Antagonistic and additive effect when combining biopesticides against the fall armyworm, Spodoptera frugiperda.

Fall armyworm, Spodoptera frugiperda (FAW) is a cosmopolitan crop pest species that has recently become established in sub-Saharan Africa and Southeast Asia. Current FAW control is almost entirely dependent on synthetic pesticides. Biopesticides offer a more sustainable alternative but have limitations. For example, pyrethrum is an effective botanical insecticide with low mammalian toxicity but is highly UV labile, resulting in a rapid loss of efficacy in the field. Beauveria bassiana is an entomopathogenic fungus that is more persistent, but there is a time lag of several days before it causes insect mortality and leads to effective control. The combination of these biopesticides could mitigate their drawbacks for FAW control. Here we evaluated the efficacy of pyrethrum and B. bassiana as individual treatments and in combination against 3rd instar FAW. Four different combinations of these two biopesticides were tested, resulting in an antagonistic relationship at the lowest concentrations of B. bassiana and pyrethrum (1 × 104 conidia mL-1 with 25 ppm) and an additive effect for the other 3 combined treatments (1 × 104 conidia mL-1 with 100 ppm and 1 × 105 conidia mL-1 with 25 ppm and 100 ppm pyrethrum). Additionally, a delay in efficacy from B. bassiana was observed when combined with pyrethrum as well as a general inhibition of growth on agar plates. These results appear to show that this particular combination of biopesticides is not universally beneficial or detrimental to pest control strategies and is dependent on the doses of each biopesticide applied. However, the additive effect shown here at specific concentrations does indicate that combining biopesticides could help overcome the challenges of persistence seen in botanical pesticides and the slow establishment of EPF, with the potential to improve effectiveness of biopesticides for IPM.

The influence of manure-based organic fertilisers on the oviposition behaviour of Anopheles arabiensis.

The rice agroecosystem provides suitable breeding habitat for many malaria vector species, and  rice-adjacent communities are consequently exposed to a greater malaria transmission risk than non-rice-associated communities. As part of efforts to expand rice production in Africa, sustainable and climate-adapted practices such as the System of Rice Intensification (SRI) are being promoted. SRI encourages the use of organic fertilisers (OFs) such as cow and chicken dung, as opposed to inorganic industrially produced fertilisers, due to their lower resource cost, apparent benefit to the rice agroecosystem and as a means to reduce the greenhouse gas emissions associated with the production of industrial fertilisers. However, the impact of OFs on mosquito fauna is not well documented and may have knock-on consequences on malaria transmission risk. Here, we demonstrate, using dual choice egg count assays, that both cow and chicken dung modulate the oviposition behaviour of Anopheles arabiensis, a major malaria vector in Sub-Saharan Africa. A significantly reduced proportion of eggs were laid in water treated with either cow or chicken dung compared to untreated water, with higher dung concentrations resulting in further reduced proportions. When presented in competition, significantly fewer eggs were laid in water treated with chicken dung than with cow dung. Moreover, there was no evidence of egg retention in any experiment, including in no-choice experiments where only dung-containing dishes were available. These results suggest both cow and chicken dung may act as oviposition deterrents to malaria vector species and that the application of manure-based OFs in rice agriculture may modulate the oviposition behaviour of An. gambiae s.l. within agroecosystems. Quantification of the ammonia present in dung-infused water showed higher concentrations were present in the chicken dung infusion, which may be one contributing factor to the difference in observed deterrence between the two dung types. Deterrence of mosquito oviposition in OF-treated farms may potentially affect the overall production of malaria vectors within rice fields and their contribution to local malaria transmission.

Live Drosophila melanogaster Larvae Deter Oviposition by Drosophila suzukii.

The worldwide invasive insect pest, Drosophila suzukii Matsumura (spotted-wing Drosophila), lays eggs in soft and stone fruit before harvest. Hatched larvae cause fruit collapse and significant economic losses. Current control methods rely primarily on foliar insecticide applications, which are not sustainable long-term solutions due to regulatory restrictions and the risk of insecticide resistance developing. We showed before that D. suzukii were deterred from laying eggs on artificial media previously visited by its sister species-Drosophila melanogaster. In the current study, laboratory choice test experiments were conducted to identify which D. melanogaster life stage (eggs, larvae, or adult) deterred D. suzukii oviposition. We demonstrated that the presence of live D. melanogaster larvae on the egg-laying media consistently deterred D. suzukii oviposition. Drosophila melanogaster cuticular hydrocarbons (CHCs) were examined as candidate for the oviposition deterrent. CHCs of larval and adult D. melanogaster and D. suzukii were analyzed. In both species, the composition of the CHCs of larvae was similar to that of adults, although quantities present were much lower. Furthermore, the CHC profiles of the two species were markedly different. However, when assayed as deterrents in the laboratory choice test experiment, CHC extracts from D. melanogaster did not deter oviposition by D. suzukii.

Components of the Female Sex Pheromone of the Newly-Described Canola Flower Midge, Contarinia brassicola.

The canola flower midge, Contarinia brassicola Sinclair (Diptera: Cecidomyiidae), is a newly-described species that induces galls on canola, Brassica napus Linnaeus and Brassica rapa Linnaeus (Brassicaceae). Identification of the sex pheromone of C. brassicola is essential to developing monitoring tools to elucidate the geographic range and hosts of this new pest, and the extent to which it threatens the $30 billion Canadian canola industry. The aim of this study was to identify and synthesize the female-produced sex pheromone of C. brassicola and demonstrate its effectiveness in attracting males to traps in the field. Two peaks were identified through GC-EAG analysis of female-produced volatiles which elicited electrophysiological responses in male antennae. These peaks were initially characterized through GC-MS and synthesis as 2,7-diacetoxynonane (major component) and 2-acetoxynonane (minor component), and the racemic compounds elicited EAG responses in male antennae. All four stereoisomers of 2,7-diacetoxynonane were synthesized and the naturally-produced compound was shown to be primarily the (2R,7S)-isomer by analysis on an enantioselective GC column, with a small amount of (2R,7R)-2,7-diacetoxynonane also present. The configuration of the minor component could not be determined because of the small amount present, but this was assumed to be (2R)-2-acetoxynonane by comparison with the configuration of the other two components. In field trials, none of the four stereoisomers of 2,7-diacetoxynonane, presented individually or as a racemic mixture, was attractive to male C. brassicola. However, dispensers loaded with a 10 µg:1 µg blend of (2R,7S)- and (2R,7R)-2,7-diacetoxynonane caught large numbers of male C. brassicola and significantly more than other blends tested. The addition of 0.5 µg of (2R)-2-acetoxynonane to this blend further increased the number of males caught. In future work, we will seek to identify the optimum trapping protocol for the application of the pheromone in monitoring and surveillance.

Identification of Components of the Aggregation Pheromone of the Guam Strain of Coconut Rhinoceros Beetle, Oryctes rhinoceros, and Determination of Stereochemistry.

The coconut rhinoceros beetle, Oryctes rhinoceros (Linnaeus 1758) (Coleoptera: Scarabaeidae: Dynastinae) (CRB), is endemic to tropical Asia where it damages both coconut and oil palm. A new invasion by CRB occurred on Guam in 2007 and eradication attempts failed using commonly applied Oryctes rhinoceros nudivirus (OrNV) isolates. This and subsequent invasive outbreaks were found to have been caused by a previously unrecognized haplotype, CRB-G, which appeared to be tolerant to OrNV. The male-produced aggregation pheromone of the endemic, susceptible strain of O. rhinoceros (CRB-S) was previously identified as ethyl 4-methyloctanoate. Following reports from growers that commercial lures containing this compound were not attractive to CRB-G, the aim of this work was to identify the pheromone of CRB-G. Initial collections of volatiles from virgin male and female CRB-G adults from the Solomon Islands failed to show any male- or female-specific compounds as candidate pheromone components. Only after five months were significant quantities of ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by males but not by females. No other male-specific compounds could be detected, in particular methyl 4-methyloctanoate, 4-methyl-1-octanol, or 4-methyl-1-octyl acetate, compounds identified in volatiles from some other species of Oryctes. Ethyl 4-methyloctanoate elicited a strong electroantennogram response from both male and female CRB-G, but these other compounds, including 4-methyloctanoic acid, did not. The enantiomers of ethyl 4-methyloctanoate and 4-methyloctanoic acid were conveniently prepared by enzymatic resolution of the commercially-available acid, and the enantiomers of the acid, but not the ester, could be separated by gas chromatography on an enantioselective cyclodextrin phase. Using this approach, both ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by male CRB-G were shown to be exclusively the (R)-enantiomers whereas previous reports had suggested male O. rhinoceros produced the (S)-enantiomers. However, re-examination of the ester and acid produced by male CRB-S from Papua New Guinea showed that these were also the (R)-enantiomers. In field trapping experiments carried out in the Solomon Islands, both racemic and ethyl (R)-4-methyloctanoate were highly attractive to both male and female CRB-G beetles. The (S)-enantiomer and the corresponding acids were only weakly attractive. The addition of racemic 4-methyloctanoic acid to ethyl 4-methyloctanoate did significantly increase attractiveness, but the addition of (R)- or (S)-4-methyloctanoic acid to the corresponding ethyl esters did not. Possible reasons for the difference in assignment of configuration of the components of the CRB pheromone are discussed along with the practical implications of these results.

Hero Turned Villain: Identification of Components of the Sex Pheromone of the Tomato Bug, Nesidiocoris tenuis.

Nesidiocoris tenuis (Reuter) (Heteroptera: Miridae) is a tropical mirid bug used as a biocontrol agent in protected crops, including tomatoes. Although N. tenuis predates important insect pests, especially whitefly, it also causes damage by feeding on tomato plants when prey populations decline, resulting in significant economic losses for growers. The pest is now established in some all-year-round tomato crops in Europe and control measures involve the application of pesticides which are incompatible with current IPM programs. As part of future IPM strategies, the pheromone of N. tenuis was investigated. Volatile collections were made from groups and individuals of mated and unmated, females and males. In analyses of these collections by gas chromatography coupled with electroantennographic (EAG) recording from antennae of male bugs, two EAG-active components were detected and identified as 1-octanol and octyl hexanoate. Unlike other mirids, both male and female N. tenuis produced the two compounds, before and after mating, and both sexes gave EAG responses to both compounds. Furthermore, only octyl hexanoate was detected in whole body solvent washes from both sexes. These compounds are not related to the derivatives of 3-hydroxybutyrate esters found as pheromone components in other members of the Bryocrinae sub-family, and the latter could not be detected in volatiles from N. tenuis and did not elicit EAG responses. Nevertheless, experiments carried out in commercial glasshouses showed that traps baited with a blend of the synthetic pheromone components caught essentially only male N. tenuis, and significantly more than traps baited with octyl hexanoate alone. The latter caught significantly more N. tenuis than unbaited traps which generally caught very few bugs. Traps at plant height caught more N. tenuis males than traps 1 m above or at the base of the plants. The trap catches provided an indication of population levels of N. tenuis and were greatly reduced following an application of insecticide.

Additive Effect of Botanical Insecticide and Entomopathogenic Fungi on Pest Mortality and the Behavioral Response of Its Natural Enemy.

Sustainable agricultural intensification employs alternatives to synthetic insecticides for pest management, but these are not always a direct replacement. Botanical insecticides, for example, have rapid knockdown but are highly labile and while biological pesticides are more persistent, they are slow acting. To mitigate these shortcomings, we combined the entomopathogenic fungus (EPF) Metarhizium anisopliae with pyrethrum and evaluated their efficacy against the bean aphid, Aphis fabae. To ascertain higher trophic effects, we presented these treatments to the parasitoid, Aphidius colemani, on an aphid infested plant in a Y-tube olfactometer and measured their preferences. Aphid mortality was significantly higher than controls when exposed to EPF or pyrethrum but was greater still when exposed to a combination of both treatments, indicating an additive effect. This highlights the potential for applications of pyrethrum at lower doses, or the use of less refined products with lower production costs to achieve control. While parasitoids were deterred by aphid infested plants treated with EPF, no preference was observed with the combination pesticide, which provides insight into the importance that both application technique and timing may play in the success of this new technology. These results indicate the potential for biorational pesticides that combine botanicals with EPF.