Mixing time and oxygen transfer characteristics of double draft tube airlift fermentor

Despite the increasing importance of airlift fermentors, very little published information is available on how the geometric configurations of the draft tubes and the air-sparging system affect the mixing and oxygen transfer characteristics of the fermentor. A 14-L air-lift fermentor was designed and build with a fixed liquid height to diameter ratio of 1.5 utilizing four equally spaced air jets at the bottom. Two jet orifice sizes were used, 1.27 and 3.81 mm i.d., and for each jet size the following four geometric configurations were used: Single inner concentric draft tube, single outer concentric draft tube, two concentric draft tubes, and no draft tubes where the fermentor was operated as a shallow bubble column. It was found that the presence of draft tubes stabilized liquid circulation patterns and gave systemically higher mixing times than those obtained in the absence of draft tubes. In addition, the double draft tube geometry resulted in higher mixing times than the single draft tubes. For the power unit volume range 20 to about 250 W/m³ the larger 3.81-mm orifices gave systemically higher k_L a values than the smaller 1.27-mm i.d. orifices. At 200 W/m³ the use of a single outer draft tube with the 3.81-mm orifices resulted in 94% increase in k_L a values over that obtained with no draft tubes. However, the effect of draft tube geometry on k_L a values when the 1.27-mm orifices were used was not significant. The air bubble formation characteristics at the jet orifices were found to be different, which reflected the differences observed in mass transfer and mixing characteristics. The power economy for oxygen transfer was found to be depend strongly on the orifice size and less on the geometric configuration of draft tubes.     

Chemosensory Responses Toward Oxygen in Euglena gracilis*

SYNOPSIS Euglena gracilis strain Z, at a concentration of 10⁶ cells/ml and in containers of ∽ 0.1-mm thickness, spontaneously forms dynamic ring patterns in the dark. These patterns are modified differentially by illumination with red and with blue light. The red light effect is abolished by treatment with an inhibitor of photosynthesis. Pattern formation is apparently the result of chemophobic responses to oxygen dissolved in the medium. Euglena can respond to both negative and positive concentration gradients, depending upon the absolute magnitude of oxygen concentration. The photo- and chemosensory transduction systems of Euglena interact at a stage which precedes the overt expression of motor responses.     

Predator exclusion experiments in an intertidal mud flat

The intensity of predation pressure exerted on the macrofauna in muddy sediments was investigated in Königshafen (island of Sylt, eastern North Sea). Tests in aquaria revealed shrimpCrangon crangon, juveniles of the shore crabCarcinus maenas, and gobiid fishPomatoschistus microps as the most important local predators. Their high abundance from July to September led to the hypothesis, that predation pressure on the infauna will be most intense during that period. Cages were set up to protect the infauna against these predators. No protective effect was achieved with cages constructed of 20-mm mesh nylon net; however, cages with screen wire and gauze of 5-mm mesh and smaller resulted in a considerable increase of the infauna. In cages of 1-mm mesh gauze, set up from March to June, the macrofauna reached an abundance of four times the control density. In the period from July to October, the factor of increase was as high as 23. Species density was 28 400 cm^–2, as compared with only 7 in the control. Species which colonized the mud flat during spring and early summer established dense settlements inside the cages, but failed to do so in uncaged mud-flat areas. It is concluded that predation by young crabs, shrimp and gobies determines to a large extent the structure and dynamics in the local intertidal macrofauna.     

Cell wall metabolism in ripening fruit. VIII. Cell wall composition and synthetic capacity of two regions of the outer pericarp of mature green and red ripe cv. Jackpot tomatoes

Pericarp discs were excised from mature green and red ripe tomato (Lycopersicon esculentum Mill. cv. Jackpot) fruit and kept in sterile tissue culture plates for 4 days, including 2 days of incubation with D-[U-¹³C]-glucose. Cell walls were prepared and differentially extracted with dimethylsulfoxide (DMSO), trans-1,2-diaminocyclohexane-N,N,N′,N′-tetraacetic acid (CDTA). Na₂CO₃, 4 M KOH and 8 M KOH. Cell wall noncellulosic neutral sugar (NS) composition and cell wall synthetic capacity (i.e. incorporation of density label into cell wall sugars) were determined by using a gas chromatograph coupled to a flame ionization detector and a mass spectrometer, respectively. In the crude cell wall, there was significantly less galactose (Gal) and glucose (Glc) in the “outer”2-mm pericarp region, including the cuticle, compared to the “inner”2-mm region immediately below it (closer to the locules). In the CDTA-soluble pectin, rhamnose (Rha), arabinose (Ara) and Gal accounted for approximately 90% of the total NS. The ratios of these sugars were very similar in the total (¹²C plus ¹³C) sugars, and also in the newly synthesized ([¹³C]-labeled) sugars, suggesting that newly synthesized NS associated with the chelator-extractable pectic fraction has a composition very similar to that of preexisting NS. In the 4 M KOH-soluble material, xylose (Xyl) and Glc accounted for approximately 70% of the total NS. The ratio of these sugars was very similar in the total sugars, but much lower in the newly synthesized portion. This suggests that the hemicellulosic polymers synthesized during the ripening process are different in type and/or proportion from those present in the developing fruit. Because the outer pericarp of tomatoes contains at least two distinct tissue types and these have a distinct cell wall composition, analysis of tomato cell wall polysaccharide composition by homogenization of the entire outer pericarp will obscure subtle changes associated with ripening/softening within specific tissue types.     

APOLAR MOVEMENT OF ZEATIN THROUGH COLEUS PETIOLES AND PISUM ROOTS AS ESTIMATED BY BIOASSAY AND RADIOACTIVE LABELLING

The movement and polarity of zeatin, a highly active, endogenous cytokinin, through petioles and roots were tested in the classical experimental arrangement using excised 5-mm sections. Zeatin in the receiver cylinders of agar was measured by soybean callus bioassay and by liquid scintillation counting of ¹⁴C that had been added in the donor cylinders as [8-¹⁴C] zeatin. Both methods agreed in showing movement, but there was no polarity in Coleus #5 petioles. The amounts moved were about one-tenth of the GA-3 movement through petioles of the third pair of leaves of the same clone. Movement of ¹⁴C-zeatin through Pisum roots was similarly statistically significant but non-polar; the amounts moved were similar to those previously observed for polar GA-3 movement through Zea roots.     

Decarboxylation and Transport of Auxin in Segments of Sunflower and Cabbage Roots. II. A Chromatographic Study Using IAA-1-14C and IAA-5-3H

The polar movement of IAA has been examined in 5-mm root segments of Brassica oleracea and Helianthus annum. The movement was studied partly with IAA-1-¹⁴C and partly with IAA-5-³H. In both plants a slight acropetal flux of ¹⁴C and IAA-³H was found through the segments. The recovered radioactivity in the agar receiver blocks and in the receiver end of the segments increased as a function of time. A large portion of the applied IAA was converted on the cut surfaces and in the tissues of the segments. Chromatographic analysis indicated different destruction products when estimated by scintillation counting and by spraying with in-dole reagent (DMCA). Chromatograms run in isopropanol: ammonia: water, 8:1:1, yielded three different substances, one spot near the starting line and one near the front, neither of which has been identified. Finally there was a spot with R_f 0.4–0.6, probably representing IAA.     

Daphnia–rotifer interactions in Patagonian communities

In fishless ponds of extra-Andean Patagonia, large Daphnia often may severely limit the abundance of small rotifers by mechanical interference (ingestion and damage after rejection). In ten laboratory experiments, 15–20 rotifers were exposed to one large daphniid for 1 day in 80 ml water with Cryptomonas erosa (5 × 10³ cells ml⁻¹). Both D. cf. pulex (3.6 mm) and D. cf. obtusa (3.4 mm) imposed significant mortality on Keratella tropica and newborn Brachionus angularis and Brachionus calyciflorus, but not on the larger neonates of Brachionus rubens or adults of B. angularis and B. calyciflorus. B. rubens neonates avoided the possibility of interference by rapidly attaching to Daphnia. The number of susceptible rotifers killed per Daphnia day⁻¹ was 2–8 for D. cf. obtusa and 8–15 for D. cf. pulex; rotifer-based clearance rates per Daphnia day⁻¹ were 10–35 and 35–65 ml, respectively. When large daphniids occurred in Laguna Los Juncos in late September and October 1997, the only common plankton rotifer was the large and probably well-protected Keratella morenoi (total length = 210 μm). Still, microscopic examination of gut contents revealed three K. morenoi in a 3-mm D. cf. obtusa and one in a 3.5-mm D. dadayana.     

Growing a stratified,cornified primary culture of rat keratinocytes with epidermis-like water permeation barrier function

Summary The culture of cutaneous keratinocytes grown on a Puropore nylon microporous membrane at the air-liquid interface has been shown to be similar to the epidermis in a number of molecular and morphologic characteristics but to exhibit a significantly greater degree of tritiated water permeation. Various culture conditions have been altered in an effort to improve the water barrier properties. A Kp value in the range of 5.5±1.6×10⁻³ has been obtained for 79% of the culturea) by plating 0.9×10⁶ viable basal cells on a piece (13-mm diameter) of membrane for 7 days of submerged growth,b) by placing two membranes on two stacked glass fiber filters (47-mm extra-thick) in a culture dish (60 mm) for 14 days of growth at the air-liquid interface,c) by replacing the growth medium, i.e., 1 ml of complete minimum essential medium (CMEM) every 24 h after lifting,d) by using 10% fetal bovine serum (FBS) in the CMEM during the submerged culture period and 15% FBS in the CMEM during the lifted culture period, ande) by adding a dialysis membrane on top and a Puropore nylon membrane below the culture when the cultures were inserted in the permeation cell for testing. The percentage of cultures with this value for Kp can be increased to 90% if only cultures with yellow, smooth, and shiny surfaces are tested. This system should be useful as a replacement for skin in testing the cutaneous permeation of some chemicals. To whom correspondence should be addressed at 1528 Public Health, The University of Michigan, Ann Arbor, Michigan 48109-2029.     

A study on annual changes in surface cover of floating-leaved plants in a lake using aerial photography

A simple method of aerial photography was used to investigate annual and between-year changes in surface cover of the floating-leaved plants, Nymphoides peltata, Nelumbo nucifera and Trapa natans, in natural stands in Lake Kasumigaura. Oblique photographs of vegetation were taken with a 35-mm camera with real color film from airplanes between 1983 and 1989. Vegetation maps were traced on the positions after projective transformation to vertical photography. The distribution of the stand of Nymphoides peltata seemed to be restricted to a region of less than 1.5 m water depth. A rise of 1.0 m in the water level after a typhoon in 1986 caused a subsequent decrease in vegetation area of Nelumbo nucifera. The vegetation recovered in shallower regions after one year in 1987. The expansion rate of Nymphoides peltata and Nelumbo nucifera in one direction was in the range 3.8–10.0 m yr^-1 and 4.1–20.8 m yr^-1, respectively. Nelumbo nucifera, Nymphoides peltata and Trapa natans, having high leaf biomass to total biomass may therefore be more vulnerable to large flood and waves. Fluctuation in water level and spatial difference in water depth may also be important factors determining the vegetation area and survival of Nymphoides peltata, Nelumbo nucifera and Trapa natans.     

Determination of cocaine and its metabolites in serum microsamples by high-performance liquid chromatography and its application to pharmacokinetics in rats

A single-solvent extraction step high-performance liquid chromatographic method is described for quantitating cocaine and its three metabolites in rat serum microsamples (50 μl). The separation used a 2.1-mm I.D. reversed-phase Brownlee C₁₈ column with an isocratic mobile phase consisting of methanol–acetonitrile–25.8 mM sodium acetate buffer, pH 2.2, containing 1.29·10⁻⁴M tetrabutylammonium phosphate (12.5:10:77.5, v/v/v). The detection limit was 2.5 ng/ml for all the compounds using an ultraviolet detector operated at 235 nm. The method was used to study the pharmacokinetics of cocaine after an intravenous (i.v.) bolus dose (4 mg/kg).     

Abscisic acid inhibition of root nodule initiation in Pisum sativum

Summary The effect of exogenous abscisic acid (ABA) on root nodule formation in Pisum sativum cv. Alaska was examined. ABA supplied to the roots at 1.9×10^-6M reduced the number of nodules/plant 61% without affecting root or shoot growth. The first noticeable inhibition of nodulation occurred at 3.8×10^-7M ABA. ABA at a concentration of 1.9×10^-6M inhibited neither root hair formation nor infection of root hairs by Rhizobium leguminosarum. Similar numbers of infection threads penetrated the cortex in both control and treated plants. ABA concentrations of 3.8×10^-6M had no effect on the doubling time or maximum density of R. leguminosarum in pure cultures. Normal nodule formation involves a polyploid cortical proliferation. This response to rhizobial infection can be imitated by culturing 1-mm pea-root segments on a medium containing 4.7×10^-6M kinetin. Under these conditions a highly significant reduction in the number of polyploid mitoses after 72 h is produced by 3.8×10^-8M ABA. A maximum inhibition of 68% was found with 3.8×10^-6M ABA. A similar range of ABA concentrations also inhibited the cytokinin-induced cell division in soybean callus. It is concluded that ABA reduces the number of root nodules/plant by inhibiting the cortical cell divisions required for root nodule formation.     

Goldfish's visual information processing patterns in food-reinforced discrimination learning between compound visual stimuli

Summary To examine how goldfish process and store information on compound visual stimuli, goldfish were trained with visual discriminative stimuli composed of varied colors and patterns using a Y-maze instrumental conditioning technique. The fish showed some very different types of information processing patterns depending upon the degree of discrimination difficulty of each constituent aspect (color and pattern). Those trained with compound stimuli with both a more easily discriminated aspect and a more difficult aspect learned the former selectively but did not learn the latter at all in spite of a high rate of training. Contrary to this, the fish trained with compound stimuli composed of two aspects with similar degrees of discrimination difficulty learned both aspects. In this case, only when fish were trained with compound stimuli composed of relatively more difficult aspects did they learn to discriminate between the stimuli more rapidly than the fish in each group trained with the constituent colored or patterned stimuli. These results were discussed in relation to visual processing patterns reported in other species and the mechanism of aspect selection.Abbreviations CP compound stimuli composed of VR and HB (7-mm ISBBs) - Cp compound stimuli composed of VR and HB (14-mm ISBBs) - cP compound stimuli composed of VG and HB (7-mm ISBBs) - cp compound stimuli composed of VG and HB (14-mm ISBBs) - HB horizontal patterns on blue background - HGra horizontal patterns on gray background - ISBBs interval spaces between bars - VG vertical patterns on green background - VGra vertical patterns on gray background - VR vertical patterns on red background     

Direct somatic embryogenesis from axes of mature peanut embryos

Summary Plant regeneration via somatic embryogenesis was obtained in peanut (Arachis hypogaea L.) from axes of mature zygotic embryos. The area of greatest embryogenic activity was a 2-mm region adjacent to and encircling the epicotyl. Somatic embryogenesis was evaluated on Murashige and Skoog media supplemented with a variety of auxin treatments. Maximum production occurred on medium supplemented with 3 mg · liter⁻¹ 4-amino-3,5,6-trichloropicolinic acid. Explant cultures were transferred to half-strength medium supplemented with 1 mg · liter⁻¹ gibberellic acid for somatic embryo germination and early plantlet growth. Plantlets, transferred to soil, were placed in a greenhouse and grown to maturity.     

Ecological variation in invasive brown trout (Salmo trutta) within a remote coastal river catchment in northern Patagonia complicates estimates of invasion impact

Salmonids were first introduced into the Chilean fresh waters in the 1880s, and c. 140 years later, they are ubiquitous across Chilean rivers, especially in the southern pristine fresh waters. This study examined the brown trout (Salmo trutta) and native taxa ecology in two adjacent but contrasting rivers of Chilean Patagonia. During spring 2016 and spring–fall 2017 we examined the variation in benthic macroinvertebrate and fish community composition and characterized fish size structure, stomach contents, and stable isotopes (δ¹³C and δ¹⁵N) to understand population structure, fish diet, and trophic interactions between S. trutta and native taxa. The native Galaxias maculatus (puye) dominated the fish community (74% of abundance). S. trutta was less abundant (16% of survey catch) but dominated the fish community (over 53%) in terms of biomass. S. trutta showed distinct diets (stomach content analysis) in the two rivers, and individuals from the larger river were notably more piscivorous, consuming native fish with a relatively small body size (<100-mm total length). Native fishes were isotopically distinct from S. trutta, which showed a wider isotopic niche in the smaller river, indicating that their trophic role was more variable than in the larger river (piscivorous). This study provides data from the unstudied pristine coastal rivers in Patagonia and reveals that interactions between native and introduced species can vary at very local spatial scales.     

Mesozooplankton structure in Dolgaya Bay (Barents Sea)

The community structure of mesozooplankton was investigated in Dolgaya Bay (southern Barents Sea), a subarctic fjord, using a Juday net (0.168-mm mesh size) in July, 2008. A total of 39 species and higher taxa were found. Average abundance, biomass and diversity (±standard error) were 153,403 ± 15,855 ind m⁻², 570 ± 61 mg dry mass m⁻², and 2.25 ± 0.09, respectively. Copepods were the most numerous species. The mesozooplankton communities were dominated by omnivores (Oithona similis and Acartia spp.). Vertical distribution of the mesozooplankton was characterized by copepod dominance at each sampled layer. There were no significant correlations among physical variables and biological parameters, except negative correlation for the mean biomass and mean water temperature.     

Hypoxia tolerance associated with activity reduction is a key adaptation for Laternula elliptica seasonal energetics

Seasonal dormancy is a widespread mechanism for reducing energy expenditure during periods of low energy availability. Seasonal variation in activity and the cost of pumping water through the siphons were investigated to estimate the importance of activity regulation to the seasonal energy budget of the Antarctic clam, Laternula elliptica. In the laboratory, a metabolic rate of 26.35 μmol O₂ h⁻¹ was estimated for a 50-mm shell length L. elliptica pumping water at −0.4 °C. In the field, the proportion of time siphons were visible at the sediment surface varied seasonally (32% visible in June/July compared to 86% in December/January). L. elliptica were actively pumping for a minimum of 19% of each 24-h period during winter (August) compared to a summer maximum when animals were actively pumping for 73% of the time (February). This resulted in a 3.7-fold seasonal difference in the calculated energy consumption of a 50-mm L. elliptica (19.2 μmol O₂ h⁻¹ in February versus 5.0 μmol O₂ h⁻¹ in August), which closely matches the 3.0-fold seasonal variation in metabolic rate found previously. Seasonal variation in activity could therefore be responsible for much of the seasonal difference in energy consumption of L. elliptica. Inter-annual variation in timing of the seasonal activity maxima (January 2004 and March 1999) was correlated with variation in the timing of the summer plankton bloom in Ryder Bay. In the laboratory, periods of extended siphon closure (133 ± 114 min, mean ± SD) were accompanied by long periods of heart arrhythmia (167 ± 135 min), during which time blood oxygen levels dropped to values close to zero. Heart arrhythmia is most likely part of a hypo-metabolic adaptation to reduce energy costs during extended periods of siphon closure. Physiological and behavioural dormancy, with associated hypoxia tolerance, appear to be key mechanisms controlling the seasonal energy budget of L. elliptica.     

Apparatus for simultaneous spectrophotometric and polarographic measurements

We have recently constructed a 10-mm, light path quartz cuvet which will accept a Clark oxygen electrode; it is temperature controlled and is suitable for use in a Unicam (Cambridge, England) SP 800 recording spectrophotometer. Several enquiries have prompted this publication, although such an apparatus was mentioned much earlier by Chance and Williams,¹ and has been used extensively. Figure 1a, b, c, and d and their legends provide sufficient detail for the construction of the cuvet and provision of the commercially available electrode, quartz faces, stirring motor and disk magnet. Circuit diagrams for temperature control (range 22–38°C., ± 0.2°C.) and stirrer speed control are shown in Figure 2a and b. The cuvet is shown situated in the spectrophotometer cell housing in Figure 3, and the cuvet with its associated equipment is shown in Figure 4.     

CdTe compact gamma camera for coded aperture imaging in radioguided surgery

The aim of this work was to assess the performance of a prototype compact gamma camera (MediPROBE) based on a CdTe semiconductor hybrid pixel detector, for coded aperture imaging. This probe can be adopted for various tasks in nuclear medicine such as preoperative sentinel lymph node localization, breast imaging with ^99mTc radiotracers and thyroid imaging, and in general in radioguided surgery tasks. The hybrid detector is an assembly of a 1-mm thick CdTe semiconductor detector bump-bonded to a photon-counting CMOS readout circuit of the Medipix2 series or energy-sensitive Timepix detector. MediPROBE was equipped with a set of two coded aperture masks with 0.07-mm or 0.08-mm diameter holes. We performed laboratory measurements of field of view, system spatial resolution, and signal-difference-to-noise ratio, by using gamma-emitting radioactive sources (¹⁰⁹Cd, ¹²⁵I, ²⁴¹Am, ^99mTc). The system spatial resolution in the lateral direction was 0.56 mm FWHM (coded aperture mask with holes of 0.08 mm and a 60 keV source) at a source-collimator distance of 50 mm and a field of view of 40 mm by side. Correspondingly, the longitudinal resolution in 3D source localization tasks was about 3 mm. MediPROBE showed a significant improvement in terms of spatial resolution when equipped with the high-resolution coded apertures, with respect to the performance previously reported with 1–2 mm pinhole apertures as well as with respect to adopting a 0.35 mm pinhole aperture.     

Single-culture aerobic granules with <Emphasis Type="Italic">Acinetobacter calcoaceticus</Emphasis>

Aerobic granules are cultivated by a single bacterial strain, Acinetobacter calcoaceticus, in a sequencing batch reactor (SBR). This strain presents as a good phenol reducer and an efficient auto coagulator in the presence of phenol, mediated by heat-sensitive adhesins proteins. Stable 2.3-mm granules were formed in the SBR following a 7-week cultivation. These granules exhibit excellent settling attributes and degrade phenol efficiently at concentrations of 250–2,000 mg l⁻¹. The corresponding phenol degradation rate reached 993.6 mg phenol g⁻¹ volatile suspended solids (VSS) day⁻¹ at 250 mg l⁻¹ phenol and 519.3 mg phenol g⁻¹ VSS day⁻¹ at 2,000 mg l⁻¹ phenol concentration. Meanwhile, free A. calcoaceticus cells were fully inhibited at phenol >1,500 mg l⁻¹. Denaturing gradient gel electrophoresis fingerprint profile demonstrated no genetic modification in the strain during aerobic granulation. The present single-strain granules showed long-term structural stability and performed high phenol degrading capacity and high phenol tolerance. The confocal laser scanning microscopic test revealed that live A. calcoaceticus cells principally distributed at 200–250 μm beneath the outer surface, with an extracellular polymeric substance layer covering them to defend phenol toxicity. Autoaggregation assay tests demonstrated the possibly significant role of secreted proteins on the formation of single-culture A. calcoaceticus granules.     

Dependence of Basipetal Polar Transport of Auxin upon Aerobic Metabolism

The movement of IAA-¹⁴C through coleoptile segments of Avena and Zea has been investigated under aerobic and anaerobic conditions. The results are as follows: Zea. Using a 5-mm segment and a 2-hour transport period anaerobic conditions reduced the total uptake of ¹⁴C from an apical donor by 74% and the proportion of the total found in the receiving block by at least 45%. Anaerobic conditions reduced total uptake from a basal donor by 58% but no ¹⁴C reached the apical receiving block in either air or N₂. Uptake from apical and basal donor blocks in N₂ is closely similar.

The presence of ¹⁴C in the basal receiving blocks, and its absence in the apical receiving blocks, in N₂ suggests that even in anaerobic conditions movement of IAA is polarized basipetally, although the movement occurs at only a fraction of the rate found in air.

Anaerobic conditions induced a similar reduction in basipetal movement of IAA in upper and lower 5-mm segments taken from the apical 10 mm of a Zea coleoptile.

Using 10-mm Zea segments no ¹⁴C was recovered in the receiving blocks at the basal end of the segment after 2 and 4 hours in N₂ whereas large amounts were recovered in air.

Avena: Using 5-mm segments and a 2-hour transport period the total uptake of ¹⁴C from an apical donor is reduced by 83%. Movement of ¹⁴C into the basal donor is totally inhibited in N₂. Total uptake of ¹⁴C from a basal donor is reduced by 61% in nitrogen and no ¹⁴C reached the apical receiving blocks regardless of the atmospheric conditions.

A time course for the movement of ¹⁴C into the basal and apical receiving blocks through 5-mm segments showed that in air the amount in the basal receivers increased for 4 hours and then remained approximately uniform. In N₂ no significant ¹⁴C reached the receivers until 6 to 8 hours after the application of donors but even then the amounts were about 12 to 14% of that in aerobic receivers. Movement of ¹⁴C into apical receivers was similar in air and in nitrogen and even after 6 to 8 hours the amount of radioactivity barely reached significant levels.