Bioaccumulation and elimination, acute toxicity analysis and risk assessment of diflubenzuron in common carp (Cyprinus carpio).

Diflubenzuron has been applied in agriculture and aquaculture, and its residues in ecological environment and food chain could result in chronic exposure and long-term toxicity effects for human health. However, limited information is available regarding diflubenzuron levels in fish and associated risk assessment. This study performed the analysis for dynamic bioaccumulation and elimination distribution of diflubenzuron in carp tissues. The results indicated that diflubenzuron was absorbed and enriched by fish body along with higher enrichment in lipid-rich tissues of fish. The peak concentration in carp muscle reached 6-fold of diflubenzuron concentration in aquaculture water. The median lethal concentration (LC50) of diflubenzuron at 96 h was 12.29 mg/L, presented low toxicity to carp. Risk assessment results showed that the chronic risk from dietary exposure to diflubenzuron through carp consumption for Chinese residents of children and adolescents, adults and elderly people were acceptable, while posed a certain risk for young children. This study provided the reference for pollution control, risk assessment and scientific management of diflubenzuron.

Potential differences in chitin synthesis ability cause different sensitivities to diflubenzuron among three strains of Daphnia magna.

Ecotoxicity testing of crustaceans using Daphnia magna has been implemented in the chemical management systems of various countries. While the chemical sensitivity of D. magna varies depending on genetically different clonal lineages, the strain used in ecotoxicity tests, including the acute immobilization test (OECD TG202), has not been specified. We hypothesized that comprehensive gene expression profiles could provide useful information on phenotypic differences among strains, including chemical sensitivity. To test this hypothesis, we performed mRNA sequencing on three different strains (NIES, England, and Clone 5) of D. magna under culture conditions. The resulting expression profile of the NIES strain was clearly different compared to the profiles of the other two strains. Gene ontology (GO) enrichment analysis suggested that chitin metabolism was significantly enriched in the NIES strain compared to that in the England strain. Consistent with the GO analysis, evidence of high levels of chitin metabolism in the NIES strain were observed across multiple levels of biological organization, such as expression of chitin synthase genes, chitin content, and chitinase activity, which suggested that the different strains would exhibit different sensitivities to chemicals used to inhibit chitin synthesis. We found that among all strains, the NIES strain was more tolerant to diflubenzuron, a chitin synthesis inhibitor, with a 14-fold difference in the 48 h-EC50 value for the acute immobilization test compared to the England strain. The present study demonstrates that the differences among strains in chitin metabolism may lead to sensitivity difference to diflubenzuron, and serves as a case study of the usefulness of comprehensive gene expression profiles in finding sensitivity differences.

The effect of chemical insecticides on the scavenging performance of Steinernema carpocapsae: Direct effects and exposure to insects killed by chemical insecticides.

Entomopathogenic nematodes are used widely in biological insect control. Entomopathogenic nematodes can infect live insects as well as dead insects (i.e., they can act as scavengers). It is important to determine compatibility of entomopathogenic nematodes with other pest management tactics such as chemical insecticides. We hypothesized that chemical insecticides have negative impact on scavenging nematodes. According to our hypothesis, we first investigated the effects of direct exposure of Steinernema carpocapsae infectivity juveniles (IJs) to three chemical insecticides, cypermethrin, spinosad or diflubenzuron in terms of nematode survival and virulence. Subsequently, using the same chemicals, we tested the effects of insecticide-killed insects on scavenger nematode penetration efficiency, time of emergence and the number of nematode progeny. Prior to our study, the impact of pesticides on scavenger nematode fitness had not been studied. Fall webworm, Hyphantria cunea, and greater wax moth, Galleria mellonella, larvae were used as host insects. The survival rate of IJs after direct exposure was 83% for cypermethrin and 93-97% for the other insecticides and control. There were no significant differences in the survival and virulence of the nematodes after 24 h exposure to insecticides. The number of nematodes that invaded the insecticide-killed host was significantly higher in cypermethrin and spinosad treated groups and live H. cunea than in the diflubenzoron treated group and freeze-killed control. However, no significant differences were observed in time of emergence. Significantly more progeny IJs emerged from Spinosad-killed insects than the freeze-killed control. In conclusion, we discovered that the fitness of scavenging IJs is not diminished by insecticides in insect cadavers. In fact, in some cases the exposure to chemical insecticides may enhance virulence.

Assessing Impacts of Insecticides on Different Embryonic Stages of the Nontarget Aquatic Insect Cheumatopsyche brevilineata (Trichoptera: Hydropsychidae).

Although the egg of aquatic insects is one of the main life stages that can be exposed to contaminants in water, little is known about the detailed impacts of contaminants on eggs of aquatic insects. The present study aimed to clarify the vulnerable embryonic stages of the caddisfly Cheumatopsyche brevilineata exposed to 2 insecticides, etofenprox and diflubenzuron, which are severely harmful to aquatic insects, and to assess the effects of exposure duration on toxicity of etofenprox to the embryonic stage. Eggs obtained from laboratory culture of the insect were exposed to etofenprox for different periods (2, 4, 6, or 8 d) and at different embryonic stages. In experiments with diflubenzuron, eggs were exposed for 2 d at different embryonic stages. These insecticides did not kill the embryos during exposure, but they inhibited hatching post exposure. Diflubenzuron also induced morphological abnormalities of hatchlings and reduced their survival. The overall median lethal concentration (LC50overall ) values varied significantly from 0.0560 to 5.19 µg/L for etofenprox among exposure durations and among embryonic stages, and from 0.442 to 2.89 µg/L for diflubenzuron between embryonic stages. The toxicity of etofenprox to the embryo was more dependent on the embryonic stage at the time of exposure than on the exposure duration. The vulnerable embryonic stage differed between the insecticides. Etofenprox more strongly inhibited hatching of embryos at later stages, whereas sensitivity to diflubenzuron was higher in the following order: stages E3 to E7 > stages E1, E2, and E8. The different responses of C. brevilineata embryos may be attributable to differences in insecticide mode of action and in functional development of insecticide target sites during embryogenesis. Environ Toxicol Chem 2019;38:1434-1445. © 2019 SETAC.

Metabolism of Diflubenzuron in Lizard ( Eremias argus) and Comparative Toxicity of Diflubenzuron and Its Metabolite.

The metabolic process of diflubenzuron in rat or fish has been well studied, but little is known about its elimination pathway in lizard. The current study predicted the metabolic route of diflubenzuron in lizard feces and compared the toxicity of diflubenzuron and 4-chloroaniline on lizard thyroid system. The amido bond cleavage was the major route for diflubenzuron elimination in lizard feces. 4-Chloroaniline as the most toxic diflubenzuron metabolite was also abundant in feces. According to liver slices, 4-chloroaniline exposure induced significant changes of nuclear shape, while diflubenzuron exposure caused significant hepatocytes clustering. On the basis of thyroid hormone and thyroid-related gene levels, triiodothyronine (T3) level in lizard liver was regulated by thyroid hormone receptors, while thyroxine (T4) concentration was modulated by dio2 and udp genes after diflubenzuron or 4-chloroaniline exposure. These results showed that both diflubenzuron and 4-chloroaniline could disrupt lizard thyroid system, which could provide evidence for lizard population decline.

Effects of insect growth-regulator insecticides on the immature stages of Harmonia axyridis (Coleoptera: Coccinellidae).

Harmonia axyridis is an important biological control predator in greenhouses and agricultural fields, and it plays a significant role in the integrated pest management (IPM) of several arthropod pests. We studied the effects of eight insect growth-regulator insecticides (IGRs) on immature stages of H. axyridis by residual toxicity (eggs and pupae) and contact toxicity (larvae) to evaluate the risk of using these IGRs in IPM systems. Diflubenzuron, hexaflumuron and lufenuron caused more than 80% mortality to H. axyridis eggs, larvae and pupae, respectively. Pyriproxyfen was also highly harmful to larvae and pupae of H. axyridis. In contrast, methoxyfenozide and buprofezin caused little mortality and were classified as slightly harmful to immature stages based on a reduction coefficient. In addition to mortality and developmental time, the fecundity, fertility and deformed eggs of offspring were affected, when the predators were exposed to IGRs. Benzoylphenylurea insecticides significantly reduced H. axyridis female fecundity and fertility and increased the number of deformed eggs. The adverse effects are closely connected with the developmental stages of the predators and types and methods of insecticides exposed. All IGRs affected, to some extent, the life-table parameters of H. axyridis when the insecticides applied on immature stages at the highest field rates. Tebufenozide, diflubenzuron, hexaflumuron and lufenuron significantly reduced the Ro, T, r and λ of beetles exposed to the insecticides. The results indicate that IGRs could disturb the population growth and biocontrol activities of H. axyridis when applied at the highest field label rates. Additional studies should be conducted to assess the effects of IGRs on H. axyridis under field conditions before incorporating them in IPM strategies.

In vitro PPARγ agonistic potential of chitin synthesis inhibitors and their energy metabolism-related hepatotoxicity.

The extensive use of chitin synthesis inhibitors (CSIs) in integrated pest management programs has a detrimental effect on the surrounding environment. Recent studies reveal that CSIs may affect non-target organisms at sublethal concentrations, highlighting the need for further ecological and health risk investigations of these compounds. In this study, we characterized the peroxisome proliferator-activated receptor γ (PPARγ) agonistic activity of fourteen CSIs in HepG2 cells using an in vitro reporter gene assay. Five of the tested CSIs showed remarkable PPARγ-mediated transactivation, and the relative agonistic potencies were diflubenzuron>chlorfluazuron>flucycloxuron>noviflumuron>flufenoxuron based on REC20 values. In addition, molecular docking indicated that different interactions may stabilize ligand binding to PPARγ. Next, we clarified that sublethal concentration of diflubenzuron caused a shift in cellular energy metabolism from the aerobic tricarboxylic acid (TCA) cycle to anaerobic glycolysis and this process was associated with the activation of PPARγ. These findings suggest that CSIs act as PPARγ agonists and exert diverse hepatotoxic effects by disrupting energy metabolism at sublethal concentrations.

Early life stages of Northern shrimp (Pandalus borealis) are sensitive to fish feed containing the anti-parasitic drug diflubenzuron.

Increasing use of fish feed containing the chitin synthesis inhibiting anti-parasitic drug diflubenzuron (DFB) in salmon aquaculture has raised concerns over its impact on coastal ecosystems. Larvae of Northern shrimp (Pandalus borealis) were exposed to DFB medicated feed under Control conditions (7.0 °C, pH 8.0) and under Ocean Acidification and Warming conditions (OAW, 9.5 °C and pH 7.6). Two weeks' exposure to DFB medicated feed caused significantly increased mortality. The effect of OAW and DFB on mortality of shrimp larvae was additive; 10% mortality in Control, 35% in OAW, 66% in DFB and 92% in OAW + DFB. In OAW + DFB feeding and swimming activity were reduced for stage II larvae and none of the surviving larvae developed to stage IV. Two genes involved in feeding (GAPDH and PRLP) and one gene involved in moulting (DD9B) were significantly downregulated in larvae exposed to OAW + DFB relative to the Control. Due to a shorter intermoult period under OAW conditions, the OAW + DFB larvae were exposed throughout two instead of one critical pre-moult period. This may explain the more serious sub-lethal effects for OAW + DFB than DFB larvae. A single day exposure at 4 days after hatching did not affect DFB larvae, but high mortality was observed for OAW + DFB larvae, possibly because they were exposed closer to moulting. High mortality of shrimp larvae exposed to DFB medicated feed, indicates that the use of DFB in salmon aquaculture is a threat to crustacean zooplankton.

The body burden and thyroid disruption in lizards (Eremias argus) living in benzoylurea pesticides-contaminated soil.

Dermal exposure is regarded as a potentially significant but understudied route for pesticides uptake in terrestrial reptiles. In this study, a native Chinese lizard was exposed to control, diflubenzuron or flufenoxuron contaminated soil (1.5 mg kg-1) for 35 days. Tissue distribution, liver lesions, thyroid hormone levels and transcription of most target genes were examined. The half-lives of diflubenzuron and flufenoxuron in the soil were 118.9 and 231.8 days, respectively. The accumulation of flufenoxuron in the liver, brain, kidney, heart, plasma and skin (1.4-35.4 mg kg-1) were higher than that of diflubenzuron (0-1.7 mg kg-1) at all time points. The skin permeability factor of flufenoxuron was more than 20-fold greater than that of diflubenzuron at the end of exposure. However, the liver was more vulnerable in the diflubenzuron exposure group. The alterations of triiodothyronine (T3) and thyroxine (T4) level after diflubenzuron or flufenoxuron exposure were accompanied with the changes in the transcription of target genes involved not only in hypothalamus-pituitary-thyroid (HPT) axis (sult, dio2, trα and udp) but also in metabolism system (cyp1a and ahr). These results indicated that flufenoxuron produced greater body burdens to lizards through dermal exposure, whereas both diflubenzuron and flufenoxuron have the potential to disturb metabolism and thyroid endocrine system.

Oral and dermal diflubenzuron exposure causes a hypothalamic-pituitary-thyroid (HPT) axis disturbance in the Mongolian racerunner (Eremias argus).

Diflubenzuron (DFB) is a potential endocrine-disrupting chemical. However, its thyroid endocrine effect on reptiles has not been reported. In this study, immature lizards (Eremias argus) were exposed to 20 mg kg-1 DFB once a week for 42 days through oral or dermal routes. Their body weight, plasma thyroid hormone levels, thyroid gland histology and the transcription of hypothalamic-pituitary-thyroid (HPT) axis-related genes in different tissues were assessed to explore the effects of DFB on the HPT axis of lizards. The body weight decreased significantly only after the dermal exposure to DFB. Triiodothyronine (T3) to thyroxine (T4) ratio in the male plasma also significantly increased after the dermal exposure. After oral exposure, the activity of thyroid gland was positively related to the thyroid hormone levels. Furthermore, the alterations in thyroid hormone levels affected the HPT axis-related gene expression, which was tissue dependent and sexually selected. The thyroid hormone receptor genes (trα and trß) in the brain and thyroid were more sensitive to oral exposure. However, only the dermal treatment affected the trα, trß and type 2 deiodinase (dio2) genes in the male liver. These results suggest that DFB exposure caused sex-specific changes in the thyroid function of lizards, and the dermal treatment may be an important route for the risk assessment of reptiles.

Mixtures of diflubenzuron and p-chloroaniline changes the activities of enzymes biomarkers on tilapia fish (Oreochromis niloticus) in the presence and absence of soil.

The insecticide Diflubenzuron (DFB), used by many fish farming, when metabolized or degraded produces the extremely toxic compound p-chloroaniline (PCA). Once in the aquatic environment, these compounds can form mixtures and their bioavailability depends on factors such as the presence of soil. The toxic effects of the isolated compounds and their mixtures in the proportions: 75%, 50%, and 25% of PCA were analyzed in tilapia (Oreochromis niloticus) in the presence and absence of soil after 96h. The enzymes catalase (CAT), acid (AcP) and alkaline (AlP) phosphatases and alanine (ALT) and aspartate (AST) aminotransferases of the liver of the tilapia (Oreochromis niloticus) were used as biomarkers. DFB and the mixture containing 75% of this compound did not present high toxicity to fish; however, 25mg/L of PCA alone and 15mg/L of the mixture with 75% of this compound promoted 50% mortality of tilapia (Oreochromis niloticus). In the presence of soil, these toxicity values decreased to 37 and 25mg/L, respectively. Independent of the presence of soil, a synergistic effect was observed when the proportion of PCA was 75% and to the mixture, with 25% PCA was observed the antagonistic effect. Different concentrations of the compounds and their mixtures induced CAT activity independently of the presence of soil. Additionally, increases in phosphatases and transaminases activities were observed. In some cases, the enzymes also had their activities decreased and the dose-dependence effects were not observed. This research showed that the presence of soil influenced the toxicity of the compounds but not altered interaction type among them. Diflubenzuron, p-chloroaniline, and mixtures thereof caused disorders in enzymes important for the health of tilapia (Oreochromis niloticus).

Side-effects of pesticides on the generalist endoparasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae).

New plant protection strategies focus on minimizing chemical pesticide use and increasing their compatibility with biological control agents. The objective was to evaluate the side-effects of glyphosate, diflubenzuron, malathion, tebuconazole and triflumuron (at 720, 45, 400, 150 and 20 g ai ha-1, respectively), pesticides authorized for soybean crops in Brazil, on the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae) reared on Anticarsia gemmatalis (Lepidoptera: Noctuidae). The emergence and female numbers produced per P. elaeisis female were higher in A. gemmatalis pupae from caterpillars fed an artificial diet treated with glyphosate. However, emergence was lower than 50% when the caterpillars were fed on soybean leaves treated with glyphosate offered ad libitum (3-5 times). Palmistichus elaeisis died before parasiting A. gemmatalis pupae treated with malathion. Diflubenzuron reduced the P. elaesis sex ratio in the second generation. Tebuconazole and triflumuron did not cause side-effects on this parasitoid. A continuous exposure to glyphosate by the host may lead to side-effects on P. elaeisis emergence, but its moderate use is acceptable for this parasitoid. Diflubenzuron had severe transgenerational side-effects. Tebuconazole fungicide and triflumuron insecticide are compatible with P. elaeisis in sustainable integrated pest management (IPM) programs, while malathion can not be included in them.

Exposing Northern shrimp (Pandalus borealis) to fish feed containing the antiparasitic drug diflubenzuron caused high mortality during molting.

Use of the chitin synthesis inhibitor diflubenzuron (DFB) as an antiparasitic drug in salmon aquaculture raises concern over its impact on marine ecosystems. Further, global drivers, such as ocean warming and acidification (OAW), may increase the toxicity of hazardous substances including DFB. The aim of the present study was to examine the combined effects of DFB-medicated salmon feed on ovigerous Northern shrimp (Pandalus borealis) under Control (pHNBS 8.0, 7.0ºC) and OAW conditions (pHNBS 7.6, 9.5ºC). DFB-exposed shrimp consumed on average 0.1-0.3 g medicated feed during the 2-week exposure period, and high mortality (61-73%) was documented at both environmental conditions. There was no significant interaction between OAW and DFB. Only 2-7% of DFB-exposed shrimp molted successfully compared to 65% in Control and 63% in OAW. The shrimp molted earlier (shorter intermolt period) and exhibited higher feeding rate at OAW compared to Control conditions. An additional experiment, where female shrimp were exposed to DFB closer to molting, noted increased mortality after only 4 d exposure, and successful molting for some shrimp after 2 to 3 weeks of depuration. High mortality of shrimp exposed to DFB-medicated feed indicates that the use of this feed in aquaculture could affect local shrimp populations.

The tissue distribution, metabolism and hepatotoxicity of benzoylurea pesticides in male Eremias argus after a single oral administration.

Benzoylurea pesticides (BPUs) are widely used to control the locust, but the toxicokinetics and hepatotoxicity of BPUs in lizards have not been investigated. In this study, the tissue distribution, metabolism and liver toxicity of diflubenzuron and flufenoxuron were assessed in the Eremias argus following a single oral exposure. Diflubenzuron preferred to accumulate in the fat and brain (>1.0 mg kg-1) and was rapidly eliminate in other tissues. In the liver, 4-chloroaniline was one of diflubenzuron metabolites, although with a concentration less than 0.05% of the accumulated diflubenzuron. No significant difference was observed in the liver histopathology between the control and diflubenzuron exposure group. The expressions of Cyp1a and Ahr gene which control the cell apoptosis were also equal to the control level. After flufenoxuron exposure, biomodal phenomenon was observed in the liver, skin, brain, gonad, kidney, heart and blood circulation was an important route for the flufenoxuron penetration. The concentrations of flufenoxuron in all tissues were greater than 1.0 mg kg-1 at 168 h. The excretion of flufenoxuron in the faeces was 1.5 fold higher than diflubenzuron. The hepatocytes in the flufenoxuron treated group showed vacuolation of cytoplasm and decreased nucleus. In addition, the Cyp1a and Ahr genes were significantly up-regulated in the flufenoxuron exposure group. These results suggested that the higher hepatotoxicity of flufenoxuron may be attributed to the higher residual level in the lizard tissues and the Cyp1a and Ahr genes can serve as biomarkers to assess the liver toxicity.

Risk assessment of various insecticides used for management of Asian citrus psyllid, Diaphorina citri in Florida citrus, against honey bee, Apis mellifera.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of citrus trees worldwide. A wide variety of insecticides are used to manage D. citri populations within citrus groves in Florida. However, in areas shared by citrus growers and beekeepers the use of insecticides may increase the risks of Apis mellifera  L. (Hymenoptera: Apidae) loss and contaminated honey. The objective of this research was to determine the environmental toxicity of insecticides, spanning five different modes of action used to control D. citri, to A. mellifera. The insecticides investigated were imidacloprid, fenpropathrin, dimethoate, spinetoram and diflubenzuron. In laboratory experiments, LD50 values were determined and ranged from 0.10 to 0.53 ng/µl for imidacloprid, fenpropathrin, dimethoate and spinetoram. LD50 values for diflubenzuron were >1000 ng/µl. Also, a hazard quotient was determined and ranged from 1130.43 to 10893.27 for imidacloprid, fenpropathrin, dimethoate, and spinetoram. This quotient was <0.447 for diflubenzuron. In field experiments, residual activity of fenpropathrin and dimethoate applied to citrus caused significant mortality of A. mellifera 3 and 7 days after application. Spinetoram and imidacloprid were moderately toxic to A. mellifera at the recommended rates for D. citri. Diflubenzuron was not toxic to A. mellifera in the field as compared with untreated control plots. Phenoloxidase (PO) activity of A. mellifera was higher than in untreated controls when A. mellifera were exposed to 14 days old residues. The results indicate that diflubenzuron may be safe to apply in citrus when A. mellifera are foraging, while most insecticides used for management of D. citri in citrus are likely hazardous under various exposure scenarios.

Cyromazine resistance in a field strain of house flies, Musca domestica L.: Resistance risk assessment and bio-chemical mechanism.

Developing resistance management strategies for eco-friendly insecticides is essential for the management of insect pests without harming the environment. Cyromazine is a biorational insecticide with very low mammalian toxicity. Resistance to cyromazine has recently been reported in house flies from Punjab, Pakistan. In order to propose a resistance management strategy for cyromazine, experiments were planned to study risk for resistance development, possibility of cross-resistance and bio-chemical mechanisms. A field strain of house flies with 8.78 fold resistance ratio (RR) to cyromazine was re-selected under laboratory conditions. After seven rounds of selection (G1-G7), the RR values rapidly increased from 8.8 to 211 fold. However, these values declined to 81fold when the cyromazine selected (CYR-SEL) strain was reared without selection pressure, suggesting an unstable nature of resistance. The CYR-SEL strain showed lack of cross-resistance to pyriproxyfen, diflubenzuron, and methoxyfenozide. Synergism bioassays using enzyme inhibitors: piperonyl butoxide (PBO) and S,S,S-tributylphosphorotrithioate (DEF), and metabolic enzyme analyses revealed increased activity of carboxylesterase (CarE) and mixed-function oxidase (MFO) in the CYR-SEL strain compared to the laboratory susceptible (Lab-susceptible) strain, suggesting the metabolic resistance mechanism responsible for cyromazine resistance in the CYR-SEL strain. In conclusion, risk of rapid development of cyromazine resistance under consistent selection pressure discourages the sole reliance on cyromazine for controlling house flies in the field. The unstable nature of cyromazine resistance provides window for restoring cyromazine susceptibility by uplifting selection pressure in the field. Moreover, lack of cross-resistance between cyromazine and pyriproxyfen, diflubenzuron, or methoxyfenozide in the CYR-SEL strain suggest that cyromazine could be rotated with these insecticides whenever resistance crisis occur in the field.

Pathological and histometric analysis of the gills of female Hyphessobrycon eques (Teleostei:Characidae) exposed to different concentrations of the insecticide Dimilin(®).

Female individuals of Hyphessobrycon eques were exposed to Diflubenzuron (Dimilin(®)) in order to determine whether exposure to sublethal levels of this insecticide causes changes in gill morphology. Fish were exposed to 0.01, 0.1 and 1.0mgL(-1) for 96h and 17 days and then submitted to pathological and histometric evaluation. Pathological lesions, such as hyperplasia, lamellar fusion, vascular congestion, secondary lamellar disarray, vasodilatation, hemorrhage and increased lamellar epithelium, were significantly more common in the gills of fish exposed to Dimilin(®) than the control. Histometric analysis documented significant changes in blood vessel diameter, primary lamellae width and secondary lamellae length, and the appearance of hemorrhage foci in all concentrations tested. Even at low Dimilin(®) concentrations, the histopathological alteration index was mild to moderate, thereby indicating that the function of this tissue was compromised. These findings indicate that indiscriminate use of Dimilin(®) can adversely affect the structural integrity of the gills of H. eques, which can cause numerous problems for fish farming systems.

Impact of insect growth regulators on the predator Ceraeochrysa cincta (Schneider) (Neuroptera: Chrysopidae).

The generalist predator Ceraeochrysa cincta (Schneider) (Neuroptera: Chrysopidae) is an important biological control agent of several arthropod pests in different agroecosystems. This study assessed the lethal and sublethal effects of six insect growth regulators sprayed on first-instar larvae of C. cincta. Lufenuron and diflubenzuron were highly harmful to first-instar larvae of C. cincta, causing 100 % of mortality before they reached the second instar. Buprofezin caused ~25 % mortality of the larvae and considerably reduced the fecundity and longevity of the insects, but substantially increased the proportion of females in the surviving population of C. cincta. Methoxyfenozide and tebufenozide did not affect the duration and survival of the immature stages, but methoxyfenozide significantly reduced the fecundity and longevity of the insects. Pyriproxyfen reduced the survival of the larval stage by 19.5 %, but did not affect the development, survival and reproduction of the surviving individuals. Based on reduction coefficient, the insecticides diflubenzuron and lufenuron were considered harmful to C. cincta, whereas buprofezin and methoxyfenozide were slightly harmful and tebufenozide and pyriproxyfen were harmless. The estimation of life-table parameters indicated that buprofezin and methoxyfenozide significantly reduced the R o , r and λ of C. cincta, whereas pyriproxyfen and tebufenozide caused no adverse effect on population parameters, indicating that these insecticides could be suitable for use in pest management programs towards the conservation and population increase of the predator in agroecosystems. However, more studies should be conducted to evaluate the compatibility of these insecticides with the predator C. cincta under semi-field and field conditions.

Subchronic exposure to diflubenzuron causes health disorders in neotropical freshwater fish, Prochilodus lineatus.

The action of diflubenzuron (DFB) was evaluated in a freshwater fish, Prochilodus lineatus, after exposure to 0.06, 0.12, 0.25, or 0.50 mg L(-1) DFB for 14 days. Erythrocyte nuclear abnormalities (ENA), the gill activity of Na(+)/K(+)-ATPase, H(+)-ATPase and carbonic anhydrase (CA), and lipid peroxidation (LPO) and histopathological changes in the gills and liver were determined. The number of micronuclei increased in fish exposed to 0.25 and 0.50 mg L(-1) DFB. Plasma Cl(-) and the CA activity decreased, while the activity of Na(+)/K(+)-ATPase and of H(+)-ATPase increased in fish exposed to 0.25 and 0.50 mg L(-1) DFB. LPO did not change in the gills but increased in the liver of fish exposed to 0.25 and 0.50 mg L(-1) DFB. In the gills, histopathological changes indicated disperse lesions and slight to moderate damage in fish exposed to 0.50 mg L(-1) DFB, whereas in the liver, these changes were significantly greater in fish exposed to 0.25 and 0.50 mg L(-1) DFB, indicating moderate to severe damage. Continuous exposure to DFB is potentially toxic to P. lineatus, causing heath disorders when the fish is exposed to the two highest DFB concentrations, which are applied to control parasites in aquaculture and to control mosquito populations in the environment.

Subacute toxicity assessment of diflubenzuron, an insect growth regulator, in adult male rats.

Diflubenzuron (DFB), an insecticide and acaricide insect growth regulator, can be used in agriculture against insect predators and in public health programs, to control insects and vectors, mainly Aedes aegypti larvae. Due to the lack of toxicological assessments of this compound, the objective of the present study was to evaluate the toxicological effects of subacute exposure to the DFB insecticide in adult male rats. Adult male rats were exposed (gavage) to 0, 2, 4, or 8 mg/kg of DFB for 28 days. No clinical signs of toxicity were observed in the DFB-treated animals of the experimental groups. However, there was an increase in serum levels of alanine aminotransferase in the group that received 8 mg/kg/DFB/day and urea at doses of 4 and 8 mg/kg/DFB/day, without altering other biochemical or hematological parameters. The subacute exposure to the lowest dose of DFB caused significant decrease in testis weight, daily sperm production, and in number of sperm in the epididymis in relation to the control group. However, no alterations were observed in the sperm morphology, testicular, epididymis, liver and kidney histology, or testosterone levels. These findings unveiled the hazardous effects of DFB on male reproduction after the subacute exposure and special attention should be addressed to the effects of low doses of this pesticide.