The influence of nerve section on the metabolism of polyamines in rat diaphragm muscle.

Concentrations of spermidine, spermine and putrescine have been measured in rat diaphragm muscle after unilateral nerve section. The concentration of putrescine increased approx. 10-fold 2 days after nerve section, that of spermidine about 3-fold by day 3, whereas an increase in the concentration of spermine was only observed after 7-10 days. It was not possible to show enhanced uptake of either exogenous putrescine or spermidine by the isolated tissue during the hypertrophy. Consistent with the accumulation of putrescine, activity of ornithine decarboxylase increased within 1 day of nerve section, was maximally elevated by the second day and then declined. Synthesis of spermidine from [14C]putrescine and either methionine or S-adenosylmethionine bt diaphragm cytosol rose within 1 day of nerve section, but by day 3 had returned to normal or below normal values. Activity of adenosylmethionine decarboxylase similarly increased within 1 day of nerve section, but by day 3 had declined to below normal values. Activity of methionine adenosyltransferase was elevated throughout the period studied. The concentration of S-adenosylmethionine was likewise enhanced during hypertrophy. Administration of methylglyoxal bis(guanylhydrazone) produced a marked increase in adenosylmethionine decarboxylase activity and a large increase in putrescine concentration, but did not prevent the rise in spermidine concentration produced by denervation. Possible regulatory mechanisms of polyamine metabolism consistent with the observations are discussed.     

Role of polyamines on de novo shoot morphogenesis from cotyledons of Brassica campestris ssp. pekinensis (Lour) Olsson in vitro

Summary The promotive effect of ethylene inhibitors (Els), i.e. AgNO₃ and aminoethoxyvinylglycine (AVG) on de novo shoot regeneration from cultured cotyledonary explants of Brassica campestris ssp. pekinensis cv. Shantung in relation to polyamines (PAs) was investigated. The endogenous levels of free putrescine and spermidine in the explant decreased sharply after 1–3 days of culture, whereas endogenous spermine increased, irrespective of the absence or presence of Els. AgNO₃ at 30 M did not affect endogenous PAs during two weeks of culture. In contrast, explants grown on medium containing 5 M AVG produced higher levels of free putrescine and spermine which increased rapidly after three days and reached a peak at 10 days. An exogenous application of 5 mM putrescine also resulted in a similar surge of endogenous free spermine of the explant. More strikingly, shoot regeneration from explants grown in the presence of 1–20 mM putrescine, 0.1–2.5 mM spermidine, or 0.1–1 mM spermine was enhanced after three weeks of culture. However, exogenous PAs generally did not affect ethylene production, and endogenous levels of 1-aminocyclopropane-1-carboxylate (ACC) synthase activity and ACC of the explant. This study shows the PA requirement for shoot regeneration from cotyledons of B. campestris ssp. pekinensis in vitro, and also indicates that the promotive effect of PAs on regeneration may not be due to an inhibition of ethylene biosynthesis.Abbreviations PAs polyamines - AVG aminoethoxyvinylglycine - SAM S-adenosylmethionine - ACC 1-aminocyclopropane-1-carboxylate - Els ethylene inhibitors     

Kinetics of polyamine synthesis and turnover in mouse fibroblasts

Kinetics of polyamine synthesis and degradation were studied in mouse fibroblasts growing in suspension culture. The approach was to prelabel cells with radioactive polyamines and to observe the rate of loss of radioactivity and the rate of decrease in specific activity of these compounds in cells. Radioactive putrescine declined with a half-life of 1.5–2h, whether derived directly from exogenous putrescine or indirectly from ornithine. Much of this turnover was due to excretion, the kinetics of which suggested that a steady-state was being established between putrescine inside and outside the cells. Within 5h of medium change, cells growing at a density of 5×10⁵cells/ml had supplied putrescine to the medium to a concentration of about 1μm. When cells were prelabelled with either putrescine or spermidine, radioactivity in cell spermidine declined with a half-life of 60h. This rate of turnover is sufficient to provide all the spermine required by the cell. Spermine synthesis was the only observed reaction of spermidine, although some excretion into the growth medium was detected. Spermine was not degraded at a detectable rate as long as cells were growing exponentially; in stationary phase, degradation to spermidine, which was excreted, became significant. The half-lives of the specific activities of spermine, spermidine and putrescine were 24, 15 and 1.5h respectively. From these values, the rate of synthesis of each was calculated. Spermidine was synthesized at 6.8 times the rate of spermine, and putrescine was synthesized at 0.46nmol/10⁶cells per h, twice the rate of spermidine. The significance of these kinetic parameters is discussed.     

Effect of carbon and nitrogen sources on growth and solasodine production in batch suspension cultures of Solanum eleagnifolium Cav.

The effect of inoculum size, carbon sources (fructose, glucose, maltose, sucrose), nitrate and ammonia on solasodine production by Solanum eleagnifolium Cav. was studied. The specific growth rate was estimated to be 0.15–0.20 d^-1 with all sugars tested at a concentration of 90 mM. Sucrose (180 mM) produced the highest biomass value (about 2.8 mg DW ml^-1) while the lowest one was produced by maltose. Although solasodine productivity values after 11 days of culture were similar for all sugars tested, the maximum values of productivity (0.9 mg g^-1 d^-1) were achieved after 6 days of culture with sucrose (180 mM). Solasodine productivity of cultures conducted with a large inoculum (20% w/v fresh material) was double that with a small inoculum (10% w/v fresh material).     

Anteroposterior gradient during nymphal-adult moulting cycle of the tropical bont tick,Amblyomma variegatum (Acarina: Ixodidae)

Summary The structure of the extensible (alloscutum) and inextensible (scutum) integument of the nymph, Amblyomma variegatum was examined during the whole bloodmeal and the nymphal-adult moulting cycle. Integumental events were tentatively correlated with the ecdysteroid levels measured by radioimmunoassay. We observed that all the integumental events were realised along an anteroposterior gradient. During the 5 days corresponding to the bloodmeal, although the hormone concentration was low, a new endocuticle was deposited on both the alloscutum and scutum. Furthermore, mitoses were initiated in the capitulum. On days 1–2 after the meal, ecdysteroid titres began to increase and reached a first peak corresponding to 4.1 ng 20-hydroxyecdysone equivalents/tick on the 4th day after the ticks dropped off their host. At this time the epidermis of the capitulum was detached and the outline of the adult capitulum was already visible. Mitotic activity in the alloscutum was initiated. On day 6 post-drop, the frontal apolysis was achieved and the ecdysteroid titres declined to basal values. A second peak much higher than the first one (maximum value of 33.7 ng/tick) and identified principally as 20-hydroxyecdysone by HPLC/RIA was noted on the 13th day post-drop. During the period of increase in the ecdysteroid levels (days 9–10 post-drop), the mitotic phase ended in the alloscutum and the apolysis began. Epicuticle was deposited after day 12 postdrop. Then, while the titre fell to low values (about 1.6 ng/tick, days 16–20 post-drop), the exocuticle was deposited and the nymphal cuticle was digested. All adult structures were functional 3 days before ecdysis. In young male as in female adults the mean value of the ecdysteroid levels corresponded to about 2.5 ng/tick. Finally, hydrolysis of tick whole extracts with esterase demonstrated a low increase of RIA-positive material, demonstrating the probable presence of natural ecdysteroid fatty-acid conjugates in this species.     

The polyamine spermine induces cystocarp development in the seaweed <Emphasis Type="Italic">Grateloupia</Emphasis> (Rhodophyta)

Polyamines such as putrescine, spermidine and spermine are ubiquitous aliphatic amines involved in reproductive events in plants and algae, and first become evident through changes in endogenous levels during reproductive development. To examine whether the differences observed in polyamines, during carposporogenesis, in the red alga Grateloupia, followed a specific pattern as is seen in other organisms, infertile axes (i.e. not showing cystocarps) were excised from the same holdfast of female fertilized individuals (i.e. showing cystocarps in other axes), and cultivated until the cystocarps became visible. Changes in the endogenous levels of free putrescine, spermidine and spermine were monitored over the 8 days of culture. The activity of enzymes related to polyamine metabolism, such as l-ornithine decarboxylase (ODC), diamine oxidase and polyamine oxidase, was measured at the beginning and end of the experimental period. Up to 50% of the infertile axes became fertile and produced cystocarps at a density of 1.91 ± 0.1 cystocarps mm⁻² after 8 days. The endogenous content of spermine increased markedly over the first 5 days of culture, then decreased to the initial level by day 8. Spermidine followed a similar pattern to spermine, whereas putrescine remained at high levels, until day 5 when it decreased abruptly. The activity of ODC was less on day 8 than on day 0, whereas the activities of diamine oxidase and polyamine oxidase increased. In parallel experiments with explants from infertile axes, exogenously added spermine (10⁻⁶ M) increased the number of cystocarps, and reversed the effect of cyclohexylamine (CHA), which is known to inhibit polyamine synthesis in Grateloupia. Serial sectioning and microscopic observation of specimens from explants cultivated in 10⁻⁶ M spermine indicated that cystocarp development was induced. The results suggest that, during transition from infertile to fertile spermine is accumulated, thus favouring the development of cystocarps, given the presumed role of spermine as an inducing agent.     

Involvement of Putrescine in the Development of Kindled Seizure in Rats

During the development of kindling by daily electrical stimulations applied to the left amygdala of rats, concentrations of the polyamines putrescine, spermidine, and spermine were measured in the left amygdala and the remainder of the cerebrum. A significant increase of putrescine concentration appeared first at the left amygdala in prekindled rats and then propagated to the remainder of the cerebrum with the development of kindling. This increase in putrescine concentration in the left amygdala was higher in prekindled rats than in fully kindled rats and lasted for at least 24 h after the final kindling stimulation. The concentrations of spermidine and spermine were slightly increased in a fully kindled state. To clarify the role of putrescine in kindling, the development of amygdaloid kindling was examined in rats after microinjections of alpha-difluoromethylornithine, a specific inhibitor of polyamine synthesis, and putrescine into the ipsilateral amygdala. Pretreatment with alpha-difluoromethylornithine (50 nmol) for 10 days accelerated both the development of behavioral kindling and the propagation of the afterdischarge from the left amygdala to the frontal cortex. In contrast, pretreatment with putrescine (200 nmol) for 10 days retarded the development of kindling. These results suggest that the increase in putrescine concentration in the kindled brain has an inhibitory effect on the development of kindling.     

Formation of Polyamines in the Rumen of Goats During Growth

Ornithine decarboxylase (E.G. 4.1.1.17) and S-adenosylmethionine decarboxylase (E.G. 4.1.1.50) and their products putrescine, spermidine and spermine were estimated in the rumen liquid from 3 groups of growing kids and 23 adult goats. Polyamines were also estimated in the feedstuff used. Marked differences in polyamine synthesis in rumen liquid were observed between the different groups of kids. Two groups of kids growing up together with adult goats had at an age of 2–4 months a peak of a few days duration in enzyme activity as well as in polyamine concentration. In these groups ornithine decarboxylase activity reached maximal values of 158±79 s (n = 4) and 100 (66–117) (n = 3) nmol[14CO2]/ml rumen liquid/h at an age of 120 and 77 days, respectively. The corresponding activity in rumen liquid from kids who were isolated from other animals was only about 1/10 of this value. By comparison ornithine decarboxylase activity in adult goats was 30.7±20 (n = 43) nmol[14CO]/ml/h. In rumen liquid from kids grown up together with adults, concentrations of the polyamines reached maximum at about the same time as ornithine decarboxylase activity. The mean maximal concentration of putrescine in the 2 groups was about 350 and 500 nmol/ml, while the corresponding value for spermidine was about 200 nmol/ml in both groups. Relatively constant and high concentration of polyamines were present in the feedstuff used. However, in growing kids the ruminai putrescine and spermidine concentration at times far exceeded those that could be accounted for by the estimated intake of polyamines by the food. The results therefore strongly indicate that polyamines are formed in considerable amounts in rumen content of kids during the phase of rapid growth. Results from a few experiments with calves also indicate that this may be true for cattle. polyamines; putrescine; spermidine; spermine; ornithine-decarboxylase; rumen liquid.     

Spermine-resistant mutants of Arabidopsis thaliana with developmental abnormalities

Fourteen spermine-resistant mutants of Arabidopsis thaliana (L.) Heynh. have been isolated from EMS-mutagenized M₂ seeds by screening for germination seedlings on a medium containing a germination inhibitory concentration of spermine. Two of these mutants have been studied initially. Genetic analysis indicated that the two mutants are allelic and the spermine resistance is due to recessive nuclear mutations at a locus we have designated SPR1. Mutant spr1-1 exhibits large, longitudinally folded-in cauline leaves whereas mutant spr1-2 is characterized by vigorous growth, large longitudinally folded-in cauline leaves, prominent flowers with 4–8 sepals, 6–8 petals, 6 (rarely 7) stamens of equal lengths, and 2–4 carpellary club-shaped pistil. Both mutants are resistant to exogenous spermine but are as sensitive as the wild-type to spermidine and putrescine.     

Metabolism of trypsin-inhibitory proteins in the germinating seeds of kidney bean (Phaseolus vulgaris)

Summary A number of proteins with trypsin-inhibitory activity was separated by isoelectric focusing and their amounts measured in the extracts of the seeds of kidney bean at various stages of germination up to 16 days.The total trypsin inhibitor content of the dormant seed, 2.2 mg per g bean rose to about 3.6 mg by the seventh day and declined slowly after the tenth day of germination. The individual trypsin inhibitors however, appeared to change independently of each other and some components disappeared almost completely with the progress of germination. The emergence of an inhibitor not found in the dormant seed was also observed. Some of the inhibitor proteins attained a maximum concentration by the 7–8th day of germination. This coincided with a similar maximum in the general protein and proteolytic enzyme content of the germinating bean seeds. The results obtained suggested that the main function during germination of these protein components might not be related to their trypsin-inhibitory activity.     

Polyamine Content in Relation to Embryo Growth and Dedifferentiation in Barley (Hordeum vulgare L.)

Putrescine, spermidine, and spermine content were analysed inzygotic embryos of barley (Hordeum vulgare L.). Changes in polyaminecontent were observed during zygotic embryo growth. In two cultivars,‘Bomi’ and ‘Golden Promise’, the totalpolyamine content in the embryos was 2.6–2.9 nmol mg^–1fresh weight 10 d after anthesis, the highest content observed.It dropped to 1.3 nmol mg^–1 fresh weight 14 d after anthesis.This drop was caused by decreases in all three polyamine concentrations.From 14 to 35 d after anthesis the putrescine content continuedto decrease while the spermidine and spermine content increased,thus the total polyamine content remained constant until 35d after anthesis. The mutant ‘Ris? 1508’ showeda constant polyamine content around 1.3 nmol mg^–1 freshweight from 14 to 35 d after anthesis. The polyamine patternwas conserved in all three lines throughout the period of investigationshowing a spermidine content higher than putrescine contentwhich was, in turn, higher or equal to the spermine content.The polyamine content measured as nmol µg^–1 proteindecreased from 14 to 21 d post anthesis in all three lines,because the protein content (µg mg^–1 fresh weight)increased during the period. In dedifferentiating zygotic embryoscultured in vitro the putrescine content (nmol mg^–1 freshweight) rose by a factor of nine and the spermidine contentdoubled within the first week of cultivation, whereas sperminecontent did not change. For embryoderived calli a repeated patternof change in polyamine content was observed throughout the subculturingperiod. Key words: Polyamines, Hordeum vulgare L., embryo development     

Determination of polyamines in digestive and reproductive tissues of adult Asterias vulgaris (Echinodermata: Asteroidea)

1. The polyamines spermine, spermidine and putrescine were extracted from tissues of Asterias vulgaris and quantitated using thin layer chromatography. 2. In the pyloric caeca, mean (+/- SE) levels of spermine, spermidine and putrescine were 138(15), 86(24) and 415(77) nmol/g wet wt tissue, respectively. In the testes, levels were 95(12), 13(6) and less than 6 nmol/g wet wt. In the ovaries, levels were 105(9), 11(0.8) and 15(8) nmol/g wet wt. 3. High levels of polyamines in the pyloric caeca may be related to secretion or excretion in this tissue. 4. We hypothesize that polyamines will be important in the regulation of cellular activity in these tissues during the annual reproductive cycle.     

Developmental pattern of poly (ADP-ribose) synthetase and NAD glycohydrolase in the brain of the fetal and neonatal rat

Poly (ADP-ribose) synthetase and NAD glycohydrolase were examined in nuclear fractions from rat brain at sequential times during late fetal and the first two weeks of neonatal life. In whole brain, both enzymes were demonstrable at all stages of development, but followed separate patterns. Activity of the synthetase which was greatest in fetal life, fell steadily with fetal maturation from 3.90±0.06 nmol/mg DNA at 16 days, to reach a nadir of 1.36±0.09 nmol/mg DNA on the 4th postnatal day. Subsequently it underwent a non sustained neonatal rise reaching a peak of 2.46±0.07 nmol/mg DNA on the 8th day. By contrast, NAD glycohydrolase activity increased steadily throughout late fetal and during the first two weeks of neonatal life, from 12.77±0.40 nmol/mg DNA on day 16 of gestation to 25.80±.95 nmol/mg DNA on neonatal day 12. In neonatal cerebellum the activity of poly (ADP-ribose) synthetase was greater at 8 than at 4 days, could be stimulated with graded concentrations of sonicated DNA up to 100 g, but was inhibited by higher concentrations of DNA and by all concentrations of exogenous histone. In an in vitro culture system of fetal rat brain cells, the activity of poly (ADP-ribose) synthetase increased steadily over six days. Cycloheximide 10^–3 M completely inhibited the activity of this enzyme. NAD glycohydrolase activity increased progressively in vitro, and after 6 days in cycloheximide (10^–3 M), the cultures contained significantly greater levels of enzyme activity. It is suggested that changing activities of poly (ADP-ribose) synthetase and NAD glycohydrolase could both provide potential markers for brain cell differentiation in this system.     

Utilisation of soil organic P by agroforestry and crop species in the field,western Kenya

A field experiment in western Kenya assessed whether the agroforestry species Tithonia diversifolia (Hemsley) A. Gray, Tephrosia vogelii Hook f., Crotalaria grahamiana Wight & Arn. and Sesbania sesban (L) Merill. had access to forms of soil P unavailable to maize, and the consequences of this for sustainable management of biomass transfer. The species were grown in rows at high planting density to ensure the soil under rows was thoroughly permeated by roots. Soil samples taken from beneath rows were compared to controls, which included a bulk soil monolith enclosed by iron sheets within the tithonia plot, continuous maize, and bare fallow plots. Three separate plant biomass samples and soil samples were taken at 6-month intervals, over a period of 18 months. The agroforestry species produced mainly leaf biomass in the first 6 months but stem growth dominated thereafter. Consequently, litterfall was greatest early in the experiment (0–6 months) and declined with continued growth. Soil pH increased by up to 1 unit (from pH 4.85) and available P increased by up to 38% (1 g P g^–1) in agroforestry plots where biomass was conserved on the field. In contrast, in plots where biomass was removed, P availability decreased by up to 15%. Coincident with the declines in litterfall, pH decreased by up to 0.26 pH units, plant available P decreased by between 0.27 and 0.72 g g^–1 and P_o concentration decreased by between 8 and 35 g g^–1 in the agroforestry plots. Declines in P_o were related to phosphatase activity (R²=0.65, P<0.05), which was greater under agroforestry species (0.40–0.50 nmol MUB s^–1 g^–1) than maize (0.28 nmol MUB s^–1 g^–1) or the bare fallow (0.25 nmol MUB s^–1 g^–1). Management of tithonia for biomass transfer, decreased available soil P by 0.70 g g^–1 and P_o by 22.82 g g^–1. In this study, tithonia acquired P_o that was unavailable to maize. However, it is apparent that continuous cutting and removal of biomass would lead to rapid depletion of P stored in organic forms.     

Optimization and maintenance of soluble methane monooxygenase activity inMethylosinus trichosporium OB3b

Soluble methane monooxygenase (sMMO) maximization studies were carried out as part of a larger effort directed towards the development and optimization of an aqueous phase, multistage, membrane bioreactor system for treatment of polluted groundwater. A modified version of the naphthalene oxidation assay was utilized to determine the effects of methane:oxygen ratio, nutrient supply, and supplementary carbon sources on maximizing and maintaining sMMO activity inMethylosinus trichosporium OB3b.Methylosinus trichosporium OB3b attained peak sMMO activity (275–300 nmol of naphthol formed h^–1 mg of protein^–1 at 25°C) in early stationary growth phase when grown in nitrate mineral salts (NMS) medium. With the onset of methane limitation however, sMMO activity rapidly declined. It was possible to define a simplified nitrate mineral salts (NMS) medium, containing nitrate, phosphate and a source of iron and magnesium, which allowed reasonably high growth rates (_max 0.08 h^–1) and growth yields (0.4–0.5 g cells/g CH₄) and near maximal activities of sMMO. In long term batch culture incubations sMMO activity reached a stable plateau at approximately 45–50% of the initial peak level and this was maintained over several weeks. The addition of d-biotin, pyridoxine, and vitamin B₁₂ (cyanocobalamin) increased the activity level of sMMO in actively growing methanotrophs by 25–75%. The addition of these growth factors to the simplified NMS medium was found to increase the plateau sMMO level in long term batch cultures up to 70% of the original peak activity.Abbreviations sMMO soluble methane monooxygenase - pMMO particulate methane monooxygenase - NMS nitrate mineral salts - TCE trichloroethene - NADH reduced nicotinamide adenine dinucleotide     

Glucose oxidase and catalase activities ofPenicillium variabile P16 immobilized in polyurethane sponge

Conidia ofPenicillium variabile P16 were immobilized in polyurethane sponge and used in repeated-batch processes in a fluidized-bed reactor. Optimal conditions for production of glucose oxidase and catalase were: inoculum size, 10%; glucose concentration, 80 g L^–1; Ca-carbonate concentration, 15 g L^–1; temperature, 28°C and aeration rate, 4 VV^–1 min^–1. In an extended repeated-batch process, glucose oxidase activity was highest after the fourth batch and catalase activity was highest after the fifth batch. Scanning electron microscopy showed that the fungus grew only in the interior of carrier particles.     

The synthesis and turnover of spermidine and spermine in mouse brain

Following the administration to mice of radiolabeled putrescine by intraventricular injection, changes in the specific radioactivity of putrescine, spermidine, and spermine have been measured. Putrescine decline was biphasic, being more rapid over the first 12 hr(t _1/2=5 hr) than over the remainder of the 48-hr period (t _1/2=11 hr) that significant labeling was detected. Spermidine was rapidly labeled during the decline in putrescine radioactivity and maximum incòrporation of label occurred at 18 hr. Subsequently, spermidine specific activity declined with a half-life of 22 days. Spermine synthesis was slower, with maximum labeling occurring after 4 days. Spermine turnover, measured at a time when spermidine radioactivity had substantially declined, was extremely slow (t _1/2=92 days). The data supports the view that putrescine is a precursor of spermidine which in turn is required for spermine synthesis.     

Growth promotion of Prunus rootstocks by root treatment with specific bacterial strains

A collection of bacterial strains obtained from a wide-range origin was screened for ability to promote growth in two types of Prunus rootstocks in a commercial nursery. Only few strains promoted growth significantly and consistently, and a strong specificity for the rootstock cultivar was observed. Irrigation of plants with Pseudomonas fluorescens EPS282 and Pantoea agglomerans EPS427 significantly increased plant height and root weight of the plum Marianna 2624 and the peach–almond hybrid GF-677, respectively. Plant height showed a higher rate of growth in early stages of development (2.6–3.5 times the non-treated controls), but the effect decreased with plant age. However, in aged plants growth promotion was more significant on root weight (1.9 times the non-treated controls) than on plant height. The efficacy of growth promotion and the persistence of strains in the root environment were dependent on the bacterial inoculum concentration applied. Increases in root development were maximum at inoculum concentrations of up to 8 log₁₀ CFU ml^–1 (ca 10 log₁₀ CFU L^–1 of potting mix). Population levels at the optimum inoculum concentration were around 7 log₁₀ CFU g f.w.^–1 root material at early stages of development and decreased to 4 log₁₀ CFU g f.w.^–1 after several months of development. The best plant growth-promoting strains were very diverse in secondary metabolite production and antagonistic ability against several plant pathogens.     

Hydrilla control with split treatments of fluridone in Lake Harris, Florida

After several unsuccessful management efforts, a split treatment of fluridone was applied to the 6700 ha Lake Harris in March and June 1987, at a rate of 3.4 kg ha^–1 (680 and 340 kg fluridone, respectively) to two 3 m deep, hydrilla-infested bays. Fluridone concentrations in the water were sampled following the June treatment. Average fluridone concentrations were 2.1 µg l^–1 prior to this second application, and a maximum concentration of 30.2 µg l^–1 was detected in the treated area on the day following application. Fluridone residues dissipated out of the plot quickly due to dilution but concentrations declined lake-wide more slowly, following a logarithmic model, with an estimated fluridone half-life of 97 days. Control of hydrilla in Lake Harris resulted from the long exposure (over 25 weeks due to the split application) to fluridone concentrations of 2 µg l^–1, well below the maximum labelled rate of 150 µg l^–1.