bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid presumptive identification of Cryptococcus neoformans by staphylococcal coagglutination
Authors: Maccani J E
Published: January 2020

A coagglutination reagent was prepared by sensitizing the Cowan I strain of Staphylococcus aureus with rabbit immune globulin directed against Cryptococcus neofromans A15 and absorbed with C. laurentii. This reagent was evaluated for its usefulness in differentiating C. neoformans from other yeast colonies rapidly. Antigen-containing extracts were prepared form Sabouraud dextrose agar cultures of 48 C. neoformans, 33 other Cryptococcus species, 21 Candida, 4 Torulopsis, 3 Saccharomyces, and 2 Rhodotorula strains. This was done by suspending a 0.001-ml loopful of colony growth in 0.5 ml of phenolized saline, mixing for 30 s, and then centrifuging. Equal volumes (50 microliters) of coagglutination reagent and yeast extract were mixed within marked circles on a glass slide and then mechanically rotated at 180 rpm for 8 min. Forty-five of the 48 strains of C. neoformans produced strong (3+ to 4+) agglutination, and 3 strains of serotype C produced weak (1+ to 2+) agglutination with the reagent. Other Cryptococcus species which reacted positively were 4 C. albidus subsp. diffluens, 7 C. albidus subsp. albidus, and 2 C. terreus strains; however, false-positive errors in identification were circumvented by performing a supplemental rapid test for nitrate utilization which differentiated these yeasts from C. neoformans. None of the other yeasts tested (including 14 C. laurentii, 2 C. luteolus, and 2 C. uniguttulatus strains) produced any degree of agglutination with the reagent. A commercial cryptococcal latex agglutination reagent (Crypto-Test, Microbiological Associates, Walkersville, Md.) proved less reliable for identifying C. neoformans yeast colonies because of cross-reactions which occurred with all other species of Cryptococcus tested.
Amino acid requirements for Legionella pneumophila growth
Authors: Tesh M J , Miller R D
Published: January 2020

The amino acids L-arginine, L-isoleucine, L-leucine, L-methionine, L-serine, L-threonine, and L-valine were essential for the growth of Legionella pneumophila in a chemically defined medium. A partial requirement for L-cysteine (or L-cystine) was also observed. A minimal medium containing only the eight required amino acids supported the growth of this bacterium only if the medium was supplemented with L-glutamic acid. This latter compound was the only amino acid capable of stimulating growth in the eight-amino acid medium.
Laboratory evaluation of the AutoMicrobic system for identification of Enterobacteriaceae
Authors: Freeman J W , Rowland R W , Overman S B , Goodman N L
Published: January 2020

The Enterobacteriaceae Biochemical Card used with the AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.) was compared with the API 20E system (Analytab Products, Plainview, N.Y.) for identifying members of the family Enterobacteriaceae. A total of 1,401 clinical isolates representing 18 species were identified by the AutoMicrobic and API 20E systems over a 5-month period. Discrepancies between these systems were resolved by conventional methods. Overall, 98.3% of the isolates were identified correctly by the AutoMicrobic system in 8 h, with 94.2% having an AutoMicrobic system confidence level greater than or equal to 90%; 92.9% of the isolates were identified correctly by the 20E system after 24 h of incubation. Discrepancies between the two systems occurred in 3.1% of the isolates. Of these, 40.9 and 59.1% proved to be correct by the AutoMicrobic and API 20E systems, respectively.
Comparison of viral isolation, direct immunofluorescence, and indirect immunoperoxidase techniques for detection of genital herpes simplex virus infection
Authors: Moseley R C , Corey L , Benjamin D , Winter C , Remington M L
Published: January 2020

Seventy-six consecutive patients presenting to a genital herpes simplex virus (HSV) clinic were enrolled in a study comparing viral isolation (VI), indirect immunoperoxidase (indirect IP), and direct immunofluorescence (direct FA) techniques for the detection of HSV antigen. Of the 76 patients, 61 (80%) demonstrated HSV by VI, compared with 66% by indirect IP and 55% by direct FA (P less than 0.05). Genital lesions from nine patients demonstrated HSV antigen by direct FA or indirect IP but were VI negative; eight of nine patients had subsequent episodes of genital HSV confirmed by VI. During the vesicular-pustular stage of the disease, VI was positive in 90%, indirect IP was positive in 76%, and direct FA was positive in 71% of the lesions, whereas with ulcerative lesions, VI was positive in 72%, indirect IP was positive in 55%, and direct FA was positive in 38%. These commercially available rapid viral diagnostic techniques are specific and useful, if adequate specimens are obtained from early genital lesions.
Rapid identification and antimicrobial susceptibility testing of gram-negative bacilli from blood cultures by the AutoMicrobic system
Authors: Moore D F , Hamada S S , Marso E , Martin W J
Published: January 2020

A procedure was developed which allows direct identification and antimicrobial susceptibility testing of fermentative and nonfermentative gram-negative bacilli from positive blood cultures. A 10-ml sample was removed from turbid blood culture bottles, and the bacteria were washed and concentrated by centrifugation. The bacterial pellet was used to inoculate an Enterobacteriaceae Plus Identification Card and a Gram-Negative General Susceptibility Card of the AutoMicrobic system. Results with these cards were compared with results obtained with standard technique for 196 blood cultures seeded with recent clinical isolates. Identification of most cultures was available in 8 h, whereas the antimicrobial susceptibility results were available in an average of 4.7 h for all organisms. Direct identification was correct for 95% of the cultures, whereas the antimicrobial susceptibility data had an average agreement of 87% with 3.8% very major and 1.4% major errors. In using this procedure it was possible to provide accurate preliminary identification and results of antimicrobial susceptibility tests for gram-negative bacilli on the same day that a blood culture was determined to be positive.
Comparative growth of Eikenella corrodens on 15 media in three atmospheres of incubation
Authors: Goldstein E J , Agyare E O , Silletti R
Published: January 2020

The abundance of growth and size of colonies of 10 isolates of Eikenella corrodens were studied using 15 commonly available media in aerobic, 5% CO2, and anaerobic atmospheres of incubation. Chocolate agar was the best medium for overall growth in all three atmospheres. Growth on Mueller-Hinton agar supplemented with 5% blood and on Brucella agar supplemented with 5% blood was equivalent to that on chocolate agar in 5% CO2 and anaerobic environments but not in an aerobic atmosphere. Strains of E. corrodens did not grow on the selective media studied. Growth on media without blood or hemin supplements occurred in anaerobic but not in aerobic or 5% CO2 environments, and was consistently poorer than growth on the same media with these supplements.
Aerobic bacterial oral flora of garter snakes: development of normal flora and pathogenic potential for snakes and humans
Authors: Goldstein E J , Agyare E O , Vagvolgyi A E , Halpern M
Published: January 2020

Garter snakes that are used for scientific laboratory studies or kept as exotic pets often become ill and die early in captivity. They may also act as reservoirs of potential human pathogens or transmit infection to man. A total of 126 strains of aerobic and facultative bacteria, most potential human and snake pathogens, were isolated from 82 garter snake oropharyngeal cultures. Coagulase-negative Staphylococcus species were the most common species isolated. Acinetobacter calcoaceticus var. anitratus, Hafnia alvei, Arizona hinshawii, Salmonella species, Shigella species, Klebsiella oxytoca, and Pseudomonas aeruginosa were among the potential pathogens isolated. The spectrum of bacteria with potential for causing oral and pulmonary infections in garter snakes is greater than has been previously appreciated. Garter snakes should also be considered reservoirs of human pathogens, and appropriate precautions should be taken by laboratory personnel and pet owners.
Improved technique for isolation of Mycobacterium kansasii from water
Authors: Powell B L , Steadham J E
Published: January 2020

A new concentration procedure, together with a new isolation medium, resulted in a 10- to 100-fold increase in the possibility of identifying Mycobacterium kansasii from water samples in comparison to a previously used procedure. In a survey which included both potable and natural water samples from many sites within the state of Texas, nine isolations of high-catalase strains of M. kansasii were obtained from 232 water samples tested. Acid-fast smear results were compared with mycobacterial isolations. An isolate from a river in central Texas is the first high-catalase strain of M. kansasii encountered in a natural water supply. Surveys of water samples from two Texas towns indicate that chlorine levels may influence the number of recoverable mycobacteria in water supplies.
Frequency of preclumped virus in routine fecal specimens from patients with acute nonbacterial gastroenteritis
Authors: Narang H K , Codd A A
Published: January 2020

A low-speed centrifugation technique for the preparation of grids after minimal purification of fecal extracts is described for examination of viruses by direct electron microscopy using negative staining. Results showed that adenovirus, astrovirus, rotavirus, and "small round" viruses were frequently shed into the gastrointestinal tract in clumps of variable size. Differential centrifugation study showed that a substantial proportion of the virus in the sample was lost in the initial pellet at the first step of clarification; this finding casts doubt on the validity of immune electron microscopy for direct typing of strains of these viruses from stools. In addition, particle counts based on conventional specimen processing are likely to grossly underestimate the true value.
Evaluation of a Charcoal Particle Test for Rheumatoid Factor
Authors: Stevens Roy W. , Hechemy Karim E. , Miller Joan H.
Published: January 2020

A rheumatoid factor charcoal card test was compared with a rheumatoid factor latex slide and tube agglutination test. Qualitative findings with 280 of 290 selected sera were the same. Quantitative results converted to standard rheumatoid factor units were similar.
Lysostaphin disk test for routine presumptive identification of staphylococci
Authors: Poutrel B , Caffin J P
Published: January 2020

The sensitivity of the lysostaphin disk test was evaluated for routine differentiation of staphylococci from micrococci. Lyophilized paper disks impregnated with 10 micrograms of lysostaphin were placed on a Mueller-Hinton agar plate inoculated with 10 species of staphylococci and 7 species of micrococci. After 16 h of incubation at 37 degrees C, a circular zone of growth inhibition was noted for all staphylococci species tested. By contrast, all the micrococci species were resistant. The activity of the impregnated disks was not diminished by storing them at 4 degrees C for 3 months. With disks impregnated by 10, 6.6, 5, and 2.5 micrograms of lysostaphin, diameters of the growth inhibition zone for S. aureus Cowan 1 NCTC 8530 strain were proportional to the logarithm of the quantity of lysostaphin present in each disk.
Modified oxidase and benzidine tests for separation of staphylococci from micrococci
Authors: Faller A , Schleifer K H
Published: January 2020

Two simple and rapid methods for the separation of staphylococci from micrococci are described. They are based on modified oxidase and benzidine tests. Micrococci and Staphylococcus sciuri yield a blue color with a 6% solution of tetramethylphenylenediamine in dimethyl sulfoxide, whereas all of the other staphylococci exhibit no coloration. Best's results were obtained with overnight cultures on blood agar. The presence of c-type cytochromes i micrococci and S. sciuri could be detected with benzidine; all noncovalently linked heme groups are removed before the addition of the benzidine reagent. The oxidase test is the simplest and most rapid method for the separation of staphylococci (except S. sciuri) from micrococci, if the nutritional requirements and the time of incubation are strictly followed. This test is especially recommended for the examination of clinical material in which S. sciuri is usually not found.
Microtest procedure for isolation of Chlamydia trachomatis
Authors: Yoder B L , Stamm W E , Koester C M , Alexander E R
Published: January 2020

Management of patients potentially infected with Chlamydia trachomatis has been hampered by the cost and time required to perform Chlamydia cultures. To isolate C. trachomatis, we developed a microtiter method that exhibited equal sensitivity but less frequent contamination than our previously used vial-cover slip culture system. In addition, costs and technician time were substantially reduced with the microtest method. Subsequent studies showed that cycloheximide-treated cells were superior to 5-iodo-2-deoxyuridine-treated cells in the microtest method and that a subpassage significantly enhanced the sensitivity of the method. The microtest method appears to be a sensitive, rapid, and economical method for isolating C. trachomatis.
Bacterial fouling of a hospital closed-loop cooling system by Pseudomonas sp
Authors: du Moulin G C , Doyle G O , MacKay J , Hedley-Whyte J
Published: January 2020

During the summer of 1979 the air-conditioning system at a hospital in Boston deteriorated, and this led to total failure of some chilling units. Patient care and operating-room areas were affected. Investigation of the problem ruled out mechanical and electrical causes, but revealed a strain of Pseudomonas sp. biofouling heat transfer tubes of the closed chilled water system. The pseudomonads apparently were stimulated to grow by low concentrations of ethylene glycol antifreeze. The proximate source of these organisms was an expansion tank located in a 33 degrees C environment. The organisms probably originated from the potable water supply of the hospital. Fouling was eventually cleaned by prolonged and expensive treatments of the closed chilled water system. Pseudomonas sp. is frequently isolated from hospital-acquired infections at our institution (Beth Israel Hospital, Boston, Mass.); however, our studies with fluorescent dye tracers indicated that organisms were prohibited from entering patient areas via contaminated water from the chillers. Microbiologists must become cognizant of seemingly unimportant microbial environments within hospitals that may indirectly contribute to hospital-acquired infections.
Branhamella (Neisseria) catarrhalis--a lower respiratory tract pathogen?
Authors: Johnson M A , Drew W L , Roberts M
Published: January 2020

Branhamella (Neisseria) catarrhalis was identified as a probable respiratory tract pathogen in seven patients, four with pneumonia and three with bronchitis. Five of the B. catarrhalis isolates produced beta-lactamase. Production of beta-lactamase correlated with penicillin resistance by the standardized disk diffusion method and also with high minimal inhibitory concentrations of penicillin for the two strains which were tested.
Screening of uropathogenic Escherichia coli for expression of mannose-selective adhesins: importance of culture conditions
Authors: Fein J E
Published: January 2020

I surveyed the prevalence of mannose-sensitive adherence factors (adhesins) in uropathogenic Escherichia coli isolates by using several rapid screening tests based on the agglutination of stained yeast cells. Mannose-sensitive adhesin activity was demonstrated best for bacteria grown in aerobic static broth cultures. Under these conditions, the majority of the strains from patients with significant bacteriuria (55[85%] of 181 isolates) or upper urinary tract infections (19[79%] incidence of mannose-sensitive adhesion activity was detected for strains cultured on solid medium. Thus, only 26 (51%) of 51 fresh isolates from cases of significant bacteriuria appeared to have mannose-sensitive adhesin activity when they were tested directly on the primary isolation medium (MacConkey agar), whereas after serial static broth subculturing, 45 (88%) of these strains were active. Many of the nonagglutinating weakly agglutinating strains detected were also nonmotile. Various other enterobacteria showed strong agglutinating activity toward the yeast cells, and in most cases these reactions were blocked by mannose. However, a few notable exceptions were found.
Selective medium for isolation of Klebsiella pneumoniae
Authors: Bruce S K , Schick D G , Tanaka L , Jimenez E M , Montgomerie J Z
Published: January 2020

Selective media for Klebsiella pneumoniae have been important in studies of hospital-acquired infections. On an agar medium which included ornithine, raffinose, and Koser citrate, K. pneumoniae strains grew as yellow mucoid colonies at 24 h and there was some increase in colony size at 48 h. Other members of Enterobacteriaceae were inhibited or produced small pink colonies on this same medium. Pseudomonas, Providencia, Acinetobacter, and Proteus species did not grow or showed very poor growth. The growth and appearance of these bacteria were not influenced by pH changes over a pH range of 5.2 to 6.4. Of 368 swabs of body sites cultured on MacConkey agar and on the test medium, 121 K. pneumoniae isolates on MacConkey agar and the same number on the test medium resulted. There were no discrepancies between the two media. Upon direct plating of stool, however, more K. pneumoniae colonies were isolated on the test medium than on MacConkey agar. Colonies on the test medium were more readily selected and identified than the colonies on MacConkey agar. There was also no inhibition of K. pneumoniae growth on the test medium compared with blood agar medium. This medium may be useful for the selective isolation of K. pneumoniae.
Detection of Bacterial Pathogens in Purulent Clinical Specimens by Immunofluorescence Techniques
Authors: Clausen Carla R.
Published: January 2020

A technique is described which may be used to identify
Haemophilus influenzae
type b,
Streptococcus pneumoniae
, group A and B streptococci, and
Staphylococcus aureus
in smears of clinical specimens.
Increased Sensitivity of a New Coagglutination Test for Rapid Identification of
Haemophilus influenzae
Type b
Authors: Grasso Robert J. , West Loyd A. , Holbrook Nikki J. , Halkias Demetrios G. , Paradise Lois J. , Friedman Herman
Published: January 2020

A newly developed rapid coagglutination test for identifying
Haemophilus influenzae
type b organisms isolated from clinical specimens correlated 100% with the slide agglutination test but was 100- to 200-fold more sensitive.
Rapid Presumptive Identification of
Gardnerella vaginalis
(
Haemophilus vaginalis
) from Human Blood Agar Media
Authors: Shaw Carol E. , Forsyth Michele E. , Bowie William R. , Black William A.
Published: January 2020

Presumptive identification of
Gardnerella vaginalis
from 48-h human blood agar cultures by using a Gram stain, hemolysis, and colonial morphology was highly accurate.
Lack of clinical relevance in routine terminal subculturing of blood cultures
Authors: Gill V J
Published: January 2020

The usefulness of performing final blind subcultures of previously negative blood cultures was evaluated over a 21-month period. From over 14,000 blood culture bottles blindly subcultured after 7 days of incubation, only 12 potentially significant organisms were found. The finding of these 12 organisms did not influence patient care since in 11 instances the same organism had already been reported from prior positive bottles and in 1 instance the patient had already died. These results suggest that blind 7-day subcultures are of minimal value. Other factors that need to be considered before eliminating the final subculture are presented.
Reagent Quality Control in Bacteriology: Cost-Effective, Easy-to-Use Methodology
Authors: Hicock Paul I. , Marshall Karen E.
Published: January 2020

A reagent quality control scheme is described which is both cost-effective and flexible enough for use with a variety of commercially available enteric identification systems.
Relationship of phospholipid chemistry to serological reactivity in the Venereal Disease Research Laboratory slide test antigen
Authors: Reeves M W , McGrew B E , McLaurin B , Pine L
Published: January 2020

A total of 13 egg lecithins, 12 beef heart lecithins, and 15 beef heart cardiolipins were assayed for the ability to function in the Venereal Disease Research Laboratory microflocculation test, as well as for purity, fatty acid composition, free amines, metals, and products of oxidation. We found that the presence of peroxides and oxidation-related ultraviolet-absorbing chromophores showed a close inverse relationship to acceptable serological activity. The degree of purity of the lipids had only a slight influence on serological activity, whereas fatty acid composition, saturation, and configuration had none at all. We did not detect contaminating iron, copper, cobalt, nickel, or free amines in these lipids. We discuss the implications of our findings for improving the chemical standards for these lipids.
Pseudomonas aeruginosa enzyme profiling: predictor of potential invasiveness and use as an epidemiological tool
Authors: Janda J M , Bottone E J
Published: January 2020

The enzymatic potential of 54 clinical and 22 environmental isolates of Pseudomonas aeruginosa from soil and water were evaluated by substrate plate assays. Clinical isolates produced substantial levels of 9 of the 11 enzymes assayed, whereas strains recovered from soil or water were relatively inert enzymatically. Elastase, deoxyribonuclease, and elevated protease activities were associated preferentially with clinical isolates of systemic origin; these activities were found twice as frequently in clinical isolates as in strains derived from sputum or the urogenital tract. Our data suggest that these factors may play an important role in the dissemination of P. aeruginosa from local or superficial sites. A comparison of the enzyme profiles of the environmental and clinical isolates indicated that colonization or infection by environmental strains of P. aeruginosa is a rare event and that environmental and clinical strains comprise separate biovars. Epidemiologically, enzyme profiles permitted the fingerprinting and differentiation of clinical strains from various sources.
Preservation and transportation of bacteria by a simple gelatin disk method
Authors: Obara Y , Yamai S , Nikkawa T , Shimoda Y , Miyamoto Y
Published: January 2020

A wide range of bacterial species, e.g., Enterobacteriaceae and Neisseria, Streptococcus, Branhamella, Haemophilus, Gemella, Pseudomonas, Flavobacterium, and Bacteroides species, were successfully preserved for 1 to 5 years by our gelatin disk drying method. The beta-lactamase activity of penicillinase-producing Neisseria gonorrhoeae was retained for more than 3 years with this method. Good results were also obtained upon airmailing many strains of N. gonorrhoeae embedded in gelatin disks from Japan to the United States. Neisseria, Branhamella, Gemella, and Haemophilus organisms suspended in the reagent used in the preparation of the gelatin disks could be preserved for 6 to 12 months after freezing the cell suspensions at -20 degrees C. Furthermore, modification of our gelatin disk preservation method made possible the safe long- and short-distance transportation of clinical isolates. Our methods can be used by any small laboratory, since they require only conventional instruments and reagents.
Tatumella ptyseos gen. nov., sp. nov., a member of the family Enterobacteriaceae found in clinical specimens
Authors: Hollis D G , Hickman F W , Fanning G R , Farmer J J , Weaver R E , Brenner D J
Published: January 2020

The name Tatumella ptyseos gen. nov., sp. nov., is proposed for a group of organisms (previously called group EF-9) isolated from clinical sources in the United States, Canada, and Puerto Rico. A total of 68% of these isolates were from sputum specimens. T. ptyseos strains are gram-negative, oxidase-negative, fermentative rods that grow on MacConkey agar. The distinctive biochemical characteristics of 44 T. ptyseos isolates were as follows: acid but no gas from D-glucose, sucrose, and, usually (71%), D-xylose (62% delayed); no acid from lactose, maltose, or D-mannitol; negative tests for indole, urea, methyl red, gelatin, L-lysine decarboxylase, and L-ornithine decarboxylase; L-arginine dihydrolase variable; phenylalanine deaminase positive; Voges-Proskauer positive by the Coblentz method but negative by the O'Meara method; nonmotile at 36 degrees C but 66% weakly motile (30% delayed) at 25 degrees C; Simmons citrate positive at 25 degrees C (89%) but Simmons citrate negative at 36 degrees C. Deoxyribonucleic acid-deoxyribonucleic acid relatedness studies on 26 T. ptyseos strains showed that they were 80 to 100% related at 60 degrees C, which indicated that they comprise a single species. The deoxyribonucleic acid relatedness to other species within the Enterobacteriaceae was 7 to 38%. This is evidence that this species belongs in this family, is distinct from all described species and is best placed in a new genus. The T. ptyseos isolates studied were susceptible to all of the antimicrobial agents tested by broth dilution; these antimicrobial agents were amikacin, ampicillin, cephalothin, chloramphenicol, gentamicin, kanamycin, tetracycline, and tobramycin. Three striking differences between T. ptyseos and other members of the Enterobacteriaceae were its large zone of inhibition around penicillin (mean diameter 24 mm), its tendency to die on some laboratory media (such as blood agar) within 7 days, and its small number (usually one) of flagella. Strain H36 (=ATCC 33301, =CDC D6168, =CDC 9591-78) is the type strain of this new species. T. ptyseos is the type species for the genus Tatumella.
Acute diarrhea and rotavirus infection in newborn babies and children in Yogyakarta, Indonesia, from June 1978 to June 1979
Authors: Soenarto Y , Sebodo T , Ridho R , Alrasjid H , Rohde J E , Bugg H C , Barnes G L , Bishop R F
Published: January 2020

A longitudinal study of acute diarrhea in children in Yogyakarta, Indonesia (June 1978 to June 1979), showed little variation throughout most months of the year in numbers of children admitted to hospital and in numbers infected with rotaviruses. Both decreased during November and December coincidentally with seasonal change from dry to wet conditions. Rotavirus particles were identified by electron microscopy in fecal specimens from 126 of 334 (38%) infants and children with acute diarrhea. Nosocomial rotavirus infections occurred in 11% of control children admitted to hospital for other reasons. Socioeconomic level and preexisting nutritional status did not influence the incidence of rotavirus excretion. Rotavirus infections were most common in children aged 6 to 24 months. There was a low incidence of infection in infants less than 6 months old. Rotavirus infection was seldom observed in newborn babies delivered in an urban hospital nursery, in a rural health center, or at home. One of 72 newborn babies with diarrhea excreted rotavirus. One of 53 healthy newborn babies excreted rotavirus. It is concluded that, in Indonesia, rotavirus infection is a major cause of childhood diarrhea throughout the year, but is an uncommon cause of diarrhea in newborn babies.
Comparison of two different enzyme immunoassays for detection of immunoglobulin M antibodies against tick-borne encephalitis virus in serum and cerebrospinal fluid
Authors: Heinz F X , Roggendorf M , Hofmann H , Kunz C , Deinhardt F
Published: January 2020

Two enzyme immunoassays for the detection in immunoglobulin M (IgM) antibodies against tick-borne encephalitis virus were compared, employing a solid phase coated either with antigen or with mu-chain-specific antiserum to human IgM. The latter IgM-capturing assay system proved to be more sensitive, and its superiority was especially prominent when high titers of tick-borne encephalitis virus-specific IgG antibodies in addition to specific IgM antibodies were present in the sample. The application of this test is a valuable extension of the diagnostic tools for the rapid diagnosis of tick-borne encephalitis by IgM detection. Since specific IgG does not interfere, diagnosis can be readily established when patients have already developed high titers of IgG antibodies relatively late after the onset of the disease.
Nosocomial transmission of Serratia marcescens in a veterinary hospital due to contamination by benzalkonium chloride
Authors: Fox J G , Beaucage C M , Folta C A , Thornton G W
Published: January 2020

During a 1-year period, Serratia marcescens was isolated from 50% of all contaminate intravenous catheters from dogs and cats in a large veterinary hospital. S. marcescens was also isolated from respiratory tracts, genitourinary tracts, skin, and other sites in hospitalized animals. A total of 55% of the clinical isolates and 66% of the intravenous catheter isolates had the same API biochemical profile. The source of the S. marcescens was determined to be aqueous benzalkonium chloride (0.025%) sponge pots located in the intensive care unit, surgery rooms, and outpatient clinic areas of the hospital. Of the 11 S. marcescens isolates submitted to the Centers for Disease Control for serotyping (6 from aqueous benzalkonium chloride sponge pots, 5 from intravenous catheters), 8 were identified as serotype O10:H11. All S. marcescens isolates tested for antibiotic susceptibilities were multiply resistant; isolates were most frequently resistant to streptomycin, cephalothin, and ampicillin. This study demonstrates that improper use of disinfectants plays an important role in the nosocomial transmission of S. marcescens.
Comparison of methods for identifying Staphylococcus and Micrococcus spp
Authors: Gunn B A , Singleton F L , Peele E R , Colwell R R , Keiser J K , Kapper C O
Published: January 2020

Three methods employed to distinguish staphylococci from micrococci were compared, using clinical and environmental strains. When these methods are used, misinterpretation of results, as well as erratic results, may occur, and suggestions for eliminating these problems are provided. The most sensitive test that combines ease of use and speed in obtaining results for distinguishing the two genera is the lysostaphin susceptibility test. Two other tests, facultatively anaerobic growth in semisolid thioglycolate agar and fermentation of dextrose, may also be used to distinguish these two genera, but results are often slow in developing, are subject to technical difficulties, and may lead to incorrect assignment of certain species of staphylococci and micrococci to their proper genera.
Comparison of acridine orange and Gram stains for detection of microorganisms in cerebrospinal fluid and other clinical specimens
Authors: Lauer B A , Reller L B , Mirrett S
Published: January 2020

Acridine orange, a fluorochrome strain, is potentially superior to the Gram stain in the direct microscopic examination of clinical specimens because it gives striking differential staining between bacteria and background cells and debris. Its value in clinical laboratories was evaluated by testing 209 cerebrospinal fluids and 288 other body fluids, tissues, and exudates by both techniques. Smears were made in duplicate, fixed with methanol, stained, and examined without knowledge of the result of the companion smear or culture. Overall, acridine orange was slightly more sensitive than the Gram stain (acridine orange, 59.9%; Gram stain, 55.8%) and equally specific in detecting microorganisms. One smear was falsely positive by the Gram stain; none was falsely positive by the acridine orange stain. We conclude that acridine orange staining is a sensitive method for screening clinical specimens and reviewing selected specimens that are purulent, but negative by the Gram stain. Bloody fluids, thick exudates, and other normally difficult-to-read specimens were easily and quickly examined. We recommend, however, that positive smears be reexamined with the Gram stain to confirm the result and determine the Gram reaction of the microorganisms.
Primary isolation of Neisseria gonorrhoeae on hemoglobin-free New York City medium
Authors: Granato P A , Schneible-Smith C , Weiner L B
Published: January 2020

New York City medium and New York City medium without hemoglobin were comparatively evaluated for their ability to support the growth of Neisseria gonorrhoeae isolated from 1,010 clinical specimens. Although hemoglobin in the form of lysed horse erythrocytes stimulated gonococcal growth, the absence of this component from New York City medium did not have a detrimental effect on the recovery of gonococci isolated from clinical specimens. Both media were comparable in their ability to cultivate gonococci from clinical material, with a total of 187 gonococcal isolates being recovered on each of the media. The results of this study showed that the preparation of New York City medium can be facilitated and that its cost can perhaps be reduced by the elimination of the hemoglobin component from the formulation without adverse effect on the recovery of N. gonorrhoeae isolated from clinical specimens.
Relationship between photochromogenicity and test temperature in mycobacteria
Authors: Tsukamura M
Published: January 2020

Although it has been recommended that photochromogenicity of mycobacteria should be tested after incubation at 25 to 30 degrees C for 2 weeks (Wayne et al., Int. J. Syst. Bacteriol. 24:412-419, 1974), our results indicate that photochromogenicity should be determined at 37 degrees C in the dark and under continuous exposure to light for 2 weeks. The test at 37 degrees C gives more stable results for Mycobacterium kansasii, Mycobacterium marinum and Mycobacterium szulgai.
Rapid Identification of Enterococci by Reduction of Litmus Milk
Authors: Schierl Elizabeth A. , Blazevic Donna J.
Published: January 2020

Of 100 strains of enterococci, 83% reduced litmus milk within 4 h. None of the 100 non-enterococcal streptococci tested were positive.
Bordetella bronchiseptica peritonitis associated with continuous ambulatory peritoneal dialysis
Authors: Byrd L H , Anama L , Gutkin M , Chmel H
Published: January 2020

Bordetella bronchiseptica has rarely been reported as an opportunistic pathogen in humans. This is the first documented report of peritonitis due to B. bronchiseptica. The organism was seen on a Gram strain of peritoneal fluid and was isolated from a culture of the fluid. It was thought that the source of infection was related to the close contact of the patient with his dog. The patient was successfully treated with chloramphenicol administered intraperitoneally and intravenously.
Serratia ficaria isolated from a human clinical specimen
Authors: Gill V J , Farmer J J , Grimont P A , Asbury M A , McIntosh C L
Published: January 2020

The first isolation of Serratia ficaria from a human source is reported. Investigation into the eating habits of the patient revealed the probable source as figs, which are one of the natural reservoirs of this organism. How this and other normally saprophytic Serratia species can be distinguished from Serratia marcescens is reviewed.
Evaluation of sucrose and magnesium sulfate as additives in aerobic blood culture medium
Authors: Eng J
Published: January 2020

Clinical blood cultures were made in duplicate in brain heart infusion broth with sodium polyanetholsulfonate and gelatin (P broth) and in the same medium with 20% sucrose (S broth). In part of the study, 0.1% magnesium sulfate was also included in the medium with sucrose (SMg broth). The results from 1,287 positive blood cultures are reported. Significant differences among the rates and speeds of isolations from these media were found in Enterobacteriaceae and Staphylococcus aureus, which were isolated more frequently from S broth and SMg broth than from P broth; in addition, recoveries were accomplished earlier (1 or more days) from S broth and SMg broth than from P broth more often than the reverse growth patterns. An additional effect of magnesium sulfate upon recoveries could not be concluded. The possible mechanisms by which sucrose promotes recoveries from clinical blood cultures are discussed.
Application of a Limulus test device in rapid evaluation of gonococcal and nongonococcal urethritis in males
Authors: Prior R B , Spagna V A
Published: January 2020

A test device incorporating Limulus amoebocyte lysate (Mallinckrodt, Inc., St. Louis, Mo.) was developed for the rapid, presumptive diagnosis of gonococcal and nongonococcal disease in males. The device, which was evaluated in 550 men with exudative urethritis, consisted of a specimen collection syringe, a dilution reservoir containing 10 ml of pyrogen-free water, and a Limulus amoebocyte lysate single-test vial. After specimen collection, the syringe was affixed to the dilution reservoir for rapid, accurate dilution of the clinical sample. Contamination of the specimen and potential biohazards to the user were prevented. The diluted sample was then transferred (via the collection syringe) to the lysate test vial for assay of endotoxin. Various incubation times at 37 degrees C were also studied in an additional 301 male patients, and time was reduced from the standard 60 to 30 min while still retaining equivalent predictability of culture results (P less than 0.05). Of the 550 males evaluated with the test device, 366 had positive cultures for Neisseria gonorrhoeae, and 184 were negative. A sensitivity of 99.2% and a specificity of 96.7% were obtained with the test device. Overall ability to predict culture results was 98.4%. Gram-stain sensitivity and specificity were 96.4% and 99.5%, respectively, with an overall accuracy of 97.5%. There were no statistical differences between the Limulus amoebocyte lysate test and Gram stain in predicting cultures (P less than 0.05). Thus, use of the Limulus amoebocyte lysate test device would enable the private physician to make an accurate, presumptive diagnosis of gonococcal and nongonococcal disease in males with exudative urethritis within 30 min without the need of a microscope and to initiate proper therapy during the patient's initial evaluation.
Detection of
Clostridium difficile
Toxin by Counterimmunoelectrophoresis: a Note of Caution
Authors: Poxton Ian R. , Byrne Marie D.
Published: January 2020

Recent methods for detection of
Clostridium difficile
toxin by counterimmunoelectrophoresis might lead to errors. False-positives may be attributable to soluble cell surface antigens reacting with impure antitoxin.
Meningoencephalitis and Compartmentalization of the Cerebral Ventricles Caused by
Enterobacter sakazakii
Authors: Kleiman Martin B. , Allen Stephen D. , Neal Patricia , Reynolds Janet
Published: January 2020

A necrotizing meningoencephalitis complicated by ventricular compartmentalization and abscess formation caused by
Enterobacter sakazakii
in a previously healthy 5-week-old female is described. A detailed description of the isolate is presented. This communication firmly establishes the pathogenicity of
E. sakazakii.
Relationship and significance of specific immunoglobulin M antibody response in clinical and subclinical melioidosis
Authors: Ashdown L R
Published: January 2020

Thirty-six hospitalized patients with circulating antibody to pseudomonas pseudomallei were classified as having either clinical or subclinical melioidosis after full clinical examinations and supplementary radiological and culture investigations. Qualitative estimations of immunoglobulin M (IgM) antibody response were determined by complement fixation, indirect hemagglutination, and immunofluorescence, and the estimations were correlated with the clinical findings in each patient. An attempt was made to appraise the importance of IgM antibody, as determined by each test, in establishing the diagnosis of clinical melioidosis. Results obtained from serological tests were of no value in differentiating between active and latent infections. However, the IgM-immunofluorescent test appeared to be relevant to the diagnosis of clinical melioidosis, since the presence and absence of IgM-immunofluorescent antibody bore a close relation to clinical and subclinical disease, respectively. Surveillance studies indicated that the IgM-immunofluorescent test may be of value in monitoring the activity and treatment of the infection, since the results of the test were generally negative 3 to 6 months after administration of chemotherapy appropriate for melioidosis.
Immunoradiometric assay for examination and quantitation of Brucella abortus-specific antibodies reactive with the antigen(s) used in the indirect hemolysis test
Authors: Tedder T F , Hoffmann E M
Published: January 2020

An immunoradiometric assay was designed to quantitate antibodies which bind to Brucella abortus antigens adsorbed to bovine erythrocytes. This allowed examination of antibodies specific for B. abortus antigens detectable in the indirect hemolysis test for bovine brucellosis. Assay parameters were optimized for measuring antigen-specific immunoglobulin G1 (IgG1), IgG2, and IgM antibodies. The immunoradiometric assay allowed examination of binding interactions which occur during the indirect hemolysis test. Affinity-purified antibovine IgG1, IgG2, and IgM were used to detect specific bovine antibodies of these classes (and subclasses). The binding of the anti-immunoglobulins was linear as a function of immunoglobulin concentration. However, the binding of bovine antibodies of the different classes and subclasses to B. abortus antigen was nonlinear. Since B. abortus-specific antibodies of all classes and subclasses were present in the "standard serum" during the immunoradiometric assays, it is possible that the non-linearity was due to competition between antibodies for antigenic sites. IgG2 and IgM antibodies specific for B. abortus antigen(s) appeared to be capable of binding independently to antigen(s). However, the binding efficiencies of IgG1 antibodies changed as the ratio of antigenic sites to antibodies was increased.
Specificity, sensitivity, and reproducibility among the fluorescent treponemal antibody-absorption test, the microhemagglutination assay for Treponema pallidum antibodies, and the hemagglutination treponemal test for syphilis
Authors: Larsen S A , Hambie E A , Pettit D E , Perryman M W , Kraus S J
Published: January 2020

Using 920 sera, we compared the specificity and reproducibility of the hemagglutination treponemal test for syphilis with those of the fluorescent treponemal antibody-absorption test and the microhemagglutination assay for Treponema pallidum antibodies; we found all three tests to be comparable. However, the hemagglutination treponemal test for syphilis, like the microhemagglutination assay for T. pallidum antibodies, lacked sensitivity in sera from patients with primary syphilis.
Nonliquid Reagent for Detecting Nitrate Reduction
Authors: Lampe Anno S.
Published: January 2020

A nonliquid reagent for detecting nitrate reduction is described. The reagent contains
N
-(1-naphthyl)ethylenediamine dihydrochloride as a substitute for the carcinogenic α-naphthylamine. The reagent was tested on 135 strains and gave reliable results.
Prevalence of
Cryptosporidium
Antibodies in 10 Animal Species
Authors: Tzipori Saul , Campbell Iris
Published: January 2020

Antibodies to cryptosporidium were detected by indirect immunofluorescence in over 80% of sera from 10 animal species, including humans.
Probing for enterotoxigenicity among the salmonellae: an evaluation of biological assays
Authors: Jiwa S F
Published: January 2020

Sixty-eight Salmonella strains representing 39 serotypes were variously screened for enterotoxigenicity by using the Chinese hamster ovary (CHO), Y1 adrenal, and Vero cell tests, rabbit skin tests for delayed permeabiltity factor (DPF) and rapid permeability factor (RPF), the rabbit ileal loop test, and the infant mouse test. An iron-sufficient medium, YT-1, and a deferrated medium, DF, were compared. Of the culture supernatant fluids of strains grown in DF medium, 66% yielded positive reactions in the CHO cell test compared with only 10% with TY-1 medium. The corresponding performances with supernatant fluids of DF medium cultures in Y1 adrenal and Vero cell tests were 85 and 69% positive, respectively. The overall agreement between the Y1 adrenal or CHO cell test and the rabbit skin test for DPF, i.e., positive or negative in both tests, was about 70%. Positivity in DPF tests was a better predictor of positivity in either the Y1 adrenal or rabbit ileal loop test than vice versa. CHO cell, DPF, and rabbit ileal loop reactivities of unheated culture filtrates were each neutralized by anticholera antitoxin. Only four strains gave positive reactions in the infant mouse test, whereas up to 66% were positive for RPF in rabbit skin, based on positivity in Ty-1 or DR medium or both. DPF and RPF were produced by 35% of the strains. Of the 28 isolates from human stools, 82 and 92% and all of 11 strains tested were positive in the DPF, Y1 adrenal cell, and rabbit ileal loop tests, respectively. The corresponding data for 17 sewage isolates, representing 17 different serotypes rarely isolated from human stools in Sweden, were 63 and 69% and 8 of 8 tested. On the basis of this investigation, rabbit skin tests for both DPF and RPF provide the most reliable means of screening for enterotoxigenicity among salmonellae.
Immunoglobulin M-immunosorbent agglutination assay for diagnosis of infectious diseases: diagnosis of acute congenital and acquired Toxoplasma infections
Authors: Desmonts G , Naot Y , Remington J S
Published: January 2020

An immunoglobulin M (IgM)-immunosorbent agglutination assay (IgM-IS-AGA) was negative in all sera from individuals negative in the Sabin-Feldman dye test, in sera from individuals with chronic Toxoplasma infection, and in cord blood samples from uninfected infants. In contrast, all sera that were obtained from individuals with a recent history of acute Toxoplasma infection and from infants with congenital Toxoplasma infection and that were positive in both the dye test and the IgM-indirect fluorescent-antibody (IgM-IFA) test were positive in IgM-ISAGA. A total of 21 (67.7%) of 31 sera that were negative in the IgM-IFA test, despite being obtained from individuals with recently acquired Toxoplasma infection, and 8 (72.7%) of 11 sera that were negative in the IgM-IFA test and obtained from infants with congenital Toxoplasma infection were positive in IgM-ISAGA. The presence of rheumatoid factor, antinuclear antibodies, or both did not cause false-positive results in the IgM-ISAGA but did so in the IgM-IFA test. Thus, IgM-ISAGA in both more sensitive and more specific than the IgM-IFA test for detection of IgM antibodies to Toxoplasma gondii and, therefore, for the diagnosis of acute congenital and acquired Toxoplasma infections.