bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antimicrobial Susceptibility Testing of
Acinetobacter
spp. by NCCLS Broth Microdilution and Disk Diffusion Methods
Authors: Swenson Jana M. , Killgore George E. , Tenover Fred C. 
Published: November 2004

ABSTRACT

Although both broth microdilution (BMD) and disk diffusion (DD) are listed by NCCLS as acceptable methods for testing
Acinetobacter
spp. for antimicrobial susceptibility, few studies have compared the results generated by the two methods. We tested 196 isolates of
Acinetobacter
spp. from nine U.S. hospitals and from the Centers for Disease Control culture collection by using BMD and DD and clinically appropriate antimicrobial agents. Categorical results for amikacin, ciprofloxacin, gatifloxacin, gentamicin, imipenem, levofloxacin, meropenem, tobramycin, and trimethoprim-sulfamethoxazole were comparable for the two methods: there was only one very major (VM) error, with tobramycin, and only one major (M) error, with meropenem, when DD results were compared with BMD results. However, VM errors were frequent with the β-lactams and β-lactam-β-lactam inhibitor combinations, while M errors were often observed with tetracyclines. For BMD, tests frequently exhibited subtle growth patterns that were difficult to interpret, especially for β-lactams. If subtle growth (i.e., granular, small button, or “starry” growth) was considered positive, error rates between BMD and DD were unacceptably high for ampicillin-sulbactam (VM error, 9.8%; minor [m] error, 16.1%), piperacillin (VM error, 5.7%; m error, 13.5%), piperacillin-tazobactam (VM error, 9.3%; m error, 12.9%), ceftazidime (VM error, 6.2%; m error, 11.4%), cefepime (VM error, 6.2%; m error, 13.0%), cefotaxime (m error, 21.2%), ceftriaxone (m error, 23.3%), tetracycline (M error, 11.4%; m error, 32.1%), and doxycycline (M error, 2.6%). When subtle growth patterns were ignored, the agreement still did not achieve acceptable levels. To determine if the problems with BMD testing occurred in other laboratories, we sent frozen BMD panels containing β-lactam drugs and nine isolates to six labs with experience in performing BMD and DD. Among these laboratories, cefepime MICs ranged from ≤8 to ≥32 μg/ml for four of the nine strains, confirming the problem in interpreting BMD results. Discrepancies between the categorical interpretations of BMD and DD tests were noted primarily with cefepime and piperacillin, for which the BMD results were typically more resistant. Clinical laboratories should be aware of these discrepancies. At present, there are no data to indicate which method provides more clinically relevant information.
Evaluation of a Candidate International Standard Preparation for Human Anti-
Toxoplasma
Immunoglobulin G
Authors: Rigsby Peter , Rijpkema Sjoerd , Guy Edward C. , Francis Janet , Gaines Das Rose 
Published: November 2004

ABSTRACT

A freeze-dried human serum preparation containing immunoglobulin G (IgG) to
Toxoplasma gondii
was assessed for its suitability as an international reference reagent in an international collaborative study by 24 laboratories from 17 countries. This candidate standard was compared with the third international standard (IS) for human anti-
Toxoplasma
serum, TOXM, with the previous second IS, TOXS, and with a range of other serum samples. Samples were tested with the Sabin-Feldman dye test and a range of agglutination assays and enzyme immunoassays. This study emphasizes the need for appropriate standards if intermethod agreement of estimates is to be achieved. On the basis of the results of this study, the preparation was established by the World Health Organization as the first IS for human anti-
Toxoplasma
IgG, with an assigned potency of 20 IU per ampoule of total anti-
Toxoplasma
antibodies.
Emergence, Spread, and Characterization of Phage Variants of Epidemic Methicillin-Resistant
Staphylococcus aureus
16 in England and Wales
Authors: Murchan S. , Aucken H. M. , O'Neill G. L. , Ganner M. , Cookson B. D. 
Published: November 2004

ABSTRACT

Epidemic methicillin-resistant
Staphylococcus aureus
16 (EMRSA-16) and EMRSA-15 are the two most important and prevalent EMRSA strains found in the United Kingdom and have also been found in a number of European countries and the United States. We describe for the first time the spread of an EMRSA strain (EMRSA-16) from its point of origin in one hospital to the surrounding hospitals and regions over the following 2 years. In the first 18 months after its original appearance, 136 hospitals referred EMRSA-16 isolates for typing, and interhospital and intraregional spread were reported: it was more prevalent in males between 60 and 80 years old and was isolated from sputum and throat more often than EMRSA-15. Important characteristics, e.g., carriage of the enterotoxin A (
sea
) and toxic shock syndrome toxin (
tst
) genes and production of urease, are described. Phage-variant strains of EMRSA-16 which share some of the characteristics of the classical strain, including toxin carriage and urease production, emerged, but without genotypic investigations, their relationship could only be inferred. A total of 129 clinical isolates from 52 hospitals, collected between March 1998 and April 1999 and representing classical EMRSA-16 (49 isolates) or phage variants (80 isolates), were compared by phage typing, pulsed-field gel electrophoresis (PFGE) following SmaI macrorestriction, antimicrobial susceptibility testing, urease production, and PCR detection of toxin gene carriage. PFGE analysis revealed 29 profiles, A1 to A29, with A1 representing the prototypic strain, NCTC 13143. All other profiles differed from A1 by 1 to 6 bands, but some differed from each other by up to 10 bands.
Effects of Intrapartum Penicillin Prophylaxis on Intestinal Bacterial Colonization in Infants
Authors: Jauréguy Françoise , Carton Mathieu , Panel Pierre , Foucaud Pierre , Butel Marie-José , Doucet-Populaire Florence 
Published: November 2004

ABSTRACT

Early-onset group B streptococcal (GBS) infections remain a leading cause of morbidity and mortality in infants. To prevent the vertical transmission of GBS and neonatal GBS infection, guidelines recommend intrapartum penicillin or amoxicillin prophylaxis. This intrapartum antibiotic prophylaxis (IAP) is suspected to favor colonization by antibiotic-resistant bacteria. However, the effects of this prophylaxis on the patterns of acquisition of gastrointestinal bacterial flora in infants have never been studied. We collected stool samples from 3-day-old infants born to mothers who received intrapartum amoxicillin (antibiotic-exposed group;
n
= 25) and to untreated mothers (non-antibiotic-exposed group;
n
= 25). The groups were matched for factors known to affect intestinal microbial colonization: gestational age, type of delivery, and type of feeding. Qualitative and quantitative differential analyses of the bacterial flora in stool samples were performed. Similar numbers of infants in the non-antibiotic-exposed and antibiotic-exposed groups were colonized by aerobic bacteria and amoxicillin-resistant enterobacteria (75 and 77%, respectively) (
P
= 0.79). In contrast, significantly fewer infants in the antibiotic-exposed group than in the non-antibiotic-exposed group were colonized by anaerobic bacteria, especially
Clostridium
(12 and 40%, respectively) (
P
< 0.05). Regarding intestinal bacterial colonization, the differences between antibiotic-exposed and non-antibiotic-exposed infants were remarkably few. The only statistically significant effect was the reduced initial bacterial colonization by
Clostridium
in the antibiotic-exposed group. In our study, the use of IAP did not favor colonization by β-lactam-resistant bacteria. However, further evaluations are required to highlight the potential risks of the widespread use of antibiotics to prevent early-onset GBS infection.
Genetic Recombination between Two Genotypes of Genogroup III Bovine Noroviruses (BoNVs) and Capsid Sequence Diversity among BoNVs and Nebraska-Like Bovine Enteric Caliciviruses
Authors: Han Myung Guk , Smiley Jeffrey R. , Thomas Christopher , Saif Linda J. 
Published: November 2004

ABSTRACT

To determine the genogroups and genotypes of bovine enteric caliciviruses (BECVs) circulating in calves, we determined the complete capsid gene sequences of 21 BECVs. The nucleotide and predicted amino acid sequences were compared phylogenetically with those of known human and animal enteric caliciviruses. Based on these analyses, 15 BECVs belonged to
Norovirus
genogroup III and genotype 2 (GIII/2) and were genetically distinct from human
Norovirus
GI and GII. Six BECVs had capsid gene sequences similar to that of the unclassified Nebraska (NB)-like BECV. The 15 bovine noroviruses (BoNVs) were more closely related to Bo/NLV/Newbury-2/76/UK (GIII/2) and other known genotype 2 BoNVs than to genotype 1 Bo/NLV/Jena/80/DE. The BoNV Bo/CV521-OH/02/US showed high nucleotide and amino acid identities (84 and 94%, respectively) with the capsid gene of Bo/NLV/Newbury-2/76/UK, whereas the nucleotide and amino acid sequences of the RNA polymerase gene were more closely related to those of Bo/NLV/Jena/80/DE (77 and 87% identities, respectively) than to those of Bo/NLV/Newbury-2/76/UK (69 and 69% identities, respectively), suggesting that Bo/CV521-OH/02/US is a genotype 1-2 recombinant. Gene conversion analysis by the recombinant identification program and SimPlot also predicted that Bo/CV521-OH/02/US was a recombinant. Six NB-like BECVs shared 88 to 92% nucleotide and 94 to 99.5% amino acid identities with the NB BECV in the capsid gene. The results of this study demonstrate genetic diversity in the capsid genes of BECVs circulating in Ohio veal calves, provide new data for coinfections with distinct BECV genotypes or genogroups, and describe the first natural BoNV genotype 1-2 recombinant, analogous to the previously reported human norovirus recombinants.
Validation of the Use of Middlebrook 7H10 Agar, BACTEC MGIT 960, and BACTEC 460 12B Media for Testing the Susceptibility of
Mycobacterium tuberculosis
to Levofloxacin
Authors: Sanders Cynthia A. , Nieda Rachel R. , Desmond Edward P. 
Published: November 2004

ABSTRACT

Levofloxacin, the active
l
-isomer of the quinolone ofloxacin, is now widely accepted for treatment of multidrug-resistant tuberculosis. Because the drug is now widely used, we sought to establish susceptibility test conditions for
Mycobacterium tuberculosis
against levofloxacin by the traditional reference method, agar proportion (AP), the commonly used BACTEC 460 radiometric system, and the newer BACTEC MGIT 960 method. To determine the stability of levofloxacin in the two newer test systems (BACTEC 460 and BACTEC MGIT 960), media containing subinhibitory levels of levofloxacin were prepared and stored at 4 and 37°C for 14 days. The stored media were inoculated with H37Rv, and the drug activity was compared to freshly prepared media. Results show that levofloxacin is stable over the course of testing. Next, optimum levofloxacin test concentrations were determined for AP, BACTEC 460, and BACTEC MGIT 960 methods. MICs were determined for 32 pan-susceptible isolates of
M. tuberculosis
obtained from presumably untreated patients and 14 quinolone-resistant isolates. The levofloxacin-resistant strains either were isolated from patients who remained culture-positive despite treatment with a quinolone agent (six strains) or contained known mutations in
gyrA
(eight strains). Levofloxacin MICs resulted in a bimodal pattern with values for resistant strains consistently higher than those for pan-susceptible strains. Results show that levofloxacin concentrations of 2 μg/ml (BACTEC 460 and BACTEC MGIT 960) and 1 μg/ml (AP) inhibited the growth of all pan-susceptible strains while permitting the growth of all levofloxacin-resistant strains. Confirmatory tests with a subset of pan-susceptible and levofloxacin-resistant isolates validated the selected test concentrations.
Type III Secretion Phenotypes of
Pseudomonas aeruginosa
Strains Change during Infection of Individuals with Cystic Fibrosis
Authors: Jain Manu , Ramirez Daniel , Seshadri Roopa , Cullina Joanne F. , Powers Catherine A. , Schulert Grant S. , Bar-Meir Maskit , Sullivan Christine L. , McColley Susanna A. , Hauser Alan R. 
Published: November 2004

ABSTRACT

Pseudomonas aeruginosa
is a frequent cause of respiratory exacerbations in individuals with cystic fibrosis. An important virulence determinant of this pathogen is its type III protein secretion system. In this study, the type III secretion properties of 435
P. aeruginosa
respiratory isolates from 56 chronically infected individuals with cystic fibrosis were investigated. Although it had been previously reported that 75 to 90% of
P. aeruginosa
isolates from patients with hospital-acquired pneumonia secreted type III proteins, only 12% of isolates from cystic fibrosis patients did so, with nearly all of these isolates secreting ExoS and ExoT but not ExoU. Despite the low overall prevalence of type III protein-secreting isolates, at least one secreting isolate was cultured from one-third of cystic fibrosis patients. Interestingly, the fraction of cystic fibrosis patient isolates capable of secreting type III proteins decreased with duration of infection. Although 90% of isolates from the environment, the presumed reservoir for the majority of
P. aeruginosa
strains that infect patients with cystic fibrosis, secreted type III proteins, only 49% of isolates from newly infected children, 18% of isolates from chronically infected children, and 4% of isolates from chronically infected adults with cystic fibrosis secreted these proteins. Within individual patients, isolates of clonal origin differed in their secretion phenotypes, indicating that as strains persisted in cystic fibrosis patient airways, their type III protein secretion properties changed. Together, these findings indicate that following infection of cystic fibrosis patient airways,
P. aeruginosa
strains gradually change from a type III protein secretion-positive phenotype to a secretion-negative phenotype.
Usefulness of Routine Epicardial Pacing Wire Culture for Early Prediction of Poststernotomy Mediastinitis
Authors: Mekontso-Dessap Armand , Honoré Stéphanie , Kirsch Matthias , Houël Rémi , Loisance Daniel , Brun-Buisson Christian 
Published: November 2004

ABSTRACT

Poststernotomy mediastinitis (PSM) is one of the most serious complications of cardiac surgery, and its associated morbidity and mortality demand early recognition for emergency therapy. In this study, we investigated the usefulness of epicardial pacing wire (EPW) cultures for the prediction of PSM. Among 2,200 patients who underwent a cardiac surgical procedure at our hospital between 1 January 1999 and 31 December 2001, 82 (3.7%) had PSM;
Staphylococcus aureus
was the organism (45.1%) most frequently isolated at the time of surgical debridement. EPWs from 1,607 (73.0%) patients, 73 (4.5%) of whom developed PSM, were cultured. EPW cultures from 466 (29.0%) were positive, most often (74.9%) for coagulase-negative
Staphylococci
. EPW cultures were truly positive in 26 cases, truly negative in 1,106 cases, falsely positive in 428 cases, and falsely negative in 47 cases (with sterile cultures in 35 cases and a culture positive for an organism different from that isolated at the time of debridement in 12 cases). EPW culture had a positive predictive value of only 5.7% and a high negative predictive value (95.9%) for the diagnosis of PSM, with an accuracy of 70.4%. However, the likelihood ratio of positive (1.27) and negative (0.89) tests indicated only small changes in pretest-to-posttest probability. Therefore, a strategy of routine culture of EPWs to predict PSM seems questionable.
Selective Discrimination of
Listeria monocytogenes
Epidemic Strains by a Mixed-Genome DNA Microarray Compared to Discrimination by Pulsed-Field Gel Electrophoresis, Ribotyping, and Multilocus Sequence Typing
Authors: Borucki Monica K. , Kim So Hyun , Call Douglas R. , Smole Sandra C. , Pagotto Franco 
Published: November 2004

ABSTRACT

Listeria monocytogenes
can cause serious illness in humans, and subsequent epidemiological investigation requires molecular characterization to allow the identification of specific isolates.
L. monocytogenes
is usually characterized by serotyping and is subtyped by using pulsed-field gel electrophoresis (PFGE) or ribotyping. DNA microarrays provide an alternative means to resolve genetic differences among isolates, and unlike PFGE and ribotyping, microarrays can be used to identify specific genes associated with strains of interest. Twenty strains of
L. monocytogenes
representing six serovars were used to generate a shotgun library, and subsequently a 629-probe microarray was constructed by using features that included only potentially polymorphic gene probe sequences. Fifty-two strains of
L. monocytogenes
were genotyped by using the condensed array, including strains associated with five major listeriosis epidemics. Cluster analysis of the microarray data grouped strains according to phylogenetic lineage and serotype. Most epidemiologically linked strains were grouped together, and subtyping resolution was the same as that with PFGE (using AscI and ApaI) and better than that with multilocus sequence typing (using six housekeeping genes) and ribotyping. Additionally, a majority of epidemic strains were grouped together within phylogenetic Division I. This epidemic cluster was clearly distinct from the two other Division I clusters, which encompassed primarily sporadic and environmental strains. Discriminant function analysis allowed identification of 22 probes from the mixed-genome array that distinguish serotypes and subtypes, including several potential markers that were distinct for the epidemic cluster. Many of the subtype-specific genes encode proteins that likely confer survival advantages in the environment and/or host.
Characterization of “
Candidatus
Piscichlamydia salmonis” (Order
Chlamydiales
), a Chlamydia-Like Bacterium Associated With Epitheliocystis in Farmed Atlantic Salmon (
Salmo salar
)
Authors: Draghi Andrew , Popov Vsevolod L. , Kahl Melissa M. , Stanton James B. , Brown Corrie C. , Tsongalis Gregory J. , West A. Brian , Frasca Salvatore 
Published: November 2004

ABSTRACT

To characterize intracellular gram-negative bacteria associated with epitheliocystis in farmed Atlantic salmon (
Salmo salar
), gills with proliferative lesions were collected for histopathology, conventional transmission and immunoelectron microscopy, in situ hybridization, and DNA extraction during epitheliocystis outbreaks in Ireland and Norway in 1999 and 2000, respectively, and compared by ultrastructure and immunoreactivity to nonproliferative gills from Ireland archived in 1995. Genomic DNA from proliferative gills was used to amplify 16S ribosomal DNA (rDNA) for molecular phylogenetic analyses. Epitheliocystis inclusions from proliferative gills possessed variably elongate reticulate bodies, examples of binary fission, and vacuolated and nonvacuolated intermediate bodies, whereas inclusions in nonproliferative gills had typical chlamydial developmental stages plus distinctive head-and-tail cells. Immunogold processing using anti-chlamydial lipopolysaccharide antibody labeled reticulate bodies from proliferative and nonproliferative gills. 16S rDNA amplified directly from Irish (1999) and Norwegian (2000) gill samples demonstrated 99% nucleotide identity, and riboprobes transcribed from cloned near-full-length 16S rDNA amplicons from Norwegian gills hybridized with inclusions in proliferative lesions from Irish (1999) and Norwegian (2000) sections. A 1,487-bp consensus 16S rRNA gene sequence representing the chlamydia-like bacterium (CLB) from proliferative gills had the highest percent nucleotide identity with endosymbionts of
Acanthamoeba
spp. (order
Chlamydiales
). Molecular phylogenetic relationships inferred from 16S rRNA gene sequences using distance and parsimony indicated that the CLB from proliferative gills branched with members of the order
Chlamydiales
. “
Candidatus
Piscichlamydia salmonis” is proposed for the CLB associated with epitheliocystis from proliferative gills of Atlantic salmon, which exhibits developmental stages different from those identified in nonproliferative gills.
Mapping of Antigenic Sites on the Nucleocapsid Protein of the Severe Acute Respiratory Syndrome Coronavirus
Authors: He Yuxian , Zhou Yusen , Wu Hao , Kou Zhihua , Liu Shuwen , Jiang Shibo 
Published: November 2004

ABSTRACT
Antigenic sites on the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) were mapped by Pepscan analysis with overlapping peptides that span the N protein sequence. Two major immunodominant epitopes located in the C-terminal region (amino acids [aa] 362 to 412) and middle region (aa 153 to 178) reacted with more than 75% of sera from SARS patients. Several minor immunodominant epitopes were reactive with about 50% of the SARS sera. Antisera from mice immunized with inactivated SARS-CoV recognized the two major immunodominant epitopes and one antigenic site located adjacent to the N-terminal region (aa 76 to 101), which did not react with the sera from SARS patients. Several monoclonal antibodies against SARS-CoV bound to the N- or C-terminal antigenic sites. These results suggest that the above antigenic sites on the N protein are important in eliciting humoral immune response against SARS-CoV in humans and animals and can be used as antigens for developing diagnostic tests.
Phaeohyphomycotic Soft Tissue Infections Caused by the Coelomycetous Fungus
Microsphaeropsis arundinis
Authors: Pendle Stella , Weeks Kerry , Priest Michael , Gill Anthony , Hudson Bernard , Kotsiou George , Pritchard Robert 
Published: November 2004

ABSTRACT

Microsphaeropsis arundinis
is an anamorphic fungal plant inhabitant belonging to the form class
Coelomycetes
. We describe two cases of
M. arundinis
soft tissue infections in immunosuppressed patients. This organism has not previously been described as causing disease in humans. It was identified on the basis of its typical ostiolate pycnidial conidiomata, ampulliform conidiogenous cells, and small, smooth-walled, brown, cylindrical conidia.
Molecular Detection of Linezolid Resistance in
Enterococcus faecium
and
Enterococcus faecalis
by Use of 5′ Nuclease Real-Time PCR Compared to a Modified Classical Approach
Authors: Werner Guido , Strommenger Birgit , Klare Ingo , Witte Wolfgang 
Published: November 2004

ABSTRACT

A nucleotide transversion from guanine to uracil in the 23S rRNA confers linezolid resistance. We describe a real-time PCR using two Taqman probes that detects a single mutated allele among the genomes of
Enterococcus faecium
and
Enterococcus faecalis
. Results were confirmed by a classical approach involving LabChip technology assayed with an Agilent Bioanalyzer 2100.
Convenience of Serum for Visceral Leishmaniasis Diagnosis by PCR
Authors: Fissore Cristel , Delaunay Pascal , Ferrua Bernard , Rosenthal Eric , Del Giudice Pascal , Aufeuvre Jean-Pierre , Le Fichoux Yves , Marty Pierre 
Published: November 2004

ABSTRACT
In this retrospective study, the usefulness of a PCR performed on serum for primary diagnosis and monitoring of Mediterranean visceral leishmaniasis (MVL) was assessed. In the case of primary diagnosis of MVL, the serum PCR showed a sensitivity of 97% and a specificity of 95%, with positive and negative predictive values of 94 and 97%, respectively.
Utility of Molecular Identification in Opportunistic Mycotic Infections: a Case of Cutaneous
Alternaria infectoria
Infection in a Cardiac Transplant Recipient
Authors: Lo Cascio G. , Ligozzi M. , Maccacaro L. , Fontana R. 
Published: November 2004

ABSTRACT

We report on a case of cutaneous infection caused by
Alternaria infectoria
in a cardiac transplant recipient. A rapid molecular diagnosis was obtained by sequence analysis of the internal transcribed spacer domain of the 5.8S ribosomal DNA region amplified from colonies developed on Sabouraud medium. Treatment consisted of a combination of systemic antifungal therapy, first with amphotericin B and then with itraconazole.