadebact and bactigen reagents for detection of Neisseria meningitidis in cerebrospinal fluid
Authors: Kurzynski T A , Kimball J L , Polyak M B , Cembrowski G S , Schell R F
Published: January 2020

The Phadebact and Bactigen reagents were evaluated for detection of Neisseria meningitidis in cerebrospinal fluids. The Bactigen test yielded stronger agglutination reactions from clinical specimens and was significantly more sensitive when used with whole-cell suspensions and purified antigens.
Response of seronegative and seropositive adult volunteers to live attenuated cold-adapted reassortant influenza A virus vaccine
Authors: Clements M L , Tierney E L , Murphy B R
Published: January 2020

The infectivity and immunogenicity of live attenuated A/Washington/897/80 cold-adapted reassortant virus vaccine was evaluated in seronegative (hemagglutination inhibition titer, less than or equal to 1:4) and seropositive (hemagglutination inhibition titer, greater than 1:4) adult volunteers. The vaccine was efficient in infecting seronegative volunteers (94%). Moreover, 51% of seropositive vaccinees were infected by the virus. After live virus vaccination, greater than 83% of both seronegative and seropositive vaccinees achieved a level of nasal wash antibody previously associated with resistance to infection with influenza A virus. These findings indicate that both seronegative and seropositive vaccinees can benefit from live virus vaccination.
Asymptomatic rotavirus infections in day care centers
Authors: Barrón-Romero B L , Barreda-González J , Doval-Ugalde R , Zermeño-Eguia Liz J , Huerta-Peña M
Published: January 2020

Rotaviruses and other enteropathogenic agents were detected in 288 (42.1%) of 684 children in day care centers of Instituto Politecnico Nacional in Mexico City. The same agents were also found in 114 (37.7%) of 302 adults directly involved in the care of the children. The study was carried out from July to December 1982 and from July 1983 to February 1984. Rotaviruses were the main enteropathogenic agents found and were detected in 169 (29.9%) of 564 children without diarrhea and in 34 (28.3%) of 120 children with diarrhea. These viruses were present in 62 (20.5%) of 302 adults without diarrhea. Of all rotavirus-positive individuals, 20% were also positive for other enteropathogens. All these observations indicate that asymptomatic rotavirus infections are not a rare event in children and that diarrhea caused by rotavirus infections is only one of the expressions of their presence.
Evaluation of xylan fermentation for the identification of Bacteroides ovatus and Bacteroides thetaiotaomicron
Authors: Cooper S W , Pfeiffer D G , Tally F P
Published: January 2020

The reliability of xylan fermentation in distinguishing Bacteroides ovatus from Bacteroides thetaiotaomicron was examined with strains previously identified by DNA homology studies or bacteriophage sensitivity testing or both. All of the 12 B. ovatus strains produced acid ranging from pH 5.0 to 5.6 in peptone-yeast-xylan broth; none of the 19 B. thetaiotaomicron strains produced acid, and the final pH was 6.0 or 6.1. Xylan fermentation and bacteriophage sensitivity-DNA homology appear to be equivalent in their ability to differentiate these two Bacteroides species.
Neuraminidase activity is not the cause of influenza virus-induced neutrophil dysfunction
Authors: Abramson J S , Wiegand G L , Lyles D S
Published: January 2020

Influenza viruses have been shown to decrease the ability of polymorphonuclear leukocytes (PMN) to respond to a variety of stimuli. This study was done to determine if viral neuraminidase was responsible for decreased PMN function. Treatment of human PMN with purified neuraminidases from influenza virus, Vibrio cholerae, or Clostridium perfringens did not significantly affect the ability of human PMN to respond to stimulation. Occasional virus preparations that lacked the ability to depress PMN function did not differ in neuraminidase activity from viruses capable of causing depression. These results demonstrate that neuraminidase activity is not the cause of influenza virus-induced PMN dysfunction.
Pseudodiffuse-type growth of a Staphylococcus aureus strain in serum-soft agar
Authors: Chomarat M , Ichiman Y , Yoshida K
Published: January 2020

Strain MC31 of Staphylococcus aureus showed diffuse-type growth in regular serum-soft agar (SSA) (pH 7.2), but in alkaline SSA (pH 8.4) the type of growth was compact. The organisms were of extra low virulence in the mouse and were unencapsulated. Based on three properties, the growth type of the S. aureus strains in SSA was designated as pseudodiffuse type.
Mechanism of cross-contamination of blood culture bottles in outbreaks of pseudobacteremia associated with nonsterile blood collection tubes
Authors: McNeil M M , Davis B J , Anderson R L , Martone W J , Solomon S L
Published: January 2020

Several studies have identified nonsterile blood collection tubes as the reservoir for contaminating microorganisms in outbreaks of pseudobacteremia. Microorganisms are thought to be transferred from the nonsterile tubes to the blood culture tubes when blood is drawn for cultures, but the validity of this hypothesis has not previously been tested. We recently investigated an outbreak of Ewingella americana pseudobacteremia in which an epidemiological association was found between pseudobacteremia and the practice of drawing blood for culturing at the same time that blood for coagulation studies is drawn. The nonsterile coagulation tubes, prepared in the hospital, were contaminated with E. americana. We studied three different procedures that personnel reported using to inoculate blood culture tubes after dispensing blood into the coagulation tubes. Although the failure to change or remove the needle before inoculating the blood culture tubes resulted in the highest percentage of cross-contamination, we found that all the techniques we simulated carried some risk of cross-contamination when coagulation tubes were filled first.
Competitive, enzyme-linked immunosorbent assay for toxic shock syndrome toxin 1
Authors: Parsonnet J , Mills J T , Gillis Z A , Pier G B
Published: January 2020

We developed a competitive, enzyme-linked immunosorbent assay for the quantitation of toxic shock syndrome toxin 1 (TSST-1). Polyvalent immunoglobulin G from immunized rabbits was used as the capture antibody, and alkaline phosphatase conjugated to purified toxin served as the indicator enzyme. A standard curve was generated with each experiment, from which the concentration of toxin in culture supernatants was extrapolated. The assay was useful for determining toxin concentrations of 0.03 to 0.5 micrograms/ml, which is a substantial, practical improvement over immunodiffusion methods. Staphylococcal enterotoxins A through E were not significantly cross-reactive in the assay, and staphylococcal protein A did not interfere with quantitation of TSST-1. By testing a variety of staphylococcal strains, we found 100% concordance between toxin determinations made with our assay and those made by the investigators from whom the strains were obtained. The competitive, enzyme-linked immunosorbent assay is a highly reproducible, inexpensive means of determining TSST-1 concentrations and may have broad applicability in the field of toxic shock research.
Comparison of the PRAS II, AN-Ident, and RapID-ANA systems for identification of anaerobic bacteria
Authors: Burlage R S , Ellner P D
Published: January 2020

Two rapid systems for the identification of anaerobes were compared to a conventional growth system aided by a computer. The rapid systems (AN-Ident and RapID-ANA) are non-growth-dependent micromethods that identify anaerobes in 4 h by the action of various constitutive enzymes on chromogenic substrates. The organisms tested were 98 anaerobes, most of which were clinical isolates. The AN-Ident system identified 76 of these to species level and 86 to genus level; the RapID-ANA system correctly identified 74 of the organisms to species level and identified 93 to genus level. The PRAS II system correctly identified 77 to species level and 96 to genus level. In most instances, adequate identification could be obtained with either of the two rapid systems, but the conventional PRAS II system remains the most accurate.
Preparation and use of cytoplasmic antigens for the serodiagnosis of paracoccidioidomycosis
Authors: McGowan K L , Buckley H R
Published: January 2020

A cytoplasmic antigen of Paracoccidioides brasiliensis (strain MTC) prepared from the yeast phase grown in the chemically defined medium of McVeigh and Morton is described. This antigen can be easily prepared, does not vary from lot to lot, and can be lyophilized without loss of activity or potency. In the immunodiffusion test, the cytoplasmic antigen demonstrated a sensitivity of 97% and a specificity of 100% when tested against 218 sera from 139 cases of paracoccidioidomycosis. When 177 sera from patients with fungal diseases other than paracoccidioidomycosis were tested by immunodiffusion, there were no false-positive reactions. In an enzyme-linked immunosorbent assay, the antigen was equally effective in identifying cases, giving a sensitivity of 100% and a specificity of 99% when a 1:640 titer was considered the threshold for clinical significance. Antigenic components in the cytoplasmic extract of P. brasiliensis were examined after fractionation by concanavalin A-Sepharose 4B column chromatography. The fraction of the cytoplasmic antigen that binds to the concanavalin A column is material identical to the specific b1 (antigen 1) precipitin band described by A. Restrepo and L. H. Moncada (Appl. Microbiol. 28:138-144, 1974).
Isolation medium for the recovery of Pseudomonas cepacia from respiratory secretions of patients with cystic fibrosis
Authors: Gilligan P H , Gage P A , Bradshaw L M , Schidlow D V , DeCicco B T
Published: January 2020

A new medium for the isolation of Pseudomonas cepacia from respiratory tract secretions of patients with cystic fibrosis (CF) is described. This medium consists of inorganic salts, 0.5% pyruvate, and 0.1% proteose peptone as nutritive components and 0.0001% crystal violet, 0.15% bile salts, 100 micrograms of ticarcillin per ml, and 300 U of polymyxin B per ml as selective agents. The medium, designated PC medium, supported superior growth of 38 of 50 stock isolates of P. cepacia after 48 h of incubation when compared with MacConkey agar (0 of 50). The medium completely inhibited the growth of 112 of 124 stock isolates of organisms commonly found in respiratory secretions of CF patients. Cultures were made on PC medium with respiratory secretions of 169 CF patients. P. cepacia was recovered from 35 patients with isolates on PC medium but from only 21 patients with isolates on MacConkey agar. Of 221 other potentially pathogenic isolates found in these specimens, only six (two Pseudomonas aeruginosa isolates, two molds, one yeast, and one Serratia marcescens isolate) grew on PC medium. PC medium should facilitate the recovery of P. cepacia from CF patients.
Comparative evaluation of three identification systems for anaerobes
Authors: Murray P R , Weber C J , Niles A C
Published: January 2020

The accuracy of two new 4-h identification systems for anaerobes, the AN-IDENT (Analytab Products, Plainview, N.Y.) and the RapID ANA (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) was compared with that of the API 20A system (Analytab Products). A total of 132 clinical isolates were tested in each of the three systems. The overall accuracies at the genus and species level for the three systems were: API 20A, 68.9 and 56.8%, respectively; AN-IDENT, 90.2 and 73.5%; and RapID ANA, 93.9 and 81.8%. Improved identification of anaerobes with the AN-IDENT and the RapID ANA systems was observed for isolates of the genus Fusobacterium, Clostridium species other than Clostridium perfringens, non-spore-forming bacilli, and isolates of the genus Peptostreptococcus. Reproducibility studies demonstrated that the results of the individual test reactions in all three identification systems were reproducible when the interpretive guidelines of the manufacturer were followed precisely.
Serogrouping of Bacteroides vulgatus by the agglutination test
Authors: Okamura N , Miyazaki K , Chida T , Niwayama K , Nakaya R , Benno Y , Mitsuoka T
Published: January 2020

The agglutination technique was used to establish a serological classification scheme for Bacteroides vulgatus strains isolated from normal human feces and clinical specimens, especially from ulcerative colitis patients. Absorbed antisera to 10 strains of B. vulgatus were prepared. These 10 absorbed antisera were species specific. Of 90 B. vulgatus strains tested, 55 (61%) were agglutinated by one or more of these 10 absorbed antisera. A total of 27 serological patterns were grouped into 18 serogroups; 10 of these serogroups contained only one group component, whereas the other 8 serogroups were composed of more than one component. This serological classification could be used to study the epidemiology of this organism.
Fine-needle aspiration biopsy in diagnosis of soft tissue infections
Authors: Lee P C , Turnidge J , McDonald P J
Published: January 2020

This study explores the efficacy of fine-needle tissue aspiration biopsy in the diagnosis of soft tissue infections that cannot be sampled satisfactorily by regular microbiological techniques. Aspiration biopsy was performed on 50 patients with presumptive soft tissue infections. The conditions investigated were decubitus, diabetic, ischemic, venous, and traumatic ulcers (2, 6, 1, 2, and 7 patients, respectively), cellulitis (23 patients), chronic osteomyelitis (5 patients), and infected surgical wounds (4 patients). Where possible, comparison with superficial cultures was made. All of the cultures obtained from aspirate samples taken from ulcers, chronic osteomyelitis, and infected surgical wounds were positive. In cellulitis, cultures from aspirates were positive in 30 and 81% of the cases, respectively, depending on the presence or absence of concomitant antimicrobial therapy. These results suggest that fine-needle deep tissue aspiration biopsy is reliable and clinically applicable for deep tissue sample collection. The procedure is simple, brief, and does not cause significant discomfort to the patient. It also plays an important role in providing a guideline for antimicrobial therapy.
Comparison of neutralization and DNA restriction enzyme methods for typing clinical isolates of human adenovirus
Authors: Fife K H , Ashley R , Shields A F , Salter D , Meyers J D , Corey L
Published: January 2020

Sixty-five adenovirus isolates collected over a 3.5-year period were typed by both standard microneutralization techniques and restriction endonuclease digestion of viral DNA. Of the 65 isolates, 47 (72.3%) representing six adenovirus types could be typed by microneutralization. Eighteen isolates demonstrated partial neutralization with standard antisera to two or more adenovirus serotypes and thus could not be definitively typed. DNA analysis permitted typing of 64 of the 65 isolates (98.5%) (including four isolates which contained mixtures of two adenovirus types), and 12 different types were identified. Neutralization and DNA typing disagreed for five isolates, and in each case, digestion with multiple restriction endonucleases and DNA hybridization studies were consistent with the type assigned by DNA analysis. In addition, the DNA analysis method allowed the identification of genomic variants (genome types) of five adenovirus types. We conclude that typing clinical isolates of adenovirus by restriction endonuclease digestion of viral DNA can be done rapidly, provides additional epidemiological and typing information, and provides fewer ambiguous results than does typing by neutralization.
Improved utility of Gonoscreen, a Limulus amoebocyte lysate assay, in the evaluation of urethral discharges in men
Authors: Prior R B , Spagna V A
Published: January 2020

The Limulus amoebocyte lysate (LAL) assay was improved for the rapid evaluation of exudative urethritis in males. Two hundred men with various quantities of urethral discharge were evaluated. Pyrogen-free Dacron swabs were used for sample collection, and a chromogenic substrate was used for visible endpoint determination after a 10-min incubation. Of the 200 patients studied, 57 (29%) had minimal urethral discharges (less than 15 microliter) and could not be evaluated with Gonoscreen (Mallinckrodt, Inc., St. Louis, Mo.), an LAL test kit for the evaluation of urethritis which involves gentle aspiration for sample collection. The improved LAL assay had a sensitivity and specificity of 95 and 97%, respectively, and an overall accuracy to predict culture results of 96%. These results were not statistically different from Gram-stained smears read by experienced microscopists or from culture results (P greater than 0.05). Prevalence of gonorrhea in the study population was 40%, and the positive predictive value of the LAL assay was 95%. Thus, the use of swabs facilitated sample collection and increased by 29% the number of patients which could be evaluated with the LAL assay. In addition, the use of a chromogenic substrate reduced incubation time by 67% (30 to 10 min) and provided an objective color endpoint.
Evaluation of Detect-A-Strep and the Culturette Ten-Minute Strep ID kits for detection of group A streptococcal antigen in oropharyngeal swabs from children
Authors: Campos J M , Charilaou C C
Published: January 2020

The Detect-A-Strep kit (DAS) (Antibodies Inc.) and the Culturette Brand 10-Minute Group A Strep ID kit (CBTMSI) (Marion Scientific) were compared with a sensitive culture method for detection of group A streptococci in oropharyngeal swabs from children. Specimens from 953 children ranging in age from 4 months to 18 years were inoculated to selective blood agar plates containing sulfamethoxazole-trimethoprim. Culture plates were incubated anaerobically for the first overnight period and aerobically in the presence of 10% CO2 for the second overnight period. The same specimens were tested within 24 h by DAS (phase 1) or CBTMSI (phase 2). One hundred thirty-five of 538 specimens (25.1%) tested by DAS and 150 of 415 specimens (36.1%) tested by CBTMSI were culture positive. DAS detected 87 of 135 (64.4%) and CBTMSI detected 93 of 150 (62.0%) culture-positive specimens. The specificities of DAS and CBTMSI were 96.5 and 99.6%, respectively. The predictive values of positive and negative results were 86.1 and 89.0% for DAS and 98.9 and 82.2% for CBTMSI. Because a reliable distinction between infected patients and carrier state patients cannot be made, we conclude that neither DAS nor CBTMSI is sufficiently sensitive to replace our culture method.
New serotypes of Treponema hyodysenteriae
Authors: Mapother M E , Joens L A
Published: January 2020

Three isolates of pathogenic Treponema hyodysenteriae, B8044, B6933, and Ack 300/8, were serotyped by double immunodiffusion using lipopolysaccharide antigen and rabbit antiserum. The precipitation reactions of these three isolates were then compared with precipitin reactions from the reference strains of serotypes 1 through 4. The data indicated that B8044, B6933, and Ack 300/8 share antigenic determinants with either serotype 1 or 2 or both. However, when the cross-reacting antibodies were removed by adsorption, all reactions were found to be monospecific. Based on these results, three new serotypes of pathogenic T. hyodysenteriae are proposed: B8044 serotype 5, B6933 serotype 6, and Ack 300/8 serotype 7.
Fungal peritonitis caused by Lecythophora mutabilis
Authors: Ahmad S , Johnson R J , Hillier S , Shelton W R , Rinaldi M G
Published: January 2020

Fungal peritonitis caused by Lecythophora mutabilis, a mold rarely isolated from humans, is described. A patient on continuous peritoneal dialysis developed clinical, microbiological, and serological evidence for peritonitis due to this fungus. In vitro susceptibility testing of the fungus revealed marked differences in the activities of various antifungal agents. Although initially responding to treatment with oral ketoconazole, intraperitoneal miconazole, and catheter replacement, the patient had a documented relapse. The patient was eventually cured following the removal of a second catheter in association with prolonged imidazole treatment.
Isolation and differentiation of herpes simplex virus and Trichomonas vaginalis in cell culture
Authors: Gentry G A , Lawrence N , Lushbaugh W
Published: January 2020

During the period January 1982 to January 1985, 2,234 specimens were cultured for isolation of herpes simplex virus (HSV). HSV was isolated from 23% of these, Trichomonas vaginalis was isolated from 1.6%, and 75.3% were negative. In 0.2% of these, HSV and T. vaginalis were isolated from the same specimen. Cytopathic effects produced by HSV were identified by their sensitivity to arabinosylthymine, whereas those produced by T. vaginalis were identified by their lack of sensitivity to arabinosylthymine and by observation of motility. Cytopathic effects produced by T. vaginalis were reproduced by trophozoites from axenic cultures of T. vaginalis as well as by lysates of T. vaginalis added to serum-free BHK cells.
Comparison of the FeKal CON-Trate system with the formalin-ethyl acetate technique for detection of intestinal parasites
Authors: Long E G , Tsin A T , Robinson B A
Published: January 2020

The ability of the FeKal CON-Trate system (Trend Scientific, Minneapolis, Minn.) to recover parasite ova, cysts, or larvae from 300 fecal specimens was compared with that of the Formalin-ethyl acetate sedimentation technique. One hundred stool samples had previously been determined to contain 129 parasite forms. The FeKal system detected 127 parasites (sensitivity, 98.4%), and the Formalin-ethyl acetate sedimentation technique detected 125 parasites (sensitivity, 96.9%). Higher counts of parasite forms were obtained for 11 of 14 species of parasites in specimens prepared by the FeKal system. Four specimens were negative by the Formalin-ethyl acetate method and two by the FeKal system. These false-negative observations occurred in specimens with low counts of parasites. There were no false-positive results.
Evaluation of the Quantum II yeast identification system
Authors: Kiehn T E , Edwards F F , Tom D , Lieberman G , Bernard E M , Armstrong D
Published: January 2020

We compared three methods for identifying clinical yeast isolates: Abbott Quantum II, API 20C, and a modified BBL Minitek system. The API 20C and modified Minitek systems agreed on the identification of 243 of 245 yeasts (99.2%). The Quantum II system correctly identified 197 (80.4%), incorrectly identified 19 (7.8%), and did not identify 29 (11.8%) of the yeasts. Most of the misidentifications with the Quantum II occurred because assimilation or biochemical results were false-positive. Sixteen different species of yeasts and 16 different Quantum II substrates contributed to the discrepancies. On retesting with the Quantum II, 31% of the discrepant strains were correctly identified, while the remaining 69% were incorrectly identified or were not identified. Erroneous biochemical and assimilation results were also noted with yeasts that were correctly identified by the Quantum II system.
Interaction between limulus amoebocyte lysate and soluble antigens from Pseudomonas aeruginosa and Staphylococcus aureus studied by quantitative immunoelectrophoresis
Authors: Baek L , Høiby N , Hertz J B , Espersen F
Published: January 2020

To investigate the interaction of Limulus amoebocyte lysate (LAL) with gram-negative bacteria, soluble antigens from sonicated Pseudomonas aeruginosa were studied by various crossed-immunoelectrophoresis methods before and after reaction with LAL. Of 64 possible, at least 7 antigens were affected, as indicated by precipitin pattern, after the reaction with LAL. The precipitates corresponding to lipopolysaccharide and Pseudomonas "common antigen" disappeared. This reaction was inhibited when LAL was pretreated with lipopolysaccharide or by heating. Several of the reacting antigens have been shown to cross-react with many other strains of both gram-negative and gram-positive bacteria. Soluble antigens from a protein A-deficient strain of Staphylococcus aureus were also studied. LAL reacted with at least four of these antigens, including the teichoic acid complex. It is concluded that LAL is highly reactive with lipopolysaccharide, but it can react with other antigens from gram-negative and gram-positive bacteria as well. It is suggested that LAL interacts with biologically important antigens from the bacterial membrane. It is proposed that the reactivity and specificity of LAL for various microbial antigens can be studied by immunoelectrophoretic techniques.
Aggregation of platelets by Fusobacterium necrophorum
Authors: Forrester L J , Campbell B J , Berg J N , Barrett J T
Published: January 2020

Broth cultures and washed cells of 13 of 24 bovine isolates of Fusobacterium necrophorum aggregated human platelets in platelet-rich plasma. The cell-free culture fluid was inactive. Bacteria stored at 4 degrees C in saline remained active for at least 3 months, but they did not release activity into the storage solution. Aggregation typically began within 1 min after the addition of 10(3) bacteria to 10(3) platelets was complete within 5.5 min. Assays for cytosolic lactic dehydrogenase revealed that platelet lysis did not occur. The release of [14C]serotonin from platelets preincubated with this amine accompanied aggregation, indicating that this was a typical aggregation-degranulation reaction. Platelet aggregation was inhibited by EDTA (88% at 2.0 mM), aspirin (75% inhibition at 1.0 mM), and quinacrine (80% inhibition at 0.25 mM). Thus the reaction was an ion-dependent, cyclooxygenase-sensitive event. Gel-filtered platelets were less sensitive to aggregation than were platelets in plasma, but this sensitivity was fully restored by the addition of plasma and partially restored with fibrinogen. Biotyping of the cultures revealed that none of the avirulent, B-type strains of F. necrophorum could aggregate platelets, whereas 13 of 16 virulent A type strains were positive. These results suggest that platelet aggregation by F. necrophorum is related to the virulence of this organism.
Subclass distribution and molecular form of immunoglobulin A hemagglutinin antibodies in sera and nasal secretions after experimental secondary infection with influenza A virus in humans
Authors: Brown T A , Murphy B R , Radl J , Haaijman J J , Mestecky J
Published: January 2020

Serum and nasal wash specimens from 13 human volunteers undergoing experimental secondary infection with influenza A/Peking/2/79 (H3N2) wild-type virus were examined for the molecular form and subclass distribution of immunoglobulin A (IgA) antibodies to the viral hemagglutinin (HA). Nasal IgA antibodies were polymeric and did not bind radiolabeled secretory component, indicating that they were secretory IgA antibodies. Both IgA1 and IgA2 antibodies were detected; however, IgA1 accounted for most of the rise in IgA anti-HA levels seen after infection. In serum virtually all of the IgA HA antibodies were of the IgA1 subclass. Furthermore, the serum antibodies were predominantly polymeric and were capable of binding radiolabeled secretory component. These results suggested that the serum IgA antibodies to HA were of mucosal origin and that influenza A virus HA preferentially stimulates an IgA1 response.
Experimental reproduction of Potomac horse fever in horses with a newly isolated Ehrlichia organism
Authors: Dutta S K , Myrup A C , Rice R M , Robl M G , Hammond R C
Published: January 2020

Potomac horse fever, a recently recognized disease of equines, characterized by high fever, leukopenia, and a profuse diarrhea, was studied for its etiology. An Ehrlichia organism was isolated in equine macrophage-fibroblast cell cultures and mouse macrophage cell cultures from the mononuclear cells of blood of infected horses. The agent was continuously propagated in mouse macrophage cell cultures. The organism multiplied in the cytoplasm of mouse macrophage cells and was identified by Giemsa staining, acridine orange staining, and by indirect immunofluorescence with convalescent sera from infected horses. The disease was experimentally reproduced in horses inoculated with Ehrlichia-infected cell culture material. The Ehrlichia organism was reisolated from the blood of these infected horses during the course of the disease. Antibody against the organism was detected in the sera of experimentally infected horses. This study confirmed that the new Ehrlichia organism is the etiological agent of Potomac horse fever.
Evaluation of a Cordia-IC enzyme-linked immunosorbent assay kit for the detection of circulating immune complexes
Authors: Landoy Z , West T E , Vladutiu A O , Fitzpatrick J E
Published: January 2020

A commercial kit (Cordia-IC) from Cordis Laboratory, Miami, Fla., was compared with the Raji cell radioimmunoassay for its ability to detect circulating immune complexes (CIC) in sera from 30 control subjects and 118 patients with infectious diseases. The 118 patients were categorized into the following groups: (i) 23 patients with bacterial endocarditis, (ii) 41 patients with bacteremia from an infected intravascular catheter or access device, and (iii) 54 patients with Staphylococcus aureus bacteremia related to a deep tissue infection. The Cordia-IC was comparable to the Raji cell radioimmunoassay in intraassay variability (4.0 versus 8.0%) and interassay reproducibility (8.7 versus 20.0%). Neither assay found CIC amounts above 12.5 micrograms equivalents (eq) of aggregated human gamma globulin (AHG) per ml in any of the 30 control individuals. In group 1, Cordia-IC detected 19 of 23 positives (mean, 73.6 micrograms eq of AHG per ml), whereas the Raji cell detected 16 of 23 positives (mean, 54.8 micrograms eq of AHG per ml). In group 2, Cordia-IC detected 19 of 41 positives (mean, 20.6 micrograms eq of AHG per ml), whereas the Raji cell detected 16 of 41 positives (mean, 15.1 micrograms eq of AHG per ml). In group 3, Cordia-IC found 38 of 54 positives (mean, 28.0 micrograms eq of AHG per ml), whereas the Raji cell found 32 of 54 positives (mean, 23.9 micrograms eq of AHG per ml). Statistically, these findings were not significantly different in any of the three patient groups (P> 0.15), and there was an overall good correlation between the results obtained by the two assays (r+0.64, P<0.001). The Cordia-IC provided a suitable assay for the detection of CIC and might find application in routine clinical laboratories.
Colonization of human wounds by Escherichia vulneris and Escherichia hermannii
Authors: Pien F D , Shrum S , Swenson J M , Hill B C , Thornsberry C , Farmer J J
Published: January 2020

In this report we present clinical descriptions of 12 Hawaiian patients from whom Escherichia vulneris or E. hermannii strains were isolated. All but two patients had soft-tissue infections with multiple bacteria, particularly Staphylococcus aureus. The other two had purulent conjunctivitis associated with S. aureus and infected malignant peritonitis with multiple organisms, respectively. In none of the cases were the Escherichia spp. found in abundant quantities or considered pathogenic. In preliminary animal pathogenicity studies, 12 strains each of E. vulneris and E. hermannii failed to cause serious symptoms in 4-week-old mice when 10(7) cells were injected intraperitoneally. When 10(6) cells were used, none of these bacterial strains injected into mouse soft tissue was capable of producing persistent wound infections. Susceptibility studies of 40 strains of these bacteria to 20 different antimicrobial agents showed that they were susceptible to third-generation cephalosporins as well as to most other cephalosporins, aminoglycosides, trimethoprim, and sulfamethoxazole-trimethoprim; these strains were only marginally susceptible or resistant to penicillin, tetracycline, chloramphenicol, and nitrofurantoin.
Clinical appearance of orofacial infections of odontogenic origin in relation to microbiological findings
Authors: Heimdahl A , von Konow L , Satoh T , Nord C E
Published: January 2020

Fifty-eight patients with acute orofacial infections of odontogenic origin were classified into two groups with respect to the severity of infection. A total of 174 anaerobic and 22 aerobic bacterial strains were isolated. Anaerobic gram-negative rods were isolated more frequently from the patients with severe infections than from the patients with infections judged as mild (P less than 0.05). The occurrence of Fusobacterium nucleatum especially appeared to be associated with the severity of the infections (P less than 0.05). Penicillin resistance among the anaerobes was rarely found, while resistance to erythromycin was a common finding. All aerobic and anaerobic bacteria were susceptible to clindamycin, and all obligate anaerobic bacteria were susceptible to nitroimidazoles.