bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genomic Comparisons and Shiga Toxin Production among
Escherichia coli
O157:H7 Isolates from a Day Care Center Outbreak and Sporadic Cases in Southeastern Wisconsin
Authors: Gouveia S. , Proctor M. E. , Lee M.-S. , Luchansky J. B. , Kaspar C. W. 
Published: December 2019

ABSTRACT

Contour-clamped homogeneous electric field pulsed-field gel electrophoresis (CHEF-PFGE) was used to compare Wisconsin isolates of
Escherichia coli
O157:H7, including 39 isolates from a 1994 day care center outbreak, 28 isolates from 18 individuals from the surrounding geographic area with sporadic cases occurring during the 3 months before the outbreak, and 3 isolates, collected in 1995, from patients with hemolytic-uremic syndrome (HUS) who were from eastern Wisconsin counties other than those inhabited by the day care center and sporadic-case individuals. The technique of CHEF-PFGE using
Xba
I identified seven highly related restriction endonuclease digestion profiles (REDPs) (93 to 98% similarity) among the 39 day care center isolates and nine
Xba
I REDPs (63 to 93% similarity) among the 28 isolates from sporadic-case individuals, including REDP 33, which was exhibited by both day care and sporadic-case isolates. PFGE analyses of sequential
E. coli
O157:H7 isolates from symptomatic day care center attendees revealed that the REDPs of 25 isolates from eight patients were indistinguishable whereas the REDPs of 2 of 6 isolates from two patients differed slightly (93 to 95% similarity). The REDPs of the three isolates from 1995 HUS patients were 78 to 83% similar, with REDP 26 being exhibited by one HUS-associated isolate and an isolate from one day care attendee who did not develop HUS. The genes for both Shiga toxins I and II (
stx
1
and
stx
2
, respectively) were detected in all but one isolate (sporadic case), and Shiga toxin production by the day care center isolates was not significantly different from that of the other isolates, including the three HUS-associated isolates. Analyses of
E. coli
O157:H7 isolates from both the day care center outbreak and sporadic cases by CHEF-PFGE permitted us to define the REDP variability of an outbreak and geographic region and demonstrated that the day care center outbreak and a HUS case in 1995 were caused by
E. coli
O157:H7 strains endemic to eastern Wisconsin.
Molecular Analysis of Non-O1, Non-O139
Vibrio cholerae
Associated with an Unusual Upsurge in the Incidence of Cholera-Like Disease in Calcutta, India
Authors: Sharma Charu , Thungapathra M. , Ghosh A. , Mukhopadhyay Asish K. , Basu Arnab , Mitra Rupak , Basu Indira , Bhattacharya S. K. , Shimada T. , Ramamurthy T. , Takeda T. , Yamasaki S. , Takeda Y. , Nair G. Balakrish 
Published: December 2019

ABSTRACT

There was an inexplicable upsurge in the incidence of non-O1, non-O139
Vibrio cholerae
among hospitalized patients admitted to the Infectious Diseases Hospital, Calcutta, India, between February and March 1996. Of the 18 strains of
V. cholerae
isolated during this period, 15 belonged to the non-O1, non-O139 serogroups (4 belonged to O144, 3 belonged to O11, 1 each belonged to O6, O8, O12, O19, O39, and O58, and 2 strains could not be typed), 2 belonged to the O139 serogroup, and 1 belonged to the O1 serogroup. Cell-free culture supernatants of 13 representative non-O1, non-O139
V. cholerae
strains evoked a distinct cytotoxic effect on CHO and HeLa cells, and the strains examined produced the nonmembrane-damaging cytotoxin. By several PCR assays, it was determined that none of the non-O1, non-O139 strains were positive for the
ctxA
,
zot
,
ace
, and
tcpA
genes and for the genes representing the heat-labile toxin, heat-stable toxin, and verotoxin of
Escherichia coli
and the various variants of these genes. Studies on the clonality of non-O1, non-O139
V. cholerae
strains by restriction fragment length polymorphism (RFLP) analysis of rRNA genes and of other genes (
hlyA
,
hlyU
,
hlx
,
toxR
, and
attRS1
) and by pulsed-field gel electrophoresis (PFGE) collectively indicate that the upsurge which occurred in February and March 1996 was caused by strains belonging to different clones. Overall, there was an excellent correlation between the results of ribotyping, RFLP analysis of various genes, and PFGE, with strains belonging to a particular serogroup showing nearly identical restriction patterns and PFGE profiles. It is clear from this study that some serogroups of
V. cholerae
can cause diarrhea by a mechanism quite different from that of toxigenic
V. cholerae
O1 and O139, and we have proposed the nomenclature of enteropathogenic
V. cholerae
to include these serogroups.
Molecular Typing of
Borrelia burgdorferi
Sensu Lato by Randomly Amplified Polymorphic DNA Fingerprinting Analysis
Authors: Wang Guiqing , van Dam Alje P. , Spanjaard Lodewijk , Dankert Jacob 
Published: December 2019

ABSTRACT

To study whether pathogenic clusters of
Borrelia burgdorferi
sensu lato strains occur, we typed 136 isolates, cultured from specimens from patients (
n
= 49) with various clinical entities and from ticks (
n
= 83) or dogs (
n
= 4) from different geographic regions, by randomly amplified polymorphic DNA (RAPD) fingerprinting with four arbitrary primers. The RAPD patterns were reproducible up to the 95% similarity level as shown in duplicate experiments. In these experiments the purified DNAs prepared on different days, from different colonies, and after various passages were used as templates. With an intergroup difference of 55%, the 136 strains could be divided into seven genetic clusters. Six clusters comprised and corresponded to the established species
B. burgdorferi
sensu stricto (
n
= 23),
Borrelia garinii
(
n
= 39),
Borrelia afzelii
(
n
= 59),
Borrelia japonica
(
n
= 1),
Borrelia valaisiana
(
n
= 12), and genomic group DN127 (
n
= 1). One strain from a patient with erythema migrans (EM) did not belong to any of the species or genomic groups known up to now. The RAPD types of
B. burgdorferi
sensu stricto,
B. garinii
, and
B. afzelii
isolates, which may give rise to human Lyme borreliosis (LB), were associated with their geographic origins. A high degree of genetic diversity was observed among the 39
B. garinii
strains, and six subgroups could be recognized. One of these comprised eight isolates from patients with disseminated LB only and no tick isolates.
B. afzelii
strains from patients with EM or acrodermatitis chronica atrophicans were not clustered in particular branches. Our study showed that RAPD analysis is a powerful tool for discriminating different
Borrelia
species as well as
Borrelia
isolates within species.
Staphylococcus lugdunensis
: Report of a Case of Peritonitis and an Easy-To-Perform Screening Strategy
Authors: Schnitzler Norbert , Meilicke Rainer , Conrads Georg , Frank Darius , Haase Gerhard 
Published: December 2019

ABSTRACT

We report on a severe case of peritonitis due to
Staphylococcus lugdunensis
. The clinical course resembled an infection due to
S. aureus
more than one due to other coagulase-negative staphylococci. Therefore, we strongly recommend identification and propose an easy-to-perform procedure for screening of this pathogen.
Haemophilus parainfluenzae
Liver Abscess after Successful Liver Transplantation
Authors: Friedl J. , Stift A. , Berlakovich G. A. , Taucher S. , Gnant M. , Steininger R. , Mühlbacher F. 
Published: December 2019

ABSTRACT

Haemophilus parainfluenzae
was isolated from a bile specimen and from an aspirate of a liver abscess in a 58-year-old liver-transplanted woman that was indicative of an invasion of the graft by an ascending route. Drug therapy, immunosuppression, rejection therapy, and Roux-en-Y choledochojejunostomy may have contributed to the septic course. Interdisciplinary cooperation was instrumental in diagnosis and successful management in this case.
Simplified Technique for Detection of Significant Bacteriuria by Microscopic Examination of Urine
Authors: Cardoso Celso Luíz , Muraro Carla Barbosa , Siqueira Vera Lúcia Dias , Guilhermetti Márcio 
Published: December 2019

ABSTRACT
A comparative study of microscopic examination of 10 μl (simplified loop technique) and 50 μl (traditional drop technique) of uncentrifuged Gram-stained urine specimens for detection of significant bacteriuria was carried out. The results demonstrated that the 10-μl loop technique can be used as an alternative to the 50-μl drop technique for presumptive diagnosis of urinary-tract infection in bacteriological practice, with the advantages of greater rapidity and ease of performance.
Evaluation of the E Test for Antimicrobial Susceptibility Testing ofPseudomonas aeruginosaIsolates from Patients with Long-Term Bladder Catheterization
Authors: Di Bonaventura Giovanni , Ricci Evandro , Della Loggia Nicoletta , Catamo Giovanni , Piccolomini Raffaele 
Published: December 2019

ABSTRACTThe E test was evaluated in comparison with reference agar methods (National Committee for Clinical Laboratory Standards) for the susceptibility testing of 248Pseudomonas aeruginosaisolates from bladder-catheterized patients against nine antibiotics. The E-test MICs correlated well with those determined by the agar dilution and disk diffusion reference methods (88 and 92.5% within 1 log2dilution step, respectively), confirming that the E test is a reliable method for the determination of MICs of antibiotics for catheterization-associatedP. aeruginosaisolates.
Enteroaggregative, Shiga Toxin-Producing
Escherichia coli
O111:H2 Associated with an Outbreak of Hemolytic-Uremic Syndrome
Authors: Morabito Stefano , Karch Helge , Mariani-Kurkdjian Patrizia , Schmidt Herbert , Minelli Fabio , Bingen Edouard , Caprioli Alfredo 
Published: December 2019

ABSTRACT

Shiga toxin-producing
Escherichia coli
O111:H2 strains from an outbreak of hemolytic-uremic syndrome showed aggregative adhesion to HEp-2 cells and harbored large plasmids which hybridized with the enteroaggregative
E. coli
probe PCVD432. These strains present a novel combination of virulence factors and might be as pathogenic to humans as the classic enterohemorrhagic
E. coli.
Detection of Staphylococcal Superantigenic Toxins by a CD69-Specific Cytofluorimetric Assay Measuring T-Cell Activation
Authors: Lina Gerard , Cozon Grégoire , Ferrandiz Josette , Greenland Timothy , Vandenesch François , Etienne Jerome 
Published: December 2019

ABSTRACTThe presence of staphylococcal superantigenic toxins in the supernatants of liquid cultures was detected by an easy and rapid method assessing the activation of T lymphocytes by cytofluorimetric measurement of CD69 expression.Staphylococcus aureuscells were grown in Eagle’s minimum essential medium supplemented with 5% heat-inactivated fetal calf serum. Supernatant fluids from allS. aureusstrains producing superantigen-related toxins, including enterotoxins A to E, toxic shock syndrome toxin, and exfoliative toxins A and B, induced CD69 expression in a significantly higher number of T cells than a cutoff of 2%. This CD69 assay might be used for initial detection of superantigens fromS. aureusstrains isolated in the context of staphylococcal toxemia or related chronic human diseases such as atopic dermatitis or Kawasaki syndrome.
Optimization of Specimen-Handling Procedures for Accurate Quantitation of Levels of Human Immunodeficiency Virus RNA in Plasma by Reverse Transcriptase PCR
Authors: Dickover Ruth E. , Herman Steven A. , Saddiq Khaliq , Wafer Deborah , Dillon Maryanne , Bryson Yvonne J. 
Published: December 2019

ABSTRACT
Human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma are currently widely used clinically for prognostication and in monitoring antiretroviral therapy. Accurate and reproducible results are critical for patient management. To determine the effects of specimen collection and handling procedures on quantitative measurement of HIV-1 RNA, we compared anticoagulants and sample processing times. Whole blood was collected from 20 HIV-1-infected patients in EDTA, acid citrate dextrose (ACD), and heparin tubes, aliquoted, and stored at room temperature. Plasma was separated from whole-blood aliquots prepared at ≤1, 3, 6, 24, and 48 h postcollection and then stored at −70°C until use. HIV-1 RNA levels were determined by the AMPLICOR HIV-1 MONITOR assay. Heparinized plasma samples, which were pretreated with heparinase prior to analysis, had the lowest baseline HIV-1 RNA levels. In the first 6 h, HIV-1 RNA levels decreased by 10, 20, and 31% in EDTA, ACD, and heparin tubes, respectively. From 6 to 48 h postcollection, HIV-1 RNA levels decreased in all anticoagulants, albeit at a slower, more consistent rate. Our results indicate that EDTA should be the anticoagulant of choice for plasma HIV-1 RNA measurement by reverse transcriptase PCR, but ACD tubes are acceptable if the plasma is separated within 6 h of blood collection. Caution must be applied in the interpretation of absolute HIV-1 RNA copy number values obtained with suboptimal specimen collection and processing procedures.
Spoligotyping Followed by Double-Repetitive-Element PCR as Rapid Alternative to IS
6110
Fingerprinting for Epidemiological Studies of Tuberculosis
Authors: Sola Christophe , Horgen Lionel , Maïsetti Jerome , Devallois Anne , Goh Khye Seng , Rastogi Nalin 
Published: December 2019

ABSTRACT

A total of 129 clinical isolates of
Mycobacterium tuberculosis
representing 91 patients were typed by a combination of direct-repeat (DR)-based spoligotyping and an inter-IS
6110
–PGRS (polymorphic GC-rich region)–PCR, also designated double-repetitive-element PCR (DRE-PCR). During the first phase of this investigation, 72 clinical strains representing 52 patients were initially typed by IS
6110
-restriction fragment length polymorphism (RFLP) and DR-RFLP, followed by spoligotyping and DRE-PCR. In the second phase of this investigation, the discriminating ability of spoligotyping plus DRE-PCR was studied for 57 isolates from 39 patients who were suspected to be epidemiologically linked, and the typing results were later confirmed by IS
6110
-RFLP and DR-RFLP analyses. The molecular clustering of the isolates remained identical irrespective of the methods used. These results show that the association of two PCR-based fingerprinting techniques for molecular epidemiology of tuberculosis has a discriminating ability similar to the IS
6110
-RFLP reference method.
Phylogenetic Relationships among Highly Virulent Newcastle Disease Virus Isolates Obtained from Exotic Birds and Poultry from 1989 to 1996
Authors: Seal Bruce S. , King Daniel J. , Locke Devin P. , Senne Dennis A. , Jackwood Mark W. 
Published: December 2019

ABSTRACTNewcastle disease virus {NDV (avian paramyxovirus type 1 [APMV1])} isolates were recovered from imported exotic birds confiscated following importation into the United States, from waterbirds in the United States, and from poultry. The exotic birds probably originated from Central and South America, Asia, and Africa. The NDV isolates were initially characterized as highly virulent because of a short mean death time in embryonated chicken eggs. The isolates were typed as neurotropic or viscerotropic velogenic by intracloacal inoculation of adult chickens. Intracerebral pathogenicity index values for the virulent NDV isolates ranged from 1.54 to 1.90, compared to a possible maximum value of 2.0. These isolates had a dibasic amino acid motif in the fusion protein cleavage site sequence required for host systemic replication. Sequence differences were detected surrounding the fusion protein cleavage site and the matrix protein nuclear localization signal, indicating evolution of highly virulent NDV. Phylogenetically, these isolates were categorized with other highly virulent NDV strains that caused outbreaks in southern California poultry during 1972 and in cormorants in the north central United States and southern Canada during 1990 and 1992. These isolates are related to NDV that may have the APMV1 strain chicken/Australia/AV/32 or a related virus as a possible progenitor. Recent virulent NDV isolates and those recovered during disease outbreaks since the 1970s are phylogenetically distinct from current vaccine viruses and standard challenge strains.
Disseminated Invasive Infection Due to
Metarrhizium anisopliae
in an Immunocompromised Child
Authors: Burgner David , Eagles Gillian , Burgess Margaret , Procopis Peter , Rogers Maureen , Muir David , Pritchard Robert , Hocking Ailsa , Priest Michael 
Published: December 2019

ABSTRACT

The first reported human case of possible disseminated infection with the insect pathogen
Metarrhizium anisopliae
var.
anisopliae
, a fungus which has been used commercially for biocontrol of insects, is described. The patient, a 9-year-old boy, had a 5-year history of pre-B-cell acute lymphoblastic leukemia and had been on chemotherapy throughout this period. After 10 days of profound neutropenia, lesions consistent with ecthyma gangrenosum appeared on his arms and legs.
M. anisopliae
was grown from specimens from three separate sites, collected at different times over a period of 1 month: a skin biopsy, a swab from the base of a lesion, and the core of another skin lesion which spontaneously discharged. The initial skin biopsy also showed histological evidence of epidermal necrosis and dermal invasion with fungal hyphae. A computed-tomography (CT) scan of the chest demonstrated a lesion in the superior segment of the lower lobe of the left lung. A CT scan of the brain revealed a lesion in the left temporoparietal region of the brain, consistent with an abscess. Despite antifungal treatment including liposomal amphotericin and 5-flucytosine, the patient eventually died. The initial portal of entry is unknown, but hematogenous dissemination to the skin appears likely because of the multiple ecthymic lesions, and the appearances of the brain lesion on the CT scan are consistent with a hematogenous fungal abscess.
Corneal Ulcer Caused by
Nocardia asteroides
in a Patient with Leprosy
Authors: Tendolkar Uma M. , Varaiya Ami , Ahuja A. S. , Motwane S. A. , Gogate A. S. 
Published: December 2019

ABSTRACT

Nocardia asteroides
is a rare cause of keratitis usually associated with trauma. We report a case of corneal ulceration caused by
N. asteroides
in a patient with leprosy. This is the first case report of nocardial keratitis from Southeast Asia. The diminished corneal sensation in a patient with leprosy could be a predisposing factor for development or exacerbation of corneal ulceration.
Nosocomial Pneumonia Caused by Three Genetically Different Strains of
Legionella pneumophila
and Detection of These Strains in the Hospital Water Supply
Authors: Lück P. Christian , Wenchel Hans-Martin , Helbig Jürgen H. 
Published: December 2019

ABSTRACT

A 44-year-old woman developed
Legionella pneumophila
pneumonia after cerebral surgery. Initially, one colony from a clinical specimen and two colonies from water samples, all belonging to serogroup 12, did not match when their DNA restriction patterns were compared. When additional colonies from the water specimens were analyzed, a serogroup 12 strain complementary to that found in the clinical specimen was identified. Other colonies from the clinical specimen were identified as serogroup 12 strains complementary to those identified from the water. In addition, the same serogroup 1 strain was isolated from the patient and the water system.
Identification and Purification of Specific
Penicillium marneffei
Antigens and Their Recognition by Human Immune Sera
Authors: Jeavons L. , Hamilton A. J. , Vanittanakom N. , Ungpakorn R. , Evans E. G. V. , Sirisanthana T. , Hay R. J. 
Published: December 2019

ABSTRACT

Disseminated infection with the dimorphic pathogenic fungus
Penicillium marneffei
is increasingly seen among patients with AIDS in southeast Asian countries. Previous studies have demonstrated the presence of humoral immune responses to this fungus in patient sera; we have confirmed this work using sera from
P. marneffei
-infected patients (
n
= 21) to develop Western blots of
P. marneffei
cytoplasmic yeast antigen (CYA).
P. marneffei
CYA was then partially purified by liquid isoelectric focusing, and fractions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Immunoenzyme development of the Western blots with pooled sera from patients with
P. marneffei
infection and with pooled sera from patients with aspergillosis (
n
= 20), candidiasis (
n
= 10), cryptococcosis (
n
= 9), and histoplasmosis (
n
= 11) revealed three antigens with relative molecular masses of 61, 54, and 50 kDa. These antigens were specifically recognized by the pooled sera from the
P. marneffei
-infected patients. The 61- and 54-kDa antigens were subsequently purified to homogeneity by preparative gel electrophoresis, and the 50-kDa antigen was partially purified by the same technique. N-terminal amino acid sequencing revealed that the 61-kDa antigen had a strong homology (87% identity) with the antioxidant enzyme catalase. The three antigens were then subjected to SDS-PAGE and Western blotting and to immunoenzyme development with individual patient sera; sera from 86% of
P. marneffei
-infected patients recognized the 61-kDa antigen, sera from 71% recognized the 54-kDa antigen, and sera from 48% recognized the 50-kDa antigen. These specifically recognized antigens are the first to be purified from
P. marneffei
and can be used either singly or in combination to detect antibody responses in a large percentage of individuals infected with
P. marneffei.
Two-Step PCR-Based Assay for Identification of Bacterial Etiology of Otitis Media with Effusion in Infected Lebanese Children
Authors: Matar Ghassan M. , Sidani Nada , Fayad Michel , Hadi Usamah 
Published: December 2019

ABSTRACT

We developed and evaluated a two-step PCR-based assay with universal primers and genus- or species-specific primers for the detection of the most prevalent bacterial etiologies of otitis media with effusion (OME) in children from Lebanese hospitals. These etiologies included
Haemophilus
,
Streptococcus
, and
Moraxella
(
Branhamella
)
catarrhalis
, which were detected in middle-ear effusion (MEE) samples taken from children with OME. A total of 47 MEE samples were aspirated from 36 patients during insertion of a tympanostomy tube performed particularly for OME. The duration of effusion in all patients was ≥2 months. DNA was extracted from MEE samples, and PCR was initially done with DNA extracts by using the universal primers RW01 and DG74, which flank an ∼370-bp fragment found in the 16S rRNA gene of all bacterial species. For the identification of specific bacteria, we used in three separate reaction mixtures the following genus- or species-specific primers: (i) a
Haemophilus
-specific probe (probe RDR125) as a primer along with DG74, (ii) a
Streptococcus
-specific primer (primer STR1; designed by us) along with DG74, and (iii) an
M. catarrhalis
-specific primer pair (primer pair MCA1-MCA2). Thirty-five MEE samples (74.5%) gave the expected 370-bp band, indicating the presence of bacterial DNA in the tested samples. Of the 35 PCR-positive samples tested, 33 (94.3%) were positive for
Haemophilus
, 3 (8.6%) were positive for
Streptococcus
, and 10 (28.6%) were positive for
M. catarrhalis
. Ten samples (28.6%) exhibited a mixed infection and were positive for both
Haemophilus
and
M. catarrhalis
. Culture was simultaneously performed for all 47 MEE samples. Ten of the 47 MEE samples (21.3%) exhibited bacterial growth. These 10 were PCR positive for bacterial DNA. The remaining 25 PCR-positive samples were negative by culture, thus showing about 53% discordance between PCR results and those of culture. The PCR assay proved to be more sensitive than culture, more rapid, less cumbersome, and more cost-effective than the available PCR-Southern hybridization-based assays.