Filial cannibalism of Nabis pseudoferus is not evolutionarily optimal foraging strategy.

Using a recursion model with real parameters of Nabis pseudoferus, we show that its filial cannibalism is an optimal foraging strategy for life reproductive success, but it is not an evolutionarily optimal foraging strategy, since it cannot maximize the descendant's number at the end of the reproductive season. Cannibalism is evolutionarily rational, when the number of newborn offspring produced from the cannibalized offspring can compensate the following two effects: (a) The cannibalistic lineage wastes time, since the individuals hatched from eggs produced by cannibalism start to reproduce later. (b) Cannibalism eliminates not only one offspring, but also all potential descendants from the cannibalized offspring during the rest of reproductive season. In our laboratory trials, from conspecific prey Nabis pseudoferus did not produce newborn nymphs enough to compensate the above two effects.

Evaluation and Selection of New Trichogramma spp. as Biological Control Agents of the Guatemalan Potato Moth (Tecia solanivora) in Europe.

The Guatemalan potato moth (Tecia solanivora) is designated a quarantine pest by the European Union, causing severe production losses in potato crops. No effective chemical control alternatives are currently available, and cultural techniques are unable to reduce harvest losses to acceptable levels. With a focus on biological control, two egg parasitoids (Trichogramma euproctidis and Trichogramma achaeae) were selected and evaluated for use under field and storage conditions. Laboratory assays (choice and no-choice) indicated the preference of both parasitoids for T. solanivora vs. Phthorimaea operculella. Trichogramma euproctidis showed the highest parasitic activity for both moths. Analysis of functional response (at 15, 20, 25 and 27 °C) confirmed the high parasitic potential of T. euproctidis. Furthermore, in assays conducted under darkness conditions, T. achaeae was unable to parasitize eggs. However, in semi-field assays, T. achaeae was more efficient in searching for eggs in the soil than T. euproctidis. Based on these results, T. achaeae was selected to be tested under field conditions, and T. euproctidis was selected for testing under storage conditions.

Effect of Relative Humidity on the Population Dynamics of the Predator Amblyseius swirskii and Its Prey Carpoglyphus lactis in the Context of Slow-Release Sachets for Use in Biological Control in Greenhouses.

Amblyseius swirskii is a predatory mite that is widely used for biological control in greenhouses. One way this predator is released is in a formulation in slow-release sachets. These sachets are prepared with the predatory mite, the factitious prey mite Carpoglyphus lactis, and a food substrate for the latter. The objective of the present study was to study the effects of microclimatic conditions in this type of formulation on the population dynamics of mites inside the sachets and on the release of predatory mites. These experiments were conducted under laboratory conditions in two trials. The ambient relative humidity affected the water content of the food substrate of the prey mite inside the sachets, with an initial value of 21.49 ± 0.42%, which was reduced to values of 4.7 ± 0.25%, 10.87 ± 1.03% and 17.27 ± 0.82% after 21 days of trials when they were exposed to low, medium and high ambient relative humidity levels, respectively. Relative humidity significantly altered the dynamics of the populations of both species inside the sachets and the exits of the predator from the sachets to the external environment.

Biological Control Options for the Golden Twin-Spot Moth, Chrysodeixis chalcites (Esper) (Lepidoptera: Noctuidae) in Banana Crops of the Canary Islands.

Chrysodeixis chalcites (Esper) (Lepidoptera: Noctuidae) is a significant pest in banana plantations in the Canary Islands. Field surveys were carried out to identify its naturally occurring parasitoids and estimate their parasitism rates between September 2007 and October 2010. Ch. chalcites was parasitized by six different larval/pupal parasitoid species: Cotesia sp., C. glomerata L. (Hym.: Braconidae), Aplomyia confinis Fallén (Dip.: Tachinidae), Hyposoter rufiventris Perez, Ctenochares bicolorus L. (Hym.: Ichneumonidae) and Aleiodes sp. (Hym.: Braconidae). Among them, Cotesia sp. was the most frequent species, accounting for 8.18% of parasitized larvae. High levels of egg parasitism were detected, with Trichogramma achaeae Nagaraja and Nagarkatti (Hym.: Trichogrammatidae) being the most widely distributed egg parasitoid. A greenhouse assay was also carried out on a commercial banana crop with the aim of evaluating the potential of T. achaeae as a biological control agent and compared with a chemical control. Five periodic inundative releases of 35 adults/m2 every 21 days were necessary to achieve an adequate parasitism level (56.25 ± 1.61%). Moreover, there was 15.75% less foliar damage in the biological control plot compared to the chemical control plot. These results indicate that T. achaeae could be a promising biocontrol agent of Ch. chalcites in greenhouse banana crops.

Effect of Temperature on Life History and Parasitization Behavior of Trichogramma achaeae Nagaraja and Nagarkatti (Hym.: Trichogrammatidae).

Trichogramma achaeae Nagaraja and Nagarkatti (Hymenoptera: Trichogrammatidae) is currently used as biological control agent for several lepidopteran pests. Knowledge of thermal requirements is essential to optimize its rearing procedures and inundative releases. The biological characteristics and two-sex life table parameters of T. achaeae were determined at five constant temperatures (15, 20, 25, 30, and 35 °C) using Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs. T. achaeae was able to develop and survive from 15 °C to 30 °C, but not at 35 °C. Temperature significantly affected the preadult development time and adult longevity, decreasing when temperature increased from 15 °C to 30 °C. Temperature significantly altered the sex ratio, being female biased between 15 °C and 25 °C. Age-stage, two-sex life table analysis indicated that net reproductive rate (R0) was highest at 20 °C. Both the intrinsic rate of increase (r) and finite rate of increase (λ) increased with increasing temperature, while the mean generation time (T) decreased significantly. In addition, functional response of T. achaeae was studied, being significantly affected by temperature and host egg density, displaying a Holling type-I at 15 °C and a Holling type-II at 25 °C. The relevance of these results is discussed for the use of T. achaeae as biological control agent.

Age-Stage, Two-Sex Life Table of Chrysodeixis chalcites (Lepidoptera: Noctuidae) at Constant Temperatures on Semi-Synthetic Diet.

The golden twin-spot moth or tomato looper, Chrysodeixis chalcites (Esper), is a polyphagous and worldwide pest that causes important aesthetic damages to banana fruits in the Canary Islands. The life history parameters of C. chalcites were determined under laboratory conditions in base on the age-stage, two-sex life table at 15, 20, 25, 30, and 35°C, 65% relative humidity (RH), and a photoperiod of 16:8 (L:D) h, when it was reared on a semi-synthetic diet. The results show that C. chalcites was able to develop and survive from 15 to 30°C, but no development occurred at 35°C. Developmental threshold temperatures of the egg, larval, pupal, and total preoviposition stages were 10.42, 11.73, 11.22, and 9.42°C, respectively, and their effective accumulated temperatures were 58.31, 265.96, 118.57, and 562.39 degree-days, respectively. The adult longevity was reduced with increasing temperature, which ranged between 16.27 and 34.85 d for females and between 14.27 and 35.21 d for males. The highest values of net reproductive rate (R0) and fecundity were observed at 25°C, with 232.70 offspring and 1,224.74 eggs, respectively. Both the intrinsic rate of increase (r) and finite rate of increase (λ) increased significantly and mean generation time (T) decreased significantly with increasing temperature. These results provide useful information that will allow predicting the impact of climate change on the distribution and population dynamics of C. chalcites and developing successful integrated management programs.

Selection of Predatory Mites for the Biological Control of Potato Tuber Moth in Stored Potatoes.

Worldwide, the potato tuber moth (PTM), Phthorimaea operculella (Zeller), is one of the most severe pests affecting potato (Solanum tuberosum L.), whether in open-air crops or during tuber storage. This work examines the potential control of this pest by two species of predatory mites, Macrocheles robustulus (Berlese) and Blattisocius tarsalis (Berlese), on pest eggs under laboratory conditions. In the two first assays, the acceptance rate of the pest eggs was assessed for each predatory mite. Then, in a third assay, the functional response of B. tarsalis was studied. The results showed that Macrocheles robustulus did not prey on the pest eggs (number of eggs surviving = 4.33 ± 0.38), whereas B. tarsalis did (number of eggs surviving = 0.5 ± 0.5). Likewise, B. tarsalis showed a type II functional response when it killed the eggs. The results showed the potential use of Blattisocius tarsalis as a biological control agent of P. operculella in potato under storage conditions.

Do Development and Diet Determine the Degree of Cannibalism in Insects? To Eat or Not to Eat Conspecifics.

Cannibalism in insects plays an important role in ecological relationships. Nonetheless, it has not been studied as extensively as in other arthropods groups (e.g., Arachnida). From a theoretical point of view, cannibalism has an impact on the development of more realistic stage-structure mathematical models. Additionally, it has a practical application for biological pest control, both in mass-rearing and out in the field through inoculative releases. In this paper, the cannibalistic behavior of two species of predatory bugs was studied under laboratory conditions-one of them a generalist predator (strictly carnivorous), Nabis pseudoferus, and the other a true omnivore (zoophytophagous), Nesidiocoris tenuis-and compared with the intraguild predation (IGP) behavior. The results showed that cannibalism in N. pseudoferus was prevalent in all the developmental stages studied, whereas in N. tenuis, cannibalism was rarely observed, and it was restricted mainly to the first three nymphal stages. Cannibalism and intraguild predation had no linear relationship with the different cannibal-prey size ratios, as evaluated by the mortality rates and survival times, although there were variations in cannibalism between stages, especially for N. pseudoferus. The mathematical model's implications are presented and discussed.

Photosynthesis Inhibiting Effects of Pesticides on Sweet Pepper Leaves.

Although a large number of pesticides of different compositions are regularly used in agriculture, the impact of pesticides on the physiology of field crops is not well understood. Pesticides can produce negative effects on crop physiology-especially on photosynthesis-leading to a potential decrease in both the growth and the yield of crops. To investigate these potential effects in greenhouse sweet peppers, the effect of 20 insecticides and 2 fungicides (each sprayed with a wetting agent) on the photosynthesis of sweet pepper leaves was analyzed. Among these pesticides, nine caused significant reductions in photosynthetic activity. The effects were observed in distinctive ways-either as a transitory drop of the photosynthetic-rate values, which was observed at two hours after the treatment and was found to have recovered after 24 h, or as a sustained reduction of these values, which remained substantial over a number of days. The results of this study suggest that the production of a crop may substantially benefit when the frequent use of pesticides can be substituted with alternative pest control methods (e.g., biological control). Our results advocate further investigation of the potential impact of pesticides, either alone or in combination, on the photosynthesis of crop plants.

Opportunistic random searcher versus intentional search image user.

We consider two types of optimal foragers: a random searcher and a search image user. A search image user can find its desired prey with higher and undesired prey with lower probability than a random searcher. Our model considers the density-dependent travelling time and the time duration of reproduction (oviposition). In the framework of optimal foraging theory for one predator-two prey systems, we find that there are ranges of prey densities in which the search image user has a higher net energy intake, and there are other ranges of prey densities in which the random searcher has higher net energy intake. The damsel bug Nabis pseudoferus Remane (Hemiptera: Nabidae) is a generalist predator rather than an omnivore. This species has a wide range of arthropod prey (predominantly insects and mites). Several aspects of the biology of this species have been studied, especially its cannibalistic behaviour, which is a quite important feature because N. pseudoferus is often used as a biological control agent against lepidopteran pests in greenhouse crops. Experimentally, we found that Nabis is a search image user in the above sense.

Can Interactions Between an Omnivorous Hemipteran and an Egg Parasitoid Limit the Level of Biological Control for the Tomato Pinworm?

Relationships between the omnivorous predator Nesidiocoris tenuis (Reuter) and the egg parasitoid Trichogramma achaeae Nagaraja and Nagarkatti were studied in the laboratory (no-choice and choice assays, and functional responses) and in a greenhouse experiment. Both natural enemies are utilized in the biological control of tomato pinworm on greenhouse-grown tomato crops. Three different food items were offered to the predator: nonparasitized prey, prey parasitized for less than 4 d by T. achaeae, and prey parasitized for more than 4 d by the parasitoid. There were significant differences in consumption of food types, with highest consumption for nonparasitized prey, followed by parasitized (<4 d) and then parasitized (>4 d), both in no-choice and choice trials. At the same time, the predator causes a significant mortality in the prey (over 80%) regardless of previous parasitism, resulting in a very coincidental intraguild predation detrimental to the parasitoid. It has also been observed that there was a change in the functional response by the predator from Type II in presence of nonparasitized prey to Type I when there was a combination of parasitized and nonparasitized prey. This represents an increase of instantaneous search rate (a') and a decrease of handling time (Th), which indicates a change in feeding behavior on the two prey types. Under greenhouse conditions, the intraguild predation reduced the percentage of parasitism by T. achaeae in just over 20%. However, when both natural enemies were present, a better control of pest Tuta absoluta (Meyrick) was achieved than in the case of application of any of them alone.

Optimal forager against ideal free distributed prey.

The introduced dispersal-foraging game is a combination of prey habitat selection between two patch types and optimal-foraging approaches. Prey's patch preference and forager behavior determine the prey's survival rate. The forager's energy gain depends on local prey density in both types of exhaustible patches and on leaving time. We introduce two game-solution concepts. The static solution combines the ideal free distribution of the prey with optimal-foraging theory. The dynamical solution is given by a game dynamics describing the behavioral changes of prey and forager. We show (1) that each stable equilibrium dynamical solution is always a static solution, but not conversely; (2) that at an equilibrium dynamical solution, the forager can stabilize prey mixed patch use strategy in cases where ideal free distribution theory predicts that prey will use only one patch type; and (3) that when the equilibrium dynamical solution is unstable at fixed prey density, stable behavior cycles occur where neither forager nor prey keep a fixed behavior.

Functional response and population dynamics for fighting predator, based on activity distribution.

The classical Holling type II functional response, describing the per capita predation as a function of prey density, was modified by Beddington and de Angelis to include interference of predators that increases with predator density and decreases the number of killed prey. In the present paper we further generalize the Beddington-de Angelis functional response, considering that all predator activities (searching and handling prey, fight and recovery) have time duration, the probabilities of predator activities depend on the encounter probabilities, and hence on the prey and predator abundance, too. Under these conditions, the aim of the study is to introduce a functional response for fighting the predator and to analyse the corresponding dynamics, when predator-predator-prey encounters also occur. From this general approach, the Holling type functional responses can also be obtained as particular cases. In terms of the activity distribution, we give biologically interpretable sufficient conditions for stable coexistence. We consider two-individual (predator-prey) and three-individual (predator-predator-prey) encounters. In the three-individual encounter model there is a relatively higher fighting rate and a lower killing rate. Using numerical simulation, we surprisingly found that when the intrinsic prey growth rate and the conversion rate are small enough, the equilibrium predator abundance is higher in the three-individual encounter case. The above means that, when the equilibrium abundance of the predator is small, coexistence appears first in the three-individual encounter model.

Optimal nutrient foraging strategy of an omnivore: Liebig's law determining numerical response.

The paper is aimed at a theoretical explanation of the following phenomenon. In biological pest control in greenhouses, if an omnivore agent is released before the arrival of the pest, the agent may be able to colonize, feeding only on plant and then control its arriving prey to a low density. If the pest arrives before the release of the agent, then it tends to reach a high density, in spite of the action of the agent. This means that according to the initial state, the system displays different stable equilibria, i.e. bistable coexistence is observed. Based on the biological situation, the explaining theoretical model must take into account the stoichiometry of different nutrients and the optimal foraging of the omnivore agent. We introduce an optimal numerical response which depends on the optimal functional responses and on the 'mixed diet-fitness' correspondence determined by 'egg stoichiometry', in our case by Liebig's Law; moreover we also study the dynamical consequences of the latter when the plant is "inexhaustible". In our model, we found that under Holling type II functional response, the omnivore-prey system has a unique equilibrium, while for Holling type III, we obtained bistable coexistence. The latter fact also explains the above phenomenon that an omnivore agent may control the pest to different levels, according to the timing of the release of the agent.

Biological control strategies for the South American tomato moth (Lepidoptera: Gelechiidae) in greenhouse tomatoes.

The South American tomato pinworm, Tuta absoluta (Meyrick) has been introduced into new geographic areas, including the Mediterranean region, where it has become a serious threat to tomato production. Three greenhouse trials conducted in tomato crops during 2009 and 2010 explored control strategies using the egg-parasitoid Trichogramma achaeae Nagaraja and Nagarkatti compared with chemical control. The effectiveness of the predator Nesidiocoris tenuis (Reuter) was also tested. In greenhouses with early pest infestations (discrete generations), periodic inundative releases (eight releases at a rate of 50 adults/m2, twice a week) were necessary to achieve an adequate parasitism level (85.63 +/- 5.70%) early in the growing season. However, only one inoculative release (100 adults/m2) was sufficient to achieve a comparatively high parasitism level (91.03 +/- 12.58%) under conditions of high pest incidence and overlapping generations. Some intraguild competition was observed between T. achaeae and the predator, N. tenuis. This mirid species is commonly used in Mediterranean greenhouse tomato crops for the control of the sweetpotato whitefly, Bemisia tabaci (Gennadius). Tomato cultivars were also observed to influence the activity of natural enemies, mainly N. tenuis (whose average numbers ranged between 0.17 +/- 0.03 and 0.41 +/- 0.05 nymphs per leaf depending on the cultivar). This may be because of differences in plant nutrients in different cultivars, which may affect the feeding of omnivorous insects. In contrast, cultivar effects on T. achaeae were less apparent or possibly nonexistent. Nevertheless, there was an indirect effect in as much as T. achaeae was favored in cultivars not liked by N. tenuis.

[Total plasma homocysteine levels. Relationship with plasmatic folic acid levels and 677C T polymorphism of 5,10-methylenetetrahydrofolate reductase]. / Concentración plasmática de homocisteína: relación con los niveles plasmáticos de ácido fólico y con el polimorfismo 677C T de la 5,10-metilenotetrahidrofolato reductasa.

BACKGROUND: Moderately increased plasma homocysteine (Hcy) in children has been associated with stroke and venous thrombosis and with a parental history of cardiovascular disease (CVD). Evaluation of Hcy concentrations during childhood and study of the factors determining its concentrations could play an important role in the primary prevention of CVD. Objective To detect cases of hyperhomocystinemia and to examine the association between Hcy levels and plasma folic acid levels and 677C T polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR). METHODS: The relationship between plasma Hcy levels, plasma folic acid levels, and the three genotypes of 677C T MTHFR polymorphism was investigated in 127 children (aged 2-18 years) and in 105 parents by multiple linear regression. RESULTS: The median Hcy levels were 5.00 mol/l in the children and 8.00 mol/l in the parents. Plasma folic acid levels were normal in all of the patients. The prevalence of the three genotypes in the children was 32.3 % for the CC genotype, 42.5 % for the CT genotype and 15.7 % for the TT genotype. Hcy concentrations were significantly higher in children with the TT genotype (p 0.018). Multiple linear regression revealed a positive direct effect of age (b 0.029, p 0.002) and a negative effect of genotype TT (b 3.886, p 0.002) on Hcy concentration. Hcy concentration was inversely correlated with folic acid levels but this correlation did not reach statistical significance. CONCLUSIONS: No cases of hyperhomocystinemia were found. To evaluate Hcy, age and plasma folic acid levels have to be taken into account in case there is a 677C T mutation. Hcy concentrations should be determined in older children with a family history of atherothrombosis and other risk factors for premature CVD.

Concentración plasmática de homocisteína: relación con los niveles plasmáticos de ácido fólico y con el polimorfismo 677 ct de la 5,10-metilenotetrahidrofolato reductasa / Total plasma homocysteine levels. Relationship with plasmatic folic acid levels and 677 CT polymorphism of 5,10-methylenetetrahydrofolate reductase

Antecedentes: El aumento moderado de la homocisteína plasmática en niños se ha relacionado con infartos cerebrales y trombosis venosas y con los antecedentes familiares de enfermedad coronaria prematura (ECP). La determinación de homocisteína en la infancia y el estudio de los factores que determinan su concentración podría ser importante para la prevención primaria de la ECP. Objetivo: Detectar algún caso de hiperhomocistinemia y valorar su relación con la concentración plasmática de ácido fólico y el polimorfismo 677C T de la 5,10-metilenotetrahidrofolato reductasa (MTHFR). Métodos: Se ha estudiado mediante la regresión lineal múltiple la relación entre la concentración plasmática de homocisteína, la del ácido fólico y los tres genotipos de la mutación 677C T de la MTHFR en 127 niños de entre 2 y 18 años y 105 de sus progenitores. Resultados: La concentración de homocisteína (mediana) fue de 5,00 y 8,00 mol/l en los niños y sus progenitores, respectivamente. Los valores plasmáticos de ácido fólico se encontraban todos en el rango de la normalidad. La prevalencia de los tres genotipos en los niños fue de 32,3% para el genotipo CC, 42,5% para el CT y 15,7% para el TT. La concentración de homocisteína era significativamente mayor con el genotipo TT (p = 0,018). En la regresión lineal múltiple se encontró un efecto directo positivo de la edad (b = 0,029; p = 0,001) y negativo del genotipo TT (b = -3,886; p = 0,002) sobre la concentración de homocisteína. El coeficiente de regresión de la concentración de ácido fólico aunque de signo negativo, no alcanzó significación estadística. Conclusiones: No se ha encontrado ningún caso de hiperhomocistinemia. Al valorar la homocisteína hay que tener en cuenta la edad y en caso de existir la mutación 677C T, los valores plasmáticos de ácido fólico. Sería conveniente determinar la homocisteína en los niños de mayor edad con antecedentes familiares de aterotrombosis y con otros factores de riesgo para la ECP (AU)

Pilot screening programme for cystinuria in the Valencian community.

Cystinuria is an autosomal recessive disorder of the kidneys and small intestine, affecting a luminal transport mechanism shared by cystine, ornithine, arginine and lysine. When cystine exceeds its solubility at low pH, the risk of stone formation increases. The data reported in the literature show a variation for the incidence of cystinuria, from 1 in 600 to 1 in 17,000, depending on the definition of cystinuria and the method used for screening the population. We set up a pilot screening programme to determine the incidence of cystinuria in the population of the Valencian Community. Urine filter paper samples submitted for the neonatal screening programme from 33,995 newborns (5-10 days old) were used for the study. Thin layer chromatography (TLC) was performed to screen cystinuric patients. To confirm positive filter paper samples, liquid samples were requested and TLC as well as the cyanide-nitroprusside test (CNT) were performed. Final diagnosis was achieved by quantifying cystine, lysine, ornithine and arginine using high-performance liquid chromatography (HPLC) in children's urine samples which remained positive for TLC and CNT for more than 1 year. We conclude that the incidence of subjects at risk for cystine stones in the Valencian Community is 1:1887. TLC is shown as a reliable method to perform newborn screening in large population to detect cystinuric subjects. Additional studies, including characterization of appropriate haplotypes, should be carried out for a more precise identification of the frequency of the different types of cystinuria in our population.

[Homocystinuria: effectiveness of the treatment with pyridoxine, folic acid, and betaine]. / Homocistinuria: eficacia del tratamiento con piridoxina, ácido fólico y betaína.

We present the results achieved with vitamin (pyridoxine and folic acid) and betaine (trimethyl-glycine) treatment of three patients with homocystinuria. Cases 1 and 2 were detected by having clinical findings suggestive of the disease (ocular and orthopedic alterations) and case 3 was diagnosed after a family metabolic screening was done. All presented a positive Brand's test and an abnormal elevation of plasma and urine homocysteine, as well as high methionine and low cystine levels in the plasma. Initially, when pyridoxine (600 mg/d) and folic acid (10 mg/d) were given for one month, a partial fall in the homocysteine levels was observed in cases 2 and 3, but not in case 1. When betaine was added (6 g/d), homocysteine disappeared from the plasma after the first month in cases 2 and 3, but only after the third month in case 1. Case 1 also showed a moderate clinical improvement in behavior and school performance. The treatment was maintained for two years in case 1, and for one year in cases 2 and 3. After betaine therapy, no disturbances were observed in the hepatic, renal and bone marrow functions, nor were there any clinically relevant ill-effects. These findings show that betaine offers a therapeutic alternative in the treatment of this disease, independent of the patient's response to pyridoxine.

Kinetochromic spectrophotometry-I Determination of fluoride by catalysis of the zirconium-xylenol orange reaction.

The reaction between slightly aged solutions of zirconyl chloride and Xylenol Orange is catalysed by the presence of substoi-chiometric amounts of fluoride, phosphate, arsenate, sulphate, citrate or oxalate. The reaction provides a method for determining fluoride in amounts from 0.5 to 5 mug in a final volume of 100 ml. This kineto-chromic reaction provides an effective molar absorptivity of 2.0 x 10(5) for the fluoride ion. The colour reaches maximal development within 90 min and remains stable thereafter for 60 min, when measured against a reaction system blank containing no fluoride. Because of its substoichiometric nature the reaction may not be used for the determination of large amounts of fluoride.