A new approach for estimating the efficiencies of the nucleotide substitution models

In this article, a new approach is presented for estimating the efficiencies of the nucleotide substitution models in a four-taxon case and then this approach is used to estimate the relative efficiencies of six substitution models under a wide variety of conditions. In this approach, efficiencies of the models are estimated by using a simple probability distribution theory. To assess the accuracy of the new approach, efficiencies of the models are also estimated by using the direct estimation method. Simulation results from the direct estimation method confirmed that the new approach is highly accurate. The success of the new approach opens a unique opportunity to develop analytical methods for estimating the relative efficiencies of the substitution models in a straightforward way.     

A new technique for long preservation of 14C-labelled Cladocerans

Three ways of preserving labelled Cladocerans fed with ¹⁴C-Chlorella for 7.5–10 min were tested. Tracer leakage in 4% formalin at room temperature is rapid and extensive (half of the label was found in the animals after 1 hour of preservation). Even when individuals are frozen and sorting is made quickly in a liquid, losses nevertheless occur (substantial decrease of animal activity after only 4–5 min in the water in one of the two experiments performed). Results obtained after freezing in 4% formalin and sorting exactly 2 hours after thawing gave consistent losses: 16 separate experiments with Daphnia pulex, Ceriodaphnia spp. and Diaphanosoma brachyurum gave apparent filtering rates underestimated from 35% to 63% for freezing periods of up to 45 days. The good agreement in in situ community filtering rates between measured values and estimated ones from individual data confirmed the validity of a correction factor of x 2 applied to animals frozen in formalin.     

A philosophy of methods development: The assimilation of new methods and information into aquatic microbial ecology

Emerging methodologies can be used to provide a strong basic understanding of the diversity of microbial behavior and interactions. However, these new methods should be thoroughly and rigorously validated under controlled conditions before being extended to uncontrolled field conditions. Data based on novel approaches are likely to provide insights that are not easily related to existing information based on conventional methodologies. As an example, measurements of the ribosomal RNA (rRNA) content of bacteria show similar spatial patterns as measurements of thymidine incorporation into DNA and leucine incorporation into protein. However, the spatial patterns are not identical, and these parameters are not equally intercorrelated nor equally predictable from basic oceanographic data. Therefore, rRNA content measurements provide a new dimension of information that can be used to explore the relationship of bacteria to their environment, complementing the information obtained from conventional methods.     

A comparison of chromium-51 and iron-59 for estimating erythrocyte survival in the cat

Erythrocyte survival studies were conducted on eight, normal, healthy, 1-year-old male specific-pathogen-free cats using both chromium-51 and iron-59 simultaneously. The chromium-51 procedure gave a half-life value of 11.1 +/- 0.9 days. This was considerably lower than would be expected on the basis of the experimentally determined iron-59 erythrocyte survival time of 51.2 +/- 14.9 days. The results of this study indicated that there was considerable loss of the chromium-51 label in the cat other than that from senescence alone. An analysis of the chromium-51 disappearance curve indicated that there were two exponential disappearance rates for the chromium-51 label and, in the absence of cell death, approximately 67% of the label was lost with a rate constant of 0.02 per day and 33% was lost with a rate constant of 0.1 per day. An equation is presented which models the loss of chromium-51 label which could be used to calculate erythrocyte survival from a chromium-51 disappearance curve. Blood volume measurements, hemograms, bone marrow differential results, and iron kinetic values also were determined and the results presented. While a reasonable approximation of the erythrocyte life span could be made by correcting the chromium-51 values for losses other than senescence, the iron-59 procedure would be the preferred method in cats.     

Sizes of isotopically labeled molecules released during lysis of tumor cells labeled with 51Cr and [14C]nicotinamide.

Previous studies of mechanisms of immune lysis have utilized the release of isotopically labeled molecules from cells labeled with Na₂⁵¹Cr0₄ and [¹⁴C]nicotinamide, among others. The interpretation of specific isotope release from such cells was hampered by the lack of knowledge about the molecular sizes of the released radioactive molecules. In the present study, P815 tumor cells were labeled with the above two isotopic compounds. The cells were then lysed by freezing and thawing, antibody and complement, or immune T-lymphocyte-mediated killing. Soluble supernatants from the lysates were chromatographed in the cold on Sephadex G-10, G-25, and G-200 using several elution media. No differences were detected due to different kinds of lysis or different eluting media. Eighty-five percent of ¹⁴C-labeled molecules released from the cells were indistinguishable from nicotinamide and clearly distinct from nicotinamide adenine dinucleotide. The results suggest that [¹⁴C]nicotinamide taken up by the tumor cells during overnight culture is released unaltered during lysis. Ninety percent of ⁵¹Cr-labeled lysate molecules had an apparent molecular weight less than 4000. Most of this material was distinct from inorganic chromate, but its chemical form is not known.     

A new method for the determination of 14C radioactivity in ketone bodies.

A reaction vessel is described consisting of a simple reaction chamber fixed to an ordinary liquid scintillation-counting vial which allows the direct determination of radioactivity in acetone as well as separately in the carboxyl and acetone moieties of acetoacetate and 3-hydroxybutyrate. Radioactivity and its distribution between carbon 1 and carbons 2 through 4 can be determined in total ketone bodies as well as sequentially in acetone, acetoacetate, and 3-hydroxybutyrate. Recoveries with labeled ketone bodies under a variety of analytical conditions are presented.     

A new sampler for stream benthos, epiphytic macrofauna and aquatic macrophytes

SUMMARY. A new sampler is described combining the advantages of both box and Surber samplers. The sample area is 200 × 250 mm (0.05 m²) and the device has been used to sample the benthos of rivers to a depth of c . 60 mm where the substratum consists of particles with diameters of 50 mm or less. The basic design may be used in water up to 0.5 m in depth, but with an optional top unit this depth may be increased to 1 m. Aquatic macrophytes can also be sampled either on their own or together with the underlying substratum. Results from sampling programmes designed to estimate macrophyte biomass, the biomass of the macroinvertebrate community and the density of Gammarus pulex are given as examples of the flexibility of the sampler on a variety of biotopes.     

New evidence for the metabolic conversion of aspartate-4-14C to glutamine-l-14C during the initial period of ammonia assimilation in the roots of barley plants

Metabolic conversion of aspartate was investigated in rootsof barley plants fed a 30-min pulse of aspartate-4-¹⁴C, followedby transfer to an ammonia or nitrate medium for 1 hr. Glutamate was the predominant labeled amino acid after the pulse;whereas, glutamine was after ammonia assimilation. Each labelappeared almost entirely at the C-1 position. The organic acidfraction was also labeled with¹⁴C. Negligible labeling of asparagineshowed that it was not a primary product of ammonia assimilation. The data demonstrated that aspartate-4-¹⁴C was transformed intoglutamate-1-¹⁴C by the tricarboxylic acid cycle mechanism, withglutamate being converted to glutamine-1-¹⁴C during ammoniaassimilation. The physiological significance of this metabolicconversion in plant metabolism is discussed. This conversionplays an important role in preventing a drop in the oxalacetatelevel when -ketoglutarate has been drained off by rapid synthesisof glutamate and glutamine during the initial period of ammoniaassimilation. This paper also presents, for die first time, evidence for theexistence of a new pathway, named the "aspartate-glutamine pathway",in higher plants. (Received August 5, 1972; )     

A new colorimetric technique for the estimation of vitamin C using Folin phenol reagent

A new colorimetric technique for the estimation of ascorbic acid by using Folin phenol reagent has been developed. The absorption maximum of the color developed by the interaction of ascorbic acid with Folin reagent is 760 nm. The technique obeys the Beer-Lambert law up to a concentration of 45 μg ascorbic acid as shown by the standard curve. The color developed has been found to be stable up to 18 h. Recovery experiments showed that the technique is almost 100% efficient. The development of the color is not obstructed by glucose, glutathione, bovine serum albumin, urea, cysteine, adenine, guanine, cytosine, uracil, sulfosalicylic acid, thymol, or oxyhemoglobin, which are compounds suspected of interfering in routine analysis. The technique is simple, quick, and efficient and can be employed for the estimation of ascorbic acid in a wide variety of biological materials.     

A substrate-induced growth-response method for estimating the biomass of microbial functional groups in soil and aquatic systems

Abstract A substrate-induced growth-response (SIGR) method is presented for estimating the biomass or density of microorganisms capable of carrying out specific metabolic functions in natural and human-made systems. The biomass of active organisms can be estimated based on the concentration of substrate needed to induce the growth of the standing population. Curves of substrate mineralization or depletion are used as indirect indicators of growth. Estimates of population size are obtained by using non-linear regression techniques to fit simple models, that contain biologically relevant parameters, to the substrate mineralization curves. In the present study, the SIGR method was used to estimate the numbers of organisms capable of mineralizing 2,4-dinitrophenol in soil and in a model waste-treatment system. The SIGR approach was tested by comparison of model estimates to estimates for the same parameters obtained by independent means.     

A comparison of purification efficiencies of various constructed ecosystems (aquatic,semi-aquatic and terrestrial) receiving urban wastewaters

Tests were carried out under controlled conditions in the Experimental Plant of Viville (Arlon, Belgium) to enhance the purification of urban wastewater by natural means. The results demonstrate the need to structure treatment systems in a series of different artificial ecosystems (or a Hierarchical Mosaic of Artificial Ecosystems — MHEA in French). The first two levels we used were made up of an unplanted aquatic ecosystem (stabilization pond) followed by a semi-aquatic ecosystem planted withTypha latifolia L. in which the water flows over the substrate. At a flow rate of 4 m²/PE (1 PE=150 1/day of typical urban wastewaters in Belgian rural zones), this first stage substantially reduces suspended solids (SS), COD and BOD₅, a significant amount of tot-N and tot-P, and reduces pathogens by 100-fold. Further, the system is easy to manage (sludge is eliminated in the first stage and biomass is collected in the second stage) and the treatment system does not clog up.Nevertheless, real and sustainable environmental protection demands even higher performance rates, and these first two stages, both in terms of design and dimension, can only be considered as a satisfactory part of a MHEA system. Artificial aquatic, semi-aquatic, and terrestrial ecosystems were systematically compared at the third and fourth stage of the system to increase the overall removal efficiency.The most complete and efficient system in our tests (i.e., the one that provides the most successful primary (SS), secondary (COD and BOD₅) and tertiary (N and P) treatment and the best pathogens removal rates) was made up of 3 sequential series of ecosystems: an aquatic ecosystem whose flow went into a plantedTypha latifolia system (surface water flow), that flowed into a terrestrial ecosystem planted withAlnus glutinosa (L.) Gaertn (vertical subsurface water flow). A total surface area (stages 1–4) of 8 m²/PE ensured a high performance level whose outflow conformed to the strictest European norms.     

First event sampling. A new technique for estimating insect attack, using the onion miner fly, Liriomyza nietzkei, as test insect

A new sampling technique called first event sampling is described that satisfies the criteria essential for any technique to be acceptable for use in insect pest management. The onion miner fly, Liriomyza nietzkei Spencer, a pest of seeded onion crops, was used as the pest insect on which this technique was developed. The technique is based on a graphical method for estimating the proportion of crop plants attacked by this pest. The statistical implications of adopting such a technique are discussed.

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Résumé Une nouvelle méthode d'échantillonnage pour estimer le taux d'attaque d'un insecte ravageur est décrite. Elle se base sur le principe du premier événement, c'est-à-dire de la première rencontre avec le ravageur ou avec son dégât.Une présentation graphique des résultats du dénombrement permet de les confronter avec le seuil économique et de décider rapidement des mesures phytosanitaires appropriées.La mouche mineuse de l'oignon, Liriomyza nietz-kei Spencer, un ravageur des cultures d'oignon de semis a servi de modèle à l'élaboration de ce type d'échantillonnage. Ce travail contient une partie statistique où les bases mathématiques sont développées.Cette méthode devra répondre aux critères d'acceptation d'un type de contrôle en protection intégrée par les conseillers techniques et par les producteurs à savoir la précision dans l'estimation de l'attaque, la facilité d'utilisation et le gain de temps.