bluuebunny/crossref_metadata_2025_split huggingface dataset:
First Identification of Autochthonous
Cryptococcus neoformans
var.
gattii
Isolated from Goats with Predominantly Severe Pulmonary Disease in Spain
Authors: Baró Teresa , Torres-Rodríguez Josep M. , De Mendoza Miguel Hermoso , Morera Yolanda , Alía Concepción 
Published: December 2019

ABSTRACT

Cryptococcus neoformans
var.
gattii
is associated with
Eucalyptus
trees growing in various tropical and subtropical regions of the world. The identification of 13 autochthonous strains of
C. neoformans
var.
gattii
in Spain is reported. These strains were isolated from lung (10 samples), liver (1 sample), and brain (2 samples) tissue specimens from six goats suffering from predominantly severe pulmonary disease that were autopsied. The animals were members of five different herds of goats grazing in rural areas of the province of Cáceres (Extremadura, Spain). Between 1990 and 1994, there were five outbreaks, in which between 2.5 and 12% of the goats were affected. Although respiratory symptoms (pneumonia) associated with cachexia were the predominant clinical picture in all outbreaks, brain and liver involvement was also documented in three of the five outbreaks. Biotyping was performed by culturing the isolates on
l
-canavanine-glycine-bromothymol blue medium and testing them for the assimilation of
d
-proline and
d
-tryptophan. Serotyping by agglutination tests confirmed the characterization of all strains as
C. neoformans
var.
gattii
serotype B. This is the first confirmation of the presence of this variety in Spain, with a peculiar ability to produce severe pulmonary and systemic disease in normal goats, particularly in the form of outbreaks of pneumonia in association with cachexia.
Development and Evaluation of a PCR-Based Assay for Detection of
Haemobartonella felis
in Cats and Differentiation of
H. felis
from Related Bacteria by Restriction Fragment Length Polymorphism Analysis
Authors: Messick Joanne B. , Berent Linda M. , Cooper Sandra K. 
Published: December 2019

ABSTRACT

The 16S rRNA gene of
Haemobartonella felis
was amplified by using universal eubacterial primers and was subsequently cloned and sequenced. Based on this sequence data, we designed a set of
H. felis
-specific primers. These primers selectively amplified a 1,316-bp DNA fragment of the 16S rRNA gene of
H. felis
from each of four experimentally infected cats at peak parasitemia. No PCR product was amplified from purified DNA of
Eperythrozoon suis
,
Mycoplasma genitalium
, and
Bartonella bacilliformis
. Blood from the experimental cats prior to infection was negative for PCR products and was greatly diminished or absent 1 month after doxycycline treatment. The overall sequence identity of this fragment varied by less than 1.0% among experimentally infected cats. By taking into consideration the secondary structure of the 16S rRNA molecule, we were able to further verify the alignment of nucleotides and quality of our sequence data. In this PCR assay, the minimum detectable number of
H. felis
organisms was determined to be between 50 and 704. The potential usefulness of restriction enzymes
Dde
I and
Mnl
I for distinguishing
H. felis
from closely related bacteria was examined. This is the first report of the utility of PCR-facilitated diagnosis and discrimination of
H. felis
infection in cats.
Separation among Species of
Mycobacterium terrae
Complex by Lipid Analyses: Comparison with Biochemical Tests and 16S rRNA Sequencing
Authors: Torkko Pirjo , Suutari Merja , Suomalainen Sini , Paulin Lars , Larsson Lennart , Katila Marja-Leena 
Published: December 2019

ABSTRACT

Fatty acids, alcohols, and mycolic acid cleavage products were determined for 13 ATCC strains and 24 clinical isolates, which were initially identified by biochemical and growth characteristics as the
Mycobacterium terrae
complex. The clinical isolates were also analyzed by partial sequencing of the 16S rRNA gene, which divided them into five genetic entities,
M. triviale
(three strains),
M. terrae
(four strains),
M. nonchromogenicum
sensu stricto (seven strains),
Mycobacterium
sp. strain MCRO 6 (seven strains), and
Mycobacterium
sp. strain 31958 (one strain). After acidic methanolysis, secondary alcohols were a characteristic feature in all members of the
M. terrae
complex but
M. triviale
. In addition to the prominent secondary alcohols, 2-octadecanol and 2-eicosanol, two previously unidentified alcohols, 2-(8,15-dimethyl)docosenol and 2-(8,17-dimethyl)tetracosenol, were detected in
M. nonchromogenicum
,
Mycobacterium
sp. strain MCRO 6, and
Mycobacterium
sp. strain 31958. Only 2-(8,17-dimethyl)tetracosenol was detected in trace amounts in
M. terrae
. Genetic differences were associated with differences in phenotypic characteristics, including growth at 42°C and pyrazinamidase production. Based on fatty acid and alcohol composition and biochemical and genetic characteristics,
M. nonchromogenicum
and
Mycobacterium
sp. strains MCRO 6 and 31958 were found to be a closely related group, named the
M. nonchromogenicum
complex. Detected genetic variations associated with phenotypic characteristics may indicate further species separation of this complex. In conclusion, the results of gas-liquid chromatography fatty acid analysis, combined with those of a Tween 80 test, enable identification of the species of the
M. terrae
complex and their separation from other nonpigmented slowly growing mycobacteria.
Evolution of an Endemic Methicillin-Resistant
Staphylococcus aureus
Population in an Australian Hospital from 1967 to 1996
Authors: Givney Rodney , Vickery Alison , Holliday Anne , Pegler Mary , Benn Richard 
Published: December 2019

ABSTRACT

The evolution over 30 years of a population of methicillin-resistant
Staphylococcus aureus
(MRSA) from a tertiary referral hospital was studied by phylogenetic analysis of
Sma
I-generated restriction fragment length polymorphisms (RFLPs). The results suggest that a new clone of MRSA appeared at the hospital in the early 1980s, which, although usually retaining its ancestral phage-type, developed four different RFLP pulsotypes in the next 16 years. This finding indicates that multiple RFLP patterns in MRSA do not necessarily represent multiple clones deriving from different
mec
gene transfer events. Such variation within a clone may be significant in the interpretation of RFLP patterns during outbreaks and emphasizes the need to use two typing methods in studies of such populations. Since the appearance of new clones of MRSA is a relatively rare event, cross-infection control is paramount in the prevention of MRSA dissemination.
Epidemiological Investigation of Vaginal
Saccharomyces cerevisiae
Isolates by a Genotypic Method
Authors: McCullough Michael J. , Clemons Karl V. , Farina Claudio , McCusker John H. , Stevens David A. 
Published: December 2019

ABSTRACT

Saccharomyces cerevisiae
is a ubiquitous, ascomycetous yeast, and vaginitis caused by this organism has been reported only very rarely. The aim of the present investigation was to assess the epidemiological relatedness of a group of vaginal and commercial
S. cerevisiae
isolates by a previously reported genetic typing method, which divided the isolates into two broad groups with numerous subtypes. Nineteen
S. cerevisiae
isolates obtained from patients suffering from vaginitis and four isolates from commercial products in the same city were analyzed. The cellular DNA from each isolate was digested with the restriction endonuclease
Eco
RI, and restriction fragment length polymorphisms were generated by horizontal gel electrophoresis. The results showed that although vaginal isolates did not cluster in any particular genetic subtype, multiple patients were infected with indistinguishable strains (there were nine distinct strains among 23 isolates). For two of three patients, all three with two episodes of
S. cerevisiae
vaginitis, different strains were isolated during the recurrence of this disease. Three other patients with indistinguishable isolates were epidemiologically related in that two were practitioners in the same clinic and the third was a patient at this clinic. We also found that one commercial strain was indistinguishable from the strain isolated from three different women at the time that they were suffering from vaginitis. The findings of the present study suggest that some
S. cerevisiae
strains may possess properties permitting persistence in the human host. Furthermore, person-to-person contact and the proliferation of the use of
S. cerevisiae
as a health-food product, in home baking, and in home brewing may be a contributing factor in human colonization and infection with this organism.
Fatal Pulmonary Infection with
Mycobacterium celatum
in an Apparently Immunocompetent Patient
Authors: Bux-Gewehr Irene , Hagen Hans P. , Rüsch-Gerdes Sabine , Feurle Gerhard E. 
Published: December 2019

ABSTRACT

Mycobacterium celatum
is a recently described mycobacterium isolated from patients who have suppressed cell-mediated immunity, such as AIDS. We present here, to our knowledge, the first report of a fatal pulmonary infection caused by
M. celatum
in a 73-year-old immunocompetent female patient. The mycobacterium was identified by a 16S rRNA sequence analysis.
Species-Specific and Ubiquitous-DNA-Based Assays for Rapid Identification ofStaphylococcus aureus
Authors: Martineau Francis , Picard François J. , Roy Paul H. , Ouellette Marc , Bergeron Michel G. 
Published: December 2019

ABSTRACTStaphylococcus aureusis the cause of serious infections in humans, including endocarditis, deep-seated abscesses, and bacteremia, which lead to toxic and septic shock syndromes. Rapid and direct identification of this bacterium specifically and ubiquitously directly from clinical specimens would be useful in improving the diagnosis ofS. aureusinfections in the clinical microbiology laboratory. A wide variety of kits based on biochemical characteristics efficiently identifyS. aureus, but the rapidity and the accuracy of each of these methods combined with testing of clinically relevant antibiotic resistance genes need to be improved. On the basis of hybridization assays with randomly selected clones from anS. aureusgenomic library, we have identified a chromosomal DNA fragment which is specific forS. aureusand which detected all 82S. aureusisolates tested. This 442-bp fragment was sequenced and was used to design a set of PCR amplification primers. The PCR assay was also specific and ubiquitous for the identification from bacterial cultures of 195 clinical strains ofS. aureusisolated from a variety of anatomical sites and obtained from hospitals throughout the world. The PCR assay that we have developed is simple and can be performed in about 1 h. This DNA-based test provides a novel diagnostic tool for the diagnosis ofS. aureusinfections.
Detection of Intimins α, β, γ, and δ, Four Intimin Derivatives Expressed by Attaching and Effacing Microbial Pathogens
Authors: Adu-Bobie Jeannette , Frankel Gad , Bain Christopher , Goncalves Azizedite Guedes , Trabulsi Luiz R. , Douce Gill , Knutton Stuart , Dougan Gordon 
Published: December 2019

ABSTRACT

Intimins are outer membrane proteins expressed by enteric bacterial pathogens capable of inducing intestinal attachment-and-effacement lesions. A eukaryotic cell-binding domain is located within a 280-amino-acid (Int280) carboxy terminus of intimin polypeptides. Polyclonal antiserum was raised against Int280 from enteropathogenic
Escherichia coli
(EPEC) serotypes O127:H6 and O114:H2 (anti-Int280-H6 and anti-Int280-H2, respectively), and Western blot analysis was used to explore the immunological relationship between the intimin polypeptides expressed by different clinical EPEC and enterohemorrhagic
E. coli
(EHEC) isolates, a rabbit diarrheagenic
E. coli
strain (RDEC-1), and
Citrobacter rodentium
. Anti-Int280-H6 serum reacted strongly with some EPEC serotypes, whereas anti-Int280-H2 serum reacted strongly with strains belonging to different EPEC and EHEC serotypes, RDEC-1, and
C. rodentium
. These observations were confirmed by using purified Int280 in an enzyme-linked immunosorbent assay and by immunogold and immunofluorescence labelling of whole bacterial cells. Some bacterial strains were recognized poorly by either antiserum (e.g., EPEC O86:H34 and EHEC O157:H7). By using PCR primers designed on the basis of the intimin-encoding
eae
gene sequences of serotype O127:H6, O114:H2, and O86:H34 EPEC and serotype O157:H7 EHEC, we could distinguish between different
eae
gene derivatives. Accordingly, the different intimin types were designated α, β, δ, and γ, respectively.
Discrimination of Strains of
Salmonella enteritidis
with Differing Levels of Virulence by an In Vitro Glass Adherence Test
Authors: Solano C. , Sesma B. , Alvarez M. , Humphrey T. J. , Thorns C. J. , Gamazo C. 
Published: December 2019

ABSTRACT

The objective of this study was the in vitro differentiation of isolates of
Salmonella enteritidis
whose virulences differed in a chick model. A total of 14 strains of
S. enteritidis
were isolated from either the environment, dairy products, or infected patients. The isolates could be divided into two groups on the basis of their virulence (50% lethal dose) in chickens infected intraperitoneally. When the strains were incubated in adherence test medium (Spanish patent 9700408), only the virulent strains produced aggregates and formed visible filaments attached to the glass tube. These results suggest, although for a limited number of strains, that aggregation in such a medium could be used as a diagnostic tool to discriminate virulent strains of
S. enteritidis.
Typhlitis Caused by Intestinal
Serpulina
-Like Bacteria in Domestic Guinea Pigs (
Cavia porcellus
)
Authors: Vanrobaeys Mia , De Herdt Peter , Ducatelle Richard , Devriese Luc A. , Charlier Gerard , Haesebrouck Freddy 
Published: December 2019

ABSTRACT

Between January 1992 and December 1996,
Serpulina
-like bacteria were demonstrated in intestinal tract lesions from 37 of 88 guinea pigs submitted to the University of Ghent in Ghent, Belgium, for necropsy because of disease and death from different unknown causes. All infected animals had a history of sudden death with minimal introductory clinical signs. Occasionally, they produced yellow, slimy feces or showed nervous signs, but the condition always had a fatal outcome within 24 h. When larger colonies of guinea pigs were involved, the disease spread very rapidly unless treatment with ronidazole was initiated. Lesions consisted of a catarrhal or hemorrhagic inflammation of the colon and cecum (typhlitis). Electron microscopy demonstrated the presence of large numbers of
Serpulina
-like organisms adhering to the cecal mucosae of these animals. Attempts to isolate the agents failed. The organisms did not stain by an immunofluorescence technique for the detection of
Serpulina hyodysenteriae
. The present data provide evidence that intestinal
Serpulina
-like organisms can be important as a cause of disease in guinea pigs.
U.S. Case Report of Cerebral Phaeohyphomycosis Caused byRamichloridium obovoideum (R. mackenziei): Criteria for Identification, Therapy, and Review of Other Known Dematiaceous Neurotropic Taxa
Authors: Sutton Deanna A. , Slifkin Malcolm , Yakulis Robert , Rinaldi Michael G. 
Published: December 2019

ABSTRACTWe report a case of cerebral phaeohyphomycosis in a 36-year-old male caused by the neurotropic fungusRamichloridium obovoideum(Matushima) de Hoog 1977 (Ramichloridium mackenzieiCampbell et Al-Hedaithy 1993). This man resided in the Middle East, where the fungus appears to be endemic and, possibly, geographically restricted, since all previous reports of brain abscesses due to this organism have been for patients indigenous to this area. As a servant of the Saudi Arabian royal family, he appeared in the United States seeking treatment for chronic weight loss, fatigue, decreased memory, and a more recent 2-week history of right-hand weakness which worsened to involve the entire right upper extremity. On the day prior to his admission, he had a focal motor seizure with rotation of the head and eyes to the right, followed by secondary generalization. A computerized tomogram showed a ring-enhancing hypodense lesion in the left parietal subcortical region with associated edema and mass effect. Diagnosis of a fungal etiology was made following a parietal craniotomy and excisional biopsy by observation of septate, dematiaceous hyphal elements 2 to 3 μm in width on hematoxylin-and-eosin-stained sections from within areas of inflammation and necrosis. Culture of the excised material grew out a dematiaceous mould which was subsequently identified asR. obovoideum. At two months postsurgery and with a regimen of 200 mg of itraconazole twice a day, the patient was doing well and returned to Saudi Arabia. His condition subsequently deteriorated, however, and following a 7-month course of itraconzole, he expired. We use this case to alert clinicians and personnel in clinical mycology laboratories of the pathogenicity of this organism and its potential occurrence in patients with central nervous system signs and symptoms who have resided in the Middle East and to review and/or compareR. obovoideumwith other neurotropic, dematiaceous taxa and similar nonneurotropic, dematiaceous species.
Practical Approach for Detection and Identification of OXA-10-Derived Ceftazidime-Hydrolyzing Extended-Spectrum β-Lactamases
Authors: Vahaboglu Haluk , Ozturk Recep , Akbal Huriye , Saribas Suat , Tansel Ozlem , Coşkunkan Figen 
Published: December 2019

ABSTRACT

A practical approach to detect and identify ceftazidime-hydrolyzing extended-spectrum mutants of OXA-10 β-lactamase is presented. Large numbers of bacteria were screened by colony hybridization, a 720-bp part of
bla
OXA
was amplified by PCR from the hybridization-positive isolates, and the products were digested by
Pvu
II and
Hae
III.
Comparison of Two Enzyme-Linked Immunosorbent Assays and One Rapid Immunoblot Assay for Detection of Herpes Simplex Virus Type 2-Specific Antibodies in Serum
Authors: Groen Jan , Van Dijk Grietje , Niesters Hubert G. M. , Van Der Meijden Willem I. , Osterhaus Albert D. M. E. 
Published: December 2019

ABSTRACT
The sensitivities and specificities of three immunoassays for the detection of herpes simplex virus type 2 (HSV-2)-specific immunoglobulin G antibodies in serum, including the one-strip rapid immunoblot assay (RIBA; Chiron Corporation) and two indirect enzyme immunosorbent assays (EIA; Gull Laboratories and Centocor), were compared by testing a panel of 1,250 serum samples from individuals attending an outpatient clinic for sexually transmitted diseases. A qualitative agreement among the three assays was observed with 1,080 serum samples (86.4%); 291 of the serum samples (23.3%) were positive, 789 samples (63.1%) were negative, and 170 serum samples (13.6%) gave a discordant result. Results were considered conclusive when a concordant result was obtained with two of three assays. The sensitivities and specificities of the RIBA, the Gull EIA, and the Centocor EIA proved to be 99.2, 99.7, and 89.9% and 97.1, 96.7, and 99.3%, respectively. These results indicate that the Chiron RIBA and the Gull EIA are especially useful and reliable for the detection of HSV-2-specific antibodies in serum.
Utility of Major Leukocyte Subpopulations for Monitoring Secondary Cytomegalovirus Infections in Renal-Allograft Recipients by PCR
Authors: Schäfer Peter , Tenschert Werner , Cremaschi Liana , Gutensohn Kai , Laufs Rainer 
Published: December 2019

ABSTRACT
The feasibility of the major peripheral blood leukocyte (PBL) subsets for use in qualitative and quantitative PCR to monitor secondary cytomegalovirus (CMV) infection and ganciclovir therapy was assessed with 188 blood samples derived from 40 CMV immunoglobulin G-positive renal-allograft recipients. In pp65 antigen-positive patients all leukocyte fractions, but only 79.5% of plasma preparations, were PCR positive. In pp65 antigen-negative samples from patients after antiviral treatment only 7.3% of polymorphonuclear cell (PMNL) samples, but 81.8% of peripheral blood mononuclear cells (PBMC), and 10.9% of plasma samples remained PCR positive. Similarly, in patients with latent infections only 5.0% of PMNL, but 51.7% of PBMC preparations, and 8.0% of plasma samples were PCR positive. Regarding patients with active CMV infection, CMV DNA copy numbers in PMNL correlated significantly with pp65 antigen-positive cell counts before and after onset of ganciclovir therapy. Significant differences in CMV DNA copy numbers in PMNL and plasma were observed (i) between patients with symptomatic infection and those with asymptomatic infection and (ii) between patients with active infection and those with latent infection. In contrast, PBMC harbored equally low CMV DNA levels both in patients with active infection and those with latent infections, and no decline of CMV DNA load in PBMC was observed during antiviral treatment. We conclude that detection of CMV DNA in PMNL, not in PBMC, is associated with active infections and is more sensitive than detection of CMV DNA in plasma. Negative PCR results for PMNL after antiviral therapy indicate recovery, and fewer unwanted positive results occur compared to PBMC and plasma. Therefore, purified PMNL should be preferred for analysis by qualitative CMV PCR to avoid unwanted positive results. The CMV DNA load in PBMC compared with that in PMNL is negligible during active infection, so mixed PBL are sufficient for use in quantitative PCR.
Detection and Identification of
Mycobacterium tuberculosis
Directly from Sputum Sediments by Ligase Chain Reaction
Authors: Moore Douglas F. , Curry Janis I. 
Published: December 2019

ABSTRACT

Sputum specimens received for the diagnosis of tuberculosis or other mycobacterial infections were tested by a ligase chain reaction (LCR)-based assay and acid-fast stain and culture techniques. Results from the LCR assay (Abbott LCx
Mycobacterium tuberculosis
[MTB] Assay) were compared to results from standard culture techniques held for 6 weeks. Four hundred ninety-three specimens from 205 patients suspected of pulmonary tuberculosis were included in the prospective study. Thirty-four (6.9%) of the specimens were culture positive for
M. tuberculosis
, and 13 (38%) of these were also fluorochrome stain positive. LCR sensitivities and specificities compared to culture were 74 and 98%, respectively. LCR sensitivity was 100% for fluorochrome stain-positive specimens and 57% for fluorochrome stain-negative specimens. Nine LCR-negative, culture-positive specimens were the result of low concentrations of
M. tuberculosis
. No inhibitors were detected in any of these specimens. Of the eight LCR-positive, culture-negative specimens, five were from patients with active tuberculosis. With these considered culture misses, final LCR sensitivity, specificity, positive predictive value, and negative predictive value were 77, 99, 91, and 98%, respectively. The same performance values for the fluorochrome acid-fast bacillus smear were 33, 98, 62, and 94%, respectively. After normal laboratory sputum processing, the Abbott LCx MTB Assay can be completed in 6 h. Thus, it is possible to have results available within 8 h of specimen submission.