bluuebunny/crossref_metadata_2025 huggingface dataset:
Pasteurella haemolytica-like bacterium from a progressive granuloma of cattle in Brazil
Authors: Ribeiro G A , Carter G R , Frederiksen W , Riet-Correa F
Published: January 2020

Three strains of a Pasteurella haemolytica-like bacterium were isolated from lesions of a progressive granuloma of cattle in southern Brazil. Their characteristics and their differentiation from P. haemolytica varieties and Actinobacillus lignieresii are described. The name "Pasteurella granulomatis" is proposed for this apparently new taxon.
Aeromonas hydrophila infection associated with the use of medicinal leeches
Authors: Snower D P , Ruef C , Kuritza A P , Edberg S C
Published: January 2020

The use of medicinal leeches (Hiruda medicinalis) is becoming more common after plastic surgery to control venous congestion of skin grafts. We describe a patient with Aeromonas hydrophila infection whose graft was treated with medicinal leeches. The infection required systemic antibiotic therapy. A. hydrophila is the predominant bacterial flora in the gut of the leech, where it plays an essential role for the animal in the digestion of blood. The potential for A. hydrophila wound infection, and appropriate antibiotic prophylaxis of the leech or patient, should be considered when medicinal leeches are used.
Pneumonia and meningitis caused by a new nonfermentative unknown gram-negative bacterium
Authors: Casalta J P , Peloux Y , Raoult D , Brunet P , Gallais H
Published: January 2020

Seven isolates of an unclassified bacterium resembling Flavobacterium spp. were characterized by growth requirements, microscopic examination, biochemical characteristics, antimicrobial susceptibility tests, protein profile analysis, and serologic data. The unclassified isolates were differentiated from Flavobacterium meningosepticum, Flavobacterium odoratum, Flavobacterium balustinum, Flavobacterium strain IIb, Chromobacterium violaceum, Aquaspirillum serpens, and Pseudomonas spp. The bacterium was a gram-negative rod with a polar flagellum. Protein profile analysis demonstrated two major protein bands present in the unclassified isolates that were absent from the Flavobacterium and Pseudomonas controls but present in the Aquaspirillum and Chromobacterium controls. However, no serologic cross-reactions were observed. Our results showed that the unclassified bacterium was distinct from any previously known genus of bacterium.
Evaluation of human immunodeficiency virus seroprevalence in population surveys using pooled sera
Authors: Kline R L , Brothers T A , Brookmeyer R , Zeger S , Quinn T C
Published: January 2020

The pooling of individual serum samples to determine human immunodeficiency virus (HIV) seropositivity was examined to assess whether testing pooled sera was technically feasible, cost-effective, and accurate for estimating seroprevalence in large population surveys. The sensitivities and specificities of three commercially available HIV enzyme-linked immunosorbent assay (ELISA) kits were tested using 65 serum pools of 15 individual serum samples each (975 total serum samples) at two different dilutions. With pooled sera, the Organon Teknika Bio-EnzaBead ELISA at half the dilution recommended by the manufacturer showed the best agreement with ELISA and Western blot results of individual sera. In subsequently testing 92 pools, each containing 15 individual serum samples from a population of American patients attending a sexually transmitted diseases clinic, the estimated seroprevalence was 5.27 compared with 4.93% in a test of 1,380 individual serum samples and 5.19% in a test of 4,028 individual serum samples from the same population. In an evaluation of 1,380 African patients using 10 serum samples per pool, the estimated seroprevalence was 5.79 compared with 6.16% in a test of individual sera. These results indicate that ELISA testing with pooled sera is highly sensitive and specific and appears to be a cost-effective means for estimating HIV seroprevalence in large population-based surveys.
Comparison of cefadroxil and cephalothin disk susceptibility test results
Authors: Barry A L , Jones R N
Published: January 2020

Diffusion susceptibility tests with 30-micrograms cefadroxil disks and 30-micrograms cephalothin disks were evaluated. For both agents, the same zone size interpretive criteria were recommended (less than or equal to 14 mm for resistance and greater than or equal to 18 mm for susceptibility). Tests were performed with 904 bacterial isolates, and the data were examined to determine whether the two cephalosporins might be used interchangeably for purposes of in vitro susceptibility testing. When Haemophilus influenzae, Listeria monocytogenes, and methicillin-resistant staphylococci were evaluated, the two agents differed significantly. For testing other species, a cephalothin disk or cephalothin MIC could be used for predicting susceptibility or resistance to cefadroxil.
Campylobacter species identification based on polymorphism of DNA encoding rRNA
Authors: Moureau P , Derclaye I , Gregoire D , Janssen M , Cornelis G R
Published: January 2020

Total DNA from five Campylobacter species was digested with a mixture of XhoI and BglII restriction endonucleases and analyzed by Southern hybridization by using a probe complementary to the DNA coding for the 16S rRNA. Each of the Campylobacter species, including C. jejuni, C. coli, C. laridis, C. fetus, and C. upsaliensis, displayed a characteristic pattern. Although some bands may be common to different species, the simplicity of the hybridization pattern enabled us to discriminate among the different species of Campylobacter.
Direct identification of Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium intracellulare from amplified primary cultures in BACTEC media using DNA probes
Authors: Peterson E M , Lu R , Floyd C , Nakasone A , Friedly G , de la Maza L M
Published: January 2020

DNA probes (Gen-Probe, San Diego, Calif.) directed at the Mycobacterium tuberculosis complex and Mycobacterium avium-M. intracellulare complex were used to identify acid-fast bacilli directly from specimens grown in BACTEC 12B bottles (Becton Dickinson and Co., Towson, Md.). Clinical specimens were inoculated directly or after decontamination into a BACTEC 12B bottle, Middlebrook 7H11 agar, and Lowenstein-Jensen medium. Conventional media were incubated at 37 degrees C in 5% CO2 and examined weekly for 6 weeks. Identification of isolates grown on conventional media by standard biochemicals, morphology, and growth characteristics served as the reference method for identification. BACTEC bottles were incubated at 37 degrees C, and a growth index was taken twice a week. When a growth index of greater than or equal to 100 was reached, 1 ml of BACTEC 12B medium was put into each of three microfuge tubes which were centrifuged for 15 min at 15,000 x g. Pellets were used in hybridization reactions with an M. tuberculosis complex probe, an M. avium probe, and an M. intracellulare probe. The results of the hybridizations of the three probes with the same sample were compared, and the highest percent hybridization was divided by the average of the lower hybridization values. If this value, the derived patient ratio (DPR), was greater than or equal to 3, then the specimen was considered positive for the organism giving the highest percent hybridization. Of the 1,988 specimens cultured, the results of conventional tests for the 190 conventional culture-positive specimens were 64 M. tuberculosis, 61 M. avium, 14 M. intracellulare, 30 other Mycobacterium spp., and 25 non-acid-fast bacilli. There were four cultures that each contained two different Mycobacterium spp. Directly probing the BACTEC 12B sediment, at DPR of >/= 3 the M. tuberculosis probe identified 83% (53 of 64) of M. tuberculosis isolates, the M. avium probe identified 92% (56 of 61) M. avium isolates, and the M. intracellulare probe identified 86% (12 of 14) of M. intracellulare isolates. There were no false-positive results at this DPR level. The false-negative results from probing the sediment from the BACTEC 12B bottle could not solely be attributed to the number of organisms present, the growth index, or antimicrobial therapy.
Antibody response to human immunodeficiency virus type 1 protease according to risk group and disease stage
Authors: Boucher C A , de Jager M H , Debouck C , Epstein L G , de Wolf F , Wolfs T F , Goudsmit J
Published: January 2020

Three groups with different routes of human immunodeficiency virus type 1 (HIV-1) transmission (homosexual men, hemophiliacs, and children) were studied for serum antibodies to a recombinant form of the HIV-1 protease using an enzyme-linked immunoassay. At 1 year after seroconversion, defined as the moment antibodies to HIV-1 proteins were first detected, 56% (34/61) of the homosexual men had antibodies to protease, and 2 years after seroconversion this percentage was 63% (24/38). Within this 2-year period these antibodies were no longer detected in 16% (9/56). A similar pattern was observed in 20 hemophiliacs who seroconverted after exposure to HIV-1-contaminated blood products. We found that 63% (160/255) of homosexual men in Centers for Disease Control stage II or III, 60% (9/15) of patients with acquired immunodeficiency syndrome (AIDS)-related complex, and 36% (14/39) of patients with AIDS had antibodies to protease. In 255 homosexual men in Centers for Disease Control stage II or III, antibodies to protease were significantly more frequently found in samples lacking HIV-1 antigen (P less than 0.001) and possessing antibodies to HIV-1 core proteins (P less than 0.001). Twenty-four persons who developed AIDS were studied longitudinally: 58% (14/24) had antibodies to protease 1 year before developing symptoms; 29% (7/24) showed a decline and 29% (7/24) showed a loss of antibodies to protease at the onset of symptoms. Within a group of 47 HIV-1-infected children, 90% (18/20) with a stable disease course were persistently protease antibody positive, versus 4 of 27 children (15%) with an unstable disease course (P = 0.0001). These data indicate that HIV-1 protease is expressed and antigenic in most HIV-1-infected individuals and that a decline or absence of antibodies to protease is strongly associated with unstable disease in children and AIDS in adults.
Francisella philomiragia comb. nov. (formerly Yersinia philomiragia) and Francisella tularensis biogroup novicida (formerly Francisella novicida) associated with human disease
Authors: Hollis D G , Weaver R E , Steigerwalt A G , Wenger J D , Moss C W , Brenner D J
Published: January 2020

Over a 12-year period, 16 human strains of a gram-negative, catalase-positive, halophilic, aerobic, nonmotile, small coccoid bacterium were received for identification. On the bases of biochemical characteristics and cellular fatty acid profiles, 14 of these strains were similar to the "Philomiragia" bacterium (Yersinia philomiragia, species incertae sedis). Additional characteristics were growth on Thayer-Martin agar but no growth or sparse, delayed growth on MacConkey agar; oxidase positive; acid production, often weak and delayed, from D-glucose, sucrose, and maltose; urease negative; no reduction of nitrates; and H2S produced but often delayed in triple sugar iron agar. Both the human isolates and the "Philomiragia" bacterium contained C10:0, C14:0, C16:0, C18:1 omega 9c, C18:0, 3-OH C18:0, C22:0, and C24:1 as major cellular fatty acids and ubiquinone eight (Q8) as the major isoprenoid quinone. These cellular acids in these relative amounts have been found previously only in Francisella tularensis and Francisella novicida, suggesting a relationship between the "Philomiragia" bacterium and Francisella species. Of the 14 human "Philomiragia"-like isolates, 9 were from blood, 3 were from lung biopsies or pleural fluid, and one each was from peritoneal fluid and cerebrospinal fluid. DNA relatedness studies (hydroxyapatite method, 50 and 65 degrees C) showed that these 14 strains were a single group that was the same species as the "Philomiragia" bacterium. Two other human strains were oxidase negative and H2S negative. They formed a single DNA relatedness group that was indistinguishable from the type strains of both F. tularensis and F. novicida. DNA relatedness of "Philomiragia" bacterium type and other strains to strains of F. novicida and F. tularensis, including the type strains, was 35 to 46%. One of the two F. novicida- and F. tularensis-like strains was isolated from blood, and the other was isolated from a cervical lymph node. On the basis of these findings, we propose transferring Y. philomiragia from the genus Yersinia to the genus Francisella as Francisella philomiragia comb. nov. Having confirmed that F novicida and F. tularensis are the same species and having shown that F. novicida is pathogenic for humans, we further propose eliminating the species F. novicida and demoting it to a biogroup of F. tularensis.
Long-term colonization of spinal cord injury patients with Klebsiella pneumoniae
Authors: Montgomerie J Z , Gilmore D S , Ashley M A , Schick D G , Jimenez E M
Published: January 2020

The duration of colonization of the human perineum, bowel, and urethra with Klebsiella pneumoniae was studied in 10 male patients with spinal cord injury while they were undergoing rehabilitation in the hospital. Colonization defined as persistence of klebsiellae of the same serotype occurred on 13 occasions in five of the patients for up to 55 days. Colonization was less marked with Klebsiella than with Pseudomonas spp. The presence of quadriplegia correlated with Klebsiella colonization and also with the presence of positive Klebsiella cultures from the urethra and perineum. Seven episodes of significant bacteriuria occurred with or after the finding of positive cultures from body sites.
Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever
Authors: Ortiz V , Isibasi A , García-Ortigoza E , Kumate J
Published: January 2020

The studies reported here were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce a humoral immune response in humans with typhoid fever. OMPs were isolated with the nonionic detergent Triton X-100 and were found to be contaminated with approximately 4% lipopolysaccharide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns showed protein bands with molecular size ranges from 17 to 70 kilodaltons; the major groups of proteins were those that correspond to the porins and OmpA of gram-negative bacteria. Rabbit antiserum to OMPs or to S. typhi recognized OMPs after absorption with lipopolysaccharide. Sera from patients with typhoid fever contained immunoglobulin M antibodies which reacted with a protein of 28 kilodaltons and immunoglobulin G antibodies which reacted mainly with the porins, as determined by immunoblotting. These results indicate that the porins are the major immunogenic OMPs from S. typhi and that the immune response induced in the infection could be related to the protective status.
Enhanced infectivity of bluetongue virus in cell culture by centrifugation
Authors: Sundin D R , Mecham J O
Published: January 2020

The effects of centrifugation of the infection of cell culture with bluetongue virus (BTV) were investigated. Baby hamster kidney cells were infected with BTV with or without centrifugation. Viral antigen was detected by immunofluorescence at 24 h in both centrifuged and noncentrifuged cultures. However, after 24 h of infection, the production of PFU in centrifuged cell cultures was 10- to 20-fold greater than that seen in cultures not centrifuged. In addition, centrifugation enhanced the direct detection of PFU from blood samples collected from a sheep experimentally infected with BTV.
Recovery of human immunodeficiency virus type 1 from supernatant fluid of routine viral cultures
Authors: Sia E O , Englund J A , Sannerud K , Hansen K , Shah N , Balfour H H
Published: January 2020

Peripheral blood mononuclear cell samples from human immunodeficiency virus (HIV) antibody-positive subjects were inoculated into cell cultures used for routine viral isolation to determine if HIV could survive in them. HIV was recovered from 5 (19%) of 26 day 1 supernatants and from four of the five samples in all three types of cell cultures. We conclude that HIV may survive for at least 24 h in standard virology cell culture systems and therefore is a potential risk for laboratory personnel.
Isolation of Neisseria canis in mixed culture from a patient after a cat bite
Authors: Guibourdenche M , Lambert T , Riou J Y
Published: January 2020

We report a case of identification of Neisseria canis in a human patient after a cat bite. This organism was isolated from a mixed culture together with Pasteurella multocida and Eikenella corrodens. It is the second case report of a human infection.
Identification of Escherichia coli that produces heat-stable enterotoxin STA by a commercially available enzyme-linked immunoassay and comparison of the assay with infant mouse and DNA probe tests
Authors: Scotland S M , Willshaw G A , Said B , Smith H R , Rowe B
Published: January 2020

By using the infant mouse test and a commercially available competitive enzyme-linked immunosorbent assay (ELISA), 100 strains of Escherichia coli carrying STA1 or STA2 genes were shown to produce the heat-stable enterotoxin STA. An additional 100 strains were negative in both tests. The ELISA was easy to perform, and results were available within 24 h. Testing strains with an enzyme-linked DNA probe kit that incorporated both STA1- and STA2-specific oligonucleotides showed that the 100 strains positive in the mouse test and ELISA also hybridized with the mixed probe. Two strains carrying STA1 genes but negative in the mouse test and ELISA also hybridized with the mixed probe.
Human papillomavirus detection in urine samples from male patients by the polymerase chain reaction
Authors: Melchers W J , Schift R , Stolz E , Lindeman J , Quint W G
Published: January 2020

Human papillomavirus (HPV) detection was performed using the polymerase chain reaction technique on urine samples from 17 male patients with condylomata acuminata in the meatus urethrae. Urine samples from 14 male laboratory volunteers were analyzed as controls. The DNA was extracted and purified from urine sediments, centrifuged at 1,800 and 100,000 x g, and subjected to 40 cycles of amplification with HPV 6 and HPV 11 type-specific anticontamination primers and the heat-stable Taq DNA polymerase. HPV was detected in the urine of 15 (88%) patients. In all positive patients the urine sediments of both the 1,800 and 100,000 x g centrifugation steps contained HPV DNA. Eight patients were found to be positive for HPV 6 DNA, six were positive for HPV 11 DNA, and one was positive for both HPV 6 and HPV 11 DNA. None of the males in the control group was positive for either HPV 6 or HPV 11 DNA. The results demonstrate that HPV can be transported by the urine, probably in exfoliated HPV-infected cells. A similar mechanism may occur during ejaculation, allowing sexual transmission of HPV viruses harbored in the cells of the male genital tract.
Ear punch biopsy method for detection and isolation of Borrelia burgdorferi from rodents
Authors: Sinsky R J , Piesman J
Published: January 2020

An ear punch biopsy method for the detection and isolation of Borrelia burgdorferi from rodents was developed. The ear punch biopsy proved to be extremely sensitive, detecting spirochetes in 100% (11 of 11) of laboratory hamsters infected by tick bite and 95.8% (23 of 24) of hamsters infected by intraperitoneal inoculation. When cultured at 4 to 6 weeks postinfection, 92 to 100% of the ear punches taken from individual hamsters yielded viable spirochetes. B. burgdorferi was detected in sequential cultures from animals as early as 4 days postinfection and as late as 20 weeks postinfection. A total of 86% (6 of 7) of field-collected white-footed mice (Peromyscus leucopus) which were positive for B. burgdorferi as determined by xenodiagnosis were also positive by the ear punch method. The ear punch biopsy method allows individual rodents to be sampled for B. burgdorferi serially over a long period and thus should prove useful for both field and laboratory experiments.
Western blot (immunoblot) assay of small, round-structured virus associated with an acute gastroenteritis outbreak in Tokyo
Authors: Hayashi Y , Ando T , Utagawa E , Sekine S , Okada S , Yabuuchi K , Miki T , Ohashi M
Published: January 2020

Small, round-structured virus (SRSV) was detected in a stool specimen of a patient during an acute gastroenteritis outbreak in Tokyo and was tentatively named SRSV-9. SRSV-9 was purified by sucrose velocity gradient centrifugation after CsCl density gradient centrifugation. The buoyant density of SRSV-9 appeared to be 1.36 g/ml in CsCl. A Western blot (immunoblot) assay using the biotin-avidin system revealed that SRSV-9 was antigenically related to the Hawaii agent but distinct from the Norwalk agent and contained a single major structural protein with a molecular size of 63.0 +/- 0.6 kilodaltons. The prevalence of SRSV-9 infection in Tokyo was surveyed by the Western blot antibody assay by using a crude virus preparation as the antigen. Seroconversion was observed in 56.5% of the patients involved in the outbreaks from which SRSV was detected by electron microscopy.
Identification of Borrelia burgdorferi and B. hermsii using DNA hybridization probes
Authors: Schwan T G , Simpson W J , Schrumpf M E , Karstens R H
Published: January 2020

Fragments of plasmid DNA from Borrelia burgdorferi and B. hermsii were cloned and tested for specificity as hybridization probes to identify these two species of pathogenic spirochetes. Three fragments from the 49-kilobase-pair linear plasmid of B. burgdorferi were tested: a 500-base-pair (bp) HindIII fragment (probe 49A), a 445-bp PstI-HindIII fragment (probe 3G), and a 320-bp HindIII fragment (probe 16H). When hybridized to purified DNA or whole spirochetes, all of the probes distinguished B. burgdorferi from the other species examined, including B. hermsii, B. parkeri, B. turicatae, B. coriaceae, B. crocidurae, and B. anserina. Probe 49A was the most useful, however, hybridizing with all strains of B. burgdorferi originating from both North America and Europe while not cross-hybridizing with B. hermsii. A 790-bp HindIII fragment of B. hermsii DNA hybridized with DNA and whole spirochetes of this species and also with B. parkeri, confirming the close relatedness of these two species. These probes provide a new method of identifying these Borrelia species once the organisms have been grown in culture.
Effect of monophosphoryl lipid A on the in vitro function of peritoneal leukocytes from uremic patients on continuous ambulatory peritoneal dialysis
Authors: Carozzi S , Salit M , Cantaluppi A , Nasini M G , Barocci S , Cantarella S , Lamperi S
Published: January 2020

We analyzed the effects of monophosphoryl lipid A (MPL), a relatively nontoxic immunostimulant derived from bacterial endotoxin, on the depressed in vitro immune function of leukocytes derived from six patients undergoing continuous ambulatory peritoneal dialysis and who had histories of recurrent bacterial peritonitis. MPL was also tested for its capacity to stimulate the proliferation of peritoneal fibroblasts, as determined by [3H]thymidine incorporation. In vitro incubation of peritoneal lymphocytes and macrophages (PM phi) with increasing amounts of MPL, up to 5 micrograms/ml, resulted in a dose-dependent enhancement of gamma interferon and interleukin-2 production by peritoneal lymphocytes and interleukin-1 release by PM phi. In vitro incubation of PM phi with MPL also resulted in an increase of PM phi bacterial killing and membrane Fc receptor number, although no change in peritoneal fibroblast proliferation was seen with any of the MPL concentrations tested. These results suggest that the peritoneal leukocyte dysfunction observed in patients undergoing continuous ambulatory peritoneal dialysis and who have high rates of peritonitis may be alleviated, to some degree, by MPL, without directly inducing a potentially deleterious fibrotic lesion.
Isolation of peplomer glycoprotein E2 of transmissible gastroenteritis virus and application in enzyme-linked immunosorbent assay
Authors: Rukhadze G G , Aliper T I , Sergeev V A
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) based on peplomer glycoprotein E2 was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). Purified preparations of E2 were isolated by solubilization of the viral membrane with nonion detergent Nonidet P-40, followed by sucrose density gradient sedimentation. ELISA optical density values with E2 antigen significantly exceeded the indices when other TGEV protein or intact virion antigen was used. It was shown that a virus protein concentration in the E2 preparation of 500 ng per well is sufficient to sensitize the solid phase of microplates. A comparison of the ELISA and the virus neutralization test for the detection of TGEV antibodies was conducted. A significant correlation between the ELISA and the virus neutralization test was shown (r = 0.97). This serological test may be successfully used for various immunologic investigations.
Characterization of Aeromonas sobria hemolysin by use of monoclonal antibodies against Aeromonas hydrophila hemolysins
Authors: Kozaki S , Asao T , Kamata Y , Sakaguchi G
Published: January 2020

Aeromonas sobria produces hemolysin in a form activable with trypsin under defined cultural conditions. In immunoblotting analyses with the culture supernatant of A. sobria, the monoclonal antibody reacting specifically to Aeromonas hydrophila CA-11 hemolysin bound to the 53,000- and 49,000-dalton bands before and after trypsinization, respectively. The monoclonal antibody reacting to A. hydrophila AH-1 hemolysin did not bind either band. A. sobria hemolysin is, therefore, related antigenically to CA-11 hemolysin, while the molecular weights before and after activation differ from those of A. hydrophila hemolysins, being 54,000 and 51,000, respectively. The hemolytic and enterotoxigenic activities of A. sobria hemolysin were both neutralized by the monoclonal antibody against CA-11 hemolysin. It seems, therefore, that the same site on A. sobria hemolysin is responsible for both biological activities.
Epidemiologic marker system for Citrobacter diversus using outer membrane protein profiles
Authors: Kline M W , Mason E O , Kaplan S L
Published: January 2020

Investigations of nursery outbreaks of Citrobacter diversus sepsis and meningitis have been hampered by lack of adequate epidemiologic markers for the organism. We studied outer membrane protein profiles from clinical isolates of C. diversus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine whether this method might be useful in the epidemiologic differentiation of strains. Paired cerebrospinal fluid isolates from each of three separate nursery outbreaks of C. diversus meningitis, paired isolates from the vagina of a postpartum woman and the cerebrospinal fluid of her newborn infant, one isolate from an infant with pneumonia and two from colonized nursery cohorts, and 30 epidemiologically unrelated clinical isolates were included. Eleven distinct profiles were differentiated by the presence or absence of five outer membrane proteins. Complete concordance of profiles was observed for epidemiologically related isolates. Unrelated epidemic strains had outer membrane protein profiles distinct from one another. Biotyping complemented determination of outer membrane protein profiles; the two markers differentiated each of the five epidemic strains from all but one of 30 unrelated nonepidemic isolates. Determination of outer membrane protein profiles is potentially useful in epidemiologic investigations of disease caused by C. diversus.
Surface properties and ultrastructure of Edwardsiella species
Authors: Wong J D , Miller M A , Janda J M
Published: January 2020

Twenty-two strains of Edwardsiella representing the three currently recognized species were evaluated for surface characteristics and ultrastructural morphology. All isolates tested possessed a high surface cell charge as detected by DEAE-cellulose chromatography; surface hydrophobicity was variable and strain but not species dependent. Two major types of adhesins detected by hemagglutination assays were identified: one was inhibited by D-mannose (mannose-sensitive hemagglutinin) and found on all three species, and a second was not inhibited by D-mannose (mannose-resistant hemagglutinin) and was principally associated with Edwardsiella tarda. The results of physiologic and ultrastructural studies suggest that both hemagglutinins are afimbrial adhesin proteins. The results of these studies suggest that there are a number of distinct surface and ultrastructural properties associated with each of the three Edwardsiella species.
Enzymatic amplification of human cytomegalovirus sequences by polymerase chain reaction
Authors: Hsia K , Spector D H , Lawrie J , Spector S A
Published: January 2020

Polymerase chain reaction (PCR) amplification was used to detect human cytomegalovirus (HCMV) sequences. The fragments selected for amplification were fragments of 130 and 152 base pairs (bp) located at two opposite ends of HCMV strain AD169 EcoRI fragment D. Amplification of the 152-bp DNA was consistently greater than that of the 130-bp DNA. At the optimal Mg2+ concentration of 5 mM, specific PCR amplification of 152-bp DNA with Taq polymerase was sensitive; only one AD169-infected fibroblast cell or 0.01 pg of AD169 fragment D DNA was needed for detection. This specific amplification was also found with various clinical HCMV isolates and peripheral blood cells and urine from patients. In 37 urine samples analyzed simultaneously by PCR and by virus cultivation, identical results were found in 35 samples, while 2 scored positive only by PCR. This suggests that specific amplification of 152-bp DNA is sensitive and can be used for rapid detection of HCMV infections.
Characterization of Aeromonas schubertii strains recently isolated from traumatic wound infections
Authors: Carnahan A M , Marii M A , Fanning G R , Pass M A , Joseph S W
Published: January 2020

Recent studies have resulted in the proposal of a new species, Aeromonas schubertii (mannitol, sucrose, and indole negative), formerly termed Enteric Group 501, on the basis of the study of seven strains isolated from the southeastern and southwestern United States and Puerto Rico. We have isolated two phenotypically similar A. schubertii strains from infected human wounds sustained in the Chesapeake Bay area. Their identification was confirmed by DNA-DNA hybridization to the Centers for Disease Control definition strain 2446-81 (ATCC 43700) for group 12. The strains were further examined for the presence of virulence-associated markers: hemolysin, hemagglutinins, cytotoxin production, agglutination in acriflavine, resistance to normal human serum, and autoagglutination phenotype. Both strains were positive for hemolysin by the plate assay, cytotoxin production at 1:10, and DNase and protease. They were resistant to human serum and negative for acriflavine agglutination, and only one of the strains was autoagglutination positive. Both strains were negative for cell-free hemolysin, hemagglutinins, pectinase, and chitinase. These isolations of A. schubertii further extend its previously described geographic distribution and reinforce its role as a primary causative agent of cellulitis with possible increased antimicrobial resistance.
Legionella tucsonensis sp. nov. isolated from a renal transplant recipient
Authors: Thacker W L , Benson R F , Schifman R B , Pugh E , Steigerwalt A G , Mayberry W R , Brenner D J , Wilkinson H W
Published: January 2020

A Legionella-like organism (strain 1087-AZ-H) was isolated from a pleural-fluid specimen from a renal transplant patient undergoing immunosuppressive therapy. Growth characteristics and gas-liquid chromatography profiles of the isolate were consistent with those for Legionella spp. The isolate fluoresced blue-white under long-wave UV light. Strain 1087-AZ-H was serologically distinct in the slide agglutination test with absorbed antisera. DNA hybridization studies placed it in a new Legionella species, Legionella tucsonensis (ATCC 49180).
Seroepidemiology of Campylobacter pylori infection in various populations
Authors: Mégraud F , Brassens-Rabbé M P , Denis F , Belbouri A , Hoa D Q
Published: January 2020

Campylobacter pylori infection has been recognized as being strongly associated with chronic gastritis and duodenal ulceration, but the prevalence of C. pylori infection in a normal population is not known. A serological survey was conducted in four countries with different geographical and socioeconomic status, in a randomly chosen population as representative as possible, by using an enzyme-linked immunosorbent assay (ELISA) with a sonic extract of two strains as the antigen. The test had a specificity of 94% when 600 ELISA units was used as the threshold. In France, few children were infected before the age of 10 years. The prevalence then increased gradually to 36.7% in the sixth decade of life. This increasing prevalence of infection with age was also observed in Algeria, Vietnam, and the Ivory Coast but at a higher rate (80 to 90%). In Vietnam, as in France, few children were infected, whereas in Africa, C. pylori infection occurred earlier. The prevalence of infection did not differ with sex for a particular age group; it also did not differ with respect to gastric symptoms, smoking and drinking habits, or urban or rural residence when these potential risk factors were studied. The epidemiological data available on peptic ulcer disease in developing countries compared with developed countries led to the speculation that infection with C. pylori is not a sufficient condition to develop this disease.
Evaluation of neutralizing antibodies to type A, B, E, and F botulinum toxins in sera from human recipients of botulinum pentavalent (ABCDE) toxoid
Authors: Siegel L S
Published: January 2020

Twenty-five serum specimens from personnel immunized with botulinum pentavalent toxoid (ABCDE) had titers of neutralizing antibodies to type A (5.7 to 51.6 IU/ml), type B (0.75 to 18 IU/ml), and type E (0.61 to 10 IU/ml) botulinum toxins. Titers for one type could not be used to predict titers for another type in individuals receiving the toxoid. Cross-neutralizing antibodies to type F botulinum toxin were not detected (less than 0.0125 IU/ml).
Neopterin and alpha and beta interleukin-1 levels in sera of patients with human immunodeficiency virus infection
Authors: Reddy M M , Grieco M H
Published: January 2020

Levels of neopterin and alpha and beta interleukin-1 (IL-1) in sera of normal controls, asymptomatic intravenous drug abusers, homosexuals, and patients with lymphadenopathy or acquired immunodeficiency syndrome were measured. Neopterin levels were elevated in the sera of human immunodeficiency virus (HIV)-seronegative intravenous drug abusers and homosexuals, as well as in the sera of HIV-seropositive patients. Alpha IL-1 was the most predominant form of IL-1 found in the sera of all groups, and its level in HIV-seronegative intravenous drug abusers was elevated compared with the level in controls, whereas its levels in asymptomatic HIV-seropositive intravenous drug abusers and asymptomatic HIV-seronegative and HIV-seropositive homosexuals were decreased relative to the level in controls. Beta IL-1 levels in sera in all groups were not significantly different from the control value, except for the HIV-seropositive homosexual group; in this group the beta IL-1 level was significantly decreased compared with the control value.
Diagnosis of Chlamydia trachomatis genital infections by cell culture and two enzyme immunoassays detecting different chlamydial antigens
Authors: Mahony J , Castriciano S , Sellors J , Stewart I , Cunningham I , Landis S , Seidelman W , Grant L , Devlin C , Chernesky M
Published: January 2020

The enzyme-amplified immunoassay IDEIA (CellTech Diagnostics), which measures lipopolysaccharide antigen, and Chlamydiazyme (Abbott Laboratories, North Chicago, Ill.), which measures several antigenic components of Chlamydia trachomatis, were compared for specimens from urethral swabs from 235 men attending a clinic for sexually transmitted diseases (culture prevalence of 14.9%) and 458 endocervical swabs from women attending planned parenthood and obstetrics-gynecology clinics (culture prevalences of 5.9 and 7.7%, respectively). Compared with cell culture, the percent sensitivites, specificities, and positive and negative predictive values for IDEIA were 62.5, 99.5, 95.2, and 94.3%, respectively, for specimens from men and 96.3, 97.9, 74.3, and 99.8%, respectively, for specimens from women; results for Chlamydiazyme for specimens from men were 81.8, 99.5, 96.4, and 97.1%, respectively, and for specimens from women, results were 85.2, 99.3, 88.5, and 99.1%, respectively. Although the specificities of IDEIA and Chlamydiazyme were comparable, the sensitivity of IDEIA appeared higher for women (96.3%) than for men (67.5%), while the sensitivities of Chlamydiazyme were similar for men (81.8%) and women (85.2%). Western blot (immunoblot) analysis of the detector reagents from the two immunoassays indicated that the differences in performance observed for the two immunoassays may be due to measurement of different antigens.
Comparative studies of the antigenic polypeptide species VP4, VP6, and VP7 of three strains of bovine rotavirus
Authors: Zheng S L , Woode G N , Melendy D R , Ramig R F
Published: January 2020

Three bovine rotavirus strains belonging to two distinct serotype groups, serotype 6 (NCDV and B641) and B223, distinct from the other six mammalian rotavirus serotypes but not yet assigned to a serotype group, were compared with each other and with canine rotavirus (K9, serotype 3) by studying the properties of their cognate polypeptide species VP4, VP6, and VP7. The three viruses showed distinct differences in the polyacrylamide gel electrophoretic migration rates of protein species VP4 and VP7, with minor differences in VP6. Differences were also observed among the migration patterns of genome segments 4, 6, and the 7-8-9 triplet, which encode VP4, VP6, and VP7, respectively. Monoclonal antibodies (MAbs) to B223, which were directed against VP4 or VP7, showed homologous specificity for neutralization and immunofluorescence (IF), although one MAb reactive with VP4 also reacted by IF and by immunoprecipitation (IP) with all four viruses and weakly neutralized B641 and K9. This MAb may react with the epitope responsible for the B223-induced one-way neutralizing and protection response of calves against B641 observed in earlier studies. MAbs reactive with VP6 by IP showed enzyme-linked immunosorbent assay and IF reactivity with all three bovine viruses and the canine virus. The two serotype 6 viruses could be distinguished by the two B641 MAbs, B641-N2b reacting by neutralization and IF with both viruses and B641-N1 reacting with B641 and the serotype 3 canine rotavirus but not with NCDV. One nonneutralizing B641 MAb reacted by IP and IF with VP7 of all four rotaviruses examined, and one B223 MAb neutralized B223 and, to low titer, B641 and K9 although reacting by IP and IF with all four viruses. Three MAb-resistant mutants were selected by passage of B223 in the presence of one of three selected B223 MAbs at concentrations which only neutralized approximately 90% of the infectious virions. The resulting mutants were 100% resistant to neutralization with their respective MAb but remained neutralizable by the same selection of MAbs as the parent B223 virus.
Diagnosis of herpes simplex virus encephalitis by detection of virus-specific immunoglobulins A and G in serum and cerebrospinal fluid by using an antibody-capture enzyme-linked immunosorbent assay
Authors: van Loon A M , van der Logt J T , Heessen F W , Postma B , Peeters M F
Published: January 2020

An enzyme-linked immunosorbent assay was evaluated for detection of intrathecal synthesis of immunoglobulin G (IgG) and IgA antibodies to herpes simplex virus (HSV) in patients with HSV encephalitis (HSVE). Since the antibody-capture principle was used and the assay was carried out at the saturation level of the anti-IgG- or anti-IgA-coated solid phase, correction for blood-brain barrier leakage was not needed. A total of 34 pairs of serum and cerebrospinal fluid specimens obtained from 20 patients with HSVE were examined. Intrathecal synthesis of HSV IgG and IgA was detected from day 7 after the onset of illness in patients with HSVE. Specimens from all 19 patients from whom paired serum and cerebrospinal fluid specimens were obtained at more than 10 days after the onset of illness were positive. Intrathecal synthesis of HSV IgG and IgA was not detected in patients with HSVE before day 7 of illness or in any of the 16 control patients with other causes of (meningo)encephalitis. Use of the antibody-capture enzyme-linked immunosorbent assay for HSV IgG and IgA allows the rapid diagnosis of HSVE during the second week of illness.
Application of quantitative immunofluorescence to clinical serology: antibody levels of Toxoplasma gondii
Authors: Kaplan D S , Picciolo G L
Published: January 2020

Methods currently used for immunofluorescent reagent standardization require subjective visual comparison of reagents with control materials. Reactivities of reagents in immunofluorescence test kits vary from manufacturer to manufacturer. To solve these problems, a quantitative immunofluorescence (QIF) method which uses a calibrated photometric system and incorporates reducing agents into the mounting medium to reduce fading was developed to replace the visual method of endpoint determination. A uranyl glass slide was used to calibrate the instrument's voltage measurements, permitting daily comparisons and measurement of the instrument reading fluctuations. The QIF method was initially tailored to the determination of serum antibodies to Toxoplasma gondii by measuring the fluorescence intensity of individual tagged organisms. The nonspecific fluorescence intensity resulting from the counterstain was eliminated by use of a red-suppressing filter. The dilution-correlated polar fluorescence component was removed by subtraction of the intensity for the matching negative control dilution from each sample dilution intensity. The QIF method showed a 94% correlation with the visual comparison method for 62 clinical specimens.
Diarrheal disease caused by enterotoxigenic Bacteroides fragilis in infant rabbits
Authors: Myers L L , Shoop D S , Collins J E , Bradbury W C
Published: January 2020

Enterotoxigenic Bacteroides fragilis caused severe, nonhemorrhagic, watery diarrhea when 10(9) CFU of a porcine or human isolate was administered orogastrically to 3-day-old rabbits. The bacterium colonized the intestinal tract with a predilection for the large intestine (10(9) CFU/g of cecal contents). Diarrhea occurred at an average of 4.6 days postinoculation, and 84% of rabbits were dead or moribund at an average of 8.8 days postinoculation. The disease was characterized by watery diarrhea and dehydration. Severe histologic lesions including inflammation, exfoliation of epithelial cells, and crypt hyperplasia were observed throughout the colon. There was no indication of bacteremia or of bacterial adherence to or invasion of intestinal epithelial cells. Rabbits inoculated with nonenterotoxigenic B. fragilis were colonized with B. fragilis but did not develop clinical disease or intestinal lesions. While the pathogenesis of this disease is undefined, clinical signs of disease and histologic changes were consistent with a mechanism of net secretion of fluid into the small intestine and decreased absorption of fluid from the large intestine. Enteric disease caused by enterotoxigenic B. fragilis in infant rabbits was similar to naturally occurring enteric disease associated with the bacterium in humans and livestock. This study established that enterotoxigenic B. fragilis is enteropathogenic in intact infant rabbits.
Enterobacter hormaechei, a new species of the family Enterobacteriaceae formerly known as enteric group 75
Authors: O'Hara C M , Steigerwalt A G , Hill B C , Farmer J J , Fanning G R , Brenner D J
Published: January 2020

The name Enterobacter hormaechei is proposed for a new species of the family Enterobacteriaceae, formerly called Enteric Group 75, which consists of 23 strains, 22 of which were isolated from humans. DNAs from 12 E. hormaechei strains tested were highly related to the type strain (ATCC 49162) by DNA hybridization, using the hydroxyapatite method (80 to 97% in 60 degrees C reactions; 80 to 90% in 75 degrees C reactions). The strains were most closely related (50 to 63%) to Enterobacter cloacae, Enterobacter dissolvens, Enterobacter taylorae, and Enterobacter nimipressuralis. E. hormaechei strains were positive within 48 h for the following: Voges-Proskauer test; citrate utilization (Simmons and Christensen); urea hydrolysis (87%); ornithine decarboxylase; growth in potassium cyanide (KCN); malonate utilization; production of acid from D-glucose, L-arabinose, cellobiose, dulcitol (87%), D-galactose, maltose, D-mannitol, D-mannose, L-rhamnose, sucrose, trehalose, and D-xylose; acid production from mucate; nitrate reduction; and o-nitrophenyl-beta-D-galactopyranoside. Delayed positive reactions were seen in tests for arginine dihydrolase, gas from D-glucose, acid from alpha-methyl-D-glucoside, and acetate utilization. E. hormaechei was negative in tests for indole production; H2S production; phenylalanine deaminase; lysine decarboxylase; gelatin hydrolysis; acid production from D-adonitol, D-arabitol, erythritol, glycerol, i(myo)-inositol, melibiose, raffinose, and D-sorbitol; esculin hydrolysis; DNase; lipase; and tyrosine clearing. Variable reactions occurred in tests for methyl red, motility, and tartrate. All strains tested were susceptible or moderately susceptible to amikacin, azlocillin, cefotaxime, ceftazidime, ceftriaxone, chloramphenicol, gentamicin, mezlocillin, moxalactam, piperacillin, trimethoprim-sulfamethoxazole, sulfisoxazole, thienamycin, tobramycin, and trimethoprim. All strains tested were resistant to nitrofurantoin; the majority were resistant to ampicillin, cefoxitin, and cephalothin. Four isolates were from blood; most other isolates were from wounds or sputum.
Three cases of neonatal meningitis caused by Enterobacter sakazakii in powdered milk
Authors: Biering G , Karlsson S , Clark N C , Jónsdóttir K E , Lúdvígsson P , Steingrímsson O
Published: January 2020

Three cases of neonatal infection caused by Enterobacter sakazakii are reported from the Department of Neonatal Intensive Care, the National University Hospital, Reykjavík, Iceland. These infections occurred during a 9-month period in 1986 and 1987. Two of the neonates, who were normal at birth, survived but were left with brain damage. The third, who had Down's syndrome and severe cardiac malformations, died. The same organism was also grown from groin and anal swabs from a healthy neonate. E. sakazakii was not isolated from any environmental sources in the neonatal wards or in the milk kitchen, but it was grown from several lots of the powdered-milk formula used in the hospital. The four E. sakazakii strains isolated from the neonates were indistinguishable from 22 strains grown from the formula. Their biotypes, plasmid DNA profiles, and antibiograms were identical.