Flor Budia

Laboratory evaluation of the novel naturally derived compound spinosad against Ceratitis capitata

Laboratory studies were conducted to determine the effect of the naturally derived compound spinosad on Ceratitis capitata Wied. (Diptera, Tephritidae). The organophosphate fenthion was used as a standard. Direct dose-dependent mortality and reduced fecundity were observed in oral treatment of adults with spinosad. The LC 90 values 14 h and seven days after treatment were 19.50 and 0.49 mg litre -1 respectively. Fenthion was less active (the LC 50 eight days after treatment was 1.17 mg litre -1 ) and did not affect the fecundity of the fly. Adults were also very susceptible to spinosad and fenthion via residual contact. For spinosad, 100% mortality was recorded 48 h after treatment for a dose of 10 mg litre -1 . Spinosad was more effective than fenthion in suppressing larval development when neonate larvae were reared on treated diet supplemented with a range of concentrations from 0.02 to 0.83 mg kg -1 diet. Last-instar larvae were much less susceptible to spinosad or fenthion when exposed via dipping or when they pupated in treated medium and both products had similar performance. A lack of ovicidal activity was observed in direct egg-treatments with spinosad but significant reductions from 1 mg litre -1 onwards were recorded for fenthion.

Toxicity and absorption of azadirachtin, diflubenzuron, pyriproxyfen, and tebufenozide after topical application in predatory larvae of Chrysoperla carnea (Neuroptera: Chrysopidae)

Abstract Susceptibility of the generalist predator, the lacewing Chrysoperla carnea (Stephens), to the insect growth regulators azadirachtin, diflubenzuron, pyriproxyfen, and tebufenozide was tested in the laboratory. Third instars were topically treated with different doses of formulated materials of each compound by direct topical exposure. At maximum field-recommended dose, pyriproxyfen and tebufenozide were harmless to C. carnea, whereas azadirachtin and diflubenzuron were harmful (respective LD90s were 24.5 and 6.9 ng active ingredient [AI] per insect). At sublethal doses of azadirachtin and diflubenzuron, females laid fertile eggs, but azadirachtin caused a slight negative effect on oviposition. Pyriproxyfen and tebufenozide had no effect on oviposition and egg fertility. As a second approach of this study, toxicity data are discussed in relation to the rate of penetration and excretion after topical application. One hour after administration, ≈80% of pyriproxyfen had penetrated; whereas for diflubenzuron and tebufenozide, only percentages of 10–20% were recorded in the same time interval. However, although pyriproxyfen penetration was fast and high, most of the compound was also quickly eliminated via excretion. Our data suggest that the use of azadirachtin and diflubenzuron in combination with C. carnea in integrated pest management (IPM) programs should be carefully evaluated. Pyriproxyfen and tebufenozide are considered to be safe for C. carnea.

Lethal and Sublethal Effects of Methoxyfenozide and Spinosad on Spodoptera littoralis (Lepidoptera: Noctuidae)

Abstract Susceptibility to methoxyfenozide ofSpodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) larvae was determined through exposure of neonate and fourth instars to dipped and sprayed pepper,Capsicum annum L., leaves. Methoxyfenozide and spinosad were tested against adults of this noctuid by oral, residual, and topical application. In larvae, we evaluated five (range, 0.001–10 mg active ingredient [AI]/liter) and six (range, 1–250 mg [AI]/liter) concentration levels by instars, respectively, by using two application methods at three different age leaf residues (0, 3, and 6 d after application). According to LC50 values, no significant differences were observed between the same age leaf residues of both application methods at 96 and 72 h after ingestion treatment on neonates and fourth instars, respectively. Nevertheless, toxicity of methoxyfenozide decreased significantly after time. For both application methods, the LC50 values of the first leaf residue (0 d after application) were significantly lower than those of 6-d-old residues. Furthermore, larval weight of fourth instars fed for 48 h with pepper,Capsicum annum L., leaves containing methoxyfenozide was significantly suppressed. Spinosad and methoxyfenozide reduced in a dose-dependent manner the fecundity and fertility ofS. littoralis adults when treated oral and residually. Likewise, when methoxyfenozide was administered orally in three different adult crosses, the fecundity was strongly affected, independently of the treated sex. We conclude that the combination of lethal and sublethal effects of methoxyfenozide and spinosad might exhibit significant effects on the population dynamics ofS. littolaris.

Influence of Azadirachtin and Methoxyfenozide on Life Parameters of Spodoptera littoralis (Lepidoptera: Noctuidae)

ABSTRACT n Effects on adult longevity, fecundity and fertility, as well as long-term effects on progeny were determined through oral exposure of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) adults to azadirachtin and methoxyfenozide. Both compounds reduced adult longevity by 2.3 d at the higher concentrations tested, but no significant differences were observed between sexes. Fecundity and fertility were significantly affected for both insecticides, although this effect was only dose-dependent for azadirachtin. The progeny from adults treated with azadirachtin or methoxyfenozide were only affected in percentage of pupation of eggs that successfully hatched, but no effects were observed in adult emergence of individuals that successfully pupated for either insecticide. In the second part of this study, each sex was exposed separately to methoxyfenozide by topical application or ingestion. Adult fecundity was more affected when moths were treated by ingestion than when treated topically, with a mean number eggs laid per female of 343 ± 89 and 932 ± 79, respectively. Finally, azadirachtin applied to pepper plants showed a significant oviposition deterrence activity on S. littoralis adults. However, when fecundity was scored for one additional day in females that had been previously exposed to pepper, Capsicum annum L., plants treated with this insecticide, the number of eggs laid per female did not differ significantly from that of controls. The effects of azadirachtin and methoxyfenozide on S. littoralis suggest changes in population dynamics of this pest in crops treated with these insecticides.

Toxicity and Pharmacokinetics of Spinosad and Methoxyfenozide to Spodoptera littoralis (Lepidoptera: Noctuidae)

Abstract The toxicity of spinosad and methoxyfenozide against neonates and fourth instars of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) was tested under laboratory conditions. According to LC50 values, no significant differences were observed between spinosad (0.50 mg [AI]/kg diet) and methoxyfenozide (0.54 mg [AI] /kg diet) after 48 h of ingestion treatment on neonate larvae, based on the overlap of 95% CL. Similarly, on fourth instars, no significant differences were observed between LC50 (2.98 and 5.17 mg [AI]/kg diet for spinosad and methoxyfenozide, respectively, at 96 h after ingestion of artificial diet) and LD50 (4.74 and 2.68 μg [AI]/g larva for spinosad and methoxyfenozide, respectively, at 144 h after topical application). In addition, spinosad and methoxyfenozide significantly suppressed weight gain of neonates and fourth instars continuously fed with artificial diet containing the insecticides. The second part of this project was focused on the effects of methoxyfenozide on the reproduction of S. littoralis by determining the effect on oviposition in three different crosses and the pharmacokinetics pattern of (14C)-methoxyfenozide in the body tissues of female and male adults after topical application in their excrement and in the deposited eggs to test transport through the female and the male adult. Methoxyfenozide negatively affected the reproduction of adults regardless the treated sex, presumably because of its presence in the adult body, accumulation in the laid eggs, and slow excretion of the product. We conclude that spinosad and methoxyfenozide represent an important choice to be used in integrated pest management where S. littoralis is a major pest.

Kaolin and copper-based products applications: Ecotoxicology on four natural enemies

Lethal and sublethal effects of kaolin clays and two copper-based products on four natural enemies found in olive orchards Anthocoris nemoralis (F.) (Hem. Anthocoridae), Chelonus inanitus (L.) (Hym. Braconidae), Chilocorus nigritus (F.) (Col. Coccinellidae) and Scutellysta cyanea Motschulsky (Hym. Pteromalidae) are described. Both kaolin and copper can be applied for controlling the olive fruit fly and the olive moth, two important pests of this crop. The products did not increase the mortality of any of the insects studied, with the exception of A. nemoralis. The sublethal effects, however, differed depending on the parameter evaluated and the insect studied. Both kaolin and coppers slightly, but significantly, reduced the life span of C. inanitus and S. cyanea. Number of eggs laid by A. nemoralis females were reduced, but not significantly compared to the controls. In the behavioural experiments, clear preference for remaining on kaolin-untreated surfaces when insects were able to choose was observed. Despite having some negative effects, the negative impact on natural enemies was lower than the impact caused by products commonly applied in this crop against the pests stated above. Therefore, both kaolin and copper can be considered as alternative products to be applied in olive orchards if an effective resistance management programme is to be developed. Furthermore, both of them are allowed in organic farming, in which the number of products that can be applied is more restricted.

Insecticidal Effects of Various Concentrations of Selected Extractions of Cestrum parqui on Adult and Immature Ceratitis capitata

Aqueous extracts of Cestrum parqui LHéritier (Solanaceae) were evaluated at different concentrations in several stages of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). For adults, the study was extended to extracts obtained with several solvents of an increasing degree of polarity. Aqueous extracts from C. parqui showed a high toxicity to neonate larvae when ingested through diet, inhibiting pupation at a concentration above 0.6%. Lower concentrations delayed the larval development and reduced the percentages of pupae formed and adult emergence. An LC50 = 0.9% after 3 d of continuous ingestion of C. parqui aqueous extracts could be calculated, whereas extracts obtained with organic compounds were nearly innocuous except with the use of the solvent methanol/water (80:20), the more polar of those tested, that killed 12.5% of adults. Aqueous extracts were also harmful to adults by diminishing the reproductive potential, which implies a significant effect on the offspring. Egg contact with insecticide or dipping third instars did not cause any adverse effect, supporting the hypothesis that only by means of ingestion can the toxic compounds of C. parqui reach the target. Our results showed that C. parqui components causing C. capitata mortality are mostly dissolved in water and not in organic solvents, which point out their polar character.

Nonsteroidal moulting hormone agonists: effects on protein synthesis and cuticle formation in Colorado potato beetle larvae

Larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say), that were orally treated with RH‐0345 at 0.1 mg l−1, RH‐5849 at 10 and 50 mg l−1, tebufenozide at 2 g l−1, and 20‐hydroxyecdysone at 2 g l−1, showed symptoms of premature moulting, followed by inhibition of ecdysis. In addition, fresh weight gain and total protein content were blocked. The effects on haemolymphal and cuticular polypeptides after PAGE were linked with premature, new epicuticle deposition as was observed under the electron microscope. These observations support the concept that the ecdysteroid‐mimicking action of the three nonsteroidal molecules is specific.

Laboratory evaluation of natural pyrethrins, pymetrozine and triflumuron as alternatives to controlCeratitis capitata adults

Three insecticides, pyrethrins + piperonil butoxide (PBO), pymetrozine and triflumuron, were tested as potential alternatives for controlling the Mediterranean fruit flyCeratitis capitata (Wiedemann) (Diptera: Tephritidae). The compounds were administered to adult flies in the laboratory by different uptake methods at the recommended rates currently used in fruit crops in Spain. Pyrethrins + PBO (80 + 320 ppm a.i.) exhibited a comparable knock-down effect to malathion (1,500 ppm a.i.) in the laboratory, irrespective of the method of application used. After these results were obtained, the number of concentrations tested was increased to carry out a dose-response analysis. Pymetrozine (300 ppm a.i.) and triflumuron (150 ppm a.i.) did not kill adults at the concentrations tested. However, pymetrozine diminished the fecundity, especially when adults were fed the insecticide; egg hatch was decreased by 59.3% compared with controls. Further experiments showed that increased period of ingestion and higher concentrations had a clear effect in reducing both fecundity and fertility. The possible use of pyrethrins + PBO and pymetrozine to reduce populations ofC. capitata is discussed.

Characterization of a new toxin from the entomopathogenic fungus Metarhizium anisopliae: the ribotoxin anisoplin.

Abstract Metarhizium anisopliae is an entomopathogenic fungus relevant in biotechnology with applications like malaria vector control. Studies of its virulence factors are therefore of great interest. Fungal ribotoxins are toxic ribonucleases with extraordinary efficiency against ribosomes and suggested as potential insecticides. Here we describe this ribotoxin characteristic activity in M. anisopliae cultures. Anisoplin has been obtained as a recombinant protein and further characterized. It is structurally similar to hirsutellin A, the ribotoxin from the entomopathogen Hirsutella thompsonii. Moreover, anisoplin shows the ribonucleolytic activity typical of ribotoxins and cytotoxicity against insect cells. How Metarhizium uses this toxin and possible applications are of interest.