bluuebunny/crossref_metadata_2025 huggingface dataset:
Variable infectivity of human-derived Giardia lamblia cysts for Mongolian gerbils (Meriones unguiculatus)
Authors: Visvesvara G S , Dickerson J W , Healy G R
Published: January 2020

To determine whether gerbils can be used as a suitable animal model for giardiasis, we attempted to infect Mongolian gerbils with cysts of Giardia lamblia isolated from the stools of 10 humans with symptomatic and asymptomatic giardiasis. We obtained 100% infection with one isolate (CDC:0284:1), as evidenced by the presence of numerous trophozoites in the intestines of the gerbil and cysts in the feces. Cysts from four patients were not infective, while cysts from the other five patients produced infections in 11 to 75% of the animals. On the basis of these and other experiments, we concluded that (i) only certain isolates of human G. lamblia infect gerbils, colonize the intestine, and complete their life cycle by undergoing differentiation into cysts; (ii) the infection could last for about 39 days, but the animals excreted maximum numbers of cysts on about day 13 postinfection; (iii) the pattern of cyst excretion was irregular, and some gerbils, like humans, excreted cysts intermittently; (iv) the minimum number of cysts needed to establish an infection in 50% of the gerbils was 100; and (v) only certain strains retained the ability to infect gerbils even after repeated animal passage.
Immune response to Escherichia coli alpha-hemolysin in patients
Authors: Seetharama S , Cavalieri S J , Snyder I S
Published: January 2020

The serum antibody response of patients infected with alpha-hemolysin (AH)-producing Escherichia coli was measured by three immunoassays: tube neutralization, microneutralization, and enzyme-linked immunosorbent assay. All three assay results showed good correlation with each other. The mean anti-AH titer in patients with E. coli infection was higher than the mean titer in noninfected patients. The hemolysin-neutralizing activity was immunoglobulin G. The amount of lipopolysaccharide (LPS) antibody did not correlate with the amount of AH antibody. LPS antibody measured by enzyme-linked immunosorbent assay was predominantly of the immunoglobulin G class. Adsorption of LPS antibody by E. coli H79 LPS did not affect anti-AH titers, indicating that LPS and AH have different antigenic determinants. AHs prepared from several different E. coli strains had identical or similar antigenic determinants at the active site. Hemolysin proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were stained identically by human sera with AH antibody and by a mouse monoclonal AH antibody.
Influence of strain characteristics and immunity on the epidemiology of Campylobacter infections in Thailand
Authors: Taylor D N , Echeverria P , Pitarangsi C , Seriwatana J , Bodhidatta L , Blaser M J
Published: January 2020

To determine how strain differences and immunity affect the clinical expression of Campylobacter infections, we conducted a study of acute diarrheal disease in Thailand in which specimens from children with Campylobacter infections were cultured weekly for up to 12 weeks to determine the serotype-specific length of time of convalescent-phase excretion and rate of reinfection. Levels of immunoglobulin G to cell-surface antigens of C. jejuni were determined in another population of healthy children who were closely related by age and location to the children in the diarrheal disease study. Campylobacter species were initially isolated from 18% of 586 children under 5 years old with diarrhea; most isolates in Thailand belonged to serotypes commonly found in developed countries. C. coli was significantly less often associated with symptomatic infections and with bloody diarrhea than C. jejuni (P less than 0.001 and P = 0.045, respectively). The peak age of isolation and the peak level of immunoglobulin G to Campylobacter species occurred before 2 years of age. The mean duration of convalescent-phase excretion was 14 +/- 2 (standard error of the mean) days for children less than 1 year old and 8 +/- 2 days for children 1 to 5 years old (P = 0.02, t test). Infection with another Campylobacter serotype was found in 34% of 105 children during the 12-week follow-up period. The rate of reinfection in these children was 15% (range, 8 to 22%) each week. Hyperendemic exposure to Campylobacter species in Thailand confers immunity to infection that is associated with an early peak in specific serum antibodies and an age-related decrease in the case-to-infection ratio and duration of convalescent-phase excretion but does not prevent asymptomatic infections.
Rapid and overnight microdilution antibiotic susceptibility testing with the Sensititre Breakpoint Autoreader system
Authors: Nolte F S , Krisher K K , Beltran L A , Christianson N P , Sheridan G E
Published: January 2020

The Sensititre Breakpoint Autoreader system (SBAS) is a broth microdilution method with one to three concentrations of each antibiotic and innovative fluorescence technology to define inhibitory endpoints. We tested 248 gram-negative bacilli and 80 gram-positive cocci using both the rapid (5 h) and overnight (18 h) SBAS procedures. Inhibitory endpoints were also determined by visual inspection of the microdilution trays after 18 h of incubation. SBAS results were compared with those obtained by a standardized disk diffusion (SDD) procedure. Agreement between the rapid SBAS and SDD results for all antibiotic-organism combinations was found in 3,730 of 4,571 (81.6%) tests, with 3.9% very major, 6.5% major, and 7.9% minor discrepancies noted. Data analysis by organism group revealed 86.8, 57.3, 71.4, and 62.3% agreement for members of the family Enterobacteriaceae, Pseudomonas spp., staphylococci, and enterococci, respectively. The results of the overnight SBAS and SDD agreed in 4,154 of 4,654 (89.2%) tests, with 2.3% very major, 1.3% major, and 7.1% minor discrepancies recorded. Concordance was noted in 90.4, 78.1, 90.6, and 83.3% of the comparisons for the members of the Enterobacteriaceae, Pseudomonas spp., staphylococci, and enterococci, respectively. The inhibitory endpoints determined with the Autoreader were as reliable as those determined by visual inspection after 18 h of incubation.
Value of direct catheter staining in the diagnosis of intravascular-catheter-related infection
Authors: Coutlée F , Lemieux C , Paradis J F
Published: January 2020

Ninety-nine intravascular catheters were evaluated by a semiquantitative culture and Gram and acridine orange direct stains. A diagnosis of catheter-related infection was determined by a retrospective review of clinical records. Compared with the culture method, direct examination of catheters lacked sensitivity. Surprisingly, for some patients, a positive stain for yeasts not recovered by culture was considered significant. The culture correlated with bacteremia but failed to predict infection in 42% of patients.
Partial characterization of a bovine group A rotavirus with a short genome electropherotype
Authors: Theil K W , McCloskey C M
Published: January 2020

A group A rotavirus (ID isolate) recovered from a diarrheic beef calf possessed a short genome electropherotype. This short genome electropherotype was a stable characteristic of the ID isolate as it remained unchanged through 3 passages in gnotobiotic calves or through 19 passages in MA104 cell cultures. Subgroup analysis with monoclonal antibodies in an enzyme-linked immunosorbent assay established that the isolate was a subgroup 1 rotavirus. Neutralization tests demonstrated that this isolate was a distinct serotype from the human group A rotavirus S2 isolate (short genome electropherotype) and the turkey group A rotavirus 174 isolate (semi-short genome electropherotype). The ID isolate was pathogenic for 5- to 21-day-old gnotobiotic calves, inducing diarrhea within 48 h postinoculation.
Urine specimen collection with external devices for diagnosis of bacteriuria in elderly incontinent men
Authors: Nicolle L E , Harding G K , Kennedy J , McIntyre M , Aoki F , Murray D
Published: January 2020

We determined the validity of using external devices for urine specimen collection from 24 elderly incontinent men residing in a nursing home by collecting three sequential specimens, two with external devices and then one by catheterization. The positive predictive value of organisms isolated in quantitative counts of greater than or equal to 10(5) CFU/ml in external devices for bladder bacteriuria was 86% for either sterile or clean collecting devices and 93% for the same organism in two consecutive specimens. The negative predictive value for organisms present in quantitative counts of less than 10(5) CFU/ml was 90% for both sterile and clean devices and 86% when the organism was present in both specimens. Contamination in external collection devices was not influenced by whether the device was clean or sterile, circumcision of the resident, or duration of time between device application and specimen collection. These data suggest that urine specimens collected by ward nursing staff with external devices are reliable for the diagnosis of bacteriuria in this patient population.
Prevalence of Gardnerella vaginalis in the urinary tract
Authors: Lam M H , Birch D F , Fairley K F
Published: January 2020

Midstream urine samples from 106 patients presenting to the Casualty Department of The Royal Melbourne Hospital with frequency or dysuria were cultured for Gardnerella vaginalis and conventional uropathogens. Urine samples collected via an open-end catheter from 70 healthy pregnant women were examined similarly. Midstream urine and other samples, including the seminal fluids and swabs of the mouths, throats, rectums, and vaginas of 33 healthy subjects, were cultured for G. vaginalis. Another 15 female patients with proven G. vaginalis bacteriuria were given a bladder washout localization test to determine the site of infection in the urinary tract. G. vaginalis in counts greater than 10(3) CFU/ml was recovered from the midstream urine of 27 of 106 patients (25%), 7 of whom also harbored conventional pathogens in counts greater than 10(4) CFU/ml. Another 11 patients with cultures negative for G. vaginalis yielded greater than 10(4) CFU of conventional pathogens per ml. G. vaginalis was cultured (greater than 10(3) CFU/ml) from catheter samples of 19 of 70 healthy pregnant women (27%), 6 of whom also harbored greater than 10(3) CFU of conventional uropathogens per ml. Two women yielded growths of conventional pathogens only. Midstream urine samples from 13 of 13 healthy males were free of G. vaginalis, whereas 5 of 20 healthy nonpregnant females yielded greater than 10(3) CFU of G. vaginalis per ml from midstream urine samples. G. vaginalis was recovered from 4 of 12 semen samples and from urethral samples from four of seven males and four of eight females. All four culture-positive females also harbored G. vaginalis in their vaginas. There was no evidence of oral or rectal carriage of G. vaginalis in 15 healthy subjects. Localization studies with 15 female patients having underlying renal disease showed that 11 patients harbored G. vaginalis in their kidneys. The result suggest that colonization or infection of the bladder and upper urinary tract by G. vaginalis is very largely a phenomenon of females, with the highest frequency in pregnant women. The prevalence of G. vaginalis in the urinary tracts of healthy females is similar to that of symptomatic subjects. However, G. vaginalis in counts greater than 10(5) CFU/ml is more likely to be associated with urinary tract symptoms. In males, this bacterial species infects the genital tract rather than the urinary tract.
Failure of charcoal-horse-blood broth with cephalexin to significantly increase rate of Bordetella isolation from clinical specimens
Authors: Hoppe J E , Weiss A , Wörz S
Published: January 2020

In a field trial conducted in cooperation with pediatricians in private practice, 198 nasopharyngeal swabs from children with suspected whooping cough were placed into charcoal-horse-blood transport medium with cephalexin (40 mg/liter). After preincubation at 36 degrees C for 1 to 2 days, the transport systems were mailed to the laboratory. There, the swabs were plated onto charcoal-horse-blood agar with cephalexin and were subsequently incubated for 48 h in cephalexin-containing charcoal-horse-blood broth which was then subcultured onto the agar. Forty-six Bordetella pertussis strains and seven Bordetella parapertussis strains were isolated (Bordetella isolation rate, 26.8%). Only three (5.7%) of the 53 Bordetella strains were detected exclusively by use of the broth. This low rate of additional isolates is probably explained by the fact that the swabs had been submitted in charcoal-horse-blood transport medium which itself acts as an enrichment medium.
Congo red binding and salt aggregation as indicators of virulence in Shigella species
Authors: Qadri F , Hossain S A , Ciznár I , Haider K , Ljungh A , Wadstrom T , Sack D A
Published: January 2020

Smooth strains of Shigella dysenteriae type 1, Shigella flexneri, Shigella boydii, and Shigella sonnei which form pigmented colonies (Pcr+) on Congo red agar were virulent in the Sereny test. Smooth variants unable to bind Congo red (Pcr-) were avirulent. Measurements of dye uptake from solution showed that S. dysenteriae type 1 bound the most dye, followed in order of uptake by S. flexneri, S. boydii, and S. sonnei. Using the salt aggregation test (SAT) to determine cell surface hydrophobicity, we found the same order of species. The SAT could not, however, detect differences in surface properties between Pcr+ and Pcr- pairs of isogenic smooth strains. Enteroinvasive Escherichia coli strains used in the study showed SAT and Congo red-binding properties which were similar to those of the S. flexneri strains. A direct correlation was found between pigment-binding ability and the presence of the large 140-megadalton plasmid in S. flexneri, enteroinvasive E. coli, and S. boydii but not in S. dysenteriae type 1 or S. sonnei strains. Congo red interacted with outer membranes and outer membrane proteins of S. dysenteriae type 1 but not with lipopolysaccharides. However, rough mutants of Shigella species deficient in lipopolysaccharides bound Congo red and formed pigmented colonies, showing that dye binding as a virulence assay may be misinterpreted in such cases. There was complete correlation of the Pcr+ phenotype with virulence in the smooth strains in this study, suggesting that Congo red binding can be utilized as a quick and reliable alternative to the Sereny test.
Pneumocystis carinii antigenemia in acquired immunodeficiency syndrome
Authors: Pifer L W , Wolf B L , Weems J J , Woods D R , Edwards C C , Joyner R E
Published: January 2020

The present study was conducted to determine the prevalence and significance of Pneumocystis carinii antigenemia in patients with acquired immunodeficiency syndrome (AIDS) and clinically or invasively diagnosed P. carinii pneumonitis. Single serum specimens from 20 AIDS patients invasively examined for P. carinii organisms and 106 AIDS patients with a clinical diagnosis only of P. carinii pneumonitis were blindly tested for P. carinii antigenemia by a counterimmunoelectrophoresis assay. In the 20 specimen-documented cases, the antigen test demonstrated a sensitivity of 75% and a specificity of 90%. The positive predictive value of the test was 90%, while the negative predictive value was 70%. In AIDS patients with specimen-documented P. carinii pneumonitis, the prevalence of P. carinii antigenemia coincided almost exactly with the prevalence of positive invasively obtained specimens (60 and 59%, respectively). In patients with a clinical diagnosis only of P. carinii pneumonitis, half as many (30%) were found to exhibit antigenemia. Sequential P. carinii antigen titers determined by a new latex agglutination technique on three AIDS patients with specimen-documented P. carinii pneumonitis demonstrated the influence of specific therapy upon P. carinii antigenemia and its potential prognostic application.
Comparison of three rapid methods for detection of antibodies to streptolysin O and DNase B
Authors: Hostetler C L , Sawyer K P , Nachamkin I
Published: January 2020

Three commercial systems were compared for ability to detect antibodies to streptolysin O (ASO) and DNase B (ADB). Streptozyme (Wampole Laboratories, Cranbury, N.J.) exhibited high sensitivity (100%) for detecting ASO but low sensitivity for ADB (22.2%). The LeapStrep (Organon Teknika, Malvern, Pa.) and Check-Spectra (Diagnostic Technology, Hauppauge, N.Y.) tests had low sensitivities for detecting ASO (35.3 and 21.4%, respectively) and ADB (22.2 and 33.3%, respectively).
Chronic Morganella morganii arthritis in an elderly patient
Authors: Schonwetter R S , Orson F M
Published: January 2020

This report describes the first case of septic arthritis caused by Morganella morganii. The elderly patient's course of pyoarthritis was atypical in its benign clinical presentation, having little inflammatory response over a prolonged period. Septic arthritis should be considered as a possible diagnosis in all elderly patients with joint effusions.
Basic properties of Tritrichomonas mobilensis hemagglutinin
Authors: Pindak F F , Mora de Pindak M , Gardner W A , Abee C R
Published: January 2020

Tritrichomonas mobilensis is a recently described enteric protozoon of squirrel monkeys. An earlier report identified one of the metabolic products of this organism as a lectinlike hemagglutinin. Its further properties were determined in this study. Culture supernatants of T. mobilensis FP4190 were concentrated by ultrafiltration through a membrane with 100,000-molecular-weight cutoff. High titers of agglutinin against human erythrocytes were obtained. Incubation at 70 degrees C for 15 min resulted in complete inactivation. Exposure to 56 degrees C for 30 min was without effect, and only partial loss of activity was obtained during incubation for up to 18 h. Maintenance at pH 4 to 9 for 4 h at room temperature had no deleterious effect. Apparent degradation of the hemagglutinin was achieved by 18 h of contact with proteinase K, but trypsin and collagenase were essentially ineffective. Papain increased the sensitivity of the test. In the presence of this enzyme hemagglutinin was demonstrated also in cultures of Tritrichomonas foetus and Tritrichomonas augusta but not in those of Pentatrichomonas hominis or Trichomonas vaginalis.
New microassay for quantitation of endotoxin using Limulus amebocyte lysate combined with enzyme-linked immunosorbent assay
Authors: Zhang G H , Baek L , Koch C
Published: January 2020

A combined method of the Limulus amebocyte lysate (LAL) test and the enzyme-linked immunosorbent assay for quantitation of endotoxin was developed based on our observation that the antigenicity of coagulogen, a major protein in LAL, was lost when LAL reacted with endotoxin as shown by immunoblotting. Determination of the residual coagulogen by an enzyme-linked immunosorbent assay system with monoclonal antibody against coagulogen revealed that the loss of the antigenicity of coagulogen was proportional to the concentration of endotoxin. An inverse linear curve was established between the endotoxin concentration and absorbance. Standard curves of the LAL enzyme-linked immunosorbent assay with different detection limits (from 0.1 to 100 pg of the control standard endotoxin per ml) were obtained from one batch of commercial LAL by adjusting incubation time and dilution of LAL. The reaction curves of various endotoxins were parallel to one another, whereas the kinetics differed from that of (1-3)-beta-D-glucan. The LAL enzyme-linked immunosorbent assay is a highly reproducible microassay, using only 10 microliter of test sample and LAL reagent; because the color and turbidity of plasma samples do not interfere with the assay, it is well suited for quantitation of endotoxins in clinical specimens.
Comparison of BACTEC 13A medium and Du Pont isolator for detection of mycobacteremia
Authors: Witebsky F G , Keiser J F , Conville P S , Bryan R , Park C H , Walker R , Siddiqi S H
Published: January 2020

BACTEC 13A medium (Johnston Laboratories, Towson, Md.) was compared with Isolator (Du Pont Co., Wilmington, Del.) concentrate for sensitivity, speed, and technical ease of isolation of mycobacteria from paired patient blood samples. Of 72 positive cultures, 63 were positive by both systems. Five positive cultures were detected by BACTEC 13A medium alone, and four were detected by Isolator alone. The median numbers of days to positivity were 12 for BACTEC 13A medium and 14 for Isolator concentrate. BACTEC 13A medium has an advantage over the Isolator in requiring less laboratory manipulation of the specimen but has the disadvantages of not providing isolated colonies or quantitation of organisms. Some technical problems with contamination in both systems are also discussed.
Biochemical, genetic, and epidemiologic characterization of Haemophilus influenzae biogroup aegyptius (Haemophilus aegyptius) strains associated with Brazilian purpuric fever
Authors: Brenner D J , Mayer L W , Carlone G M , Harrison L H , Bibb W F , Brandileone M C , Sottnek F O , Irino K , Reeves M W , Swenson J M
Published: January 2020

Brazilian purpuric fever (BPF) is a recently recognized fulminant pediatric disease characterized by fever, with rapid progression to purpura, hypotensive shock, and death. BPF is usually preceded by purulent conjunctivitis that has resolved before the onset of fever. Both the conjunctivitis and BPF are caused by Haemophilus influenzae biogroup aegyptius (formerly called H. aegyptius). Isolates from 15 BPF cases, mainly from blood or hemorrhagic cerebrospinal fluid, case-associated isolates from 42 persons in towns where BPF cases occurred, and control strains from 32 persons in towns without BPF cases were characterized biochemically, genetically, and epidemiologically. Results indicated that a single clone was responsible for all BPF cases identified in six Brazilian towns from 1984 through 1986. All of 15 (100%) case strains were the same clone as was 1 of 32 (3%) control strains (P = less than 10(-8). Isolates of the clone were preferentially intrarelated by DNA hybridization (99% relatedness, hydroxyapatite method at 60 and 75 degrees C) and were separable from other H. influenzae biogroup aegyptius strains (approximately 90% relatedness at 60 degrees C and 82% relatedness at 75 degrees C). All isolates of the BPF clone and no other strains contained a 24-megadalton plasmid of restriction endonuclease type 3031, were of a single multilocus enzyme mobility type, were of a single sodium dodecyl sulfate-polyacrylamide gel electrophoresis type, and were in one of two ribosomal DNA restriction patterns. All BPF clone isolates reacted with monoclonal antibodies produced from a case strain; only 3 of 62 (5%) other strains reacted with this monoclonal antibody. Ninety percent of BPF clone strains and 27% of other strains were relatively resistant to sulfamethoxazole-trimethoprim.
Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin type II
Authors: Handl C , Rönnberg B , Nilsson B , Olsson E , Jonsson H , Flock J I
Published: January 2020

The gene for Escherichia coli heat-stable enterotoxin type II (STII) was fused to the genes for protein A from Staphylococcus aureus and beta-galactosidase in two different expression systems. Antibodies raised in rabbits against the protein A-STII fusion protein recognized the beta-galactosidase-STII fusion protein. The latter fusion protein was used as the immobilized antigen in an enzyme-linked immunosorbent assay (ELISA) for detection of STII. The correlation between the results of the ELISA and the intestinal loop test in piglets was 95%, suggesting that the ELISA can be used to reliably detect STII.
Alcaligenes piechaudii from chronic ear discharge
Authors: Peel M M , Hibberd A J , King B M , Williamson H G
Published: January 2020

A recently described bacterium, Alcaligenes piechaudii, was isolated repeatedly from the ear discharge of a diabetic man. This appears to be the first report of human infection in which there is clinical evidence of a pathogenic role for this species.
Evaluation of autoscan-4 for identification of members of the family Enterobacteriaceae
Authors: Gavini F , Husson M O , Izard D , Bernigaud A , Quiviger B
Published: January 2020

A study was performed to compare the Autoscan-4 (MicroScan, Inc., Mahwah, N.J.) with conventional biochemical methods for identifying clinical isolates of the family Enterobacteriaceae. The Autoscan-4 yielded correct identification of 95.4% of the isolates at the species level and 98.4% at the genus level. Only one misidentification was observed. The identification of both common and less-common isolates of Enterobacteriaceae makes this system highly efficient.
Bartholin's gland abscess caused by Neisseria sicca
Authors: Berger S A , Gorea A , Peyser M R , Edberg S C
Published: January 2020

Neisseria sicca has been reported to cause bacterial meningitis, endocarditis, pneumonia, and spondylitis. Surgical drainage from a Bartholin's gland abscess contained N. sicca in pure culture. Neisseria species recovered from cerebrospinal fluid and gynecologic specimens must be carefully examined to avoid misidentification as meningococci or gonococci.
Remarkably homogeneous population of adenovirus type 3 and 7 genome types
Authors: Arens M , Dilworth V
Published: January 2020

A total of 270 frozen adenovirus isolates, obtained from January 1981 to December 1986, were grown in KB cells. DNA was successfully prepared from 248 of these isolates, and the prepared DNA was digested initially with SmaI restriction endonuclease and then analyzed by agarose gel electrophoresis. Restriction fragment patterns showed that 68 of these DNAs were either adenovirus type 3 (Ad3) or Ad7. Further analysis by digestion with BamHI showed that 24 isolates were Ad3a and 44 isolates were Ad7b. When Ad3 isolates were digested with SmaI or PstI, the restriction fragment patterns of the 24 isolates were identical. Of these 24 isolates, 20 (83%) also gave identical patterns when digested with HindIII, and the patterns of the remaining 4 isolates were different from that of the 20 but identical to each other. All Ad3 isolates were obtained in 1982, 1983, and 1986. All the 44 Ad7b isolates gave identical fragment patterns when digested with BglI or PvuII, and 43 of 44 (98%) isolates gave identical patterns when digested with SstII. We obtained at least two Ad7 isolates during each of the 6 years studied. Although considerable genome heterogeneity within adenovirus serotypes and subtypes has been reported, our population of isolates is remarkably homogeneous. This homogeneous set of genome types was obtained from diverse anatomical sites and was associated with a broad spectrum of clinical syndromes.
Clinical evaluation of diagnostic hemoperfusion for in vivo enrichment of bacteria and fungi in comparison with a conventional blood culture technique
Authors: Kühnen E , Schaal K P , Keller F , Bartels F
Published: January 2020

The hemoperfusion module, a newly developed technique for recovering pathogenic microorganisms from patients suffering from septicemia, was compared with conventional blood cultures. The module was interconnected with the extracorporeal circulation of 92 predominantly hemodialyzed patients. Nearly 12 liters of flowing blood (up to 200 ml min-1; mean running time, 60 min) came in contact with the coated charcoal. Of 99 modules examined, 44 (44.7%) yielded positive cultures and contained 54 potentially pathogenic bacteria or fungi (22 species). Only 32 of 190 (16.8%) conventional blood cultures were positive and contained 37 microorganisms (10 species). Even when patients were receiving antibiotic treatment, the frequency of isolation was significantly higher in hemoperfusion (21 of 44 modules positive, 47.7%) than in conventional blood cultures (10 of 88 cultures positive, 11.4%). In contrast, 23 of 55 modules (41.8%) and 22 of 102 conventional blood cultures (21.6%) were positive when patients were not treated with antibiotics prior to blood sampling. Altogether, hemoperfusion modules appeared to be superior to and more sensitive than the conventional blood cultures used and seemed to be a valuable tool for detecting septicemia.
DNA probes to identify Shiga-like toxin I- and II-producing enteric bacterial pathogens isolated from patients with diarrhea in Thailand
Authors: Seriwatana J , Brown J E , Echeverria P , Taylor D N , Suthienkul O , Newland J
Published: January 2020

When Shigella species, Escherichia coli, and five other bacterial enteric pathogens isolated from children with diarrhea in Thailand were tested for hybridization under stringent conditions with probes for Shiga-like toxins I and II, only 30 Shigella dysenteriae 1 hybridized with the Shiga-like toxin I probe.
Use of murine monoclonal antibodies for laboratory diagnosis of varicella-zoster virus infection
Authors: Gleaves C A , Lee C F , Bustamante C I , Meyers J D
Published: January 2020

The laboratory diagnosis of varicella-zoster virus (VZV) infection was reevaluated by direct immunofluorescent-antibody staining (DFA) and centrifugation culture with newly available murine monoclonal antibodies. Specimen smears were examined by DFA using monoclonal antibodies to VZV and to herpes simplex virus types 1 and 2. Specimens were also inoculated into shell vials for centrifugation culture and into standard tube cell culture. Of 68 specimens tested from 60 patients, 39 (57%) were positive for VZV by at least one method. DFA was positive in 36 of 39 (92%); centrifugation culture was positive at 24 h in 23 of 39 (59%) and at 48 h in 31 of 39 (79%); and standard culture was positive in 25 of 39 (64%). Twenty-three of the 39 positive specimens (59%) were positive by all three techniques. Forty-three of the 60 patients were considered to have VZV by clinical criteria, and 35 of these 43 (81%) had laboratory confirmation of the diagnosis. These data confirm that DFA is the method of choice for the rapid laboratory confirmation of VZV infection. The centrifugation culture assay can provide an alternative method to DFA for the laboratory diagnosis of VZV infection.
Cross-reactive monoclonal antibodies for diagnosis of pneumococcal meningitis
Authors: Waltman W D , Gray B , McDaniel L S , Briles D E
Published: January 2020

A diagnostic test for the detection of Streptococcus pneumoniae meningitis was developed using monoclonal antibodies (MAbs) to phosphocholine (PC) and non-PC determinants of pneumococcal teichoic acids. These MAbs do not recognize other bacteria that commonly cause meningitis. By using a dot blot assay, these MAbs were compared with a polyvalent pneumococcal capsular omniserum and an antiserum made to whole cells for their ability to detect pneumococci in infected spinal fluids. An immunoglobulin M (IgM) anti-PC antibody gave a positive reaction with 16 of 22 (73%) pneumococcal culture-positive spinal fluids. One false-positive result out of 45 pneumococcal culture-negative spinal fluids was also observed. D3114/63, an IgM MAb to non-PC determinants of teichoic acids, detected 15 of 22 of the pneumococcal culture-positive spinal fluids with one false-positive result. IgG2b and IgG3 anti-PC MAbs were less efficient than the IgM anti-PC MAb at detecting pneumococci in spinal fluids. Like the IgM anti-PC MAb, omniserum detected 73% of the culture-positive pneumococcal spinal fluids, with one false-positive result. The use of anti-PC or D3114/63 MAbs instead of a pooled serum such as omniserum has several advantages: (i) use of a single cross-reactive antibody rather than 83 pooled antibodies; (ii) possibility of a higher concentration of reactive antibody, which may increase the sensitivity of the test; (iii) a standardized antibody preparation; (iv) ease of preparation of the antibody; and (v) less expense.
Comparison of blood agar, ampicillin blood agar, MacConkey-ampicillin-Tween agar, and modified cefsulodin-Irgasan-novobiocin agar for isolation of Aeromonas spp. from stool specimens
Authors: Kelly M T , Stroh E M , Jessop J
Published: January 2020

The performance of four media for the isolation of Aeromonas strains from stool specimens, the importance of ampicillin-susceptible Aeromonas strains in the selection of culture media, and the usefulness of beta-hemolysis in screening blood-containing media for Aeromonas strains were evaluated in two phases. In the first phase, 36 of 1,672 stool specimens yielded Aeromonas isolates. Ninety-seven percent of the isolates were detected on blood agar containing 20 micrograms of ampicillin per ml (ABA), and 47% were detected on MacConkey agar containing 100 micrograms of ampicillin per ml and 1% Tween 80. In the second phase of the study, 43 of 1,924 stool specimens yielded Aeromonas isolates. Fifty-one percent of the isolates were detected on blood agar and on modified cefsulodin-Irgasan-novobiocin agar, and 84% were detected on ABA. The combination of ABA and modified cefsulodin-Irgasan-novobiocin agar provided 100% recovery of the Aeromonas isolates encountered. All of the Aeromonas isolates detected on blood agar were also detected on ABA, and 89% of the Aeromonas isolates detected on these media were beta-hemolytic. These results suggest that ABA is superior to the other media evaluated for the isolation of Aeromonas strains from stool specimens, but optimal recovery of the organism may require the use of more than one medium. The results also suggest that the occurrence of ampicillin-susceptible strains is not a limitation on the use of ABA, but at least 10% of Aeromonas isolates will be missed if beta-hemolysis is used to screen ABA plates for these organisms.
Incidence of enteric adenoviruses among children in Thailand and the significance of these viruses in gastroenteritis
Authors: Herrmann J E , Blacklow N R , Perron-Henry D M , Clements E , Taylor D N , Echeverria P
Published: January 2020

In countries with temperate climates, enteric adenoviruses have been shown to be a substantial cause of pediatric gastroenteritis. To determine the incidence of adenovirus infection in a tropical climate, stools were collected from children under age 7 during a 1-year period at an outpatient clinic in Bangkok, Thailand. Stools from 1,114 children with gastroenteritis and from 947 children without gastroenteritis were tested. Each stool was tested for adenovirus group antigen and for specific enteric adenovirus types (Ad40 and Ad41) by monoclonal antibody enzyme immunoassays. We found that 4.4% (49 of 1,114) of children with gastroenteritis and 1.8% (17 of 947) of children without gastroenteritis were positive for adenovirus group antigen. In tests for specific enteric adenovirus types, 2.0% (22 of 1,114) of the tests were positive in children with gastroenteritis and 0.6% (6 of 947) were positive in children without gastroenteritis. There was a significant correlation (P less than 0.02) of gastroenteritis with nonenteric adenovirus types (27 of 1,114) as well as with specific enteric adenovirus types (P less than 0.01). By comparison, 19.7% of children with gastroenteritis and 0.7% of those without gastroenteritis were positive for rotavirus infection. In the adenovirus-infected children with gastroenteritis, there were coinfections with rotavirus only in those with nonenteric adenovirus infection (7 of 27 children). There were no significant differences in the association of bacterial or parasitic infections with either enteric or nonenteric adenovirus infections in either group of children studied. These data demonstrate that Ad40 and Ad41 are causes of gastroenteritis in this population, but among the spectrum of diarrheal etiologies, they may be proportionately less important than they are in countries with temperate climates.
Safety of refrigerated storage of admixed parenteral fluids
Authors: Weil D C , Arnow P M
Published: January 2020

Many hospital pharmacies are reluctant to store admixed parenteral fluids longer than 24 to 48 h because of concern about possible microbial contamination. We evaluated the safety of prolonged refrigerated storage of admixtures by culturing mixtures in 471 bags prepared routinely in a hospital pharmacy and stored at 4 degrees C for up to 15 days. Low-level contamination (1 CFU per bag) was found in 3 of 253 solutions of saline and/or glucose into which nonantibiotic additives had been injected. None of 171 saline and/or glucose solutions with antibiotic additives or 47 parenteral nutrition fluids was culture positive. The risk of contamination did not increase with duration of storage, and none of 107 bags stored greater than or equal to 5 days was culture positive. Laboratory studies to measure growth of bacteria and fungi in glucose infusate and parenteral nutrition solutions confirmed that storage at 4 degrees C suppresses growth. Eight of twelve bacterial isolates grew in glucose solutions at 25 degrees C, while none grew at 4 degrees C. Of 13 species of bacteria and fungi inoculated in parenteral nutrition fluids, 8 proliferated at 25 degrees C and none proliferated at 4 degrees C. We conclude that refrigerated storage of parenteral fluids for up to 1 week following admixture, as permitted by Centers for Disease Control guidelines, does not appear to increase the risk of microbial contamination when standard aseptic procedures for admixing and storage are followed.
Anti-Trypanosoma cruzi and anti-laminin antibodies in chagasic patients after specific treatment
Authors: Gazzinelli R T , Galvão L M , Cardoso J E , Cançado J R , Krettli A U , Brener Z , Gazzinelli G
Published: January 2020

The antibody response to Trypanosoma cruzi epimastigote and trypomastigote stages, as well as to laminin, was studied in several groups of chagasic patients. In six patients who were cured of the parasite, the serum antibody titers as revealed by indirect immunofluorescence and hemagglutination tests against epimastigotes (conventional serology) and a complement-mediated lysis test with living trypomastigotes did not differ from those of normal individuals. In seven presumably cured patients, although the complement-mediated lysis test turned negative, conventional serology remained positive. Sera from this group of so-called "dissociated" patients presented significant lower mean antibody titers against epimastigote but not trypomastigote stages than did sera from 14 untreated patients (P less than 0.01). Most of the antibodies against trypomastigotes, including the residual levels found in cured patients, were absorbed by mouse laminin. In fact, significantly higher titers of anti-laminin antibodies were observed in sera from untreated chagasic patients (1.131 +/- 0.458) and cured patients (1.103 +/- 0.572) than in sera from eight normal individuals (0.459 +/- 0.402) (P less than 0.01). The anti-laminin titers were higher in sera of patients of blood group A or O than in those of patients of group B or AB. In Western blotting (immunoblotting) analysis against trypomastigotes, sera from chronic untreated patients recognized many polypeptide bands ranging from 26 to 160 kilodaltons, whereas no protein bands were observed with sera from cured patients. Only faint bands of parasite proteins were observed with sera of dissociated patients. In conjunction, the above data suggest that the anti-trypomastigote antibodies which persist after parasitological cure of patients with Chagas' disease are due mainly to cross-reactive epitopes from mouse laminin.
Evaluation of serologic assays for diagnosis of whooping cough
Authors: Granström G , Wretlind B , Salenstedt C R , Granström M
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for the immunoglobulin G (IgG), IgM, and IgA response to Bordetella pertussis filamentous hemagglutinin (FHA) and pertussis toxin (PT) and a neutralization test (NT) in a microplate tissue culture assay for neutralizing antibodies to PT were evaluated in paired sera from 90 patients with culture-confirmed pertussis. Eighty patients were children (age, less than 15 years), and 6 of 80 children had been immunized with three doses of diphtheria-tetanus-pertussis vaccine as infants. A significant titer rise (greater than or equal to twofold), determined by ELISA, of IgG, IgM, and IgA to FHA was recorded in 75 (83%), 28 (31%), and 47 (52%) of the patients, respectively. A significant titer rise to PT in IgG was found in 83 (92%), IgM in 29 (32%), and IgA in 44 (49%) of the patients. A significant titer rise to FHA or PT in IgG was found in 88 (98%) of the patients, in combination with a significant rise in the titer of IgA to FHA. These data were obtained in a single serum dilution of 1:500. Titrations performed later showed that the titer rise to FHA in IgG was a mean of 6.5-fold, which was significantly lower than the mean 67.0-fold rise in IgG to PT (P less than 0.001). The mean titer of IgG to FHA in convalescent-phase serum was 270, which was also significantly lower than the mean PT titer of 2,943 (P less than 0.001). A significant rise (greater than or equal to fourfold) in PT titer by NT was found in 58 of 83 (70%) of the patients. The NT was significantly less sensitive than the ELISA for the determination of the IgG titer to PT ( P< 0.001). Results showed that a 100% (90 of 90) sensitivity in terms of titer rises was achieved in the serologic diagnosis of pertussis by ELISA in a single-point determination of the IgG and IgA responses to FHA and of the IgG response to PT.
Similar DNA restriction endonuclease profiles in strains of Legionella pneumophila from different serogroups
Authors: van Ketel R J
Published: January 2020

DNA of strains of Legionella pneumophila serogroups 1, 3, 4, and 6, isolated from patients and environmental sources, was examined by restriction endonuclease analysis (REA). Major differences in profiles enabled subtyping in many strains with the same serogroup antigen. However, a cluster of L. pneumophila strains, originating from all the examined serogroups, had similar restriction endonuclease profiles, sometimes with minor differences. This suggests that the genetic similarity between strains of L. pneumophila of different serogroups is sometimes closer than in strains with the same serogroup antigen. Seven environmental sources harbored two L. pneumophila strains with various serogroup antigens; six sources had similar restriction endonuclease profiles. The resolution of small differences in profiles is hampered in REA by the great magnitude of DNA fragments; even upon extensive analysis, these differences are not always readily visualized. Double digestions with the restriction enzymes HpaI and HpaII showed the best results and sometimes revealed differences not evident by digestions with a single endonuclease. REA has a great capacity for accurate epidemiological typing of L. pneumophila, in addition to classical serogrouping; it appeared that the results of the two techniques do not necessarily correlate. On the other hand, it should be stressed that small differences in profiles are not easily detected by REA.
Persistence of elevated levels of galactosyl-alpha(1-3)galactose antibodies in sera from patients cured of visceral leishmaniasis
Authors: Avila J L , Rojas M , García L
Published: January 2020

Using rabbit erythrocyte-derived neutral glycosphingolipids enriched for a ceramide pentasaccharide as the antigen, we detected elevated anti-galactosyl-alpha(1-3)galactose (anti-G alpha G) antibody levels in 76% of children with active visceral leishmaniasis (kala-azar [KA]) and in 42% of clinically cured patients with KA who had been treated about 5 years previously with meglumine antimonate (30 mg/kg in a series of 15 daily injections). The long-term persistence of elevated G alpha G antibodies was also found in 56% of children living in the same geographic zone who, at the time of the initial clinical examination, had fever and evident splenomegaly with hyperglobulinemia but a negative bone marrow aspirate for leishmanial bodies. Five years after antimonate treatment, these clinically cured children with presumptive KA were studied serologically. Their mean G alpha G antibody values were slightly lower than those in patients with active KA but were still abnormal. Using different biochemical and immunological approaches, we found that elevated G alpha G antibodies present in patients with KA bound specifically to glycoconjugates with an alpha(1-3)-terminal galactose residue. G alpha G antibodies were mainly distributed between immunoglobulin classes G and M in patients with active KA and in antimonate-treated patients with clinically cured KA. The possibility of the existence of remnant living parasites or the persistence of inserted G alpha G epitopes in parasitized macrophages was proposed as a mechanism to explain the long-term persistence of abnormal G alpha G antibodies in patients apparently cured of KA.
Microbial inhibition on hospital garments treated with Dow Corning 5700 antimicrobial agent
Authors: Murray P R , Niles A C , Heeren R L
Published: January 2020

We evaluated the efficacy of the antimicrobial activity of cotton-polyester fabric treated with 3-(trimethoxysilyl)propyldimethyloctadecyl ammonium chloride (DC 5700), a quaternary ammonium compound bound irreversibly to the material. Significant antimicrobial activity was observed with 58 of 61 gram-positive cocci but with only 1 of 35 gram-negative bacilli and 0 of 5 yeasts. No inhibition of bacilli or yeasts was observed when the DC 5700 concentration ranged from 0.05 to 1.0% or when antimicrobial activity was assayed by the agar overlay bioassay or broth agitation methods.
Naturally acquired rabies in an armadillo (Dasypus novemcinctus) in Texas
Authors: Leffingwell L M , Neill S U
Published: January 2020

The first case of rabies in an armadillo is reported. The rabies fluorescent-antibody test and mouse inoculation procedure were used to substantiate the presence of the virus. The Centers for Disease Control authenticated our findings and was able to determine the source of infection by monoclonal antibody typing.
Passive hemagglutination test for detection of antibodies to human immunodeficiency virus type 1 and comparison of the test with enzyme-linked immunosorbent assay and Western blot (immunoblot) analysis
Authors: Vasudevachari M B , Uffelman K W , Mast T C , Dewar R L , Natarajan V , Lane H C , Salzman N P
Published: January 2020

A passive hemagglutination test (PHA) was developed for detecting antibodies to human immunodeficiency virus type 1 (HIV-1) utilizing sheep erythrocytes cross-linked with purified envelope glycoprotein (gp160) of HIV-1. In an analysis of 216 human serum samples, 100% correlation was observed in 86 reactive and 124 nonreactive serum samples between PHA and commercial enzyme-linked immunosorbent assays and Western blot (immunoblot) analysis. Serum samples from gp160-immunized chimpanzees also reacted equally well in PHA. The test is simple, rapid, and inexpensive, thus providing an alternate, quick method of detecting HIV antibodies. These advantages and the thermal stability of the reagents that are used make this an attractive alternative for detecting prior exposure of individuals to HIV-1.
Detection of herpes simplex virus by 8 h in shell vial cultures with primary rabbit kidney cells
Authors: Tse P , Aarnaes S L , de la Maza L M , Peterson E M
Published: January 2020

Shell vial (SV) cultures for herpes simplex virus using primary rabbit kidney cells stained at 8 h after inoculation were compared with 20-h SV cultures as well as conventional tissue culture. Of the 326 clinical specimens examined, conventional culture detected 67, and of these, 61 (91%) and 42 (63%) were detected by 20-h SV cultures and 8-h SV cultures, respectively.