Consumption of sugars by Anastrepha suspensa (Diptera: Tephritidae).

We were interested in determining the feeding response of the Caribbean fruit fly, Anastrepha suspensa Loew (Diptera: Tephritidae), to various sugar concentrations to develop an improved bait for adults. We compared the consumption of 0.01-1.00 M concentrations of glucose, fructose, raffinose, and sucrose in no-choice tests for 24-h- and 6-d-old male and female flies. Sucrose was the most consumed sugar or within the most consumed group of sugars at 0.02-0.20 M concentrations. There were no differences in consumption among sugars at 0.01, 0.40, and 1.00 M. Consumption generally increased with increasing sugar concentration except that sucrose consumption peaked at 0.20 M. Twenty-four-hour females consumed less fructose than other sugars; 24-h males consumed more sucrose than fructose or raffinose, with an intermediate response to glucose. Females in the 6-d group consumed more sucrose than the other three sugars, whereas 6-d males exhibited no difference in consumption among sugars. In choice tests, flies consumed more sugar solution than water, but the difference between 0.20 M fructose and water was not significant for 24-h males or 24-h females. In choice tests between 0.20 M fructose and 0.20 M sucrose, both 24-h and 6-d females showed a preference for fructose. Males of both age classes showed no preference. These results indicate that the responses of flies to different sugars can vary by sugar, gender, and age.

Field comparison of chemical attractants and traps for Caribbean fruit fly (Diptera: Tephritidae) in Florida citrus.

Field studies in citrus were conducted to compare the following as attractants for the Caribbean fruit fly, Anastrepha suspensa (Loew): torula yeast-borax; propylene glycol (10%); a two-component lure consisting of ammonium acetate and putrescine; a two-component lure consisting of ammonium bicarbonate and putrescine; and a three-component lure consisting of ammonium bicarbonate, methylamine hydrochloride, and putrescine. Various combinations of these attractants in glass McPhail, plastic McPhail-type (Multi-Lure), and sticky panel traps were investigated in two replicated studies. In one study on wild flies, the most effective and least complex trap-lure combination tested was the Multi-Lure with propylene glycol baited with ammonium acetate and putrescine. This trap-lure combination captured significantly more female and male flies than the standard glass McPhail baited with torula yeast-borax in water. All of the trap-lure combinations were female biased, with an overall average of 80.8% (SEM 1.4) flies captured being female. A second study on laboratory-reared, irradiated flies indicated no significant differences among these trap-lure combinations with respect to number of flies recaptured, although rankings based on mean number of flies recovered per trap per day supported results of the first study. The percentage of flies recaptured that were female (83.0%, SEM 0.9) was statistically the same as in the first study. Weekly percentage recovery of flies during the second study was low, possibly due to our fly release strategy. Future release/recovery studies with laboratory-reared flies would benefit from some basic research on release strategies by using different trap densities and on relating recapture rates of laboratory-reared flies (nonsterile and sterile) to capture rates of wild flies.

Blood cholinesterases as human biomarkers of organophosphorus pesticide exposure.

The organophosphorus pesticides of this review were discovered in 1936 during the search for a replacement for nicotine for cockroach control. The basic biochemical characteristics of RBC AChE and BChE were determined in the 1940s. The mechanism of inhibition of both enzymes and other serine esterases was known in the 1940s and, in general, defined in the 1950s. In 1949, the death of a parathion mixer-loader dictated blood enzyme monitoring to prevent acute illness from organophosphorus pesticide intoxication. However, many of the chemical and biochemical steps for serine enzyme inhibition by OP compounds remain unknown today. The possible mechanisms of this inhibition are presented kinetically beginning with simple (by comparison) Michaelis-Menten substrate enzyme interaction kinetics. As complicated as the inhibition kinetics appear here, PBPK model kinetics will be more complex. The determination of inter- and intraindividual variation in RBC ChE and BChE was recognized early as critical knowledge for a blood esterase monitoring program. Because of the relatively constant production of RBCs, variation in RBC AChE was determined by about 1970. The source of plasma (or serum) BChE was shown to be the liver in the 1960s with the change in BChE phenotype to the donor in liver transplant patients. BChE activity was more variable than RBC AChE, and only in the 1990s have BChE individual variation questions been answered. We have reviewed the chemistry, metabolism, and toxicity of organophosphorus insecticides along with their inhibitory action toward tissue acetyl- and butyrylcholinesterases. On the basis of the review, a monitoring program for individuals mixing-loading and applying OP pesticides for commercial applicators was recommended. Approximately 41 OPs are currently registered for use by USEPA in the United States. Under agricultural working conditions, OPs primarily are absorbed through the skin. Liver P-450 isozymes catalyze the desulfurization of phosphorothioates and phosphorodithioates (e.g., parathion and azinphosmethyl, respectively) to the more toxic oxons (P = O(S to O)). In some cases, P-450 isozymes catalyze the oxidative cleavage of P-O-aryl bonds (e.g., parathion, methyl parathion, fenitrothion, and diazinon) to form inactive water-soluble alkyl phosphates and aryl leaving groups that are readily conjugated with glucuronic or sulfuric acids and excreted. In addition to the P-450 isozymes, mammalian tissues contain ('A' and 'B') esterases capable of reacting with OPs to produce hydrolysis products or phosphorylated enzymes. 'A'-esterases hydrolyze OPs (i.e., oxons), while 'B'-esterases with serine at the active center are inhibited by OPs. OPs possessing carboxylesters, such as malathion and isofenphos, are hydrolyzed by the direct action of 'B'-esterases (i.e., carboxylesterase, CaE). Metabolic pathways shown for isofenphos, parathion, and malathion define the order in which these reactions occur, while Michaelis-Menten kinetics define reaction parameters (Vmax, K(m)) for the enzymes and substrates involved, and rates of inhibition of 'B'-esterases (kis, bimolecular rate constants) by OPs and their oxons. OPs exert their insecticidal action by their ability to inhibit AChE at the cholinergic synapse, resulting in the accumulation of acetylcholine. The extent to which AChE or other 'B'-esterases are inhibited in workers is dependent upon the rate the OP pesticide is activated (i.e., oxon formation), metabolized to nontoxic products by tissue enzymes, its affinity for AChE and other 'B'-esterases, and esterase concentrations in tissues. Rapid recovery of OP BChE inhibition may be related to reactivation of inhibited forms. AChE, BChE, and CaE appear to function in vivo as scavengers, protecting workers against the inhibition of AChE at synapses. Species sensitivity to OPs varies widely and results in part from binding affinities (Ka) and rates of phosphorylation (kp) rather than rates of activation and detoxif

Pesticide protective clothing.

In looking into future directions of "protective clothing" for pesticide users, the final EPA ruling scheduled for release in June 1992 (Smith personal communication) places the majority of its recommendations on the label. Therefore, it will be up to pesticide manufacturers to provide protective clothing information on their products labels. Research on protective clothing continues to show variations due to fabric characteristics of fiber content, fabric construction and finish. These variabilities are compounded by variation s due to pesticide formulation and pesticide chemical and are further compounded by field studies vs laboratory simulations. With hundreds of fabric variations compounded by the thousands of chemical compounds and formulation variations, the consumer cannot be expected to make proper decisions regarding protective clothing. The user of the product does not have the knowledge to select the appropriate clothing. Manufacturers must supply this information, but they must also have data to support recommendations of their clothing for pesticide protection. This shifts the responsibility for recommending adequate protective clothing, as well as the concern for product liability, to the manufacturer.

Saliva as a monitoring medium for chemicals.

Saliva has been used to estimate plasma levels in humans for a variety of analytes. The potential for estimating the internal dose of pesticides in humans has been demonstrated with the rat. The utility and problems of saliva-sampling techniques remain to be investigated. Measurement of pesticides and other environmental toxicants in saliva has great potential because of sampling simplicity and the potential accuracy of using the saliva concentration of a chemical to estimate its concentration in human tissues.

Phototoxic coumarins in limes.

Coumarins in the rind and pulp of Persian and Key limes were quantified. In the rind of Persian limes, coumarin concentrations were in the order: limettin > bergapten > isopimpinellin > xanthotoxin > psoralen. In the rind of Key limes, psoralen and xanthotoxin were analytically absent; limettin was 10 times more concentrated than either bergapten or isopimpinellin, which were equal in concentration. Coumarin content in Persian lime pulp was in the order: isopimpinellin > limettin > bergapten > xanthotoxin > psoralen. For Key lime pulp, the concentrations of limettin, isopimpinellin and bergapten were equal; psoralen and xanthotoxin were not detected. Coumarins in lime pulp were 13 to 182 times less concentrated than those in the peel. Based on the amounts and types of coumarins, Persian limes appear to be potentially more phototoxic than Key limes. Although bergapten may be the main component of limes responsible for phytophotodermatitis, dermatological interaction assays with psoralen, bergapten, xanthotoxin and limettin should be conducted.

Sampling methods as abundance indices for adult Diaprepes abbreviatus (Coleoptera: Curculionidae) in citrus.

Beat sampling and two type of traps, cup traps and Tedders traps, were evaluated as sampling methods to detect and estimate population densities of adult Diaprepes abbreviatus L. weevils newly colonizing young citrus trees. The study was conducted over a 65-wk period across a 0.25-ha area of 80 citrus trees [Citrus sinensis (L.) Osbeck] (1.2-1.5 m tall). Beat samples were taken weekly to determine the number of trees infested and number of new adult weevils per tree. Sixteen of the 80 trees studied were each monitored weekly using one of the following trapping methods: cup traps in trees, cup traps on a stake in the ground within the tree drip line, cup traps on a stake in the ground outside of the drip line, Tedders traps on the ground within the drip line, and Tedders traps on the ground outside of the drip line. Weevils collected each week from trees and traps were removed from the study site. Based on the coefficients associated with Taylor's power law, the optimum numbers of trees to sample for an SEM equal to 25% of the mean estimate decreased from 50 trees at a mean of 0.5 new weevils per tree to 30 trees at a mean of 0.8 new weevils per tree. A significant relationship was found between the weekly mean number of new weevils per tree and the proportion of trees infested, a binomial relationship that could be further explored in the search for a sampling program for adult D. abbreviatus. Regression analyses indicated that three of the trapping methods served at least as weak indices of the presence and abundance of new weevils: cup traps in trees, Tedders traps inside the dripline and Tedders traps outside the dripline. Cup traps in trees and Tedders traps inside the dripline captured the most weevils and most frequently detected weevils. Although relatively inefficient as abundance indices of populations of new weevils, these two trapping methods appeared to have some value with respect to signaling when weevils first appeared in trees during the spring.

Quantifying individual fruit fly consumption with Anastrepha suspensa (Diptera: Tephritidae).

We needed a technique to compare the consumption of baits by individual Carribbean fruit fly, Anastrepha suspensa (Loew). By improving consumption and determining individual dose, we could lower pesticide concentration while retaining bait/pesticide efficacy and potentially reduce the environmental impact of fruit fly bait/pesticide eradication methods. We report here a precise dye-based technique for the quantification of consumption by individual adult A. suspensa fruit flies. Fluorescein, measured at 491 nm, and cresol red, measured at 573 nm, were efficiently extracted with 0.1 M NaOH and quantified with a spectrophotometer. The lower limit for this method with 0.1% dye concentration is 300 nl consumed by an individual fly. Dye movement to the hindgut and possible defecation occurred in approximately 4 h; maximum ingestion occurred in approximately 1 h. Maximum experimental time is limited to 4 h. Flies preferred feeding upside down compared with right side up when given a choice; consumption was equal when flies were given no choice of feeding position. Thus, maximum bait/pesticide efficacy might be achieved with an upside-down presentation. Regurgitation led to a 100% overestimation of actual consumption with the J-tube presentation of food. Our individual fly consumption technique will be useful in comparing consumption in phagostimulant studies, estimating dose in oral toxicity tests, differentiating behavioral and physiological resistance in toxicant studies, ultimately leading to improved bait/pesticide methods and reduced environmental impact of area wide fruit fly eradication programs. This technique could be applied to studies of tephritid consumption, to the consumption of other insects, and to regurgitation studies.

Abundance of Diaprepes abbreviatus (L.) (Coleoptera: Curculionidae) neonates falling to the soil under tree canopies in Florida citrus groves.

The purpose of these experiments was to estimate the number and distribution of Diaprepes abbreviatus (L.) neonate larvae dropping from the canopy of infested citrus trees. The number of neonates was monitored in the field using passive funnel traps in two simultaneous experiments and a separate experiment for an additional year. In one experiment, traps were placed from trunk to dripline in the cardinal directions under each of five trees (132 traps total). In a second experiment, eight traps were placed under each tree in the cardinal directions, one trap 30 cm from the trunk and one trap 30 cm from the dripline/direction for 25 trees (200 traps total). Larvae were collected weekly for 50 wk in conical tubes containing ethylene glycol as a preservative. Traps closer to the tree trunk captured more larvae than traps nearer the dripline. The area under the tree canopy was positively correlated with the total estimated number of larvae captured per tree. The estimated number of total larvae/tree over the course of our experiments ranged from 955 to 7,290. The highest number of neonate larvae observed in 1 wk was 67 +/- 6/m2. There was an inverse relationship between the number of traps beneath a tree and the number of trees that needed to be sampled to estimate mean population density with a given precision. However, there was a direct relationship between number of traps/tree and the total number of traps needed for a given precision. This passive technique could be used to quantify the destructive larval stage and to assess D. abbreviatus management strategies.

Sodium tetraborate effects on mortality and reproduction of Anastrepha suspensa (Diptera: Tephritidae).

When flies were treated with 0- 0.5% sodium tetraborate by feeding for 24 h, mortality in treatments was not different from controls. Fecundity and fertility were reduced by 0.5% sodium tetraborate. When flies were fed for 48 h, mortality of both males and females increased in the 0.5% sodium tetraborate treatment; oviposition was eliminated for 20 d after treatment. When treatment was extended to 168 h, 0.1% sodium tetraborate caused increased mortality and decreased fecundity and fertility. Fed for 168 h, 0.2 and 0.5% sodium tetraborate killed almost all flies within the 7-d treatment. Oviposition of survivors in 0.1 and 0.2% sodium tetraborate treatments was arrested for 20 d after treatment.

Prediction of agricultural worker safety reentry times for organophosphate insecticides.

Concepts and current methods of determining worker safety reentry times are reviewed. Comparison of human monitoring studies, factors in a worker reentry episode and exposure estimation methods illustrate the advantages of estimation methods. Research needs for worker reentry times are discussed. A systematic view of the worker reentry situation is suggested.

Scientific misconduct in environmental science and toxicology.

Scientific misconduct easily occurs in environmental science and toxicology; we encountered four such cases. Only one case was discovered by editors; three were reported by other authors. All guilty authors were eventually banned from future publication in the Bulletin of Environmental Contamination and Toxicology. Cases in the Bulletin indicate that scientific misconduct may occur undetected across phyla, genera, and species; that distance from the publishing source makes detection more difficult; that editors and reviewers are not organized to take action against scientific misconduct; that plagiarized authors are likely to report plagiarism; and that there is only a small risk of censure from any source for authors engaging in scientific misconduct.

Comparative disappearance of dioxathion, malathion, oxydemetonmethyl and dialifor from Florida citrus leaf and fruit surfaces.

Surface residue disappearance rates of dioxathion, malathion, oxydemetonmethyl, and dialifor were the same for fruit and leaves, and they increase with temperature. Disappearance rates were in the order of malathion greater than or equal to oxydemetonmethyl greater than dialifor greater than or equal to dioxathion. Malaoxon was present in relatively constant amounts under dry, cool, clear conditions but was not detected under wet, hot, cloudy conditions.

HPLC identification of allelopathic compounds fromLantana camara.

Aqueous extracts ofLantana camara L. leaves inhibited ryegrass (Lolium multiflorum Lam.) germination and seedling growth. Phytotoxic compounds were fractionated from crude aqueous extracts and fractions were evaluated for their phytotoxicity. Inhibition was most pronounced with the alkaline and acid hydrolysates. Plant inhibition by the crude extract reflected a complex interaction of numerous individual components of diverse chemical compositions and potencies. Presumptive identification of the individual components was accomplished with high-performance liquid chromatography (HPLC). Thirteen phenolic compounds were identified, and most of these compounds were phytotoxic to ryegrass seedlings. Radicle elongation was more sensitive to the toxins than shoot elongation.