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L Raskin

Publications and source records attributed to L Raskin.

32 records · Page 2Linked to original sources
PubMed1998

A Comparison of the Use of In Vitro-Transcribed and Native rRNA for the Quantification of Microorganisms in the Environment.

Abstract Nearly full-length, small subunit (SSU) rRNA was transcribed in vitro from clones of SSU rDNA genes. Comparing the use of in vitro-transcribed and native rRNA indicated that, when in vitro-transcribed rRNA was used as a standard for quantitative hybridizations with oligonucleotide probes, the population was consistently underestimated. The population abundance was expressed as a percentage of specific target SSU rRNA (determined with a specific oligonucleotide probe), relative to the total SSU rRNA (measured with a universal probe). Differences in hybridization signals could be related to specific probe target locations and rRNA denaturation conditions, suggesting that higher order structure is important in quantitative membrane hybridizations. Therefore, in vitro-transcribed rRNA cannot always be used for the absolute quantification of microbial populations, but can be employed as a standard to quantify shifts in population abundance over time, and to compare community structure in various environments.

Journal Article↗
PubMed1998

Quantification of Gordona amarae strains in foaming activated sludge and anaerobic digester systems with oligonucleotide hybridization probes.

Previous studies have shown the predominance of mycolic acid-containing filamentous actinomycetes (mycolata) in foam layers in activated sludge systems. Gordona (formerly Nocardia) amarae often is considered the major representative of this group in activated sludge foam. In this study, small-subunit rRNA genes of four G. amarae strains were sequenced, and the resulting sequences were compared to the sequence of G. amarae type strain SE-6. Comparative sequence analysis showed that the five strains used represent two lines of evolutionary descent; group 1 consists of strains NM23 and ASAC1, and group 2 contains strains SE-6, SE-102, and ASF3. The following three oligonucleotide probes were designed: a species-specific probe for G. amarae, a probe specific for group 1, and a probe targeting group 2. The probes were characterized by dissociation temperature and specificity studies, and the species-specific probe was evaluated for use in fluorescent in situ hybridizations. By using the group-specific probes, it was possible to place additional G. amarae isolates in their respective groups. The probes were used along with previously designed probes in membrane hybridizations to determine the abundance of G. amarae, group 1, group 2, bacterial, mycolata, and Gordona rRNAs in samples obtained from foaming activated sludge systems in California, Illinois, and Wisconsin. The target groups were present in significantly greater concentrations in activated sludge foam than in mixed liquor and persisted in anaerobic digesters. Hybridization results indicated that the presence of certain G. amarae strains may be regional or treatment plant specific and that previously uncharacterized G. amarae strains may be present in some systems.

Anaerobiosis↗
PubMed1997

Group-specific small-subunit rRNA hybridization probes to characterize filamentous foaming in activated sludge systems.

Foaming in activated sludge systems is characterized by the formation of a thick, chocolate brown-colored scum that floats on the surface of aeration basins and secondary clarifiers. These viscous foams have been associated with the presence of filamentous mycolic acid-containing actinomycetes. To aid in evaluating the microbial representation in foam, we developed and characterized group-, genus-, and species-specific oligonucleotide probes targeting the small subunit rRNA of the Mycobacterium complex, Gordona spp., and Gordona (Nocardia) amarae, respectively. The use of a universal base analog, 5-nitroindole, in oligonucleotide probe design was evaluated by comparing the characteristics of two different versions of the Mycobacterium complex probe. The temperature of dissociation of each probe was determined. Probe specificity studies with a diverse collection of 67 target and nontarget rRNAs demonstrated the specificity of the probes to the target groups. Whole-cell hybridizations with fluorescein- and rhodamine-labeled probes were performed with pure cultures of various members of the Mycobacterium complex as well as with environmental samples from a full-scale activated sludge plant which experienced foaming. Quantitative membrane hybridizations with activated sludge and anaerobic digester foam showed that 15.0 to 18.3% of the total small-subunit rRNAs could be attributed to members of the Mycobacterium complex, of which a vast majority consisted of Gordona rRNA. Several G. amarae strains made up only a very small percentage of the Gordona strains present. We demonstrated that group-specific rRNA probes are useful tools for the in situ monitoring and identification of filamentous bacteria in activated sludge systems.

Corynebacterium↗
PubMed1996

Critical evaluation of membrane supports for use in quantitative hybridizations.

The quantification of 16S rRNA by oligonucleotide probe hybridization was investigated with MagnaGraph (Micron Separation, Inc. [MSI]), Magna Charge (MSI), Magna (MSI), Immobilon-N (Millipore Corporation), and Nytran (Schleicher & Schuell, Inc.) membranes as supports for nucleic acid immobilization. The levels of detectability provided by the Magna Charge and Immobilon-N membranes were 20 to 50 times better than those obtained with the MagnaGraph, Magna, and Nytran membranes. The variability of the signal response for individual membranes ranged from 10 to 50%, with the Magna Charge and Immobilon-N membranes demonstrating the lowest variability.

Journal Article↗
PubMed1996

Competition and coexistence of sulfate-reducing and methanogenic populations in anaerobic biofilms.

The microbial population structure and function of natural anaerobic communities maintained in laboratory fixed-bed biofilm reactors were tracked before and after a major perturbation, which involved the addition of sulfate to the influent of a reactor that had previously been fed only glucose (methanogenic), while sulfate was withheld from a reactor that had been fed both glucose and sulfate (sulfidogenic). The population structure, determined by using phylogenetically based oligonucleotide probes for methanogens and sulfate-reducing bacteria, was linked to the functional performance of the biofilm reactors. Before the perturbation, the methanogenic reactor contained up to 25% methanogens as well as 15% sulfate-reducing bacteria, even though sulfate was not present in the influent of this reactor. Methanobacteriales and Desulfovibrio spp. were the most abundant methanogens and sulfate-reducing bacteria, respectively. The presence of sulfate-reducing bacteria (primarily Desulfovibrio spp. and Desulfobacterium spp.) in the absence of sulfate may be explained by their ability to function as proton-reducing acetogens and/or fermenters. Sulfate reduction began immediately following the addition of sulfate consistent with the presence of significant levels of sulfate-reducing bacteria in the methanogenic reactor, and levels of sulfate-reducing bacteria increased to a new steady-state level of 30 to 40%; coincidentally, effluent acetate concentrations decreased. Notably, some sulfate-reducing bacteria (Desulfococcus/Desulfosarcina/Desulfobotulus group) were more competitive without sulfate. Methane production decreased immediately following the addition of sulfate; this was later followed by a decrease in the relative concentration of methanogens, which reached a new steady-state level of approximately 8%. The changeover to sulfate-free medium in the sulfidogenic reactor did not cause a rapid shift to methanogenesis. Methane production and a substantial increase in the levels of methanogens were observed only after approximately 50 days following the perturbation.

Journal Article↗
PubMed1996

Characterization of universal small-subunit rRNA hybridization probes for quantitative molecular microbial ecology studies.

Universal oligonucleotide hybridization probes targeting the small-subunit rRNA are commonly used to quantify total microbial representation in environmental samples. Universal probes also serve to normalize results obtained with probes targeting specific phylogenetic groups of microorganisms. In this study, six universal probes were evaluated for stability of probe-target duplexes by using rRNA from nine organisms representing the three domains of Bacteria, Archaea, and Eucarya. Domain-specific variations in dissociation temperatures were observed for all probes. This could lead to a significant bias when these probes are used to quantify microbial populations in environmental samples. We suggest lowering the posthybridization wash stringency for two of the universal probes (S-*-Univ-1390-a-A-18 and S-*-Univ-1392-a-A-15) examined. These two probes were evaluated with traditional and modified hybridization conditions to characterize defined mixtures of rRNAs extracted from pure cultures and rRNA samples obtained from anaerobic digester samples. Probe S-*-Univ-1390-a-A-18 provided excellent estimations of domain-level community composition of these samples and is recommended for future use in microbial ecology studies.

Archaea↗
PubMed1995

Characterization of microbial communities in anaerobic bioreactors using molecular probes.

The microbial community structure of twenty-one single-phase and one two-phase full-scale anaerobic sewage sludge digesters was evaluated using oligonucleotide probes complementary to conserved tracts of the 16S rRNAs of phylogenetically defined groups of methanogens and sulfate-reducing bacteria. These probe results were interpreted in combination with results from traditional chemical analyses and metabolic activity assays. It was determined that methanogens in "healthy" mesophilic, single-phase sewage sludge digesters accounted for approximately 8-12% of the total community and that Methanosarcinales and Methanomicrobiales constituted the majority of the total methanogen population. Methanobacteriales and Methanococcales played a relatively minor role in the digesters. Phylogenetic groups of mesophilic, Gram-negative sulfate-reducing bacteria were consistently present at significant levels: Desulfovibrio and Desulfobulbus spp. were the dominant sulfate-reducing populations, Desulfobacter and Desulfobacterium spp. were present at lower levels, and Desulfosarcina, Desulfococcus, and Desulfobotulus spp. were absent. Sulfate reduction by one or more of these populations played a significant role in all digesters evaluated in this study. In addition, sulfate-reducing bacteria played a role in favoring methanogenesis by providing their substrates. The analysis of the two-phase digester indicated that true phase separation was not accomplished: significant levels of active methanogens were present in the first phase. It was determined that the dominant populations in the second phase were different from those in the single-phase digesters.

Bacteria, Anaerobic↗
PubMed1994

Group-specific 16S rRNA hybridization probes to describe natural communities of methanogens.

Eight oligonucleotides which are complementary to conserved tracts of 16S rRNA from phylogenetically defined groups of methanogens were designed and characterized for use as hybridization probes for studies in environmental and determinative microbiology. The target-group specificity and temperature of dissociation for each probe were characterized. In general, the probes were very specific for the target methanogens and did not hybridize to the rRNAs of nontarget methanogens. Together, the eight probes circumscribe methanogens now represented in pure culture (with the exception of members of the family Methanothermaceae). Three probes are order specific; two identify members of the order Methanobacteriales, and one is specific for the order Methanococcales. The fourth probe encompasses three families belonging to the order Methanomicrobiales, the third order within the current classification. The fifth probe is specific for the remaining family within this order (Methanosarcinaceae). Three additional probes encompass different genera within the Methanosarcinaceae.

Ecology↗
PubMed1994

Quantification of methanogenic groups in anaerobic biological reactors by oligonucleotide probe hybridization.

The microbial community structure of anaerobic biological reactors was evaluated by using oligonucleotide probes complementary to conserved tracts of the 16S rRNAs of phylogenetically defined groups of methanogens. Phylogenetically defined groups of methanogens were quantified and visualized, respectively, by hybridization of 32P- and fluorescent-dye-labeled probes to the 16S rRNAs from samples taken from laboratory acetate-fed chemostats, laboratory municipal solid waste digestors, and full-scale sewage sludge digestors. Methanosarcina species, members of the order Methanobacteriales, and Methanosaeta species were the most abundant methanogens present in the chemostats, the solid-waste digestors, and the sewage sludge digestors, respectively.

Anaerobiosis↗
PubMed1981

Relationship between intellectual status and reading skills for developmentally disabled children.

The relationship between WISC-R Full Scale IQ and scores on the Woodcock Reading Mastery Tests were explored for 80 developmentally disabled children. While the children's reading skills correlated moderately and significantly with intellectual status, abstract reading skills, e.g., word comprehension, correlated more highly with Full Scale IQ than did concrete ones, e.g., word identification. The development of concrete learning patterns by such children was discussed, with an emphasis on the emotional importance of these learning styles to the children and their families.

Child↗
PubMed1975

[Attenuation and smoothness of injected signals in a Rijlant network].

The V.C.G. methods must handle the E.K.G. information through weighing systems, because of the volume and the eccentric location of the heart in the chest, of the inhomogeneity of the surrounding tissues and of the irregular shape of the body surface. Most of these weighing systems use compensating networks. On the other hand, Rijlant simulates an homogeneous medium where the cardiac currents spread and where remote observation would eliminate much of the distorsions. In order to understand how such a simulated medium operates, we drive it by means of sinusoidal waves and we observe how the amplitude and the phase of the signals change when we switch on the different leads. Each value recorded at a same level in the network may be considered as the sum of a true proximity effect added to an other component depending upon the lead orientation. The latter is referred to a mixed result of the driving waves that we call "the modulus effect". From the injection level to the other levels, the proximity effect rapidly vanishes while the modulus effect does not disappear until grounded level is reached. The decrease of the modulus effect either to zero (grounded level) or to a finite value (free end of the network) is not expressed as a formula because Rijlant's network does not simulate an euclidian space. We conclude that, at least as the proximity effect is concerned, the instrument performs what it is expected to do.

Heart↗