Sexual selection in true fruit flies (Diptera: Tephritidae): transcriptome and experimental evidences for phytochemicals increasing male competitive ability

In male tephritid fruit flies of the genus Bactrocera, feeding on secondary plant compounds (sensu lato male lures = methyl eugenol, raspberry ketone and zingerone) increases male mating success. Ingested male lures alter the male pheromonal blend, normally making it more attractive to females and this is considered the primary mechanism for the enhanced mating success. However, the male lures raspberry ketone and zingerone are known, across a diverse range of other organisms, to be involved in increasing energy metabolism. If this also occurs in Bactrocera, then this may represent an additional benefit to males as courtship is metabolically expensive and lure feeding may increase a fly's short-term energy. We tested this hypothesis by performing comparative RNA-seq analysis between zingerone-fed and unfed males of Bactrocera tryoni. We also carried out behavioural assays with zingerone- and cuelure-fed males to test whether they became more active. RNA-seq analysis revealed, in zingerone-fed flies, up-regulation of 3183 genes with homologues transcripts to those known to regulate intermale aggression, pheromone synthesis, mating and accessory gland proteins, along with significant enrichment of several energy metabolic pathways and gene ontology terms. Behavioural assays show significant increases in locomotor activity, weight reduction and successful mating after mounting; all direct/indirect measures of increased activity. These results suggest that feeding on lures leads to complex physiological changes, which result in more competitive males. These results do not negate the pheromone effect, but do strongly suggest that the phytochemical-induced sexual selection is governed by both female preference and male competitive mechanisms.

Responses of fruit flies (Tephritidae: Dacinae) to novel male attractants in north Queensland, Australia, and improved lures for some pest species

Male fruit fly attractants, cue-lure and methyl eugenol (ME), have been successfully used for the last 50 years in the monitoring and control of Dacini fruit flies (Bactrocera and Dacus species). However, over 50% of Dacini are non-responsive to either lure, including some pest species. A new lure, zingerone, has been found to weakly attract cue- and ME-responsive species in Malaysia. In Australia it attracted a weakly cue-responsive minor pest Bactrocera jarvisi (Tryon) and three non-responsive' species. Similar compounds were tested in Queensland and attracted cue- and ME-responsive species and two non-responsive' species. In this study, 14 novel compounds, including raspberry ketone formate (RKF) (Melolure) and zingerone, were field tested in comparison with cue-lure and ME at 17 sites in north Queensland. The most attractive novel lures were isoeugenol, methyl-isoeugenol, dihydroeugenol and zingerone. Several non-responsive' species responded to the new lures: Bactrocera halfordiae (Tryon), a species of some market access concern, was most attracted to isoeugenol; B.barringtoniae (Tryon), B.bidentata (May) and B.murrayi (Perkins) responded to isoeugenol, methyl-isoeugenol and dihydroeugenol; two new species of Dacus responded to zingerone. Bactrocera kraussi (Hardy), a cue-responsive minor pest in north Queensland, was significantly more attracted to isoeugenol than cue-lure. The cue-responsive D.absonifacies (May) and D.secamoneaeDrew were significantly more attracted to zingerone than cue-lure. Bactrocera yorkensisDrew & Hancock, a ME-responsive species was significantly more attracted to isoeugenol, methyl-isoeugenol and dihydroeugenol than ME. The preferential response to RKF or cue-lure was species specific. Six species were significantly more attracted to RKF, including the pests B.tryoni (Froggatt), B.frauenfeldi (Schiner) and minor pest B.bryoniae (Tryon); eight species were significantly more attracted to cue-lure including the pest B.neohumeralis (Hardy). These findings have significance in the search for optimal male lures for pest species elsewhere in the world.

Low-dose methyl bromide fumigation as a quarantine disinfestation treatment for nectarines against Queensland fruit fly (Diptera:Tephritidae)

White nectarines (Prunus persica var. nucipersica) were fumigated with methyl bromide (MB) at a nominal treatment dose of 18 g m-3 at 18°C for 5 h and 30 min as a quarantine disinfestation treatment against Bactrocera tryoni, the Queensland fruit fly. Three large scale trials were conducted against each of the four immature lifestages, eggs and first, second and third instars. There were no survivors from the estimated 43,614 eggs, 41,873 first instars, 41,345 second instars and 33,549 third instars treated, thereby resulting in an efficacy of GROTERDAN99.99% mortality at the 95% confidence level for each lifestage. Of the 12 trials reported herein, the highest concentration of MB, sampled from the chamber headspace analysed by gas chromatography, was 18.7 g m-3. The maximum chamber temperature from 5 min readings was 19.7°C and the maximum fruit core temperature was 19.5°C. The treatment time for all trials was exactly 5.5 h. Thus the recommended treatment dose to disinfest nectarines from B. tryoni is 19.0 g m-3 MB at 20.0°C for 5.5 h. Fruit quality trials were conducted on white nectarines at three combinations of treatment parameters: 15 g m-3 MB at 19°C for 5.25 h; 18 g m-3 MB at 19°C for 5.5 h and 21 g m-3 MB at 19°C for 5.5 h. The fruit were stored at 0, 4 and 8 days at 4°C and 8 days at 4°C followed by 4 d at 22°C. They were then were assessed for skin colour, flesh colour, skin defects, flesh defects, fruit weight loss, flesh firmness, total soluble solids, titratable acidity and rots. There was no significant difference between untreated control and MB treated fruits in any of the parameters measured. Thus the treatments did not have adverse effects on fruit quality.

Evaluation of tomato production systems at Bowen in mitigating the risk of fruit fly infestation: a systems approach for market access

Queensland fruit flies Bactrocera tryoni and B. neohumeralis are considered major quarantine pests of tomato, a major crop in the horticultural production district around Bowen, North Queensland, Australia. Preharvest and/or postharvest treatments are required to meet the market access requirements of both domestic and international trading partners. The suspension from use of dimethoate and fenthion, the two insecticides used for fruit fly control, has resulted in the loss of both pre and postharvest uses in fresh tomato. Research undertaken quantitatively at Bowen evaluated the effectiveness of pre-harvest production systems without specific fruit fly controls and postharvest mitigation measures in reducing the risk of fruit fly infestation in tomato. A district-wide trapping using cue-lure baited traps was undertaken to determine fruit fly seasonal patterns in relation to the cropping seasons. A total of 17,626 field-harvested and 11,755 pack-house tomatoes were sampled from ten farms over three cropping seasons (2006-2009). The fruit were incubated and examined for fruit fly infestation. No fruit fly infested fruit were recorded over the three seasons in either the field or the pack-house samples. Statistical analyses showed that upper infestation levels were extremely low (between 0.025 and 0.062%) at the 95% confidence level. The trap catches showed a seasonal pattern in fruit fly activity, with low numbers during the autumn and winter months, rising slightly in spring and peaking in summer. This seasonal pattern was similar over the four seasons. The main two species of fruit fly caught were B. tryoni and B. neohumeralis. Based on the results, it is clear that the risk of fruit fly infestation is extremely low under the current production systems in the Bowen region.

A generic individual-based spatially explicit model as a novel tool for investigating insect-plant interactions: A case study of the behavioural ecology of frugivorous Tephritidae

Computational modelling of mechanisms underlying processes in the real world can be of great value in understanding complex biological behaviours. Uptake in general biology and ecology has been rapid. However, it often requires specific data sets that are overly costly in time and resources to collect. The aim of the current study was to test whether a generic behavioural ecology model constructed using published data could give realistic outputs for individual species. An individual-based model was developed using the Pattern-Oriented Modelling (POM) strategy and protocol, based on behavioural rules associated with insect movement choices. Frugivorous Tephritidae (fruit flies) were chosen because of economic significance in global agriculture and the multiple published data sets available for a range of species. The Queensland fruit fly (Qfly), Bactrocera tryoni, was identified as a suitable individual species for testing. Plant canopies with modified architecture were used to run predictive simulations. A field study was then conducted to validate our model predictions on how plant architecture affects fruit flies’ behaviours. Characteristics of plant architecture such as different shapes, e.g., closed-canopy and vase-shaped, affected fly movement patterns and time spent on host fruit. The number of visits to host fruit also differed between the edge and centre in closed-canopy plants. Compared to plant architecture, host fruit has less contribution to effects on flies’ movement patterns. The results from this model, combined with our field study and published empirical data suggest that placing fly traps in the upper canopy at the edge should work best. Such a modelling approach allows rapid testing of ideas about organismal interactions with environmental substrates in silico rather than in vivo, to generate new perspectives. Using published data provides a saving in time and resources. Adjustments for specific questions can be achieved by refinement of parameters based on targeted experiments.

Efficient Management of Fruit Pests by Pheromone Nanogels

Environment-friendly management of fruit flies involving pheromones is useful in reducing the undesirable pest populations responsible for decreasing the yield and the crop quality. A nanogel has been prepared from a pheromone, methyl eugenol (ME) using a low-molecular mass gelator. This was very stable at open ambient conditions and slowed down the evaporation of pheromone significantly. This enabled its easy handling and transportation without refrigeration, and reduction in the frequency of pheromone recharging in the orchard. Notably the involvement of the nano-gelled pheromone brought about an effective management of Bactrocera dorsalis, a prevalent harmful pest for a number of fruits including guava. Thus a simple, practical and low cost green chemical approach is developed that has a significant potential for crop protection, long lasting residual activity, excellent efficacy and favorable safety profiles. This makes the present invention well-suited for pest management in a variety of crops.

柑桔小实蝇生物学特性及其防治研究

柑桔小实蝇 Dacus (bactrocera) dorsalis (Hendel)在云南景谷1年发生5代,以老熟幼虫和蛹在土壤中越冬或越夏。2月中至8月中旬,成虫产卵于芒果果实内,致使芒果腐烂。室内用40%氧化乐果、50%杀螟松和80%的敌敌畏1 000、1 500倍液,对成虫的毒杀率达100%,幼虫达88.73%,蛹为10.0%。大田防治用1 000倍液,同时用性信息素进行诱杀,保果效果达95.74%。

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 December 2009-31 January 2010

This article documents the addition of 220 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Allanblackia floribunda, Amblyraja radiata, Bactrocera cucurbitae, Brachycaudus helichrysi, Calopogonium mucunoides, Dissodactylus primitivus, Elodea canadensis, Ephydatia fluviatilis, Galapaganus howdenae howdenae, Hoplostethus atlanticus, Ischnura elegans, Larimichthys polyactis, Opheodrys vernalis, Pelteobagrus fulvidraco, Phragmidium violaceum, Pistacia vera, and Thunnus thynnus. These loci were cross-tested on the following species: Allanblackia gabonensis, Allanblackia stanerana, Neoceratitis cyanescens, Dacus ciliatus, Dacus demmerezi, Bactrocera zonata, Ceratitis capitata, Ceratitis rosa, Ceratits catoirii, Dacus punctatifrons, Ephydatia mülleri, Spongilla lacustris, Geodia cydonium, Axinella sp. Ischnura graellsii, Ischnura ramburii, Ischnura pumilio, Pistacia integerrima and Pistacia terebinthus. © 2010 Blackwell Publishing Ltd.

Entomofauna associated with the olive tree in southern Portugal

Since 2004 several studies have been carried out in order to identify the main insect species that usually inhabiting the olive ecosystem. The field trials have taken place in two olive groves, one situated in Olhão and the other one in Loulé, both in Algarve and also under Integrated Pest Management (IPM). The sampling techniques used differ according to their purpose (sticky traps, pheromone traps, pitfall traps and samples of aerial parts of the trees such as inflorescences, leaves, fruits and branches). Results showed that the main insect pests of olive tree in southern Portugal were the olive fruit fly Bactrocera oleae Gmelin (Diptera: Tephritidae) and the olive moth Prays oleae Bernard (Lepidoptera: Hyponeumetidae). Other insect pests were also found in our olive groves namely the olive psyllid Euphyllura olivina Costa (Homoptera: Psyllidae), the olive dark beetle Phloeotribus scarabaeoides Bernard (Coleoptera: Curculionidae), the mediterranean black scale Saissetia oleae (Olivier) (Homoptera: Coccidae) and the olive thrip Liothripes oleae Costa (Thysanoptera: Phlaeothripidae). Concerning the auxiliary insects that were found in our olives groves they belong to the following orders and families: Diptera (Syrphidae), Coleoptera (Carabidae, Coccinelidae and Staphylinidae), Hemiptera (Anthocoridae and Miridae), Neuroptera (Chrysopidae) and Hymenoptera (Braconidae, Encyrtidae, Eulophidae, Formicidae and Trichogrammatidae).

Estudo de implementação da NP EN ISO 22000:2005 na Cooperativa de Olivicultores de Fátima, C. R.L.

O conceito de “Segurança Alimentar” faz sentido no momento em que o ser humano tem consciência de que o que ingere deve ser inócuo para a sua saúde e bem-estar. Para dar resposta a esta preocupação global, a indústria alimentar teve necessidade de adotar diretrizes orientadoras para a produção de alimentos seguros. Em consequência disso, surgiram as normas para os Sistemas de Gestão da Segurança Alimentar (SGSA) e a sua implementação é hoje em dia um elemento fundamental para a competitividade da empresa e dos produtos que comercializa. O trabalho desenvolvido na presente dissertação visa o estudo da implementação de um Sistema de Gestão da Segurança Alimentar com base no referencial de certificação NP EN ISO 22000:2005, na Cooperativa de Olivicultores de Fátima C. R. L.. A implementação de um sistema de gestão e a sua posterior certificação, é uma mais-valia para uma empresa. Ao obter a certificação, obtêm-se reconhecimento e satisfação dos clientes e outras partes interessadas, melhoria da imagem e prestigio, acesso a novos mercados, redução de custos de funcionamento através da melhoria do desempenho operacional e uma nova cultura com a sensibilização e motivação dos colaboradores, orientada para a melhoria continua e para a satisfação dos clientes e outras partes interessadas. Tendo por base os Códigos de Boas-práticas e HACCP existentes, fez-se a revisão e atualização de todo o sistema. Reapreciou-se o processo de fabrico e reavaliaram-se os pontos críticos. Reviram-se os Programas Pré-requisitos e melhorou-se o sistema HACCP. Os SGSA têm uma elevada relevância nas organizações associadas ao sector alimentar também nas indústrias produtoras de azeite, uma vez que estas devem implementar metodologias capazes de assegurar que os perigos para a saúde dos consumidores são eliminados ou reduzidos a níveis aceitáveis. Os principais perigos na indústria do azeite numa fase inicial são a presença de microrganismos patogénicos e parasitas da azeitona (Ex.: Bactrocera oleae) que se podem desenvolver devido ao excessivo tempo de espera até à operação ou por temperaturas inadequadas, podendo este perigo ser resolvido através do controlo na receção, de fornecedores, e do tempo de operação; boas práticas de fabrico e formação do pessoal. A presença de folhas, terra, pedras e metais diversos é outro dos perigos mais frequentes que são contornados através do controlo na receção; inspeção visual; avaliação dos fornecedores, limpeza e lavagem das azeitonas. Ao longo da produção e embalamento os principais perigos são os resíduos de produtos de higienização, incorporação de partículas estranhas ou restos de sujidade e a contaminação química por resíduos de massas e óleos de lubrificação, perigos estes que são colmatados através de boas práticas de fabrico; plano de higiene e de manutenção dos equipamentos e do uso de lubrificantes homologados para a indústria alimentar.

Sex differences in insect immune function : a consequence of diet choice?

Males often have reduced immune function compared to females but the proximate mechanisms underlying this taxonomically widespread pattern are unclear. Because immune function is resource-dependent and sexes may have different nutritional requirements, we hypothesized that sexual dimorphism in immune function may arise from differential nutrient intake (acquisition hypothesis). To test this hypothesis, we examined patterns of phenoloxidase (PO) activity in relation to nutrient consumption in Queensland fruit flies (Q-flies). In the first experiment, flies were allowed to choose their preferred nutrient intake. Compared with males, female Q-flies had higher PO activity, consumed more calories, and preferred a higher protein:carbohydrate (P:C) diet, suggesting that differential acquisition could explain sex differences. In the second experiment, we restricted flies to one of 12 diets varying in protein and carbohydrate concentrations and mapped PO activity for each sex onto a nutritional landscape. Counter to our hypothesis, females had higher PO activity than males at any given level of nutrient intake. Both carbohydrate and protein intake affected PO activity in females but only protein affected PO activity in males. Our results indicate that sex differences in Q-fly immune function are not solely explained by sex differences in nutrient intake, although nutrition does contribute to the magnitude of these sex differences.

Cost of reproduction in the Queensland fruit fly: Y-model versus lethal protein hypothesis

The trade-off between lifespan and reproduction is commonly explained by differential allocation of limited resources. Recent research has shown that the ratio of protein to carbohydrate (P : C) of a fly's diet mediates the lifespan–reproduction trade-off, with higher P : C diets increasing egg production but decreasing lifespan. To test whether this P : C effect is because of changing allocation strategies (Y-model hypothesis) or detrimental effects of protein ingestion on lifespan (lethal protein hypothesis), we measured lifespan and egg production in Queensland fruit flies varying in reproductive status (mated, virgin and sterilized females, virgin males) that were fed one of 18 diets varying in protein and carbohydrate amounts. The Y-model predicts that for sterilized females and for males, which require little protein for reproduction, there will be no effect of P : C ratio on lifespan; the lethal protein hypothesis predicts that the effect of P : C ratio should be similar in all groups. In support of the lethal protein hypothesis, and counter to the Y-model, the P : C ratio of the ingested diets had similar effects for all groups. We conclude that the trade-off between lifespan and reproduction is mediated by the detrimental side-effects of protein ingestion on lifespan.