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PubMed1976

[Indication of halprowia (chlamydia) antigens by the direct immunoperoxidase method].

The authors modified and approbated the direct immunoperoxidase method for the indication in the infected L cells of the halprowia (chlamydia) antigens--etiological agents of trachoma, ocular and urogenital forms of paratrachoma, arthritis, meningopneumonia and enzootic abortion of sheep. Different types of localization of the group-specific halprowia antigen were revealed in the cytoplasm of the affected cells by light microscopy; electron microscopy demonstrated localization of this antigen in the membrane of the cell wall of the elementary and initial bodies. Specificity of the test-system used, no lesser sensitivity than the sensitivity of direct immunofluorescent method of indication of the same microbies, with a possibility of detection of stably stained antigens by widely accessible light microscopy underly practical usefulness of direct immunoperoxidase method of the diagnosis of halprowiosis (chlamydiosis) of man and animals.

Antigens, Bacterial↗

PubMed1975

Bacillus subtilis GSY 1057 assay for aflatoxin B activation by rainbow trout (Salmo gairdneri).

A rapid and sensitive microbial assay was developed to detect lethal products of aflatoxin B metabolism by rainbow trout (Salmon gairdneri) Mt. Shasta strain. Bacillus subtilis GSY 1057 (hisA1, uvr-1, metB4), a DNA repair deficient strain, was incubated for 20 min in the 20,000 times g supernate from trout liver homogenates which had been preincubated for 10 min with various levels of aflatoxin B. Serial dilutions of the incubation mixture were plated in triplicate on tryptose blood agar base plates and colonies were counted after 12 hr at 37 degrees C. One mumole aflatoxin B in 3.2 ml incubation mixture reduced viability 60%.

Aflatoxins↗

PubMedDate not supplied

Natural propolis extract NIVCRISOL in the treatment of acute and chronic rhinopharyngitis in children.

In the work there are shown the results of a "case control" study carried out in a children collectivity (preschool children and school-children), regarding the action of an aqueous propolis extract, NIVCRISOL, in acute and chronic inflammatory diseases of the upper airways. The preparation, which had a rich content of flavonoids, was administered to a group of preschool children and school-children treated during the whole cold season 1994-1995. The monitoring of the subgroups investigated was performed by clinical observation of the health state and recording of the incidence of symptoms characteristic to acute or chronic rhinopharyngeal diseases, as well as by a periodical laboratory examination for the detection and characterization of viral, microbial and fungal germs carriage. The analysis of the data obtained pointed out the favourable effects of this local treatment, expressed by lowering of the number of cases with acute or chronic symptoms, and decrease and sometimes suppression of the viral-microbial flora carriage of the upper airways. These positive results, the good tolerance of the preparation, the advantages of the therapy with natural products and more economic criteria entitle us to propose the administration of this preparation as an adjuvant medication in the local treatment of some clinical forms of acute or chronic rhinopharyngeal diseases.

Acute Disease↗

PubMed2005

Evidence of a large novel gene pool associated with prokaryotic genomic islands.

Microbial genes that are "novel" (no detectable homologs in other species) have become of increasing interest as environmental sampling suggests that there are many more such novel genes in yet-to-be-cultured microorganisms. By analyzing known microbial genomic islands and prophages, we developed criteria for systematic identification of putative genomic islands (clusters of genes of probable horizontal origin in a prokaryotic genome) in 63 prokaryotic genomes, and then characterized the distribution of novel genes and other features. All but a few of the genomes examined contained significantly higher proportions of novel genes in their predicted genomic islands compared with the rest of their genome (Paired t test = 4.43E-14 to 1.27E-18, depending on method). Moreover, the reverse observation (i.e., higher proportions of novel genes outside of islands) never reached statistical significance in any organism examined. We show that this higher proportion of novel genes in predicted genomic islands is not due to less accurate gene prediction in genomic island regions, but likely reflects a genuine increase in novel genes in these regions for both bacteria and archaea. This represents the first comprehensive analysis of novel genes in prokaryotic genomic islands and provides clues regarding the origin of novel genes. Our collective results imply that there are different gene pools associated with recently horizontally transmitted genomic regions versus regions that are primarily vertically inherited. Moreover, there are more novel genes within the gene pool associated with genomic islands. Since genomic islands are frequently associated with a particular microbial adaptation, such as antibiotic resistance, pathogen virulence, or metal resistance, this suggests that microbes may have access to a larger "arsenal" of novel genes for adaptation than previously thought.

Bacteriophages↗

PubMed1994

Evaluation of affirm VP Microbial Identification Test for Gardnerella vaginalis and Trichomonas vaginalis.

A commercial system (Affirm VP Microbial Identification Test; MicroProbe Corp.) for detection of vaginal pathogens was evaluated with 176 consecutive women attending a sexually transmitted disease clinic for genital complaints. Vaginal swab specimens were used for culture of Gardnerella vaginalis and Trichomonas vaginalis, preparation of a vaginal smear for Gram stain interpretation, and wet mount evaluation. An additional swab was used to evaluate the 30-min nonisotopic oligonucleotide probe test. The automated probe system detected G. vaginalis in 69 (95%) of 73 women having > 5 x 10(5) CFU of G. vaginalis per ml by culture, and 20 (43%) of 47 specimens with < or = 5 x 10(5) CFU of G. vaginalis per ml. There were three false positives and four false negatives for the Affirm VP test compared with > 5 x 10(5) CFU of G. vaginalis per ml. The probe system detected G. vaginalis in 57 (90%) of 63 vaginal specimens from women having clue cells on wet mount examination, and in only 3 (3%) of 113 women without clue cells, suggesting that the Affirm probe for G. vaginalis could be used as a surrogate for wet mount examination for clue cells. The T. vaginalis probe was positive for 12 of 12 specimens positive by wet mount and 12 of 15 specimens positive by culture. There were no false positives and three false negatives for the Affirm VP test compared with culture and/or wet mount for T. vaginalis. The Affirm VP Microbial Identification System is a rapid, objective, and automated test for the detection of T. vaginalis and clinically significant levels of G. vaginalis that is comparable to wet mount examination for clue cells and is superior to wet mount examination for the detection of trichomonads.

Animals↗

PubMed2007

In vitro and in vivo effects of hydrolysates from conglycinin on intestinal microbial community of mice after Escherichia coli infection.

AIMS: To detect the effect of pepsin-hydrolysate conglycinin (PTC) on the growth of Escherichia coli O(138)in vitro, and investigate the effect of PTC on intestinal microbial community of mice after E. coli infection. METHODS AND RESULTS: Serial dilution method was used to detect the antibacterial activity of PTC in 96-well cell-cultivated plates. Fifty-five KM mice were randomly assigned to five groups: normal, feeding-E. coli control, HCl-full hydrolysis of conglycinin, conglycinin and PTC. Orally administrated with hydrolysates from conglycinin for 21 days, each mouse was fed with 2 x 10(8) CFU ml(-1) of E. coli O(138) on the 22nd day. The mice activities were monitored and polymerase chain reaction-denaturing gradient gel electrophoresis was used to analyse the microbial community in mice faeces. The results showed that PTC could inhibit growth of E. coli O(138) at nitrogen concentrations of more than 520 mg l(-1). There was high similarity of intestinal microbial community in mice between PTC and normal groups. CONCLUSION: PTC inhibits growth of E. coli O(138), keeps mice healthy following oral administration of E. coli infection and maintains a balanced active microbial community in their gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the antibacterial activity of PTC against E. coli and its ability to maintain healthy intestinal microbial community in mice even after they were infected with E. coli. This observation is significant in the application of PTC to prevent gastrointestinal diseases caused by E. coli and unbalanced intestinal microflora.

Animals↗

PubMed2003

Novel liquid chromatography-electrospray ionization mass spectrometry method for the quantification in human urine of microbial aromatic acid metabolites derived from dietary polyphenols.

An HPLC-ESI-MS-MS method was developed to quantify in human urine fourteen aromatic acids known as metabolites of dietary polyphenols. These metabolites were determined simultaneously in a single 20-min chromatographic analysis with multiple reaction monitoring detection. The inter- and intra-day precisions, calculated from quality control samples were 8.8 and 5.3%, respectively, and the mean accuracy was 2.3%. The method was tested on urine samples collected from one healthy volunteer who consumed a polyphenol-rich diet for 3 days. Increased levels of several aromatic acid metabolites were observed, demonstrating that the method can be used to detect changes in the excretion of microbial metabolites induced by the consumption of polyphenol-containing foods in humans.

Acids↗

PubMed2000

Monitoring the microbial contamination of beef carcass tissue with a rapid chromogenic Limulus amoebocyte lysate endpoint assay.

A chromogenic Limulus amoebocyte lysate (LAL) endpoint assay was found to be an accurate and rapid means of gauging levels of beef carcass microbial contamination within 10 min. The assay demonstrated a high correlation with the total mesophilic bacterial and coliform surface populations from inoculated beef carcass surface tissues. This assay was tested on a set of actual beef carcass surface samples (n = 121) demonstrating the utility of the chromogenic LAL test as a means of monitoring carcass microbial contamination in a near real-time fashion. Classifying the chromogenic LAL results into four contamination groups was found to be a sound means of utilizing the resultant chromogenic LAL data for detecting carcasses with high levels of microbial contamination. For beef carcass testing, this assay can be used with no instrumentation other than the required 37 degrees C incubator and, as an option, a microplate reader.

Abattoirs↗

PubMed2005

Effect of sdiA on biosensors of N-acylhomoserine lactones.

Many gram-negative bacteria synthesize N-acylhomoserine lactones (AHLs) and then use transcription factors of the LuxR family to sense and respond to AHL accumulation in the environment; this phenomenon is termed quorum sensing. Bacteria produce a variety of AHLs, and numerous bacterial reporter strains, or biosensors, that can detect subsets of these molecules have been constructed. Many of these are based on Escherichia coli because this species does not produce AHLs. However, both Escherichia and Salmonella spp. contain a LuxR homolog named SdiA that can detect exogenous AHL synthesized by other microbial species. In this study we have determined that sdiA of E. coli and Salmonella spp. can activate an RhlR-based biosensor plasmid in response to AHLs other than what the biosensor was designed to detect. SdiA does not activate LuxR-, LasR-, or AhyR-based biosensor plasmids, although the presence of sdiA in E. coli does interfere with the function of the AhyR-based biosensor. Because sdiA interferes with the function of two of the four reporters, we have constructed a set of E. coli biosensor strains that lack sdiA. The set includes control reporters that allow the luxR dependence of responses to be determined.

4-Butyrolactone↗

PubMed1983

Natural atmospheric microbial conditions in a typical suburban area.

Ambient outdoor concentrations and size distributions of airborne microbial particles were measured approximately weekly for 2 years in a Washington, D.C., suburban area. The study objective was to characterize microbial air quality in the vicinity of a proposed sewage sludge composting facility. During the study, 379 samples were taken at 17 stations, using Andersen microbial samplers. Concentration ranges (in viable particles per cubic meter) were as follows: airborne mesophilic fungi, 0 to 7,220 with a geometric mean of 273; thermophilic fungi, 0 to 193 with a median of 2.1; Aspergillus fumigatus, 0 to 71 with a median of 1.0; aerobic bacteria, 4.2 to 1,640 with a geometric mean of 79; and fecal streptococci, 0 to 5.7 with a median of 0. No fecal coliforms were recovered. The potentially respirable fraction (less than 8 microns) averaged 34% for total bacteria, 56% for mesophilic fungi, 91% for thermophilic fungi, and 95% for A. fumigatus. The specific sampling location was not a major factor affecting microbial particle concentrations or size distributions. Conversely, the time of year was an important determinant of viable particle concentrations for all groups of microorganisms studied. The highest concentrations were observed in summer and fall, with significantly lower levels detected in winter. In general, the microbial data did not correlate with other variables, including weather conditions, measured in this study.

Air Microbiology↗

PubMed1991

Near on-line detection of enteric bacteria using lux recombinant bacteriophage.

The potential for a revolution in microbial testing can be perceived with the near on-line detection of indicator microorganisms. By definition, these are microorganisms present in significant numbers within a food which, while not pathogenic, can be related through increasing count to the increased probability of pathogen contamination. We have used recombinant lux+ bacteriophage to detect enteric indicator bacteria without recovery or enrichment in 50 min, provided that they are present at levels greater than 10(4) g-1 or cm-2. After a 4-h enrichment, samples having enteric counts of 10 g-1 or cm-2 can be distinguished from background.

Animals↗

PubMed1998

Volatile compounds originating from mixed microbial cultures on building materials under various humidity conditions.

We examined growth of mixed microbial cultures (13 fungal species and one actinomycete species) and production of volatile compounds (VOCs) in typical building materials in outside walls, separating walls, and bathroom floors at various relative humidities (RHs) of air. Air samples from incubation chambers were adsorbed on Tenax TA and dinitrophenylhydrazine cartridges and were analyzed by thermal desorption-gas chromatography and high-performance liquid chromatography, respectively. Metabolic activity was measured by determining CO2 production, and microbial concentrations were determined by a dilution plate method. At 80 to 82% RH, CO2 production did not indicate that microbial activity occurred, and only 10% of the spores germinated, while slight increases in the concentrations of some VOCs were detected. All of the parameters showed that microbial activity occurred at 90 to 99% RH. The microbiological analyses revealed weak microbial growth even under drying conditions (32 to 33% RH). The main VOCs produced on the building materials studied were 3-methyl-1-butanol, 1-pentanol, 1-hexanol, and 1-octen-3-ol. In some cases fungal growth decreased aldehyde emissions. We found that various VOCs accompany microbial activity but that no single VOC is a reliable indicator of biocontamination in building materials.

Carbon Dioxide↗

PubMed1998

Detection of ethanol in a two-component glucose/ethanol mixture using a nonselective microbial sensor and a glucose enzyme electrode.

Chemometric theory was applied to a microbial sensor for determinations of ethanol in the presence of glucose. Microbial sensors, consisting of Gluconobacter oxydans cells immobilized on Clark-type amperometric oxygen electrodes, exhibited good sensitivity but low selectivity toward ethanol and glucose. An Eksan-G commercial glucose analyzer was used as a second sensor for multivariate calibration and analyses. Microbial sensors exhibited nearly complete additivity for total glucose plus ethanol concentrations from 0.0 to 0.6 mM. Within this linear range, chemometric analyses provided estimates of ethanol concentration with measurement errors of less than 8%. Multivariate calibration thus is a promising approach to enhance the usefulness of microbial sensors.

Acetobacteraceae↗

PubMed1999

Detection of antimicrobial substances in individual cow and quarter milk samples using Delvotest microbial inhibitor tests.

The use of antibiotic therapy to treat and prevent udder infections of cows during the dry period is a key component of mastitis control in many countries. At the same time, the general public is becoming increasingly aware of potential hazards from antibiotic residues in foods. Consequently, Delvotest Cow Test (Royal Gist-brocades NV, Delft, The Netherlands), an on-farm version of Delvotest P, a microbial inhibitor test for antimicrobials, is being increasingly used by farmers to assess that milk from individual cows is fit for consignment to the bulk tank. Occasional reports of unexplained positive test results have led to suggestions of possible false-positive reactions in milk from individual cows. To investigate the potential causes of such positive test results, three separate investigations were undertaken. In a field survey of unexplained positive reports from farmers, 14 milk samples from six farms that tested positive were all found to contain antibiotic residues. In more formal investigations of individual quarter milk samples from an experimental herd, none of 134 milk samples from midlactation cows yielded positive reactions; for cows that had just calved, 16 of 144 milk samples were positive, and, of those, 13 had somatic cell counts > 4,000,000/ml. Natural inhibitors were responsible for 1 positive reaction, 8 positive reactions were related to incomplete milking, and 7 samples contained beta-lactam antibiotics. Positive reactions caused by antibiotic persisted in individual quarter samples for up to 7 d postcalving compared with 4 d for milk samples from the whole udder. Delvotest was sensitive to cephalonium, the active ingredient of Cepravin Dry Cow (Mallinckrodt Veterinary Ltd., Uxbridge, United Kingdom), which is the market-leading product in the United Kingdom. Test results yielded a partial purple color reaction in the presence of 8 micrograms/kg of cephalonium and a completely purple reaction at 16 micrograms/kg. These results confirm the validity of Delvotest when used to examine composite milk samples from individual cows supplying the United Kingdom dairy industry and suggest that, with proper attention to milk withdrawal periods and complete milking, there is no obvious risk of antibiotic contamination of milk.

Animals↗

PubMed2000

Rapid detection and quantification of members of the archaeal community by quantitative PCR using fluorogenic probes.

We describe a rapid, reproducible, and sensitive method for detection and quantification of archaea in naturally occurring microbial communities. A domain-specific PCR primer set and a domain-specific fluorogenic probe having strong and weak selectivity, respectively, for archaeal rRNA genes (rDNAs) were designed. A universal PCR primer set and a universal fluorogenic probe for both bacterial and archaeal rDNAs were also designed. Using these primers and probes, we demonstrated that detection and quantification of archaeal rDNAs in controlled microbial rDNA assemblages can be successfully achieved. The system which we designed was also able to detect and quantify archaeal rDNAs in DNA samples obtained not only from environments in which thermophilic archaea are abundant but also from environments in which methanogenic archaea are abundant. Our findings indicate that this method is applicable to culture-independent molecular analysis of microbial communities in various environments.

Archaea↗

PubMed1992

Microbial resistance surveillance techniques. Blood culture versus multiple body site monitoring.

The continuing problem of nosocomial bacterial infections has resulted in the development of new techniques to monitor their prevalence and detect the emergence of drug resistance. "Focused microbial surveillance," in which bacterial incidence and antimicrobial susceptibilities are assessed by specific hospital units, provides unique information in the evaluation of emerging outbreaks of resistant Gram-negative pathogens. Within a given unit, however, pooled microbiologic data may be misleading, since they reflect only the average of multiple susceptibilities and may represent either colonization or infection. Formerly, at the University of Iowa Hospitals and Clinics, bacterial surveillance within a particular unit had typically been performed using blood-sample analysis, which was viewed as an effective index of the activity of most classes of antimicrobial agents. However, specific body site surveillance (wounds, sputum, urine, and so forth) may offer advantages over blood-culture analysis with some forms of resistance (or types of bacteria), such as stably derepressed type-I beta-lactamase production, which seems to develop rather quickly. Site-specific surveillance may enable earlier detection of rapidly emerging resistant strains and the identification of virulent new serotypes or specific causes. Our data demonstrated that there is a greater probability of detecting resistant bacterial strains of Enterobacter colonizing and subsequently producing infections in the respiratory tract, urinary tract, or wound site. There was a time delay of almost 3 years between first observing resistant strains in sputum, urine, or pus and encountering them in an alarming incidence among blood cultures. Other studies confirm this particular phenomenon. In a survey of penicillin-resistant enterococci, the majority of clinical isolates were obtained from sites other than blood: seven from urine; five from wounds; two each from rectal swabs, ascitic fluid, and blood; and one each from a peritoneal catheter, Bartholin's cyst abscess, and pancreatic abscess. Effective techniques for selective surveillance should be both sensitive and cost effective. Current evidence suggests that site-specific monitoring (wounds, sputum, pleural effusions, urine, and feces) offers the advantages of more rapid identification of resistance trends prior to their appearance in the bloodstream cultures.

Bacteria↗

PubMed2003

Sol-gel column technology for capillary isoelectric focusing of microorganisms and biopolymers with UV or fluorometric detection.

The sol-gel surface modification is used for capillary isoelectric focusing of microorganisms and biopolymers. The coating strongly decreases the electroosmotic flow so that it enables the use of the short capillaries down to 100 mm in the separation length. The examples of capillary isoelectric focusing of the low-molecular-mass pI markers and mixed cultures of microbial populations of Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Enteroccocus faecalis with UV detection are shown. It is possible to quantify bacterial cells according to their peak areas; the minimum detectable number of microbial cells was 5 x 10(2)-1 x 10(3). The compatibility of sol-gel capillaries with sensitive fluorometric detection of fluorescent pI markers together with fluorescein labeled proteins is demonstrated.

Bacteria↗

PubMed2006

daime, a novel image analysis program for microbial ecology and biofilm research.

Combinations of microscopy and molecular techniques to detect, identify and characterize microorganisms in environmental and medical samples are widely used in microbial ecology and biofilm research. The scope of these methods, which include fluorescence in situ hybridization (FISH) with rRNA-targeted probes, is extended by digital image analysis routines that extract from micrographs important quantitative data. Here we introduce daime (digital image analysis in microbial ecology), a new computer program integrating 2-D and 3-D image analysis and visualization functionality, which has previously not been available in a single open-source software package. For example, daime automatically finds 2-D and 3-D objects in images and confocal image stacks, and offers special functions for quantifying microbial populations and evaluating new FISH probes. A novel feature is the quantification of spatial localization patterns of microorganisms in complex samples like biofilms. In combination with '3D-FISH', which preserves the 3-D structure of samples, this stereological technique was applied in a proof of principle experiment on activated sludge and provided quantitative evidence that functionally linked ammonia and nitrite oxidizers cluster together in their habitat. This image analysis method complements recent molecular techniques for analysing structure-function relationships in microbial communities and will help to characterize symbiotic interactions among microorganisms.

Biofilms↗